The Handbook of Mummy Studies: New Frontiers in Scientific and Cultural Perspectives 9811533539, 9789811533532

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Dong Hoon Shin Raffaella Bianucci Editors

The Handbook of Mummy Studies New Frontiers in Scientific and Cultural Perspectives

The Handbook of Mummy Studies

Dong Hoon Shin • Raffaella Bianucci Editors

The Handbook of Mummy Studies New Frontiers in Scientific and Cultural Perspectives

With 368 Figures and 25 Tables

Editors Dong Hoon Shin Laboratory of Bioanthropology Paleopathology and History of Diseases Department of Anatomy and Cell Biology Institute of Forensic and Anthropological Science Seoul National University College of Medicine Seoul, South Korea

Raffaella Bianucci New Mexico Health Enhancement and Marathon Clinics Research Foundation Albuquerque, NM, USA Warwick Medical School, Biomedical Sciences University of Warwick Coventry, UK Legal Medicine Section, Department of Public Health and Paediatric Sciences University of Turin Torino, Italy

ISBN 978-981-15-3353-2 ISBN 978-981-15-3354-9 (eBook) ISBN 978-981-15-3355-6 (print and electronic bundle) https://doi.org/10.1007/978-981-15-3354-9 © Springer Nature Singapore Pte Ltd. 2021 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors, and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, expressed or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. This Springer imprint is published by the registered company Springer Nature Singapore Pte Ltd. The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore

Preface

Mummies, either natural or embalmed, ancient or modern, are the bodies of once living organisms – human and animals – which are “temporarily immortalized.” These bodies and their associated artifacts (e.g., coffins, shrouds, clothes, ceramic and wooden objects, cosmetics, texts, weapons, games) have attracted generations of people of all ages. Why? Because they represent a visible medium between life and death, past and present, human fallacy and eternity. Independently from the region of the world they originated from, these bodies open a window to their past. Through the application of refined investigation techniques as well as philological studies, scholars are challenged to reconstruct the life habits of the individuals of the past, their health state, the environments in which they lived, their beliefs, and their interactions with other cultures. Thanks to the contribution of mummy experts with different backgrounds, both the scientific and cultural aspects of the populations of the past can be explored. To do so, the Handbook of Mummy Studies has been structured into different parts to allow readers to approach the theme in a logical way. The “General Aspects of Mummy Studies” allows familiarization with the history of mummy studies and the various types of mummification and embalming techniques used in the different regions of the world. Ethical guidelines linked to the treatment of ancient remains from the burial site to their exhibitions either in Museums or in cemeterial areas (i.e., catacombs and crypts) follow. Particular attention is paid to the application of autoptic procedures in mummies based on their provenance, religious background, and on the different laws of each country. Within this part, two chapters are dedicated to major pathological conditions such as cancer and cardiovascular diseases, which affected ancient human populations. The second part (“Research Techniques of Mummy Studies”) is dedicated to the modern noninvasive (medical imaging/radiology/craniofacial reconstruction) or minimally invasive techniques (endoscopy and stable isotope analysis); noninvasive techniques allow scientists to identify the age at death, sex, stature, presence/absence of organs (eventually removed during the embalming procedures), postmortem dislocation, and the presence/absence of pathognomonic lesions of both soft and hard tissues; specific interest is paid to the reconstruction of the facial features. This part covers statistical methods used both in anthropological and mummies researches. v

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Preface

The third part of the book focuses on ancient DNA studies which look into the fascinating world of human, animal, and wildlife genetics; this topic has consistently developed with the introduction of next-generation sequencing. By using soft mummified tissues, new information on the genetic history of humankind and past genomic diversity of ancient parasites is proposed. Cutting-edge techniques such as next-generation sequencing, low copy number DNA analysis, and SNP-based DNA phenotyping are further described. The concept of pathoecology, that is the ecological approach to reconstruction of infection patterns in past and present populations, is presented in the fourth part “Archaeoparasitology.” The diversity of parasites (whipworms, hookworms, Ascaris, pinworms, etc.) found in mummies is explored. These ancient bodies are unique sources of data to look at changing parasitological conditions over time. Hence the pathoecological approach to mummy studies provides outlooks relevant to current and future trends in parasitology. Analysis of mummy digestive tracts, taphonomy and arthropod associates of decay, and coprolite analysis for diet ingestion are also discussed in this part. The parts from five to eight (“Egyptian Mummies,” “Mummies in Europe,” “Mummies in South America and the Pacific Region,” “Mummies in Asia”) are dedicated to a detailed description of the mummies (human but also animal) from all continents. While there is more familiarity with mummies from Western world, special relevance is given to the description of mummified bodies from Siberia, South Korea, China, and Japan. This approach allows expansion of knowledge of the cultural heritage of the Eastern world, the posthumously treatment of the bodies, and religious beliefs. Since the process of active preservation of the bodies is generated and maintained by culture, a specific part is dedicated to the “Cultural Aspects of Mummy Studies.” Notwithstanding the recent explosion of pioneering researches in mummy studies, the fora to discuss their cultural aspects are unfortunately still insufficient. The contributors to this part discuss recent trends and novel research outcomes in cultural aspects of mummy studies. Over the past several decades, many achievements have been reported by pioneering researches on mummies from all over the world. This book provides a definitive text for both the established researcher and the student wanting to learn about the broad scope of mummies and the methods used in their study. It brings together a broad range of experts in their fields to provide a synthesis of the subject. We hope that it will help promote the subject and lead to a wider readership in both the scientific and general populations. Seoul, Korea Coventry, UK September 2021

Dong Hoon Shin Raffaella Bianucci

Acknowledgments

I am grateful to my lab members past and present for their great work on ancient peoples’ health and diseases: Drs. Chang Seok Oh, Jong Ha Hong, Hyejin Lee, and Ms. Jieun Kim. I must also point out the generous help of my collaborators: Drs. Soong Deok Lee, Myeung Ju Kim, Jaehyup Kim, Dong Soo Yoo, In Sun Lee, Won Joon Lee, Yi Suk Kim, Do Seon Lim, Shiduck Kim, Myung Ho Shin, Eun Jin Woo, Tae Sup Cho, Ho Chul Ki, Taeshik Kim, Mi Kyung Song, and Eun Joo Lee. As for archaeology, I am grateful to Drs. Yongjun Kim, Soon Chul Cha, Sang Yuk Shim, Yuri Bang, Young Moon Shin, Jin Og Ju, In Uk Kang, Jun Bum Park, and Sori Min. I also remember the friendship of Mark Spigelman, who helped me at the beginning of this research. My special thanks to Drs. Karl Reinhard, Jong-Yil Chai, and Min Seo for their archaeoparasitological collaboration. For my work in India and Siberia, Drs. Vasant Shinde and Sergey Slepchenko were my best friends/collaborators. I would like to express my respect also to the two professors who were my mentors: Drs. Sang Ho Baik and Sa Sun Cho. Without their teaching, I would not have been the researcher I am today. Sincere gratitude must be given to my mother (Young Ja Lee), father (Dr. Young Chul Shin), and sister (Dr. Eun-Kyoung Shin). I don’t know how to thank them, because without their faith in me, this book would not have been possible. Finally, if I have to express my gratitude to only two people, it is my beloved wife Eunju and daughter Minjae. I think they’ve sacrificed too much to support me and my research so far. With my infinite love and respect, I want to dedicate this book to them. Dong Hoon Shin This book is lovingly dedicated to my parents Mario and Graziella for their endless support. I am most grateful to my mentor and dearest friend, Professor Otto Appenzeller, whose constant presence in my life made it all possible. A special thought is dedicated to the late Professor Adauto Aráujo; his beautiful mind and graceful personality are sorely missed. Raffaella Bianucci

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Contents

Volume 1 Part I

General Aspect of Mummy Studies . . . . . . . . . . . . . . . . . . . . .

1

1

History of Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Jane E. Buikstra and Kenneth C. Nystrom

3

2

Dried, Tanned, Frozen, Embalmed, Smoked: A Glimpse into Mummification Mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dario Piombino-Mascali and Haley Carr

41

Showing Respect to the Dead: The Ethics of Studying, Displaying, and Repatriating Mummified Human Remains . . . . . . . . . . . . . . . Heather Gill-Frerking

59

3

4

Ethical Considerations of Anthropologists’ Fieldwork . . . . . . . . . . Katherine I. Placher and Christopher J. Bae

5

Radiological and Ethical Considerations of Autopsy in Mummy Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Yi-Suk Kim, Myeung Ju Kim, Chang Seok Oh, Jong Ha Hong, and Dong Hoon Shin

6

Paleo-Oncology and Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . Andreas G. Nerlich and Raffaella Bianucci

7

The Burden of Arteriosclerotic Cardiovascular Disease in Ancient Populations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Andreas G. Nerlich, Francesco Maria Galassi, and Raffaella Bianucci

Part II

Research Techniques of Mummy Studies

..............

89

111

131

147

163

8

Craniofacial Reconstruction in Mummy Studies . . . . . . . . . . . . . . Won Joon Lee and Dong Hoon Shin

165

9

Endoscopy in Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mark Spigelman and Dong Hoon Shin

179 ix

x

10

Contents

Stable Isotope Analysis in Archaeological Science and Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mai Takigami and Minoru Yoneda

197

11

Radiology Applications in Mummy Science . . . . . . . . . . . . . . . . . . Ronald G. Beckett and Gerald J. Conlogue

211

12

Medical Imaging in Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . Robert D. Loynes and Raffaella Bianucci

253

Part III

Ancient DNA Analysis and Mummy Research

..........

269

........

271

.....................

285

15

Ancient DNA Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Choongwon Jeong

301

16

Ancient DNA and Paleoparasitology in Brazil . . . . . . . . . . . . . . . . Alena Mayo Iñiguez

317

17

Analysis of Low Copy Number DNA and Degraded DNA . . . . . . . Hwan Young Lee

351

18

A Primer on the Population Genetic Analysis of Ancient Genomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Choongwon Jeong

371

Application of SNP-Based DNA Phenotyping to Archaeological and Forensic Cases . . . . . . . . . . . . . . . . . . . . . . . . . Hwan Young Lee

387

13

History of Ancient DNA Analysis in Mummy Research Jong Ha Hong, Chang Seok Oh, and Dong Hoon Shin

14

Current Trends in Ancient DNA Study Choongwon Jeong

19

Part IV 20

Archaeoparasitology

..............................

Mummies, Parasites, and Pathoecology in the Ancient Americas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Adauto Araújo and Karl J. Reinhard

21

Archaeoparasitology of Korean Mummies . . . . . . . . . . . . . . . . . . . Min Seo, Jong Ha Hong, Karl J. Reinhard, and Dong Hoon Shin

22

Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice . . . . . . . . . . . . . . . . . . . Karl J. Reinhard, Julia Russ, Isabel Teixeira-Santos, You Zhou, and Vaughn M. Bryant

409

411 439

461

Contents

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23

Mummy Taphonomy and Arthropod Associates of Decay . . . . . . . Jessica Smith, Lauren Gipson, Dario Piombino-Mascali, and Rimantas Jankauskas

24

Ancient Parasites Seen in the Archaeology and Medical Contexts in the Han Dynasty, China . . . . . . . . . . . . . . . . . . . . . . . . Xiaoya Zhan, Wuyun Qi, and Hui-Yuan Yeh

25

26

Dietary Stress in Combat: Coprolite Analysis of a Korean War Marine Killed in Action . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Karl J. Reinhard, Marina Milanello do Amaral, Gregory E. Berg, and Brianna Neu Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent . . . . . . . . . . . . . . . . . . . . . . . . . . . Sergey Mikhailovich Slepchenko, Min Seo, Jong Ha Hong, Chang Seok Oh, and Dong Hoon Shin

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513

533

547

Volume 2 Part V 27

Egyptian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Daniel Antoine and Marie Vandenbeusch

28

Animal Mummies in Ancient Egypt and South America . . . . . . . . Salima Ikram

29

Trends in Use of Organic Balms in Egyptian Mummification Revealed Through Biomolecular Analyses . . . . . . . . . . . . . . . . . . . Richard P. Evershed and Katherine A. Clark

Part VI

Mummies in Europe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

563

565 629

653

717

30

Life and Diseases of the Neolithic Glacier Mummy “Ötzi” . . . . . . Andreas G. Nerlich, Angelika Fleckinger, and Oliver Peschel

719

31

Mummies in Crypts and Catacombs . . . . . . . . . . . . . . . . . . . . . . . Andreas G. Nerlich and Raffaella Bianucci

741

32

The Vác Mummy Project: Investigation of 265 Eighteenth-Century Mummified Remains from the TB Pandemic Era . . . . . . . . . . . . . Helen D. Donoghue, Ildikó Pap, Ildikó Szikossy, and Mark Spigelman

33

Bog Bodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ronald G. Beckett

777 807

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Contents

34

Bog Bodies and Natural Mummification of Siberia . . . . . . . . . . . . Sergey Mikhailovich Slepchenko, Igor Konstantinovich Novikov, Jong Ha Hong, Do Seon Lim, Chang Seok Oh, Jieun Kim, Hye Jin Lee, and Dong Hoon Shin

833

35

Vladimir Il’ič Lenin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Raffaella Bianucci, Francesco Maria Galassi, and Andreas G. Nerlich

851

36

Ancient Greece and Mummies: The Primacy of the Soul over the Body . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Francesco Maria Galassi and Elena Varotto

37

Guanche Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conrado Rodríguez-Martín

38

Embalming in France (from Twelfth to Nineteenth Century): Principle and Development of Techniques . . . . . . . . . . . . . . . . . . . Philippe Charlier

Part VII

Mummies in South America and Pacific Region . . . . . . . .

877 883

907

929

39

South American Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Guido Lombardi and Bernardo Arriaza

931

40

Brazilian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Shênia Patrícia Corrêa Novo

945

41

Fire Mummies of the Kabayan Region of Benguet Province, Luzon, the Philippines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ronald G. Beckett

42

Smoked Bodies of Papua New Guinea . . . . . . . . . . . . . . . . . . . . . . Ronald G. Beckett

Part VIII

Mummies in Asia

................................

957 983

1009

43

Mummies in Siberia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1011 Sergey Mikhailovich Slepchenko, Jong Ha Hong, and Dong Hoon Shin

44

Archaeological Findings of the Tarim Basin Graves and Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1033 In Uk Kang, Jinseong Han, Jong Ha Hong, Jieun Kim, Dong Hoon Shin, and Victor H. Mair

45

Joseon Dynasty Mummies of Korea . . . . . . . . . . . . . . . . . . . . . . . . 1049 Dong Hoon Shin, Chang Seok Oh, Jong Ha Hong, Myung Ho Shin, Myeung Ju Kim, and Hye Jin Lee

Contents

xiii

46

Mummies in China: Mawangdui and Other Related Mummies . . . 1073 Kyeongmi Joo and Dong Hoon Shin

47

Mummies in Japan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1103 Hisashi Fujita, Jun Koike, Hiroki Sugimori, and Dong Hoon Shin

48

Mummies of Song-Ming Dynasty in China . . . . . . . . . . . . . . . . . . . 1117 Dong Hoon Shin, Chang Seok Oh, and Jong Ha Hong

Part IX

Cultural Aspects of Mummy Studies . . . . . . . . . . . . . . . . . .

1137

49

Fake and Alien Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1139 Guido Lombardi and Conrado Rodríguez Martín

50

Mummy Clothing Found in East Asia . . . . . . . . . . . . . . . . . . . . . . 1153 Dong Hoon Shin, Chang Seok Oh, Jong Ha Hong, and Mi Kyung Song

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1171

About the Editors

Dong Hoon Shin graduated from Seoul National University College of Medicine in South Korea and received his certification of geriatrician. After serving as a public physician for three years, he earned his Ph.D. in Anatomy. He currently teaches and researches as a full professor and chairperson in the Department of Anatomy and Cell Biology at Seoul National University. Over the years, his research interests have mainly focused on acquiring scientific information on premodern peoples’ health and disease status. Using research techniques and tools ranging from the anatomical and histological to the molecular, he has worked to reveal the physical and/or pathological traits of ancient peoples and their patterns at different historical stages. He has conducted viral, bacterial, and helminthic pathogen investigations with archaeologically obtained human remains such as Korean mummies. These have afforded vivid glimpses of premodern peoples’ health and disease status and, thus, of their actual lives, in significant detail from both medical and sociocultural perspectives. To enrich his understanding of human diseases of the past, he is also actively joining in fieldwork at archaeological sites outside South Korea. Currently, they include the Indus Valley Civilization ruins of presentday India and the permafrost burial grounds of West Siberia. Based on his academic achievements, he was inducted as a National Geographic Explorer and Kavli Frontiers of Science Fellow.

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About the Editors

Raffaella Bianucci has specialized in biological anthropology, palaeopathology, and medical history with an emphasis on reconstructing the impact of historical plague pandemics in Europe between the sixth and eighteenth centuries. She is known for her pioneering work on the identification of Yersinia pestis F1 protein in ancient skeletal remains. Her researches have contributed significant insights into the systematic identification of putative plague victims uncovered from several plague pits scattered around Europe. She currently is investigating the historical impact of plague on Medieval populations and the biological role of rats and rat fleas in the interhuman transmission of the infection. As a palaeopathologist, her research has focused on the reconstruction of illnesses of several members of important families in Renaissance Italy (the Medici and the Aragonese). By applying the most advanced techniques in medicine and biochemistry, she is investigating the cause of death and the embalming techniques used to preserve ancient Egyptian mummies. Currently she is the group leader of a multidisciplinary project on Spanish church mummies from Quinto (Zaragoza) and on bog bodies from the Netherlands. From 2015, she has specialized in Medical Humanities. Together with Prof. Francesco Maria Galassi (Flinders University, Adelaide, Australia), she has introduced in Italy a new branch of investigation, namely palaeopathography, in classical palaeopathology. Palaeopathography encompasses both the philological and clinical analysis of ancient documental sources and archives and the clinical investigation of ancient works of art with the purpose of identifying the historical presentation and evolution of diseases throughout history. She has published numerous peer-reviewed articles in leading scientific journals such as Nature Communications, The Lancet, The Lancet Oncology, The Lancet Respiratory Medicine, The Lancet Neurology, The Lancet Diabetes and Endocrinology, the American Journal of Medicine, Chest, the American Journal of Physical Anthropology, the Journal of Archaeological Science, PlOS Pathogens, and PlOS ONE. Currently, she is the coeditor and author of a book on Art and Medicine in the

About the Editors

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Uffizi Gallery (Pontercoboli Editore) which will be published in December 2021. Raffaella Bianucci is Vice President of the New Mexico Health Enhancement and Marathon Clinics Research Foundation, Albuquerque (NM, USA) and Honorary Research fellow at Warwick Medical School, Biomedical Sciences, The University of Warwick (UK). She gained her Ph.D. at the University of Florence and completed her postdoctoral training at the universities of Turin (Italy) and Oslo (Norway).

About the Section Editors

Andreas G. Nerlich Institute of Pathology Academic Hospital Munich-Bogenhausen Munich, Germany

Choongwon Jeong School of Biological Sciences Seoul National University Seoul, Republic of Korea

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About the Section Editors

Karl J. Reinhard School of Natural Resources University of Nebraska-Lincoln Lincoln, NE, USA

Ronald G. Beckett Biomedical Sciences, Bioanthropology Research Institute Quinnipiac University Hamden, CT, USA

Contributors

Daniel Antoine Department of Egypt and Sudan, The British Museum, London, UK Adauto Araújo Departamento de Endemias Samuel Pessoa, Escola Nacional de Saúde Pública, Oswaldo Cruz Foundation, Rio de Janiero, Brazil Bernardo Arriaza Instituto de Alta Investigación, Universidad de Tarapacá, Arica, Chile Christopher J. Bae Department of Anthropology, University of Hawaii at Manoa, Honolulu, HI, USA Ronald G. Beckett Biomedical Sciences, Bioanthropology Research Institute, Quinnipiac University, Hamden, CT, USA Gregory E. Berg Defense POW/MIA Accounting Agency (DPAA), Joint Base Pearl Harbor-Hickam, HI, USA Raffaella Bianucci New Mexico Health Enhancement and Marathon Clinics Research Foundation, Albuquerque, NM, USA Warwick Medical School, Biomedical Sciences, University of Warwick, Coventry, UK Legal Medicine Section, Department of Public Health and Paediatric Sciences, University of Turin, Torino, Italy Vaughn M. Bryant Department of Anthropology, Texas A&M University, College Station, TX, USA Jane E. Buikstra Center for Bioarchaeological Research, Arizona 900 S. Cady Mall, Mail Code 2402, School of Human Evolution and Social Change, Arizona State University, Tempe, AZ, USA Haley Carr Forensic and National Security Sciences Institute, Syracuse University, Syracuse, NY, USA

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Contributors

Philippe Charlier Laboratoire Anthropologie, Archéologie, Biologie (LAAB), Université Paris-Saclay (UVSQ), UFR des Sciences de la Santé, Montigny-LeBretonneux, France Fondation Anthropologie, Archéologie, Biologie (FAAB) – Institut de France, Paris, France Département de la Recherche et de l’Enseignement, Musée du quai Branly – Jacques Chirac, Paris, France Katherine A. Clark Organic Geochemistry Unit, School of Chemistry, University of Bristol, Cantock’s Close, Bristol BS8 1TS, UK Gerald J. Conlogue Bioanthropology Research Institute, Quinnipiac University, Hamden, CO, USA Shênia Patrícia Corrêa Novo Escola Nacional de Saúde Pública Sergio Arouca – ENSP, Fundação Oswaldo Cruz – Fiocruz, Rio de Janeiro, Brazil Marina Milanello do Amaral Superintendência da Polícia Técnico-Científica, São Paulo, Brazil Helen D. Donoghue Centre for Clinical Microbiology, Division of Infection and Immunity, University College London, London, UK Richard P. Evershed Organic Geochemistry Unit, School of Chemistry, University of Bristol, Cantock’s Close, Bristol BS8 1TS, UK Angelika Fleckinger South Tyrol Museum of Archaeology, Bolzano, South Tyrol, Italy Hisashi Fujita Doshisha University, Kyotanabe, Japan Francesco Maria Galassi Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia FAPAB Research Center, Avola, Italy Heather Gill-Frerking NTK, Frankford, ON, Canada Observatory for the Mummified Heritage of Sicily, Santa Lucia del Mela, Italy Lauren Gipson Department of Forensic Science, University of Nebraska-Lincoln, Lincoln, NE, USA Jinseong Han Department of History (Archaeology), Kyung Hee University, Seoul, South Korea Jong Ha Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea Salima Ikram American University in Cairo, New Cairo, Egypt Alena Mayo Iñiguez Laboratório de Biologia de Tripanosomatídeos with Instituto Oswaldo Cruz, Fundação Oswaldo Cruz – FIOCRUZ, Rio de Janeiro, RJ, Brazil

Contributors

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Rimantas Jankauskas Institute of Biomedical Sciences, Vilnius University, Vilnius, Lithuania Choongwon Jeong School of Biological Sciences, Seoul National University, Seoul, Republic of Korea Kyeongmi Joo Department of Archaeology, Chungnam National University, Daejeon, Korea In Uk Kang Department of History (Archaeology), Kyung Hee University, Seoul, South Korea Jieun Kim Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul, Korea Myeung Ju Kim Department of Anatomy, Dankook University College of Medicine, Cheonan-si, Chungcheongnam-do, Republic of Korea Yi-Suk Kim Department of Anatomy, Catholic Institute for Applied Anatomy, The Catholic University of Korea, Seoul, South Korea Jun Koike Department of Nursing, Daito Bunka University, Higashimatsuyama, Japan Hwan Young Lee Department of Forensic Medicine, Seoul National University College of Medicine, Seoul, Korea Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Korea Hye Jin Lee Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Republic of Korea Ministry of National Defense Agency KIA Recovery and Identification, Seoul, Republic of Korea Won Joon Lee National Forensic Service Seoul Institute, Seoul, South Korea Do Seon Lim Department of Dental Hygiene, College of Health Science, Eulji University, Seongnam, Korea Guido Lombardi Cátedra Pedro Weiss, Universidad Peruana Cayetano Heredia, Lima, Perú Robert D. Loynes KNH Centre for Biomedical Egyptology, University of Manchester, Manchester, UK Victor H. Mair Department of East Asian Languages and Civilizations, University of Pennsylvania, Philadelphia, PA, USA Andreas G. Nerlich Institute of Pathology, Academic Hospital MunichBogenhausen, Munich, Germany

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Contributors

Brianna Neu Department of Anthropology, University of Nebraska, Lincoln, NE, USA Igor Konstantinovich Novikov Kurgan State University, Kurgan, Russian Federation Kenneth C. Nystrom Department of Anthropology, State University of New York at New Paltz, New Paltz, NY, USA Chang Seok Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea Ildikó Pap Department of Anthropology, Hungarian Natural History Museum, Budapest, Hungary Oliver Peschel Institute of Legal Medicine, Ludwig-Maximilians-University Munich, Munich, Germany Dario Piombino-Mascali Department of Anatomy, Histology and Anthropology, Faculty of Medicine, Vilnius University, Vilnius, Lithuania Katherine I. Placher Department of Anthropology, University of Hawaii at Manoa, Honolulu, HI, USA Wuyun Qi Institute of Archaeology, Chinese Academy of Social Sciences, Beijing, China Karl J. Reinhard School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE, USA Conrado Rodríguez-Martín Instituto Canario de Bioantropología and Museo Arqueológico de Tenerife, OAMC-Cabildo de Tenerife, Santa Cruz de Tenerife, Spain Julia Russ Morrison Microscopy Core Research Facility, Center for Biotechnology, University of Nebraska-Lincoln, Lincoln, NE, USA Min Seo Department of Parasitology, Dankook University College of Medicine, Cheonan, South Korea Dong Hoon Shin Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea Myung Ho Shin Department of History, Pukyong National University, Busan, Republic of Korea Sergey Mikhailovich Slepchenko Institute of the Problems of Northern Development, Tyumen Scientific Center of the Siberian Branch of the Russian Academy of Sciences, Tyumen, Russia Surgut State University, Surgut, Russian Federation

Contributors

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Jessica Smith Department of Anthropology, University of Nebraska-Lincoln, Lincoln, NE, USA Mi Kyung Song Department of Fashion Design and Marketing, Seoul Women’s University, Seoul, South Korea Mark Spigelman The Kuvin Center for the Study of Infectious and Tropical Diseases and Ancient DNA, Hadassah Medical School, The Hebrew University, Jerusalem, Israel Division of Infection and Immunity, Centre of Clinical Microbiology Royal Free hospital UCL London, London, UK Department of History and Archaeology, Macquarie University, NSW, Australia Hiroki Sugimori Department Higashimatsuyama, Japan

of

Nursing,

Daito

Bunka

University,

Ildikó Szikossy Department of Anthropology, Hungarian Natural History Museum, Budapest, Hungary Mai Takigami Yamagata University, Yamagata, Japan Isabel Teixeira-Santos School of Natural Resources, University of NebraskaLincoln, Lincoln, NE, USA Marie Vandenbeusch Department of Egypt and Sudan, The British Museum, London, UK Elena Varotto Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia FAPAB Research Center, Avola, Italy Hui-Yuan Yeh School of Humanities, Nanyang Technological University, Singapore, Singapore Minoru Yoneda Laboratory of Radiocarbon Dating, The University Museum, The University of Tokyo, Tokyo, Japan Xiaoya Zhan School of Humanities, Nanyang Technological University, Singapore, Singapore You Zhou Morrison Microscopy Core Research Facility, Center for Biotechnology, University of Nebraska-Lincoln, Lincoln, NE, USA

Part I General Aspect of Mummy Studies

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History of Mummy Studies Jane E. Buikstra and Kenneth C. Nystrom

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Egypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Egyptian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . European Tourism Begins: ~450 BCE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Influence of Egyptian Mummification Upon Egyptian and Greek Medical Practice . . . The Medieval Period (476–1500 CE), the Renaissance (1300–1600 CE), and the Emergence of the “Mummy” . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Era of European Exploration and Discovery (15th–17th Centuries) . . . . . . . . . . . . . . . . . . . . . . . . The Spanish Conquest of the South American Andes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The 17th, 18th, and early 19th Centuries: Collecting, Spectacles, and Stirrings of Science . . . Unwrapping Egyptian Mummies in Europe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Napoleon and the Savants (1798–1801) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Nineteenth- and Early Twentieth-Century Spectacles and Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Egyptomanie and Unrolling Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Nineteenth-Century Scientific Study of Egyptian Mummies in the UK . . . . . . . . . . . . . . . . . . . . . Eighteenth to Twentieth Centuries: Scientific Observations in South America Begin and the Earliest Prepared, Artificial Mummies in the World Are Discovered . . . . . . . . . . . . . . . Collecting and Viewing Mummies in North America . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Nineteenth-Century American Scientific Contributions to the Egyptian Question . . . . . . . . . . Early Twentieth Century . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Summary, Conclusions, and Fast Forward to the Twenty-First Century . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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J. E. Buikstra (*) Center for Bioarchaeological Research, Arizona 900 S. Cady Mall, Mail Code 2402, School of Human Evolution and Social Change, Arizona State University, Tempe, AZ, USA e-mail: [email protected] K. C. Nystrom Department of Anthropology, State University of New York at New Paltz, New Paltz, NY, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_2

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Abstract

The recorded history of mummy studies begins with Herodotus’s insightful observations of Egyptian mummification procedures, including socioeconomic distinctions between elaborate and less intricate forms. Such differences were confirmed by Diodorus, writing a few centuries later, and then again by the savant P. C. Rouyer, who traveled with Napoleon to Egypt at the turn of the nineteenth century. The Bonaparte incursion, an enormously unsuccessful military effort, ignited an eighteenth-century simmering interest in Egypt into a remarkable Egyptomania that coursed across museums and lecture halls, eventually leading to professional Egyptology with cultural and biological interest in the mummies as people. A world away, in the coastal South-Central Andes, sixteenth-century Spanish explorers encountered Inca mummies within a landscape that included the living and the ancestors. By the end of that century, the “extirpation of idolatries” led to the destruction of the Royal Incas, including their mummies, whose preparation methods remain a source of disputation. As mummies crossed the seas to grace private and public collections, study initially focused upon them and their accompaniments as cultural materials, as scientists described preparation procedures, along with associated artifacts. Scientific study of the cadavers themselves, while earlier moments occurred, began with the development of imaging procedures in the very late nineteenth century. This chapter will follow the histories of discovery and interpretation of mummies, focusing upon the contrastive Egyptian and Andean chronologies and ending in the early twentieth century, where the chapters in this volume begin their contemporary treatments. Keywords

Human mummies · Animal mummies · History · Egypt · Andes · Guanches · Canary Islands

Introduction Recorded encounters with human and animal mummies began in Egypt with observations made by Herodotus during the mid-fifth century BCE. Initial reports from the Americas were made by Spanish conquistadors and padres during the sixteenth century CE. A growing desire to see these exotic, ancestral figures, beginning in the Renaissance, was ignited as the passion of Egyptomania by the Napoleonic military and intellectual invasion of the Nile Valley at the turn of the nineteenth century. Mummies from across the globe were sought for displays by individuals, museums, and celebrations of Western accomplishments such as the World’s Columbian Exposition in Chicago (1893) and the Panama-California Exposition (1915– 1916). Beginning in the late nineteenth century and extending to today, a tradition of scientific study has flourished, frequently aided by imaging devices and other techniques borrowed from the biomedical sciences.

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Stock-taking syntheses of mummy studies began with Pettigrew’s (1834) summary of his public “unrolling” events featuring Egyptian mummies, then extending to locations other than Egypt where ancient and recent mummies had been reported. While imaging methods, along with tissue rehydration and histology developed at the turn of the twentieth century, physical studies of mummies concentrated in the creation of collections, such as those of the Manchester Museum (UK).These would be followed with renewed interest in mummy research stimulated by the dissections of holdings from the Pennsylvania Museum (Cockburn and Cockburn 1980; Cockburn et al. 1998; Roberts et al. 2007) during the latter part of the twentieth century. One of the early twenty-first century’s exquisitely detailed offerings, The Scientific Study of Mummies (Aufderheide 2003), reflects the author’s lifelong commitment to and excellence in joining mummy science with state-of-the-art biomedical and historical observations. Historical treatments have also focused upon figures and regions prominent in the study of mummified remains (Buikstra and Roberts 2012). The biomedical approach, which typically, but not always (Thompson et al. 2013) focuses upon case studies, has been enhanced by bioarchaeological perspectives (Nystrom 2019). This is the landscape of mummy studies surveyed here. This chapter can only briefly sketch a history of mummy studies, which truly requires a book-length treatment (Aufderheide 2003) or encyclopedic review, such as that presented in the ensuing chapters. Therefore, this prefatory statement will focus upon historical sequences of observations for two key, distinctively different regions – Egypt and the South-Central Andes – where embalming or similar purposeful treatments enhanced the social significance of ancestral figures, whose remains figured importantly in the political, spiritual, and social spheres of the living at scales ranging from the community to the state. This history focuses upon discovery and interpretation by Western eyes, ranging from historians to travelers, to colonists, and to scholars. Included are chronological summaries, followed by historical treatments written primarily by European and Euro-American writers, extending into the twentieth century. Discussion also touches upon hypothesized impacts of mummification knowledge especially in relationship to medical knowledge in ancient Greece and the diffusion of culture, as proposed by Elliot Smith. The closing section reflects upon twentieth- and twenty-first-century trends in mummy studies.

Egypt Egyptian Mummies Egyptian Mummies play a central role in any discussion of purposefully desiccated human remains. Time depth for anthropogenic treatment and written records of both embalming practices and the recovery, display, and study of mummified Egyptian people have excited European reporting, beginning with the Greek historian, Herodotus, considered below. Further discussion is prefaced by a broad brush of Egyptian

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history pertinent to mummification, based primarily upon Aufderheide (2003: 212– 250; 395–404), beginning with “spontaneous” or natural mummification and extending through increasingly elaborate treatments, followed by the decline in adherence to standards promoted during the Pharaonic Period. This overview provides essential background information for the ensuing discussions. The Egyptian mummification tradition is typically traced to the unification of upper and lower Egypt at the beginning of the Archaic Period, which includes Dynasty 1 (3050–2813 BCE) and Dynasty 2 (2813–2663 BCE). By this time, there was also evidence of wrapped animal burials, subsequently associated with mummification procedures that paralleled those of human remains, including evisceration and brain removal, along with elaboration of bundled forms. Bulls and rams were the earliest thus treated, reflecting the herding tradition developed in the Nile Valley during the 6th millennium BCE. Such animal glorification is typically glossed as cult behavior, though it may also be understood in terms of the shifting relationship of humans to animals formerly hunted and now herded. Ingold (2000) emphasizes the partnership that typically forms between hunters and their prey, which becomes asymmetrical as more power is exerted over herds during the process of domestication. Narratives that emerge in the course of this shifting symbiotic pattern, Ingold says (2000: 61), “tell us as much about how the narrators view their own humanity as they do about their attitudes and relations to non-human animals.” So it was as the ancient Egyptian animal “cults” developed. Most discussions emphasize natural, spontaneous mummification in Egypt prior to the end of Dynasty 2 or the beginning of the Old Kingdom Period, which includes Dynasties 3–6 (2663–2395 BCE). Recent analyses of predynastic burials have, however, identified the antibacterial applications with a chemistry much like that of more recent periods on funerary wrappings predating the Old Kingdom Period by 1000–1500 years (Jones et al. 2014).During the Old Kingdom Period, the predominant mastabas (subfloor tombs, with flat-roofed single-storey aboveground structure) gave way in Dynasty 4 to the monumental pyramids. Mummification became more sophisticated, with evisceration, followed by desiccation with natron and the application of resin to wrapping materials to resist microbial infestation and rehydration. The subsequent First Intermediate Period (Dynasty 11a; 2160–2066 BCE) is poorly known, as this was a time of political turmoil and strife. Generally consistent is the absence of features molded into the external surface of the resin that covered the body wrappings. The Nile Valley was reunited during the Middle Kingdom (Dynasties 11b–14; 2066–1650 BCE), a period of some variation in embalming treatment. In general, tissue preservation exceeds that of the Old Kingdom Period, perhaps because resin was applied directly to the body. Mummification was extended to elite individuals other than the ruling family, and transnasal craniotomy was introduced as a method for removing the brain via a channel driven through the cribriform plate from the nasal passages. Similar to the earlier period of political divisions, the Second Intermediate Period (Dynasties 15–17; 1650–1550 BCE) witnessed separate control of the Upper and Lower Nile valleys. Embalming procedures for this period are not well documented. The following New Kingdom Period (Dynasties 18–20; 1550–1069 BCE) is considered that of paramount

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achievement in the art and craft of mummification, perfecting the process begun at the end of the Middle Kingdom Period for royal individuals, including transnasal craniotomy, evisceration, desiccation, waterproofing with resin, cosmetic applications, and wrapping. Other elite individuals were treated more simply, without evisceration and craniotomy, but they were desiccated and often accompanied by quantities of grave goods. During the Third Intermediate Period (Dynasties 21–25; 1064–656 BCE), there was an initial effort to improve the physical appearance of the mummies’ subcutaneous packing. Soon, however, efforts began to simplify and the quality of treatment and thus preservation decreased. During the Late Period (Dynasties 26–31; 664–332 BCE), the quality of treatment was quite uneven, with most of the information coming from Nubia as a result of the Sudan Archaeological Surveys of 1907–1911, mitigating the impact of Aswan Dam elevation near the head of the First Cataract. The subsequent Greek (Ptolemaic) Period lasted from 332 BCE until the initiation of the Roman Period in 30 BCE, ending in 395 CE. Mummification processes shifted to place greater emphasis upon the appearance of the external aspect of the mummy bundle, rather than the person wrapped within. Also during this period, intensive mummification of all manner of nonhuman animals occurred in remarkable numbers, including ibises and other birds, crocodiles, mongooses, baboons, wolves, and domestic dogs. Cats, previously, taken to Bubastis for mummification, as noted by Herodotus, also witnessed increased mummification volume. Perhaps, as the political control shifted from the local to the external, Egyptians conspicuously endorsed traditional methods for evoking the power of familiar gods to maintain balance within their world. During the subsequent Roman period, portraits were introduced for mummies and their coffins. Christianity, with its belief in bodily resurrection, was not in theory opposed to mummification and certainly is not today. An edict by the Roman Christian Emperor Theodosius in 392 CE, however, forbade mummification, and it subsequently disappeared, leaving a remarkable record of shifting political, religious, social, and technological efforts to achieve a balanced world for both human and nonhuman persons living within the Nile Valley and surrounds. This extraordinary achievement, unsurprisingly, has excited no small interest within subsequent generations. Discussion now turns to efforts to understand ancient Egypt through the medium of their mummies.

European Tourism Begins: ~450 BCE To appreciate the power of mummy studies, one must indeed drill down into investigations of individual lives, but this is only a beginning. The simple or elaborate ceremony of interring the dead within a preserving environment, as well as any measures taken to ensure the eternal integrity of the corpse, provides insights into technology, scientific knowledge, and religious beliefs of the deceased individual’s world. More than that, the subsequent history of discovery, archaeological inferences, and public interest must be appreciated within their historical contexts, which in Egypt covers centuries, beginning with the historian-tourist, Herodotus

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(ca. 424–485 BCE), who visited North Africa during the Persian occupation, in the 27th Dynasty, mid-fourth century BCE. While early twentieth-century writers such as Grafton Elliott Smith (1914: 189) dismissed Herodotus’s account as a “strange jumble of dragoman’s tales,” today’s scholars generally accept his narrative, which describes three distinctive procedures for mummification, ranging from the elaborate removal of internal organs to less invasive dissolution by a solution injected through the anus. The final, least expensive technique involved purging the digestive system and then enclosing the body in desiccating natron, a procedure followed in all three cases. Herodotus also spoke of removing the brain through the nasal passages, partly through the action of the tool and through the introduction of an apparently emulsifying liquid. While most domestic and wild animals were simply buried, Herodotus notes that cats were taken to Bubastis and embalmed there (Herodotus 2006, Macaulay translation). This observation of animal embalming at a limited scale would conform to the time he was visiting Egypt, during the Late Period, but prior to Ptolemaic times, when effusive displays of animal mummification occurred. Diodorus Siculus (ca. 90–30 BCE), a Greek historian born in Sicily, compiled a series of volumes between 60 and 30 BCE that recounted events extending from mythic history to approximately 60 BCE or the beginning of Julius Caesar’s Gallic Wars. Volume 91 records Diodorus’s visit to Egypt, wherein he generally corroborated Herodotus’s characterization of the embalming process. He elaborated upon the roles of those engaged in body treatments, including a “scribe” who drew the location of the first incision on the left side of a corpse. The “slitter” who then first cut the body was said to run away rapidly to escape those who pursued, due to their dismay at the violence perpetrated upon the body. He also reported that animals, such as the cat, ibis, hawk, ichneumons, dogs, wolves, and crocodiles, among others, were also embalmed and buried with ceremony, as they would have been at the time of his visit (Diodorus Siculus 1933: 83, 91).

The Influence of Egyptian Mummification Upon Egyptian and Greek Medical Practice A number of writers have asserted that the knowledge of anatomy gained by practitioners in the course of mummification led to advances in human anatomical knowledge and in medical science by Egyptian physicians (Smith 1914).Ancient Greek writers glossed as “pseudo-Galen” argued that “it is from the dissection of dead bodies when they are embalmed that many treatments used in surgery came to be discovered by the first doctors” (Jouanna 2012: 16). Speculation has also emerged that the knowledge acquired through the Egyptian evisceration and embalming process may have influenced the practice of medicine in Greece either prior to or during existence of the intellectual center or Museum in Alexandria, founded by Alexander the Great in 331 BCE (Bernal 1987, 1991, 2006; Ghalioungui 1968, 1980; but see Lefkowitz and Rogers 1996; Lefkowitz 1997). Homer (eighth century BCE) in Book 4 of the Odyssey cited Egyptian medical practitioners as being “the wisest of men,” particularly in their knowledge of

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medicinal “drugs” (Jouanna 2012:8). In considering the possible transfer of anatomical knowledge from the Egyptian embalmers to Egyptian medical practitioners, however, it should be emphasized that the two were in very different priestly castes. Further, there is no real dissection or morphological observation implied in the methods begun during the Old Kingdom Period by which the embalmers cleared the viscera from the abdomen, and most frequently the excised brains were nearly or completely liquefied if extracted through the transnasal route (Rouyer 1809; Herodotus 2006). There is no indication that the removal of organs through the small apertures encouraged dissection or careful observation, with emphasis being placed instead upon the integrity of the body rather than the extracted tissues. The organization of the Egyptian medical profession, as witnessed by the relevant papyri (e.g., Ebers, Edwin Smith, and Chester Beatty VI), appears to be focused upon individual organs and the integration of incantations with the administration of medical procedures or decoctions. This magico-religious medical practice contrasted with the Greek system, which tended to be of a generalist nature and less infused with religious overtones. Egyptian doctors were legally bound to mandated procedures, which could lead to severe penalties including execution if not followed. The Greeks were freer to deviate from received wisdom and therefore innovate. Even in Alexandria, the Greeks associated with the Museum did not interact extensively with the Egyptian medical priesthood (von Staden 1989; Palter 1996). While it is clear that Greek anatomist-physicians, such as Herophilus (335– 280 BCE) and Erasistratus (310–250 BCE), benefited from their experience in Alexandria, influence from either Egyptian medical specialists or embalmers is uncertain, excepting the Egyptian Pharmacopoeia (Palter 1996). Von Staden (1989), in his Herophilus: The Art of Medicine in Early Alexandria, makes a compelling case for the lack of influence of Egyptian medicine, with its combination of religion, magic, and medical practice on both Alexandrian and earlier Greek medical practices.Further, Alexandria and its Greek inhabitants were not integrated with the Egyptian community; early Alexandria was Hellenistic, not Egyptian. Dissection was sanctioned in Greece and may have enjoyed only an ephemeral popularity, if that, in Alexandria. Greek theories of blood flow, for example, clearly differentiated venous and arterial flow and failed to posit an anal anastomosis, as did the Egyptians (von Staden 1989). Schwabe (1978) has asserted that the earliest Egyptian medical sophistication, as witnessed in the Kahun papyrus, occurred in the course of sacrificing animals, including dissections associated with embalming procedures. He argued that there were no real distinctions between Egyptian healers of animals and humans and that the identification of privileged knowledge with embalmers of humans is a Western construct built by the medical doctors who have been the profession most closely aligned with mummy studies across the centuries. When identical treatments of human and animal bodies are termed dissection and butchery, respectively, the bias is obvious. Also underscoring the link between animal and human medicines, Cave (1950: 569) has emphasized that the Egyptians wrote specific names for “heart, lung, kidney, bladder, stomach, bowel, uterus, vagina, diaphragm, spinal cord, brain, cerebral convolutions, and meninges,” writing “these terms with animal

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determinatives.” While Cave used this as an argument for Egyptians as early comparative anatomists, it seems more likely that the terms initially developed in the course of sacrificing wild and domestic animals.

The Medieval Period (476–1500 CE), the Renaissance (1300–1600 CE), and the Emergence of the “Mummy” While the first Greek and Roman historian-tourists reported the embalming process itself during the dynastic times, the term “mummy” was yet to be generated. Writing about the popular embalmed ancient bodies of Egypt, historian Karl H. Dannenfeldt (1916–2001) reported that: Unfortunately, the great interest in mummies and the money expended on their procurement were not directed to the acquisition of specimens for study and display, because by the time of the Renaissance mummy had become a highly prized drug. In medical tradition pissasphalt from the near east had long been recognized as a curative drug. In trade it usually was called mummia. Since the appearance of this natural pissasphalt was similar to that of the bituminous materials used by the ancient Egyptians in the mummification process, it became the practice to substitute the materials found in the bodies of the Egyptian mummies for the natural product. Abd Allatif, the Arabian historian and physician, writing in Cairo in 1203, remarks, ‘The mummy found in the hollows of corpses in Egypt, differs but immaterially from the nature of mineral mummy; and where any difficulty arises in procuring the latter, may be substituted in its stead.’ The next step was to substitute the dried flesh of the mummy for the hardened bituminous deposits found in the cavities of the body. The reputation of mummia, of any kind, as a drug was considerable. Avicenna (ibi Sina, 980–1037) found it useful in abscesses, eruptions, fractures, concussions, paralysis, hemicrania, epilepsy, vertigo, spitting of blood from the lungs, affections of thethroat, coughs, palpitations of the heart, debility of the stomach, nausea, disorders of the liver and the spleen, internal ulcers, and cases of poison. It was especially good for contusions. It was to be administered in decoctions of marjoram, thyme, elder, barley, roses, lentils, jujubes, cummin seed, caraway, saffron, cassia, parsley, oxymel, wine, milk, butter, castor, syrup of mulberries, etc. Dannenfeldt 1959: 17

Dannenfeldt (1959) went on to describe a large number of Egyptian mummies changing hands during the Renaissance, although antiquities were somewhat preserved by an active fraud introduced by entrepreneurs, who both manufactured mummies and remarked upon the cannibalistic inclinations of Christian Europe. In 1564, a merchant obviously engaged in fraud remarked to Guy de Fontaine, a physician to the King of Navarre, that he “marveled that the Christians, so daintymouthed, could eat the bodies of the dead” (Dannenfeldt (1959: 19). Writing more recently than de Fontaine, one of Napoleon’s savants, Vivant Denon (1747–1825; see below), also remarked upon trafficking in mummies. He reported that he had “promised an unlimited reward to any who should procure me one [mummy] whole and untouched; but the cupidity of the Arabs deprived me of this satisfaction; for they sell at Cairo the resin which they find in the belly and skull of these mummies and there is no preventing them from committing this violence to

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them” (Vivant Denon 1803: 242). Thus, the use of dark exudate or even tissues from mummies occupied a valued place in the Medieval, Renaissance, and early Modern Pharmacopoeia, adding to the mystique of Egypt, which was to be crystalized by the intellectual and material byproducts of Napoleon’s invasion of Mamluk Egypt in 1798. Theories that advanced the healing properties of “mummia” apparently persisted into early twentieth-century medicine. According to Babraj (2001: 13), “as late as1924 a respected German pharmaceutical company had on its price list a drug called Mumia cera Aegiptiaca. . . which was sold at 12 marks in gold per kilogram.” Such was the pervasive link of health to exotic Egypt.

Era of European Exploration and Discovery (15th–17th Centuries) The Colonization of the Canary Islands The economic, political, and religious reasons that drove Europeans to conquer the globe led to the discovery of places, such as the Canary Islands and Western South America, where mummies and artificial mummification procedures were compared to those of ancient Egypt, frequently assuming similarities and upon occasion cultural links that overstated or fabricated evidence. In the Canary Islands, for example, Pettigrew (1834: 232–233) reported a passage (1) from Glass’s 1764 History of the Canary Islands that cites an ancient Spanish document. This document reports a Guanche embalming procedure from the Island of Palma, one of the 13 Canary Islands, apparently settled by the Berbers in antiquity and then conquered with considerable effort by the Spanish in the fifteenth century. This is followed (2) by an 1802 statement drawn from the essays of the French Naturalist, Jean Baptiste Bory de St. Vincent (1778–1846): 1. . . . they opened it, and took out the bowels; then twice a day they washed the porous parts of the body, viz. the arm-pits, behind the ears, the groin, between the fingers, and the neck with cold water: after washing it sufficiently, they anointed those parts with sheep’s butter, and sprinkled them with a powder made of the dust of decayed pine-trees, and a sort of brush-wood, which the Spaniards call Bressos, together with the powder of pumice-stone; then they let the body remain till it was perfectly dry, when the relations of the deceased came and swaddled it in sheep or goats’ skins. . . 2. . . ..it was done by removing the intestines, washing the body with salt water, filling the large cavities with aromatic plants, and then drying the body either in the sun or by means of a stove. In some cases it is stated that corrosive liquids were merely poured down throat prior to the process of desiccation. This process is said to require fifteen or sixteen days, at the expiration of which time the body was bolded in goats’ skins, and then put in a kind of coffin, made out of one solid piece of wood, and this was deposited in a grotto excavated in the solid rock. . . these mummies or xaxos, as they are called are found in an upright position.. . . Pettigrew 1834: 232–233

More recent studies of the few Guanche mummies that remain after the worldwide dispersal that accompanied the virtual extinction of their culture note that not

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all the mummies appear to have been anthropogenically prepared; some mummification occurred naturally in the caves of the Canary Islands (Rodríguez-Martín 1995; Aufderheide 2003). This echoes Pettigrew’s (1834) observation that while “his” Guanche showed evidence of artificial embalming, an example observed by Blumenbach (1794) did not. Pettigrew (1834: 237) also underscored the “singular” similarity of Guanche mummification practices in relationship to those of the ancient Egyptians, despite the distance separating the two locations.

The Spanish Conquest of the South American Andes The Era of Exploration led to Spanish conquests within the Western Hemisphere, where the conquistadors encountered mummies and wrote about them with wonder and in reference to knowledge derived from the well-known Egyptian examples. The Andean landscape inhabited by mummies is exceptionally broad and deep, extending from some of the world’s highest mountains where Capacocha (“Qhapaq hucha” in Quechua) children and chosen women were sacrificed, by strangulation, suffocation, blows, or being drugged and entombed alive (Ceruti 2015), to coastal regions where the world’s earliest prepared bodies reflect the complex ideologies associated with fisherfolk and their small-scale (Chinchorro) communities that flourished millennia before Egyptian desert dwellers began to mummify their dead (Guillén 1992; Arriaza 1995; Arriaza et al. 2005). Mummified ancestors abound across the Andes, many simply preserved by arid environments associated with the world’s driest desert, the Atacama, or by dry cave microenvironments, found even within the humid and rainy cloud forests of Amazonas Province. Although the time depth of Andean mummification long predates Egyptian examples, recorded history is much more recent, beginning with the conquests and explorations of Spanish conquistadors, which led to the first early sixteenth-century encounters with mummies. Following failed expeditions in 1524 and 1526, Francisco Pizarro (1471/6–1541) began in 1532 his successful conquest of the Inca Empire (1438–1533). His forces arrived at the capital, Cuzco, in 1533, having executed the Inca Atahualpa in the northern highland city of Cajamarca the previous year. The Royal Inca mummies impressed the invading Spanish as embalmed “Lords,” who were accorded all the amenities they had enjoyed in life, including material and human support. Upon reaching Cuzco, Pedro Pizarro (1515–1602) – a younger first cousin of Francisco – reported surprise when he believed that he was going to speak to a living Indian and instead “was taken to a bundle, [like] those of these dead folk, which was seated in a litter, which held him and on one side was the Indian spokesman who spoke for him. . .”(Pizarro 1921 [1571]: 205). Pedro Pizarro also commented upon many people in Cuzco who served the mummified Lords, “for each day they took them all out into the plaza and sat them down in a row, each one according to his antiquity, and there the men and women servitors ate and drank, and . . . they burned here everything which they had placed before the dead in order that he might eat of the things which they eat, and here in this fire they consumed it” (Pizarro 1921 [1571]: 251).

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Most of the chroniclers assumed that the mummies had been “embalmed” in some manner. The indigenous nobleman-chronicler Felipe Guamán Poma de Ayala (ca. 1535 – 20 years of age) and for approximately 10 years in young individuals (5–15 years old) (Hedges et al. 2007). As for teeth, whereas their formation and eruption periods vary with tooth type, they are fully formed in people by approximately 15 years of age and are not remodeled and thus retain their isotopic information from that young age (Hilson 1996). Soft tissues with rapid turnover rates are useful in examinations of short-term diets. For instance, the remodeling rates of skeletal muscle and skin collagen in young men are 0.016  0.002%/h and 0.037  0.003%/h, respectively (Babraj et al. 2005). The time spans of isotopic information are estimated as 8–9 months for skeletal muscles and 3–4 months for skin collagen, although this information varies by age, sex, and diet. Hair has a more short-term temporal resolution, because it grows at a rate of approximately 1 cm/month. Additionally, hair is not resolved and absorbed by turnover but is retained outside the body, thus maintaining a sequential isotopic transition. However, time is needed for any dietary change to be reflected in the hair, due to the amino acid pool in the body (Zlotkin 1985; Sandford and Kissling 1993). In fact, it takes about 12 months for the carbon isotope and about 5 months for the nitrogen isotope to achieve an isotopic equilibration following a dietary change (O’Connell and Hedges 1999).

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Isotopic Analyses of Mummies The Ice Man (Ötzi) is one of the most famous mummies. In 1991, he was found at the Hauslabjoch pass, located between Ötztal (a valley in North Tyrol) and Vinschgau (a valley in South Tyrol). The male mummy, who had lived sometime between about 3370 and 3100 BC (Kutschera and Müller 2003), has retained various information on historical and ancient lifestyles. To decipher his life history, researchers have conducted many investigations, including isotopic analyses. A hair analysis using carbon and nitrogen isotope ratios revealed his C3-resource-dependent diet and low animal-protein consumption (Macko et al. 1999). A bone analysis using carbon, nitrogen, and strontium isotope ratios and transition elements reported the possibility that his domicile had been located in the southern elevated areas of the Ötztal Valley and that he seasonally migrated between the two regions through the Hauslabjoch pass (Hoogewerff et al. 2001). Additionally, analyses of tooth enamel, bone, and the contents of his intestines via the isotope ratios of oxygen, strontium, and lead suggested that he had spent his childhood in the southeast area of the region where he was found (Kutschera and Müller 2003). Studies such as these provide fascinating clues to the lifestyles of peoples who lived approximately 5000 years ago. Egyptian mummies also have a long research history. Iacumin et al. (1996) performed carbon, nitrogen, and oxygen isotope analyses and reported an ancient diet of high C3-resource consumption based specifically on bioapatite and collagen from a mummy’s bone excavated in the Asyut region. They also analyzed the hair and skin of Egyptian mummies whose excavation regions were uncertain. They concluded, based on the isotopic difference between the No. 8 mummy’s soft tissues and bone, that he had lived in another country and died upon his arrival in Egypt. In the meantime, analyses of mummies of the Roman-Byzantine period (400–700 AD) from the Kharga Oasis indicated a characteristic C3-resource-dependent diet with few seasonal variations (White et al. 1999). They noted that this result was in contrast to the findings from Nubian mummies of Wadi Halfa, whose diet showed a seasonal change between C3 and C4 resources. A recent study using carbon, nitrogen, and sulfur isotope analyses of tooth enamel and the bone suggested a C3resource-dependent diet, a dietary change associated with the introduction of olive oil, and a low consumption rate of marine and freshwater fishes (Touzeau et al. 2014). Studies entailing immigration estimations also have been reported. For instance, strontium, carbon, and oxygen isotopic analyses for the estimation of migration using tooth enamel suggested that some individuals found in Nubia had been immigrants during the New Kingdom period (Buzon et al. 2007; Buzon and Bowen 2009) (Fig. 2). The soft tissues of Andean mummies, especially the hair, have been maintained in good condition due to the fact that those mummies had been naturally formed in a dry and/or icy environment. Therefore, many relevant studies on isotopes have been reported. In the coastal areas of Peru and Chile, for example, there are many desiccated mummies that were formed by burial in a desert environment. Hair analyses on mummies from the Ilo area and Moquegua Valley in the southern coastal region of Peru (1000–1450 AD) showed a temporal dietary change (Knudson et al.

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Fig. 2 Migration estimation for Egyptian mummies based on strontium isotope ratio (Buzon et al. 2007). Some tombs show different 87Sr/86Sr values (estimated via ancient and modern animals and the burial matrix) from the local ones

2007). The isotopic data shows that while some individuals had maintained a stable diet, some others had temporarily increased or decreased their marine resource consumption. Moreover, some individuals had moved between the river mouth and inland areas 50 km away from the sea, suggesting that there was active exchange between inland and coastal groups. Compared with hair samples showing short-term diets, the carbon and nitrogen isotope ratios of bone samples reflective of long-term diets have indicated diversified intra-site diets and a positive correlation between marine resource consumption and distance from the sea (Tomczak 2003). This dietary variety also was reported based on δ88/86Sr, which is the indicator of discrimination between marine resources and/or terrestrial resources (Knudson et al. 2010). This 87Sr/86Sr ratio suggested that there had been nonlocal individuals who came from outside of the Moquegua Valley (Knudson and Buikstra 2007), which fact would be indicative of active movement of people from within the valley as well as from distant regions. A mummy of an Andean traveler found in the Atacama Desert, northern Chile, also indicated that he had taken a round trip between inland and coastal areas (Knudson et al. 2012). Although high marine resource consumption was proved based on bone samples, a temporal decline of carbon and nitrogen isotope ratios in hair samples implied a short-term trip to the inland areas. Meanwhile, dried mummies also have been discovered in graves on cliffs and in caves in mountainous areas. In these cases, most of the mummies had lost their hair

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in the years since their burial but retained skin and muscles. Finucane (2007) investigated six mummies found in a cave of the Ayacucho region in the southern highlands of Peru. He analyzed bone, skin, and muscle samples with reference to carbon and nitrogen isotope ratios in order to estimate their diets. The mummies showed a high contribution of C4 resources and the possibility of fertilizer utilization for maize agriculture, although the dietary variations from different tissue types were unclear. One of the most interesting topics of Andean mummy analyses is sacrificed children that were offered in ritual events by the Inca. The child mummies had been frozen for 500 years on the top of a mountain. Therefore, they were perfectly preserved, looking almost as if they were alive; consequently, hair samples were available for analysis (Fernández et al. 1999). Wilson et al. (2007) investigated five samples: three child mummies and one hair bunch in a bag discovered at the Llullaillaco Volcano and a mummy from the Sara Sara Volcano. They simultaneously analyzed carbon, nitrogen, oxygen, and hydrogen isotope ratios, revealing that the Llullaillaco mummies had undergone extreme isotopic transitions a year before their death (Fig. 3). Increases in the carbon and nitrogen isotope ratios could be interpreted as enhanced consumption of C4 resources and terrestrial animals, given the long distance from the sea. The oxygen and hydrogen isotope ratios also were found to have risen at 4–5 months prior to death. The authors considered that this could be attributed to the isotopic difference in the available water associated with high altitude. Additionally, they noted that the isotopic transition might reflect changes in cooking or the consumption of maize cultivated in the lowlands. The girl mummy at the Sara Sara volcano showed no change in the carbon and nitrogen isotope ratios, while the sulfur isotope ratio in her hair had fallen a year before her death. This might be explained by a decrease in marine resource consumption. Similarly to the Llullaillaco mummies, the Sara mummy also showed increased oxygen and hydrogen isotope ratios, suggesting the possibility that children consumed specially cooked food as part of their ritualistic sacrifice. This suggests that whereas common burial mummies reflect typical ancient diets and life history, ceremonial mummies can indicate ritual dietary changes. Both data types of course are essential information for any relatively comprehensive understanding of historical lifestyles.

Conclusion As discussed above, isotopic analyses have succeeded in reconstructing human life histories for both short and long periods of time. Researchers face the challenge of determining where and how historical (e.g., ancient) humans lived and why they died in specific places, using multiple isotopes such as carbon, nitrogen, sulfur, oxygen, lead, and strontium. Carbon, nitrogen, and sulfur are applied mainly for dietary reconstruction, and oxygen, lead, strontium, and sulfur are used in estimations of birthplace locations and migrations. When one or more isotope ratios are added to an analysis, additional means of revealing the life history of historical

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Fig. 3 Isotopic analysis of hair samples of Inca-sacrificed child mummies (Wilson et al. 2007; Copyright 2007 National Academy of Sciences, USA). The isotopic values exhibit temporal changes in their diet, lifestyle, and living environment

people are at hand. For example, it is possible to learn an ancient society’s detailed resource strategies, such as animal husbandry, agriculture, tool production, trading, and trading routes, by combining isotopic information from animal bones, food

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residue, and ceramics. As novel applications of new elements continue to be attempted, the future of this field will expand as additional types of elements and of analytical artifacts become available.

Cross-References ▶ Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection

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Radiology Applications in Mummy Science A Practical Approach

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Ronald G. Beckett and Gerald J. Conlogue

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Brief History of Radiology as Applied to Mummy Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification Processes and Preservation of Bony Material and Soft Tissue: A Justification for Paleoimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Paleoradiology/Paleoimaging: Applications in Bioarchaeological Studies . . . . . . . . . . . . . . . . Radiological Modalities and Complementary Paleoimaging Methods . . . . . . . . . . . . . . . . . . . . . . . . Field Imaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Advanced Imaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Value-Added Data: What Can be Learned . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Practical Guide to the Application of Radiological Methods to Mummy Science . . . . . . . . . . . . Study Designs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Limitations and Opportunities in Paleoimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Challenges Related to Technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Challenges to Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Interpretational Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Medical Versus Bioarchaeological Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Diagnosis by Consensus Comments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Training Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Looking Forward: Opportunities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . New Emerging Technologies/Instrumentation/Tele-Interpretations . . . . . . . . . . . . . . . . . . . . . . . . Standards in Paleoimaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Bioarchaeology of Care Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chapter Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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R. G. Beckett (*) Biomedical Sciences, Bioanthropology Research Institute, Quinnipiac University, Hamden, CT, USA e-mail: [email protected] G. J. Conlogue Bioanthropology Research Institute, Quinnipiac University, Hamden, CO, USA © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_46

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R. G. Beckett and G. J. Conlogue

Abstract

Beginning with a brief historical perspective, this chapter introduces the subject of radiology and how it has been applied in mummy science. Radiology is examined regarding its current value in bioarchaeological studies including imaging data collection from mummified remains, archaeological contexts, and associated artifacts. The chapter differentiates between radiography (image gathering) and radiology (image interpretation) in terms of how each is critical to making sense of past lives. A brief review of the varied radiologic modalities is discussed with a consideration of “value-added data” regarding a given mummy study. Each radiologic method is framed in what it can do in terms of what can be learned, as well as what it cannot do with regards to technological and methodological limitations. The value of individual mummy case studies as compared to larger population studies is compared and contrasted. The chapter presents the reader with a practical guide for the application of radiological studies ranging from field data collection and interpretation to the use of advanced imaging modalities using brief case examples. The use of triage as a decision-making strategy regarding specific steps of medical imaging and the methodologies used is discussed. Often the use of medical imaging employs an approach approximating a medical model of application. This chapter offers an argument for expanding the conceptual basis for imaging studies of mummified remains to employ an anthropological/archaeological approach. Further, the chapter describes varied paleoimaging methods complementary to radiology from a value-added perspective. The limitations of technologies used in radiology as well as the limitations associated with data collection and data interpretation are revisited and are explored in greater depth. The diagnosis by consensus construct is presented arguing that many perspectives improve our vision and thus the interpretability of the data. The chapter finishes with a look to the future of radiology in mummy science studies from (1) a technological point of view, (2) the continued development of standards, as well as (3) from a data application perspective including the application of imaging data to the Bioarchaeology of Care model regarding the lived experience of past peoples and populations. Keywords

Radiology · Paleoimaging · Paleoradiology · Medical imaging · Bioarchaeology · Mummies

Introduction Brief History of Radiology as Applied to Mummy Science Radiology is often defined as the science dealing with X-rays and other high-energy radiation as applied in the diagnosis and treatment of disease. When radiologic methodology is applied to archaeological remains, it is referred to as paleoradiology.

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Paleoradiology applied to mummified human remains is a powerful adjunct to data collection and therefore to the understanding of the lived experience of past peoples. Paleoradiology is one of many nondestructive imaging methods belonging to a larger group of imaging approaches known as paleoimaging. Among those imaging methods under the paleoimaging umbrella are photography, videography, radiology, endoscopy, ground penetrating radar, LiDar, X-ray fluorescence (XRF), and advanced medical imaging including computed tomography (CT), multidetector computed tomography (MDCT), dual energy computed tomography (DECT), micro-CT, and magnetic resonance (MR). All are key contributors to the understanding of past peoples and their cultures (Lynnerup 2010). This chapter will focus primarily on medical imaging modalities and their application in mummy science studies. From a historical perspective, every paleoimaging modality has been applied to bioarchaeological contexts soon after its technological development. One of the first and the most useful tools in preserving data and presenting a representation of the original context in bioarchaeological studies is photography. Photography and anthropology have a very long relationship with field scientists bringing images of cultural and archaeological significance to scientific meetings, journals, and to the world (Collier and Collier 1986; Edwards 1992). Paleoimaging continued to expand based on the invention of newer technologies in the fields of photography, industry, and medicine. As in the past, as soon as new imaging technology is developed, it is applied to bioarchaeological research. Not long after the discovery of the X-ray in 1895 by William Roentgen, X-rays were soon used to image mummified remains. In March of 1896, Carl George Walter Koenig, a German physicist, published a paper “14 Photographs with X-rays Taken by the Physical Society of Frankfurt am Main.” Koenig presented the first X-rays involving mummies, an Egyptian mummified cat and the knees of an Egyptian child mummy (Beckett and Conlogue 2010). In 1977, the first CT scans were conducted on mummified remains (Lewin and Harwood 1977). Not long after the introduction of the flexible fiberoptic endoscope in the early 1970s, reports related to its application in bioarchaeology began appearing (Tapp et al. 1984). Early reports on endoscopy followed the medical model by which the application was limited to targeted biopsy, much like procedures in living patients. As with other paleoimaging modalities, endoscopic applications later began following the anthropological and archaeological research approach (Beckett and Conlogue 1998; Beckett and Guillén 2000) enhancing its contributions. Magnetic Resonance Imaging (MR), first thought to be useless in dry mummified subjects (Notman 1983), has found its way into the paleoimaging arsenal (Conlogue et al. 2008, 2010; Öhrström et al. 2013). Each new generation of current imaging technology is soon applied to mummified remains. It is beyond the scope of this chapter to offer detail regarding each of the many paleoimaging modalities. Rather a brief presentation of the modern paleoimaging technologies available to researchers will be described followed by a discussion of current applications and challenges within the bioarchaeological context. For the reader desiring further in-depth descriptions of each paleoimaging modality, the authors refer them to Beckett and Conlogue’s Paleoimaging: Field applications for Cultural Remains and Artifacts (CRC Press 2010) as well as in upcoming texts on the subject (Beckett and Conlogue 2020; Beckett et al. 2020).

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As described, paleoimaging then is a broad field of study utilizing the application of various imaging modalities intended to capture images of the context, external appearance and internal context of cultural remains and artifacts. An initial broad data collection approach or survey is employed to assure that the data collected can be assessed within the overall bioarchaeological context. The findings from the imaging survey often inform the decision to move on to advanced imaging modalities that can further explore bioarchaeological questions. Paleoradiology is a key modality in the paleoimaging data collection and interpretation continuum. It is important to note that in mummy science studies, radiography is a separate entity within the paleoradiology definition. Radiography is a critical part of paleoradiology, in that it refers to the process of data collection which involves awareness and expertise regarding the manipulation of the various parameters and approaches to obtaining useful images from complex instrumentation that would result in reliable and interpretable data. Radiography, or paleoradiography in this context, requires a specialized set of skills, knowledge, and critical thinking in order to provide information that will inform later interpretations. We will discuss this aspect of data collection in some detail later in this chapter. Mummy science is an important aspect of the broader concept of bioarchaeology. Bioarchaeology is defined as a reconstruction of lives using the available evidence that supports an understanding of individuals and groups of past peoples within the context of their lives and their interactions with their environments. Further, bioarchaeology is informed by a variety of sciences and specializations including biology, anthropology, anatomy, nutrition, paleopathology, epidemiology, forensics, medicine, paleoimaging, and others. It is the intent of bioarchaeological studies to form as complete a representation of humankind’s journey on earth using reliable evidence and considering as many aspects as possible in the analysis phase. In order to better understand the application of medical imaging modalities in bioarchaeological research, specifically mummified remains and artifacts, it is important to have a clear understanding of the subjects.

Mummification Processes and Preservation of Bony Material and Soft Tissue: A Justification for Paleoimaging Conventional belief is that mummies are a product of ancient Egypt, while in fact mummies have been found on every continent except Antarctica. The oldest known intentional mummies are of the Chinchorro culture of the Atacama Desert in what is now Southern Peru and Northern Chile. Chinchorro mummification pre-dates Egyptian efforts by around 2000 years (Cockburn et al. 1998). In the global context, mummification processes are highly varied. Arthur Aufderheide (2004) in his book, the Scientific Study of Mummies, offers an organizational scheme for the classification of mummification processes. His mummification taxonomy includes four distinct categories. These categories are spontaneous, anthropogenic, spontaneousenhanced, and intermediate mummification. Spontaneous mummies are those that are produced as a result of nature, more specifically, the impact of the environment where the body was interred. There are

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many environments and conditions that will halt or greatly impede the normal process of decomposition resulting in mummified remains. Spontaneous mummies are also referred to as natural, unintentional, or accidental mummies. Anthropogenic mummification, also called artificial mummification, is represented by individuals who were intentionally mummified with a concerted effort to maintain the human shape and form. While the Egyptians are perhaps the most well-known for this classification of mummification, cultures from across the globe used various methods to preserve their dead. Cultures such as the Ibaloy of the Philippines and the Anga of Papua New Guinea used techniques that produced excellent body preservation and which were very different from the Egyptian method (Beckett et al. 2011). The next classification is called spontaneous-enhanced mummification. Recall that in the spontaneous group, chance played the biggest role. In spontaneousenhanced mummification, however, a culture realized that their deceased were being mummified by their burial practices in terms of where they were buried. The culture then intentionally buried their dead in that or a similar environment in an attempt to mummify them, thus reducing chance and enhancing the spontaneous forces. Though there was intentional placement in the right environment, there were no additional treatments as seen in the anthropogenic group. The final classification on Aufderheide’s taxonomy is intermediate mummification. Here, there is a temporary preservation of the body allowing for ceremonial rituals to be planned and conducted. The intermediate type of mummification was often practiced to allow relatives at a distance to travel to the ceremonial site and participate in burial traditions (Aufderheide 2004). One can even consider modern funerary embalming practices that include a wake period to fall into this classification. Within each of these four broad categories, various processes have been used to produce preserved human and animal remains. We will review several of these processes as each method creates specific challenges and dictates approaches to the application of medical imaging. Spontaneous mummification results as a product of the environment in and around the deceased individual. Basically, any environment that will impede or stop the forces of decomposition has the potential to produce a mummy. The environment may bring about rapid desiccation or create a chemical alteration of the remains that will be hostile to the processes of decomposition. When imaging mummies of this category, one may find varied degrees of decomposition. In most cases, soft tissues have desiccated often losing their morphology and spatial relationship within otherwise well preserved integument and skeletal systems. The interpretational challenges are great when analysis is made in the context of normal hydrated and living individuals. Caves often exhibit the conditions that will produce a spontaneous mummy. Caves can be warm or cool dry environments and, when coupled with ventilation, can stop the forward progress of decomposition by enhancing the rate of desiccation leaving mummified remains. Individuals who were buried in the ground in direct contact with certain soil conditions may also lead to natural desiccation.

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Subterranean crypts and tombs as seen throughout Europe may produce conditions that are ideal for the rapid desiccation of humans and animals. The tomb environment is often cool and dry and if the walls are made from limestone, the dehydration impact can be much greater. Excellent examples of crypt-induced spontaneous mummification can be seen in the mummies of Guanajuato, Mexico and in Urbania, Italy, at the Church of the Good Death (Fornaciari 1982; Aguilar 2009). Special environments can also produce spontaneous mummies. Persons buried in soils rich in alkaline constituents such as those produced with ash as a part of the soil matrix can produce a spontaneous mummification process known as saponification. In this setting, the fats of the body coming into contact with the lye produced from water and ash are converted to adipocere. The formation of adipocere halts any decomposition that has begun. Two well-known examples of saponification are the Soap Man at the Smithsonian Institution in Washington DC and the Soap Lady at the Mutter Museum in Philadelphia (The Soap Lady 2001). Another special environment that can produce natural mummies is that of burials in acidic soil conditions such as those found in peat bogs. Decomposition cannot proceed when the environment is anaerobic, acidic, and under pressure. Temperature also plays a role in the preservation process. These conditions create an environment in which microbial activity associated with decomposition cannot function. Generally speaking, the skin is “tanned” and soft tissues are preserved, but the acidic environment leeches calcium from the skeletal structures (Lynnerup 2010). Upon scientific study, the decalcified bones, in particular the skull, have caused interpretational challenges and create particular challenges for paleoimaging methods. Additional special environments that can produce mummified remains include soils high in heavy metals, such as arsenic, and extreme cold conditions. The extreme cold can produce either frozen or freeze dried human remains. Each of these categories of mummies creates different challenges for medical imaging. Great care must be taken to not allow any thawing of the individuals and great care must also be taken to avoid contamination of the remains with microbial flora. In the case of the presence of certain potentially toxic substances like arsenic, care must also be taken to avoid contamination of the imaging suite. Anthropogenic intentional mummification presents in various forms. The most widely known is that of the Egyptian method of mummification where the individual is eviscerated with the body being systematically desiccated by using Natron, a saltlike substance that draws the body fluids into itself. Once saturated, the Natron is discarded with a fresh volume reapplied until full desiccation is achieved. Packets of the desiccant have also been placed inside the body cavities of these mummies. In each of the cultures that practice intentional mummification, materials native to their environment are employed. In the fringe highlands of Papua New Guinea, the Anga use local plants and a smoking method to preserve their honored dead (Beckett et al. 2011; Beckett and Nelson 2015). The Ibaloy of the Kabayan region of Luzon, Philippines, also use a smoking method that is different from the Anga and uses materials endemic to their environment. Medical mummies have been produced using a variety of substances including heavy metals (Conlogue et al. 2005), waxes, and varnishes and silicone in the more modern plastination.

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In many cases, mummies are interred with cultural artifacts that have a wide variety of intended meanings and purposes. Food may be entombed with the deceased so that they may have nourishment in the afterlife. Objects may be placed that represent who the individual was in a given culture or what their occupation was. When medical imaging is applied, particularly in the case of CT and MR scanning, metallic objects can pose obvious problems, such as streak artifact in the case of CT imaging, and must be considered during data collection and interpretation. These artifacts and their spatial orientations hold great meaning for the bioarchaeologist and care must be taken to gather imaging data that will allow a thorough analysis. Consideration of these varied preservation processes and the resultant mummification along with the many cultural practices is critical to the application of medical imaging. Understanding of these and other variables help to develop an imaging strategy that allows for the collection of data that can best serve the science of bioarchaeology.

Paleoradiology/Paleoimaging: Applications in Bioarchaeological Studies Paleoradiology then is the application of imaging methods to collect and record visual data of cultural remains and artifacts intended to allow multidisciplined interpretations within the bioarchaeological context. Those who participate in such studies need to be well versed in the varied mummification methods and the meanings of the cultural data collected. Paleoradiology interpretations are best conducted using a variety of scientists whose knowledge contributes to the broad understanding of the meaning of the imaging data collected within the context of a given culture and their environment. This will be discussed later in this chapter. Recall that while paleoradiology is a term used to describe radiographic imaging modalities as they are applied in the bioarchaeological context (Chhem 2006; Chhem and Brothwell 2008; Beckett 2014), other medical imaging modalities, such as endoscopy, complement radiographic data collected and may add meaning to the interpretation of the images. The relationship is a critical one and will be described later in this chapter. Imaging methodologies are very helpful in answering specific anthropological and archaeological questions. The data derived from imaging methods are of significance particularly when the mummified remains are within a burial chamber, enclosure, or wrapped in textiles, bandages, or other presentations that decrease the efficacy of macroscopic visual inspection alone. Given that imaging can “see what can’t be seen,” a variety of questions can be answered adding to collected data and assisting in a reconstruction of lives. Paleoradiology is used to answer a multitude of bioarchaeological questions about cultural remains including assessment of the age at death, sex, dental condition, pathological conditions, trauma, biomechanical stress, burial and funerary practices, artifacts, and other burial “inclusions” and associations. For the most

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part, paleoradiology is a powerful nondestructive data collection methodology. If these data are collected within the construct of bioarchaeology and employing complementary imaging methods, considerable meaning can be made that can greatly enhance our understanding of humankind’s journey on earth. Regarding the archaeological perspective, imaging can provide data that may allow interpretations related to the use of and type of artifacts present, evidence of both funerary and burial practices, as well as any medical interventions and potentially assist with understanding and defining the temporal context of the remains. Imaging may also guide additional research by assisting in target biopsy for pathological or genetic analysis and/or artifact retrieval. Paleoradiologic data collection methods are ideal for the bioarchaeological setting. The methods are nondestructive and can help avoid the need to conduct a destructive autopsy on the remains which would render them unavailable to future researchers and to future nondestructive technologies. Documentation of ancient remains and artifacts through medical imaging methods is essential. Collecting and archiving accurate data from a subject or group of subjects is crucial to current and future study. The significance in data archiving is that mummified remains are not always stable and are at risk of deterioration over time. In some cases, the images are all that remain. Thus paleoradiology, along with complementary paleoimaging methods, employ advanced medical technologies to capture images in order to provide evidence to help bioarchaeologists answer simple and complex questions about specific individuals or populations. These images hold great potential in our attempt to enhance understandings about ancient lives and their relationships to anthropology.

Radiological Modalities and Complementary Paleoimaging Methods We have established that medical imaging in the bioarchaeological context has a rich history and that the continued evolution of this relationship is demonstrated in the fact that when new or updated imaging technology is developed it is soon applied to ancient materials. Medical imaging in such contexts can be subdivided into two broad categories: field (contextual) imaging and advanced imaging. The distinction is an important one in that field imaging has the potential to preserve the original bioarchaeological context while advanced imaging generally requires moving the object of study away from the original bioarchaeological context (Conlogue et al. 2004b). It is important to make informed decisions about applying advanced imaging modalities in that the potential additional data that may be derived must be considered in light of the potential risks of moving the object of study. As we examine the varied modalities on imaging sciences in the context of bioanthropology, bioarchaeology, and paleopathology, it is important to understand some of the fundamental differences in how these modalities capture an image. In this way we can understand the value of each method as each has the potential to “bring to light” images that the others do not. Photography and endoscopy primarily

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utilize the visible light portion of the electromagnetic, EM, spectrum. Thus, they provide images of what can be seen provided there is access to those targets. However, there are a number of applications where both of the modalities employ alternate light sources that are on either side of the visible light spectrum, infrared and ultraviolet, to identify structures beneath the surface. This allows us to see a bit deeper and broader as we focus on those targets of special interest. Beyond ultraviolet is a portion of the spectrum identified as X-ray. The science dealing with the production and utilization of that segment of EM is termed radiology. Radiologic methods then allow us to “see” even deeper than what is provided by photography or endoscopy in that with radiology we are able to see through objects to varying degrees depending on radiographic parameter manipulation. Advanced imaging methods such as multidetector computed tomography (MDCT), dual energy computed tomography (DECT), and microcomputed tomography (micro CT), all utilize the X-ray section of the EM. There are lower energies within that spectrum as well that prove useful in our study of mummies and associated artifacts. The lower energies in that region make possible technologies such as scanning electron microscopy (SEM) and X-ray fluorescence (XRF), in determining structure of objects or analysis of composition. Here, paleoimagers are able to determine what an object of interest is made up of from an elemental analytic perspective. Beyond the X-ray portion of the spectrum is the highest, most penetrating energy segment identified as gamma, utilized in clinical applications for treatment of cancers and in industry to image very dense materials. While in mummy science we are not “treating” the subjects of a given study, there are settings in which powerful gamma imaging can provide information as to what may be inside a very dense archaeologic object. Recall that in medical and industrial applications, the individuals who operate the equipment are known as radiographers. In the former, the individuals who interpret the resulting images are known as radiologists, medical doctors with specialized training.

Field Imaging Field imaging is defined as that medical imaging that can be applied at or near to the original context. Thus, any imaging applied outside of a standard imaging facility such as a hospital or free standing imaging facility is considered field imaging. Such locations may include but are not limited to the point of discovery, cultural interment practices such as tombs, caves, and niches, as well as museums and private collections. While such modalities as photography and handheld XRF analysis can clearly be used in the field, we will focus our discussion on those technologies most often associated with medical applications. The two main technologies that fall into this category are radiography and endoscopy. These two technologies have proven to be well adapted to collect specific data at the point of origin of the subject negating the need to risk damage to the object by moving it to an imaging center (Beckett and Conlogue 2010). Onsite analysis can answer a wide variety of the bioarchaeological

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Fig. 1 Photograph of a gas-powered generator enabling portable X-ray application to mummies within the caves of the Ibaloy culture (Credit: Ronald Beckett)

questions previously described. In order for the technology to be applicable to a variety of field settings, such as tombs, caves, and remote environments, it is necessary to modify the instrumentation in a manner that renders it portable and flexible in order to meet the requirements of fieldwork (Fig. 1).

Radiography in Field Settings Ever since the discovery of mummies, scientists have been finding ways to better understand peoples from past cultures. It is no surprise that medical imaging plays an important role in the nondestructive analysis of mummified remains and skeletal material. At this point, it is important to recall the historical application of imaging to mummified remains as well as a consideration of its evolution (Beckett and Conlogue 2010; Beckett 2014). Generally speaking, most of these early mummy studies were accomplished by transporting the remains to a clinical imaging facility. Moving remains of what are often fragile and unstable any distance may result in irreversible damage to the individual. Although the external features might not indicate any change, structures not visible but that are within the remains, such as residual brain material or even the heart, may shift from its original position. Therefore, the safest approach would be to radiograph the remains close to the recovery site or at a location, such as a museum, where the remains are conserved. If the studies are conducted outside of an established imaging facility, it is appropriate to consider them field radiographic studies.

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There are two studies that provide excellent examples for approaches to field radiography. The first involved transporting the imaging equipment to a Museum. Harris and Weeks began their studies of the Royal Mummies in the Cairo Museum in the 1960s (Harris and Weeks 1973). Initially they used an isotope source to obtain the images, but later switched to a conventional dental X-ray tube because of possible radiation hazard to personnel as well as lower image resolution from the isotope source. Since the equipment was mobile, there was very little movement of the mummies to acquire the images. Since they were working in the large city of Cairo, if there was anything that might be needed like additional X-ray film or chemicals to process the images, it could be located and procured relatively easily. The second case illustrates establishing a field facility in a remote location. Notman and Anderson planned to radiograph the mummified remains of three individuals that died during the Franklin Expedition in 1845 (Beattie et al. 2004). However, the bodies were buried on Beechey Island in the Canadian arctic. Therefore, everything that would be required would need to be brought with them. There would be no opportunity to “pick-up” something they might suddenly need. In addition, they would need to improvise or be creative in solving problems that might arise. Since the bodies were radiographed only a few meters from where they were interred, there was minimal disruption of the remains. In both cases, but particularly with the latter, the use of film as a recording media presented major problems. Not only were darkrooms a necessity for loading and unloading film holders or cassettes, but lightless enclosures were also required to chemically process the exposed films. Film had the added disadvantage of not being able to correct an exposure once it was taken. Under or overexposed radiographs required repeat exposures following the re-adjustment of setting. Between 1998 and 2001, Conlogue et al. (1999, 2004b) established a field radiographic facility in the cloud forests of north central Peru to radiograph 208 mummy bundles. Similar to the expedition into the Canadian arctic, everything including film and chemicals were transported to the site. During the study period, 1080 radiographs were taken of the 208 mummies, but with an average of less than four films per hour. The productivity measure of “images per hour” is an important aspect to consider as time in the field is precious and the more information that can be collected in the shortest period of time allows for a greater degree of potential interpretability. By the 1990s radiographic film was being replaced in the clinical setting by one of two types of digital recording media. The first was direct digital radiography (DR) where the digital image receptor was directly connected by a cable to the processing unit of the computer. Within seconds of the exposure the image appeared on the monitor. With the second type of system, computed radiography (CR), a photostimulable phosphor plate is placed into a holder that resembled the typical X-ray cassette. Once the exposure is taken, the plate is placed into to device called a reader. Within the device, the plate is removed from the cassette and scanned by a laser which releases the energy trapped in the phosphor layer. As the laser scanning releases the energy captured during the exposure it transforms the data into an image that appears on a monitor in approximately 2 min or less from when the

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plate went into the reader. The plate is then “cleared” by fluorescent light, returned into the cassette and ready for the next X-ray exposure. The common advantage of both DR and CR versus film was that the digital media had much wider latitude or dynamic range than X-ray film. A wide latitude means that, for example, on an image of a mummy, a broader range of densities, including bone, skin, and even the foam supporting the remains would be demonstrated (Conlogue et al. 2011a). While on a radiograph using film as the recording media, only the bone may be visualized. In addition, with the digital system the X-ray exposure setting did not need to be as precise, almost totally eliminating the need for repeat exposures. Without the need to spend time repeating exposures and the 6–10 min required to hand process the X-ray film, the total number of X-rays taken per hour dramatically increased. Between 2010 and 2013, Conlogue et al. (2011b) and Conlogue and Viner (2014) established a temporary imaging facility in the crypt under St Bride’s Church on Fleet Street, London, England (Fig. 2). Using a DR system, the skeletal remains of 211 individuals were radiographed with an average rate of 15 images per hour or almost 4 times as many exposures that were possible with film in the Peruvian study. Using a CR system to radiograph, a collection of 34 mummies at the Museo Jacinto Jijón y Caamaño in Quito, Ecuador, in January 2013, Beckett et al. (2013a, b), over a five-day period, totaling 31 h, acquired 131 images or an average of about 6 radiographs per hour. On closer inspection, the average per hour imaging rate increased from approximately 4 per hour on the first day to about 8.5 X-rays per

Fig. 2 Digital X-ray system used in the crypts under St. Bride’s Church, London, UK, demonstrating the applicability and flexibility of field imaging (Credit: Ronald Beckett)

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hour on the fourth day. The increased rate of throughput can be attributed to the newly assembled team’s familiarity with the overall operation, including a system to create a queue of remains ready to be examined. A novel approach to field imaging using DR methodology is seen in a field imaging study conducted in 2015 of 60 mummified priests from the early eighteenth through late nineteenth centuries in the burial chambers of the Mother Church of Saint Nicolas of Bari in Gangi, Sicily, Italy. This study provides an example of using two DR imaging stations in a field setting at the same time (Beckett et al. 2015). The approach allowed one station to collect anterior–posterior projected images while the other captured laterally projected images. Due to the fragility of many of the mummies, a total of 36 mummies could be moved from their niches and were radiographed. Between the stations, 602 images were produced in 14.25 h, thus the productivity achieved was 42 images per hour (about 21 per station). In a 2013 CR study of skeletal remains at the Museum of London, 683 images were obtained, or approximately 10 radiographs per hour, over a 13-day period (Conlogue and Viner 2014). Because individual skeletal components are easier to position for each projection than regions of a mummified body, the number of exposures per hour will always be slightly greater. The difference in the throughput between the DR and CR is due to the longer waiting period for the CR plate to be “read and cleared.” As seen in the Gangi study, two DR units greatly increased the productivity. In addition, both DR and CR provided the opportunity to change the appearance of the image once it has been displayed. This process, known as postprocessing, permitted operations, such as edge enhancement, that improved the quality of the image. From a single exposure, several images can be produced with different appearances. The resulting images can be transmitted electronically to specialists, such as radiologists, paleopathologists, and bioarchaeologists worldwide almost instantaneously. Standard X-ray film would require a photograph to be taken of the radiograph before it could be electronically transmitted. One critical note regarding conventional radiography in general is that the resultant image is two dimensional. This means that there will be a superimposition of shadows (Conlogue et al 2004a). In the field setting, some of the superimposition may be lessened with skillful positioning on the part of the radiographer yet must always be considered when interpreting the image.

Endoscopy as a Complementary Field Method Endoscopic methodologies have been applied in both medical and industrial settings. Ridged endoscopy has been utilized in medical practice since as early as 1806 (Bozzini 1806). A great leap forward in medical endoscopy was seen with the development of the flexible fiberoptic endoscope. Since its inception in the 1970s, flexible fiberoptic endoscopy has been a powerful tool in the visualization and recording of images of pathological conditions in various body cavities, organ passages, and vessels in living humans. Industrial uses of endoscopic visualization have also evolved. As with radiography, endoscopy was applied to mummified remains soon after its development. These early uses of the endoscope followed

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the medical model of application such as a targeted biopsy (Tapp et al. 1984). As technological advances made endoscopy more portable and available to use at a patient’s bedside or in an emergency procedure, the tool became a valuable asset to bioarchaeological research in the field (Beckett and Conlogue 1998, 2010; Beckett et al. 1999a, b; Beckett and Guillén 2000). When used in concert with field radiographic methods, the endoscopic image has the potential to overcome some of the problems associated with radiography. Specifically, the endoscope has the potential to visualize and record images that provide detail of anatomical characteristics or artifacts obscured by the superimposition of shadows seen in a standard radiographic representation (Conlogue et al 2005). While softer tissues and other structures of lower density are difficult to see on a radiograph, endoscopic imaging may provide added detail to these target structures thus complementing the radiograph in the field setting. Using endoscopy to visualize anatomical or pathological targets in ancient remains provides for the potential of biopsy of those structures for further analysis. Artifacts associated with remains may also be examined by endoscopy with great detail and without the distorting effects of advanced imaging methods such as 3D rendering used with MDCT data. These artifacts and their association within the remains among the wrappings or textiles of a mummified body, or within a burial vault or tomb can be visualized and recorded. Artifact removal may be accomplished as well as removal of artifacts within artifacts that are within or otherwise associated with the remains. The endoscope may also provide a visual evaluation of the interior of an enclosed space, such as a tomb, giving researchers pre-excavation data upon which to make decisions regarding excavation approach and an assessment of the stability of the structure (Beckett and Conlogue 2010). Stereoscopic 3D endoscopy has been applied in anthropological contexts as well (Seiler et al. 2013). Field endoscopic imaging in concert with radiography and as a stand-alone method has been used to help answer a wide variety of bioanthropological questions associated with ancient remains such as sex, dental condition, age at death, burial and funerary practices, and paleopathological features (Beckett and Conlogue 2010). While we tend to consider the technology of endoscopy within the medical context, it is important to note that industrial endoscopic instrumentation often offers greater durability and flexibility in scope and lens configurations adding to its utility in bioarchaeological contexts (Beckett and Conlogue 2010). Considering its portability and application beyond the medical model, endoscopy has become a well-established field imaging methodology. Standards for the use of endoscopy in bioarchaeological research have been recommended and applied in various bioarchaeological research settings (Fig. 3) (Beckett and Guillén 2000; Beckett and Conlogue 2010; Beckett et al. 2013a, b).

Field Imaging: A Conceptual Framework In field imaging, it is important to consider all that can be learned from the various methodologies. Here we need to also consider the additional complementary

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Fig. 3 Endoscopy used to collect anthropological data from a mummy in a remote village of the Anga culture in the fringe highlands of Papua New Guinea (Credit: Ronald Beckett)

paleoimaging methods of photography and XRF as contributors as well. A conceptual framework may be as follows: 1. 2. 3. 4.

Looking at (visual inspection, photography, alternate light illumination). Looking through (field radiography – DR, CR). Looking within (endoscopy). Determining what it is made of (XRF elemental analysis).

While each of these methods have their strengths and weaknesses, combining methods can synergistically enhance the meaning that can be made from the images and data collected.

Advanced Imaging Advanced imaging in the bioarchaeological context provides the opportunity to use the most modern and up to date medical imaging modalities on ancient remains and artifacts. Advanced imaging requires that the remains or artifacts be removed from the context and transported to a facility equipped with the required technology. In bioarchaeology, as with forensics, the context is a key factor in the interpretation of the gathered data. In the case of mummy science, the internal environment also provides context of the internal structures, be they anatomic or artifactual, offering key information regarding the study subject(s). Thus moving the remains or artifacts form their context devalues the interpretability of observed relationships. Some studies have used truck-mounted advanced imaging systems, yet the remains still

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need to be moved into the vehicle. With these cautions in mind, these powerful tools can add a great deal of understanding of ancient peoples and their interactions with their environments providing an enhanced understanding of our ancestors’ experience. The primary advanced imaging modality utilized in bioarchaeological research is multi-detector computed tomography, or MDCT. Other advanced imaging methods such as dual energy CT scanning (DECT), micro-CT scanning, and magnetic resonance imaging (MR) have found their way into the imaging arsenal for bioarchaeological studies.

MDCT During the 1970s, computed tomography (CT) was developed and revolutionized medical imaging by eliminating a number of disadvantages associated with plane radiography. In 1977, the first CT scans were conducted on mummified remains. A complete discussion of the physical principles involved and the continuing evolution of the modality can be found elsewhere, however, the advantages require elucidation to fully appreciate the significance to paleoimaging. As discussed above, there are a few drawbacks when using film as a recording media. This concern includes the necessity for repeat exposures, magnification, superimposition of shadows, and, in some cases, the need for liquid chemical processing. CT can address many of this issues. Axial CT images eliminate superimposition of structures and make it possible to take measurements directly from the images. CT also permits the differentiation of tissues of very similar densities. MDCT scanners collect data in a volumetric manner allowing the spatial relationships of structures to be examined. The data may be further reformatted into various viewing planes and can also be reconstructed to create 3D rendering of the structures. For the medical interpreter, the axial sections and the viewing planes are perhaps the most valuable. For the bioarchaeologist, 3D rendering is of great value in that they are more accustomed to working with “bone in hand.” The most significant achievement was the assignment of a numerical value, the CT or Hounsfield number, for different types of tissue. Using CT technology, the researcher can select a region of interest (ROI), an unusual density for example, and measure that density against known density standards using the CT or Hounsfield number. Hounsfield numbers are used in mummy imaging to assist in the accuracy of the interpretation of the densities. While this is a valid application of the Hounsfield number approach, the inter-manufacturer variation in Hounsfield numbers readings of the same target density (ROI) with the same exposure variables renders the method problematic in terms of reliability. With that said, this advancement has transformed radiography from merely a qualitative to a quantitative modality. Dual-Energy-Computed Tomography Dual-energy-computed tomography (DECT) captures data at two separate exposure settings in a single pass. DECT was developed for contrast media injections. The maximum “absorption” of X-ray for contrast media is 80 kVp – making it

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very “white” on the x-rays. The other kVp setting is to penetrate the other tissues including bone. By simultaneously imaging the body with the two kVp setting and “superimposing” the sections, the contrast filled vessels are more prominently demonstrated. This allows the images to be viewed as a single presentation enhancing the interpretation of spatial relationships and the density variations seen in mummified remains. These images can be post processed allowing a more detailed analysis of both dense and less dense structures (Donnino et al. 2009; Wanek et al. 2011). If two passes at different exposures were conducted with MDCT, the ability to “overlay” those images is greatly reduced. Thus the DECT image provides certain interpretational advantages when compared to MDCT data alone.

Multiplanar Reformatting and 3D Postprocessing MDCT and DECT scans provide critical data to bioarchaeological studies materials considering their ability to examine spatial relationships, provide a means to measure and quantify data, present the data in any view or plane, create curve-linear reconstructions (Fig. 4), and provide 3D representations of the data. Combining the data, particularly DECT images, 3D reconstructions allow researchers to perform animations such as digital unwrapping (Fig. 5) or autopsy (Rühli et al. 2004). Threedimensional reconstructions are also useful in directing next steps in the research. For example, the detection and exact location of paleopathologies, such as calcified lymph nodes (Fig. 6), allows for a directed biopsy. Further, the presence and spatial location of artifacts and the analysis of internal anatomical features may be conducted (Conlogue et al. 2005).

Fig. 4 A curve–linear reconstruction from CT data showing dental disease in an ancient Egyptian mummy with bioarchaeological implications related to diet and dental care (Credit: Ronald Beckett)

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Fig. 5 CT data used to conduct a virtual “unwrapping” of an Egyptian Ibis mummy. Note the wrappings seen in the top panel are digitally removed from the image in the bottom panel (Credit: Ronald Beckett) Fig. 6 Three-dimensional reconstruction of CT data showing calcified lymph node targeted for biopsy using endoscope and biopsy tool. Note also the remnants of desiccated thoracic organs (Credit: Ronald Beckett)

Micro-CT When an image of the microanatomy of small structures is required to answer very specific research questions, micro-CT technology is of great value. Micro-CT can be

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Fig. 7 Micro-CT image of an excised calcified hilar lymph node formatted to show internal features (Credit: Ronald Beckett)

applied to an examination of the bones of the inner ear, muscle tissue, or smaller objects associated with the mummified remains. Micro-CT scans typically involve small samples in benchtop or preclinical scanners, although industrial micro-CT scanners can accommodate objects as large as a skull. However, the best resolution is achieved with a small sample. In a study of an excised calcified lymph node (Fig. 7), micro-CT was used to examine the specimen’s pathological features leading to a better understanding of the internal anatomy. This in turn helps to suggest further diagnostic studies such as sampling the node for aDNA. Figure 8 demonstrates the anatomical detail possible with micro-CT imaging when compared side-by-side to a clinical scan. In a study using micro-CT data, researchers (Du Plessis et al. 2015) demonstrated the ability of the detailed images to be used to create a 3D print producing an accurate 1:1 physical replica of a falcon mummy without needing to unwrap the subject. Micro-CT has also been used to enhance our understanding of the processes associated with mummification. In one such study, microscopic tissue alterations occurring in mummification were examined with micro-CT (Wanek et al. 2011). Micro-CT has been shown to enhance the detail of images when compared to standard CT in fetal mummies (Schanandore 2018). Clearly as more and more research is conducted the use of micro-CT in bioarchaeological settings will continue to grow.

Digital Volume Tomography (DVT) DVT is a small CT scanner designed to create high-resolution 3D images of the head and neck. Its use in bioarchaeological studies is limited to detailed study of pathologies and anomalies associated with the skull or mummified head (Schmidt et al. 2013). DVT may also be employed to create detailed 3D images of the distal extremities such as the hands or feet. Small mummified animals may also “fit” into the scanner. DVT may be applied to artifacts and may prove to have field applications.

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Fig. 8 Micro-CT showing a mid-sagittal slice of the cranium of an anencephalic fetal mummy from ancient Egypt as compared to a clinical CT scan. Left is the clinical scan – slice thickness 0.625mm. Right is the micro-CT scan – voxel size 63.8 μm. Specimen EA 493, Maidstone Museum, Maidstone, UK – Samantha Harris, Collections Manager. Clinical CT data: courtesy of M. Garrad, KIMS Hospital, Maidstone. Micro-CT data: courtesy of Andrew Ramsey and Michael Hadland, Nikon Metrology, Tring UK. (Image courtesy: Andrew Nelson)

Magnetic Resonance Imaging (MR) MR “measures” mobile hydrogen content in tissue. The highest mobile hydrogen content is in water. However, there is also lower mobile hydrogen content within fat. At a specific magnetic field intensity, mobile hydrogen protons will “precess” or rotate at a specific frequency. If a radio frequency (RF) is sent in at that precessional frequency, it will tip the hydrogen proton out of its axis of orientation. The longer the RF is sent in, the further it is tipped, i.e., 90
or 180
. It acts like compressing a spring. Once the RF is turned off, the proton returns to its original orientation. As long as the process continues, an RF of the same frequency is generated by the proton. The proportion of the signal generated is proportional to the hydrogen content. MR protocols are based on factors such as how long the RF is sent in (how far the proton is tipped over) and the lengths of time the receiver “listens” for the return signal after the RF is turned off. Thus, high signal is white on the image; low signal is gray on the image. Two very basic protocols are identified as T1 and T2. With T1, there is a longer “listening” time, and the images are “great” for anatomy considering that water has lower signal ¼ gray and fat has a high signal ¼ white. With T2, there is a shorter “listening” time which is great for tumors because water, particularly areas of edema, is white and fat is gray. In a very general sense, mummies are nearly or completely desiccated. Dehydrated remains lack the mobile hydrogen ions from water seen in living patients. As described, these ions are necessary to attain an MR signal. Thus, MR was considered inapplicable to mummy science (Notman 1983). More recently researchers have been finding ways to enhance technique and technology enabling them to achieve signal (Fig. 9) and image acquisition from dehydrated mummified

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Fig. 9 MR image showing signal likely from hydrogen ions present in the fats of this ~1000-year-old female mummy from Pachacamac, Peru (Credit: Andrew Nelson)

remains (Beckett and Conlogue 2010; Conlogue et al. 2010; Öhrström et al. 2013). The MR images produced from mummified remains and seen in the T1 images are likely lipids in various states of change. Even in individuals embalmed with arsenic or formaldehyde, proteins are “fixed” but fats are not affected. MR will continue to be explored as a means of data collection in bioarchaeological research with the hopes of adding value to the study of past peoples. On a final note, synchrotron medical imaging (SMI) is finding its way into bioarchaeological research. This application is in its infancy and coupled with poor accessibility to the technology, the use of and contribution to mummy studies at present is minimal (Bertrand et al. 2003).

Value-Added Data: What Can be Learned Paleoradiology has great potential for providing data to address a wide variety of bioarchaeological question. The nondestructive nature of imaging methodology makes it ideal for getting at information that would otherwise require more invasive measures such as unwrapping a mummy bundle or opening a burial coffin. Imaging methods can in essence, “see what cannot be seen.” Imaging data can help to answer some basic bioanthropological questions such as sex determination, age at time of death, and possibly the stature of the individual. Sociocultural questions may also be addressed. Funerary and burial practices can be assessed through documentation of

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burial inclusions and artifact identification and analysis. Other cultural practices like intentional/cultural cranial modification and medical practices can be assessed. Imaging data can offer clues as to the social status of the individual when findings are compared to larger groups within a given culture. Evidence of intercultural trade and contact can be seen in the radiologic data. Imaging data can provide clues as to the mummification method (if any) employed by a culture as well as changes to those methods over time. These data can then inform the placement of cultural mummification within the taxonomic classification framework described in this chapter. Radiologic imaging data can provide information regarding the health and lived experiences of past peoples. Evidence of paleopathologic abnormalities as well as biomechanical stress assessment, dental condition, and trauma may all be documented with imaging modalities. Paleoradiology has great potential in answering broader bioarchaeological questions as well. The imaging data can challenge or validate earlier views of methods and cultural practices that have been considered the norm for many years. Consider that Herodotus recorded the mummification practices in ancient Egypt with his writings driving most of our impressions and understandings of Egyptian mummification. Wade et al. (2011) and Wade and Nelson (2013a, b) challenged these centuries old assumptions related to Herodotus’ writings about Egyptian mummification practices by reviewing 125 published reports as well as assessing data from first hand MDCT studies related to the route of excerebration as well as evisceration practices. Herodotus suggested that the brain was removed through an opening made in the cribriform plate allowing access to the cranial vault. Wade’s imaging research demonstrated that there was in fact great variability in the route of excerebration among ancient Egyptian mummification practices and the method was not as homogeneous as the long held beliefs. Regarding evisceration, Herodotus stated that the heart was left in the body for magic-religious purposes. Wade and Nelson’s work reviewing 150 cases reported in the literature and again using first hand MDCT with 3D reconstructions on 7 mummies demonstrated that the heart was removed from the bodies during embalming at a greater frequency than previously believed. These two cases demonstrate the importance of revisiting and challenging long held notions regarding ancient practices. These imaging studies help bioarchaeological researchers understand the mummification practices of past peoples while providing a guide as to what may be expected moving forward with regards to variabilities among such practices. While some of the questions posed seem straight forward, imaging studies may face several challenges when trying to provide answers. Taphonomic changes over long periods of time influence the morphology of anatomical structures appearance as well as their spatial orientation within the mummified remains making interpretations more difficult. Determining the sex of an individual mummy can also be challenging. Visual inspection may be rendered useless when the external genitalia are desiccated beyond identification. Paleoradiographic images can assist with sex determination by demonstrating those skeletal markers associated with a given sex. With that said, at times, especially in very young individuals, skeletal development is not yet mature enough to make a clear sex determination. If the individual is wrapped, radiographs can reveal burial inclusions that may suggest the sex on the

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individual. Artifacts such as weaving tools are seen as female related while maleoriented artifacts, such as tweezers used for shaving, are helpful clues offered by imaging in wrapped mummy bundles. Age at the time of death is difficult to answer in mummified remains if the individual is beyond the age of typical dental and skeletal development. Researchers rely on biostress markers, bone infrastructure, cranial suture fusing, dental wear, and wear on the auricular surface of the ilium and pubic symphysis. It is often difficult to attain the paleoimaging images required to visualize these specific bony regions. 3D reconstructions from CT data tend to “smooth” surface appearance reducing the interpretability. Bioarchaeological features make age at death determinations difficult since cultural practices impact target structures. Dental wear and attrition of ancient populations often show premature wear patterns as a result of sand in their food. Ancient work environments and practices influence the rate and location of arthritic changes giving a younger individual the appearance of being older at the time of death. Age at death remains a challenge yet paleoimaging can provide some otherwise unobtainable data. Villa et al. (2013) explored the trabecular patterns of the pelvic bone with CT scans using a population of recently deceased individuals. Results provided reasonable accuracy regarding age estimations. Trabecular patterns seen radiographically in the femoral neck and calcaneus among subjects from ancient populations have also been used to assist with age at death estimations (Beckett and Conlogue 2010), yet the concerns of taphonomic change make such applications problematic. To refine the reliability of age at death determinations through paleoimaging, more research needs to be conducted on the impact of taphonomic change on bony infrastructures in varied environments. There are many unique uses for radiologic imaging in ancient materials. These applications include imaging to search for fossils enmeshed in a rocky matrix, anatomical or material targeted biopsies, coprolite detection and retrieval, and mummified animals to list a few. Another valid use of radiography and other paleoimaging modalities is their application to museum collections. Here as well as in the field, imaging can be used to assess the stability of mummified remains and artifacts with the images helping to determine conservation needs and direction. Many museums have very large collections that were brought to the institution in the eighteenth, nineteenth, or twentieth century when resources to study these collections in terms of funding, personnel, and technology were scarce. Paleoimaging modalities play an important role in helping museums understand their collections.

Practical Guide to the Application of Radiological Methods to Mummy Science Study Designs Given the fragility and rarity of the subjects, mummified human remains, it is imperative that careful thought be given to the study design in order to maximize the interpretable data collected as many of the “subjects” may not be available in an

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ongoing basis. When designing a radiology, or broader paleoimaging study, whenever possible and as dictated by the case, it is important to include multiple imaging methods. As discussed earlier in this chapter, multimodal imaging significantly increases not only the quantity and variability of data collected but the interpretability of that data is greatly enhanced allowing researchers to make more informed interpretations regarding the study subject(s) at hand. Once the methods have been selected and arranged a recommended step-wise approach may be referred to as SADP (Survey, Assessment, Discussion, and Proceed). The SADP approach recognizes that mummy research begins with observation derived hypotheses, yet the initial data collected is always descriptive in nature and may yield new and more complex research questions. In the SADP framework the first step, S, represents an initial survey. Those methods best suited for the initial survey include conventional radiography (CR, DR, or film if digital recording media are not available), endoscopy, photography, and XRF when appropriate. Each of these methods are portable and easily transported to remote locations and applied at the point of discovery preserving the context of the case. The A in SADP represents the next step and refers to the initial assessment of the data collected during the survey. It is in this step where initial interpretations are made. These initial assessments should involve all the players associated with the research. The use of teleradiology, discussed later, can enhance the assessment stage. The next step is one of the most critical steps as the D represents discussion. It is in this step where the research team considers critical decision-making regarding the case at hand. The concept of triage discussed in this section and ethical decision-making are key components of this step as researchers weigh the need for, as well as the pros and cons of moving the case forward. And finally, the last step is P for proceed. Once the discussion has yielded informed decisions regarding the continued direction of the research, arrangements can be made for further analyses or in some cases, conservation. As suggested in the SADP approach, it is critical that any study design lets the data lead the research.

Workflow: Multistation Design An important factor associated with any radiographic imaging study of mummified remains and associated artifacts is that of time management. Often research is conducted in locations that may be somewhat remote requiring travel and coordination of the team of researchers. It becomes important then to include a workflow design for an imaging project that will allow for efficient “throughput” and data flow. This is particularly important when the study involves a large number of mummies. Key factors of a workflow plan include a consideration of the team players, the physical layout where the research is to be conducted, a plan for subject movement into and through the imaging stations, an organized and systematic plan of radiographic projections, sequencing of the project including movement, and time for a methodological audit in order to find ways to improve the process (study the study design). The research team needs to be made up of at least one professional radiographer or other paleoimager who will help to establish the initial plan regarding the instrumentations required, the movement of the equipment to and from the research

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location, and who is able to troubleshoot both the technology as well as the process. This individual should be well versed in nontraditional application of radiologic methodology. The research team “captain” must be available to help direct and make decisions while the study is in process. The additional team members may include other professionals who will operate research data collection stations other than the radiographer, such as a bioarchaeologist, endoscopist, and so on. Lab assistants or students of medical imaging, anthropology, archaeology, or other related fields are essential to the make-up of the remainder of the team and often serve as record keepers for the documentation of procedures and methods. In this way, any publications will include details of the study which will allow for a replication of method. Perhaps most important is having knowledge of the physical layout where the research is to be conducted prior to selecting equipment and personnel. Electrical requirements should be considered as in some locations a power generator may be required. It is important to know as much about the working conditions as possible including the times that the subjects will be available. In all cases, utmost respect must be given to the established norms and protocols of the research location. Once the characteristics of the physical location are understood and the equipment issues are considered, a workflow scenario can then be devised. In particular, the workflow design will include how the mummies (in the case of large groups) will be brought to and pass through the imaging stations. This stage must include a systematic plan for moving the mummies which may require removing them from an exhibited or stored location and transporting them to the imaging area. Transport plans may include the need to devise special harnesses or slings designed specifically for that research setting with care for the subjects always at the forefront of thinking. Once the workflow scheme is designed, the team must determine exactly which radiographic projections will be employed taking care the most useful data is collected that will serve the initial survey concept. Not only which projections, but the sequence of those projections is key to maximizing workflow. Additionally, the exposure parameters should be standardized. When multiple modalities are used, the stations and workflow are designed within a systems approach. The systems approach is both structured and dynamic at the same time in that while there are assigned stations for each modality, the physical layout allows for free flowing communication between and among the stations creating an organic system in which feedback and input from the involved modalities is encouraged and enhanced. The systems approach is in contrast to a simple multidisciplinary design where each communication flows in one direction. Multidisciplinary designs are those in which each specialization is compartmentalized creating an impediment to communication and information flow. In the systems design feedback is in real time. As described earlier, knowing the physical layout is crucial to developing a systems approach workflow environment. In the systems approach, the stations are generally on a fixed location with some being both fixed and fluid. The fluid station is dependent of the portability of a given modality. What has been described thus far is an approach to the survey aspect of the SADP concept. With that in mind it is important that assessment of the data be reserved for the discussion step of the SADP model. However, it is important to make some

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preliminary assessments of the imaging data at the point of collection. There are two main intents of this cursory assessment within the initial survey step. One, to assure that the image quality with regards to projection, positioning, and exposure provide interpretable data. And two, to bring attention to a feature that will inform another station or modality within the systems approach. The remaining steps of in-depth assessment, discussion, and proceeding will be addressed in section “Triage and Ethical Decision Making.”

Teleradiology Whenever possible, a study design is enhanced when it includes as many participants as possible. Given that most imaging data is now digital in nature, it is feasible to employ teleradiology in the study design. Teleradiology is simply the transmission of radiographic and other images through the internet. In this way, and in real time, professional many miles away can provide input into the research while it is in process. In the past, there have been times where a study has been completed, all the equipment were packed and transported back to the country of origin, only to discover that an additional view would have been beneficial to the interpretation of the case at hand. While teleradiology can be an important if not critical aspect of the assessment and discussion aspects of the SADP model, it can serve the survey step as well. Images can be sent via teleradiology to offsite professionals who can inform the team regarding positioning of the subject with respect to the image produced or request another projection that may highlight a suspected lesion or other anomaly. Triage and Ethical Decision-Making During the assessment and decision-making steps of the SADP model, discussions and decisions must be informed by researcher-established guidelines that follow both practical and ethical criteria. In an imaging study, the concept of triage assists the researchers in deciding what the next steps might be. Triage is a construct that is used in medical practice typically in emergency or catastrophic situations that would otherwise exceed the capabilities of a given institution. Using measurable criteria, medical practitioners decide which patients will get immediate medical treatment. This is often a scoring system of sorts. In the assessment and discussion stage of an imaging study of mummified remains, the triage concept is utilized to rank research priorities and decisions based on a variety of factors. One of the major decisions following the initial survey is to determine if the study would benefit from a more advanced and aggressive form of imaging or other analyses of an individual mummy or group of mummified remains. This usually involves decisions to move a mummy or group of mummies to an advanced imaging facility for MDCT or MR scans. Additionally, the question may be one of unwrapping a mummy for further study or taking a biopsy of a target lesion within a mummy. It might even be a decision about conducting a full or partial autopsy on the mummified remains. When conducting a triage discussion, many factors need to be considered. First and foremost is an honest assessment of the risk to benefit ratio. Questions that the team must address are:

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Benefits 1. What is the benefit that may be derived from advanced imaging, unwrapping, biopsy? 2. What is the compelling need to move the subject to collect these data? 3. What is the potential value-added data to the bioarchaeological understanding of a culture? Risks 1. What risks are involved? 2. What do the initial images taken on site tell us about the stability of the subject? 3. Is it reasonable to assume that the mummy can be moved without causing damage? 4. How will damage be evaluated? (Pre and post transport radiographs). Transport Concerns/Costs 1. If apparently stable and transport judged important, how will it be conducted? 2. How will the subject be packed and secured for transport? 3. What are the costs associated with transport and imaging? Data Management 1. If advanced imaging is conducted, what will become of the data? 2. Where will the mummy and the new data be stored? 3. How will the data be used? 4. In the case of biopsy, what labs will conduct the analysis and what are the objectives of collecting tissue? All of these questions can be summarized into three broader categories: Should we do it (is there a clear research objective?), can we do it (is it safe, cost effective, transportable etc.?), and what is the value of doing it (how will the additional data contribute to the case?). Another ethical issue is that research in the mummy sciences often attracts media coverage. The integrity of the mummified remains and the research value must be guarded against sensationalistic media coverage. We see too often media coverage in which a mummy is brought to an advanced imaging facility for a CT scan and while the facility and the researchers are given “screen” time, no discernable contribution to the understanding of the case with regards to research objectives or related image interpretations is reported. During the assessment and discussion steps, the team must rigorously evaluate the positive and negative potentials of moving the remains. If the body is to be moved for advanced imaging, the team must first establish clear and measurable objectives that will demonstrate the value added of advanced imaging with regards to a richer understanding of both the individual mummy as well as the broader bioarchaeological context. In all cases, group consensus and objective decisionmaking is the goal. The major risk in moving a mummy is that of not only damaging the remains, but also altering the spatial relationship of anatomic features within the body as well as

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the relationship of artifacts associated with the burial. Such variations can render interpretations invalid. Whenever possible, imaging should be conducted in the field. Even advanced procedures, such as a tissue or targeted lesion biopsy can be conducted in the field without the need for advanced imaging. Several techniques have been developed to localize biopsy targets within a 3D structure using radiographs and spinal needles placed at 90
from one another. Endoscopic-guided biopsy with radiographic correlation is also a very effective technique. We suggest that the primary objective of an imaging project should be to keep it simple and do the least possible harm. The portability of imaging and other paleoimaging instrumentation offers a powerful data collection method for conducting sophisticated research in the field. This premise goes against many in paleoradiology who consider a CT scan of mummified remains to be the Gold Standard for imaging in mummy sciences. One cannot argue that advanced imaging can bring a tremendous amount of data to bear on an objective driven research project, yet CT scanners are not readily accessible to researchers in remote settings. In fact, standard onsite radiography, complemented with additional paleoimaging modalities, can readily answer many biological and archaeological questions. Most of the mummies are discovered in very remote locations where scanners simply are not available. Nor would it be practical to move the mummy lashed to the back of a truck across poor roads. When a mummy is housed in a museum, it is very likely that the remains have already been disrupted through movement, display, and travel. In these cases, moving to an advanced imaging facility may be more practical. The use of a CT scan can certainly assess conservation needs as well as provide useful bioarchaeologic information with the understanding that the movement to the museum from its country of origin or its handling while in the museums care may have altered the spatial relationships of anatomic structures or associated artifacts rendering the data questionable. Each case is unique and the assessment and discussion steps of the SADP model must serve to objectively inform the P how to proceed. A very positive and sometimes overlooked outcome from advanced imaging studies on mummified remains is that the digital data can be stored for future analysis. These data can then be added to larger data bases where future researchers can call on that data to make new meanings through comparative analysis on a much broader scale. Even if a single case did not yield little or no diagnostic results the images may serve another researcher who is looking at different questions, such as a comparison of excerebration routes. Digital data archiving will be discussed in more detail later in this section. When the advanced study is completed, the research team should be certain to leave the facility with the data in hand as many medical scanning facilities purge their system memory.

Case and Population Imaging Studies Given the vast array of imaging modalities that are currently applied to bioarchaeological material, and considering that as advances are made in these technologies and as newer technologies emerge, it is safe to surmise that they will continue to be applied to ancient remains and artifacts. With each study, the derived data contributes to our knowledge of lifestyle, health, and disease in ancient times.

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Two primary study approaches currently employed in bioarchaeological research that use imaging data are the case study and population studies designs. Earlier in this chapter we described the use of field radiology, and several large population studies were described. However, one need only to search the popular media and scientific literature to realize that there are a great number of case studies being conducted as well. As in medicine, both the single case study and the larger population studies hold value and provide us with a broader understanding of the lives of past cultures. Further, those data also hold meaning to our lives today. The case study often brings us a snapshot of an interesting or unusual case, which becomes an important point of information to be recorded and associated with data from future studies. In contrast, larger population studies can provide us with a more significant understanding of such characteristics as the patterns of disease or trauma among the culture being studied and can then be related to the bioarchaeological profile of that culture. Recent imaging research of ancient cultures has helped modern physicians understand the history of particular diseases. A large CT scanning study that considered primarily mummified remains from Egypt but also made comparisons from mummies around the globe focused on the diagnosis of atherosclerosis (Thompson et al. 2013). They discovered that there was a high incidence of atherosclerosis even among royal mummies who were thought to have a more quality diet than the general population in ancient Egypt. These findings not only add clarity to our understanding of ancient disease patterns, but also offer insight into our understanding of these specific diseases in modern populations. In this case, the authors suggest that genetics may play a larger role in the development of atherosclerosis than previously thought and that diet, while important, may play a lesser role. It is important to note that in a diagnosis-targeted study design, vast amounts of data are captured in each CT scan. A great amount of information is held within those scans and researchers who employ the diagnosis targeted model are encouraged to review the scans and conduct a more extensive or global analysis of the data collected. Given that CT scans are expensive and time consuming these data other than the target diagnosis are extremely valuable and may provide critical insight into past peoples, their conditions and their artifacts. The study sample size in this case was 137 mummies and while this was a targeted diagnosis model, the data can then be archived so that other conditions, like osteoarthritis, excerebration methods, or trauma patterns, to name a few, can be studied as the data is revisited by future research teams.

Data Archiving There is ever-increasing volume of bioarchaeological data being created by both field imaging and advanced imaging technologies. These procedures create massive volumes of data. Archiving these large quantities of data is being addressed. There are several examples of digital archives that are making the data available to researchers from across the globe. Two notable examples include the IMPACT project (Internet Mummy Picture Archiving and Communication Technology) (Nelson and Wade 2015) and the Digitised Disease Project

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(http://www.digitiseddiseases.org/preview/#). In this way, the data from an individual case study can be placed into a broader context allowing for comparisons and contrasts to other imaging studies. There are several examples of the use of these archival systems appearing in the current literature, which demonstrate the research potential of being able to cross-reference vast amounts of information. In one such example, the impressions of Herodotus as he described the ancient Egyptian mummification procedure have been clarified (Wade and Nelson 2013a). In question was the retention of the heart as reported by Herodotus. Through archival cross-referencing digital images, it was determined that the heart was in fact removed in the majority of those Egyptian mummies studied. Studies such as these help to refine our understanding of the preparation and burial practices of ancient cultures (Wade and Nelson 2013b). These archival digital data opportunities will certainly help establish bioarchaeological contexts for larger numbers of seemingly unassociated case studies. Each archive system has its potential limitations. Some of the archives are disease specific while others are more broad in scope. Not only should the imaging data be stored for future review, but supportive contextual information like previous related studies, provenance, and up to date results and interpretations should also be made available. In the spirit of reproducibility of study characteristics the technical variables associated with the image acquisition should be made available. Further, a network of experts in imaging analysis as well as contextual experts such as bioarchaeologists and radiologists should be available to support interpretations in new studies. Finally, data archives have great potential in the training and preparation of professionals less familiar with the unique nature of paleoradiology and its relationship to bioarchaeological questions.

Limitations and Opportunities in Paleoimaging There are a variety of factors that impact the appropriate application of medical imaging in bioarchaeological research. It is important to recognize the technical limitations and associated challenges. In addition, two additional major categories of limitations must be considered. The first of these is challenges of data collection. Once the data has been collected, there is the issue of interpreting the data in the context of the many variables associated with ancient remains and artifacts. When interpreting, two primary approaches are employed, the medical approach and the bioarchaeological approach. While each of these approaches has much to offer, using one without the other results in incomplete consideration of the sophisticated data collected. Another major issue associated with paleoimaging is that of proper training of the data collectors and the data interpreters. Yet given the powerful potential of paleoimaging in the bioarchaeological context, many opportunities exist for enlightened collaborations and technological creativity.

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Challenges Related to Technology Each imaging modality has unique limitations. For example, the endoscope, to be applicable to the imaging on internal structures or artifacts, requires a route of entry. It is beyond the scope of this chapter to delve into the limitations of each imaging method and the reader is referred to Paleoimaging: Field Applications for Cultural Remains and Artifacts (Beckett and Conlogue 2010) for a more in-depth treatment of these concerns. Generally speaking, the greatest challenge to imaging in the bioarchaeological context is that of instrumentation accessibility. In field applications, getting the right technology coupled with the right individuals who are able to transport, operate, and trouble shoot those technologies is a critical consideration. Even though the instrumentation for standard X-ray, endoscopy, photography, and XRF is quite portable, the varying conditions in different global locations requires that the individual possess experience and knowledge not only in their imaging specialization but also in the bioarchaeological model of scientific study. Access to advanced imaging modalities is limited as well. Advanced imaging is usually found in medical or free standing imaging facilities. While there are mobile CT units and even “portable” CT units as used by the military, it is difficult to make arrangements for the transportation and operation of these units. When access can be procured at an imaging facility or hospital, the studies are often conducted in the after-hours when there is less potential for a living patient to need a scan thereby reducing access. There are a few imaging research facilities associated with universities but this is the exception rather than the rule. Even when arrangements can be made there are the challenges associated with the physical act of transporting the mummies to the facility. The logistics of ethical transport of human remains not only creates the need for careful planning, but the transport itself creates additional risks to the remains or artifacts. Subjects may be damaged or subjected to environmental shock. The internal context may be disrupted, decreasing the interpretability and the bioarchaeological significance. If the mummy is to be moved, standard imaging must be conducted prior to and after the moving the remains. In this way, researchers will be better able to gauge the impact of the packing and transport process on the internal context of the subject under study. The concept of triage discussed earlier in this chapter is useful to the decision-making process regarding moving forward with additional digital imaging data collection. Another concern in advanced imaging is storage of large volumes of digital data. On site data storage capabilities at medical facilities are reserved for living clients. Transferring the data to data archiving efforts described earlier in this chapter is an attempt to overcome this data storage issue. While accessibility, portability and archiving the data are clear challenges to paleoimaging; another issue is that advanced imaging systems use preset algorithms that establish exposure variables designed for living humans. These algorithms have improved the productivity and efficiency of imaging facilities among the living, yet the preset variables are not well suited for mummified remains. Overriding the algorithms or using them in a creative manner in order to capture the useful data

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for bioarchaeological studies requires a skilled paleoimager willing to spend the time during the scan and in postprocessing efforts (Conlogue 2015). This reality presents as an additional challenge to paleoimaging projects.

Challenges to Data Collection An interpretation can only be as good as the data collected. In paleoimaging, this requires an individual skilled in not only the standard application of the instrumentation, but one who is creative in their thinking and who can stretch the limits of that instrumentation and apply that skill to the task at hand. This may require modification of either the equipment itself or of overriding instrumentation built-in parameters. The quality of the data collected is dependent upon the knowledge of what data is to be collected, why, what it may mean, and what additional data collection is needed after reviewing the initial images. The individuals must therefore know their instrumentation, know how to troubleshoot that instrumentation in the field, and know what data is critical to the particular study at hand. This requires knowledge about ancient cultural practices and potential diseases associated with that culture. Paleoimagers require knowledge of the importance of context, taphonomic forces, mortuary and burial practices, and mummification methods. They need to appreciate the potential for transport artifact that may potentially disrupt spatial relationships of internal objects or anatomical structures. Knowledge of complementary modalities is critical. While attempts at standardization have been made, there remains a lack of consensus regarding technique and procedure in paleoimaging applications. A vast array of bioarchaeological factors may dictate what needs to be imaged and in what manner in order for the images to be useful in interpretational efforts.

Interpretational Challenges The primary challenge to the interpretation of data rests in an understanding of the potential taphonomic changes potentially associated with the study at hand. Understanding the taphonomic potentials requires a knowledge of the taxonomy of mummies described earlier in this chapter, how the remains were mummified within that taxonomic classification, and the associated burial and funerary practices that would impact the continued decomposition over many years. Knowledge of how unique burial environments will impact the stages of decomposition, the degree to which the remains decompose, and the morphologic alteration brought on by the specific conditions is critical. In a recent experiment designed to emulate the mummification process associated with Chiribaya subterranean tombs, various stages of decomposition and mummification were seen on the same individual (Garcia et al. 2014). These stages were directly associated with gravity dependence and included dry mummification, caseation, and saponification. These findings are similar to a report by Guillén (2004) regarding the taphonomy of the actual mummified remains of the Chiribaya in the Atacama Desert.

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There are many taphonomic variables that will influence the morphology, anatomical position, and the spatial relationship among and between anatomical structures and associated artifacts. Among these are the impacts of predation, watershed, weather patterns, temperature, pressure, atmosphere, and soil characteristics. Interpretations are further complicated by post-depositional changes associated with the handling of the remains or artifacts. If advanced imaging is deemed critical to the case at hand, the interpretation of data collected may be compromised by transport artifact. Such transport impacts include further positional changes altering spatial relationships among and between anatomical structures and artifacts and potential trauma to the subject. Such trauma may include disruption of wrappings and possible fractures to skeletal parts. To account for these potentials, it is advised that the subjects or objects first undergo a field imaging examination in their original context or as close to that context as possible. Once the subject arrives at the advanced imaging facility, the images can then be evaluated in context of the pre-transport images. It is advised that the subject is then reimaged once returned to the field site or place of conservation. Additional challenges to interpretation rest in the experience a researcher has with mummified ancient remains and the specific approach to imaging interpretation. The two major approaches used for interpretations are the medical approach and the bioarchaeological approach.

Medical Versus Bioarchaeological Approach Given that the imaging data for the most part is collected by medical specialists such as radiographers and interpreted by other medical specialists including radiologists, cardiologists, orthopedists, pulmonologists, medical dentists, and many others, the medical approach to data collection and interpretation seems a logical one. Indeed, these medical experts bring with them a vast knowledge of diagnostic variation and the skills to make those interpretations. The radiographer is certainly expert in setting the imaging parameters of their instruments to collect the needed data. However, most of this expertise of data collection and interpretation is based on their experience with living patients rather than ancient and desiccated subjects. Further, among the living the medical physician has the advantage of gathering a medical history and may conduct a physical exam in order to assist with their diagnosis. Modern CT scanners have preset exposure variables or algorithms to assure proper image gathering for the targeted body structure. In modern medicine this helps with “throughput” resulting in a more time-efficient approach. These algorithms are established based on hydrated human bodies and are not well matched to the imaging requirements of dehydrated human remains. Various protocols are being developed for all imaging modalities applied to bioarchaeological materials. Even if the data is collected in a manner that will maximize interpretability from the images, there are a variety of temporal factors that may influence the interpretation. Desiccation, as well as additional taphonomic forces and mortuary practices or rituals, can alter normal morphologic appearance of skeletal and organ systems.

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Anatomical position of these structures is often altered making diagnosis challenging for the uninitiated medical physician. Often a radiologist will make an interpretation based on their modern experience not taking into account the potential of the pre-antibiotic impact of varied pathological processes on the human organism. Diagnoses can be skewed by modern etiologies that may give the same appearance on the medical image, when in fact those conditions did not likely exist in ancient times. One example would be Harris growth arrest lines seen on the long bones of an inner city child in more recent times being interpreted as brought on by lead paint ingestion, while the same lines seen on the remains of an ancient human child may indicate a variety of body wide processes that would have arrested growth, such as diet or repeated seasonal system wide infections. A paleoradiologist, one who is skilled in both the imaging variables associated with ancient remains and the importance of the bioarchaeological context, is a specialist within a specialty. The learning curve associated with the ability to collect and interpret imaging data within the bioarchaeological context has yet to be defined. Yet, it is very apparent that such preparation is necessary. In contrast, the bioarchaeological approach to interpretation or diagnosis considers the varied environmental constraints and cultural characteristics of the associated group. All of these varied factors may influence the disease process and the degree of care the individual may have received while living. The bioarchaeological diagnosis also considers specific aspects of the cultural systems in place that may influence a disease process. Examples of aspects that may impact imaging findings include any potential buffering from disease development such as living conditions, medical practices, food sources, trade, family structure, proximity related to population density, and social stratification (if any). There may also be genetically based host resistance factors that reduce the risk of disease contraction (Martin et al. 2013).

Diagnosis by Consensus Comments Recognizing that there is likely interpretational bias among interpreters of imaging data necessitates the adoption of the broader bioarchaeological model. Without all of the evidence required to make accurate and valid determinations, a consensus based on varied inputs is required. This process is called a diagnosis by consensus. The diagnosis by consensus model of data interpretation implies that many views and many eyes are better than a few (Wade et al. 2019). Results derived from a diagnosis by consensus offers a broader group of differentials that are contextually relevant to the study at hand. A key factor of any diagnosis is the attainment of usable imaging data. The radiographer is responsible for the quality of the images used for interpretations. The skill on the part of the radiographer to attain usable images is paramount. Accurate interpretations require images that demonstrate the target area or anatomy in a clear and reproducible manner. This requires special attention to the superimposition of shadows associated with plane radiography.

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The diagnosis by consensus team would include perspectives from bioanthropology, bioarchaeology, medical specialties, and those involved with image acquisition. The diagnosis by consensus model supports the team approach to science and can dissuade unilateral or uninformed interpretations. Diagnosis by consensus provides a more accurate and informed differential interpretation which will better serve our understanding of past cultures within the bioarchaeological construct. In cases where such diagnoses can be “ground-truthed” by microsampling, biopsies, or prosections, these procedures will further enhance diagnostic accuracy.

Training Challenges Currently, no specific paleoimaging training programs exist for imagers, medical physicians, or for bioarchaeologists. This serves as an impediment to the maximization of paleoimaging data collection and interpretation. Ideally, a paleoradiographer should be a radiologic technologist expert in the technical aspects of all modalities of medical and industrial systems and who has been mentored by a seasoned paleoradiographer and a bioarchaeologist familiar with imaging data. This is true for the paleoendoscopist as well. Familiarity with all endoscopic instrumentation, both medical and industrial, and their potentials and limitations are a must. Experience in interpreting endoscopic images in a variety of mummified conditions is also critical. This researcher needs to be skilled in biopsy procedures and artifact retrieval and the ability to recognize pathological conditions is critical. The endoscopist must be mentored by an experienced endoscopist along with a bioarchaeologist familiar with endoscopic images. Similar to the image collectors, image interpreters having a medical background must be mentored by those with experience within the bioarchaeological context. Conversely, bioarchaeologists must become familiar with the application potentials and the limitations associated with each of the imaging modalities. In practice, the medical practitioner and the bioarchaeologist must mentor one another if the paleoimaging data is to be used to its fullest potential.

Looking Forward: Opportunities With the varied limitations to paleoimaging described above, there are many opportunities that would move paleoimaging forward and expand its contributions to bioarchaeological research. An article (Beckett 2014) identified several opportunities for research regarding paleoimaging. Among them are conducting research to discover how to best match the paleoimaging approach (a triage approach versus “scan every mummy”) to answering specific bioarchaeological questions. Another opportunity in paleoimaging is to find ways to share expertise in the form of formal mentoring programs for image collectors and image interpreters. As we look forward, there are additional opportunities for paleoimaging as well. Technologic

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advances, standardization, and expanding the application of imaging data to the emerging bioarchaeology of care framework are all opportunities waiting to be explored.

New Emerging Technologies/Instrumentation/Tele-Interpretations Many opportunities exist in finding new methods of combining imaging data with the available software reconstruction systems currently available and in the development of new combinations of systems designed specifically for bioarchaeological research. One important concept is that of the bioarchaeologist’s skill of working with the “bone in hand.” While 3D printing has become popular to demonstrate paleopathological conditions in bony structures, more can be done to create 3D representations of structures showing associations and spatial relationships assisting in the understanding of bioarchaeological constructs. Recently, there have been several start-up companies that can produce 3D printing of paleopathological conditions of the skeleton from either laser surface scanning or Dicom ® data. As new imaging modalities make their way into medical practices, they must be evaluated for their application in the bioarchaeological context from a value-added perspective. Newer does not necessarily equate to better in terms of what data can be obtained. An opportunity does exist for the development of a portable paleoimaging suite, or package that would include portable instrumentation proven to be useful in obtaining bioarchaeological data. Such “kits” may include DR, endoscopy, photography, and XRF instrumentation to name a few. Emerging telemedicine models are a technological development that can be applied to the interpretational challenges associated with imaging data. We previously discussed the use of teleradiology in filed settings to get real time input into interpretations and guidance on needed imaging views. Here we are referring to teleconferencing that can serve the diagnosis by consensus efforts of study groups. Experts from around the globe can bring their experience to bear on a given study strengthening the interpretations of imaging data.

Standards in Paleoimaging In the spirit of reproducible science, standards help establish guidelines for future research as well as provide an historical record of what did and what did not work. Standards are an evolutionary process with improvements in radiologic applications being built on past progress. Standards serve to validate methodology as well as instrumentation. Paleoimaging standards have been addressed but are yet to be firmly established creating an opportunity for continued development. The reader is referred to Beckett and Conlogue (2010) where proposed standards for instrumentation, procedure, and documentation are described in great detail. Each of the varied imaging methods possess nuances associated with that particular manufacturer or with their method of application. For example, CR and DR

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manufacturers will take a slightly different approach to image acquisition. Given this variability, it is not prudent to suggest specific standards related to instrumentation and application since each instrument will be different. However, some standards can be recommended. Positioning of the mummy, minimal required views, exposure variables, and data storage are examples of those characteristics of instrumentation and application that can be standardized. Procedural standards and documentation standards become critical in developing application guidelines that will enhance the reproducibility of imaging studies (Beckett 2017). A standardized procedural approach would include the imaging team establishing objectives as related to a given study. These objectives should be written down with imaging approaches established to respond to those objectives. All procedures should begin with a complete visual inspection complemented with preliminary photography. The next step would be to select the imaging modalities appropriate for the case. Recalling the SADP model described earlier in this chapter, the next standard procedure would be to start with an imaging survey using standardized exposure settings, positioning, and views. This would be followed by and initial assessment of the original data produced allowing for recalibration of approach and a refinement of focus to improve the images in terms of quality and imaging targets. All of the preliminary standardized steps would include a standard method of documentation. This documentation would record and assure that there was consistency in the views, exposure variables, and instrumentation used to gather the initial survey images. Such documentation would include photographs of instrumentation setups, procedures, handling of the mummies, and the physical context of the study. Postprocedural conference follows allowing for assessment and discussion of the initial images with the goal of establishing new objectives, and the identification of any additional imaging targets for analysis. If the assessment and discussion suggests that advanced imaging would be able to contribute to the study outcomes or to new objectives based in the initial readings, a risk/benefit dialogue should be an intentional and separate piece of this step. The discussion must carefully consider the need for and potential value added from advanced imaging. Finally, after the assessment and discussion step, the research team will proceed with the project informed by the analysis if the initial data.

Bioarchaeology of Care Applications The bioarchaeology of care (BOC) is a relatively new approach (Tilley 2015) within the larger bioarchaeology umbrella. As you recall, bioarchaeology attempts to reconstruct the lives of past people and cultures using all available evidence including archaeological, bioanthropological, as well as environmental sciences. The BOC approach seeks to determine whether there is enough evidence to suggest that care was rendered to those who showed signs of a pathological condition. The approach looks at the potential for care giving that would have been required for an individual to function within a culture or even to survive at all. Evidence that may suggest that care was given include a clear pathological diagnosis and possibly archaeological

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findings that demonstrate supportive technologies. The BOC approach does not consider care giving as something surprising among ancient human populations, rather it provides a framework for assessing the plausibility of care provision in any given instance and adding these probabilities to the overall understanding of that population. Given that the BOC approach requires the diagnosis of a specific or combined condition, imaging data can provide insight into what that individual or group of individuals may have been inflicted with. A clear diagnosis based on interpretation of radiographic and other imaging data is the cornerstone of the BOC model. Images can help determine if there was evidence of a condition that would have required care. Further, imaging can help determine if that individual lived with that condition beyond what would be expected without care. Employing the BOC approach affords the research team the opportunity to expand the cast of “interpreters.” A pathologic condition causes physiologic challenges which in turn bring on functional limitations. Through the individual’s life span there is progression of the disease and thus a progression of the pathophysiological impact and resultant impairments (Conlogue et al. 2017; Beckett and Conlogue 2019). These changes over time must be considered in the overall assessment and inform the probability that care was given. Thus, experts in the care and treatment of living individuals afflicted with similar conditions can add valuable information regarding not only the lived experience of the study subject but also suggest the types of care needed to survive. Some of these professionals may include rehabilitation specialists; occupational, physical, respiratory therapists; and/or physiatrists.

Chapter Summary As we consider characteristics of the bioarchaeological context as being similar to forensic sciences by trying to piece together an event or relationships among varied points of evidence from some time in the past, it becomes apparent that the most useful imaging data is collected at or as near to the original context as possible. Careful consideration must be given to moving those cultural remains or artifacts to an advanced imaging facility. If it is determined that advanced imaging can answer questions unresolved by the field imaging efforts, a pre- and posttransport study must be conducted to gauge the impact, if any, to the remains with regards to shifts in the anatomical features or inclusions within the internal context and to determine if any additional post-depositional trauma is attributed to the transport. It is clear that medical imaging methods have and will continue to play a critical role in the evaluation of bioarchaeological material. In this role, medical instrumentation operators and image interpreters will be challenged to flex their conventional skills to embrace the constructs of bioarchaeology. Conversely, bioarchaeologists must flex to know what can and cannot be done with medical imaging and function as a team to arrive at an interpretation or diagnosis by consensus utilizing the vast expertise of both disciplines.

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As newer imaging tools and software become available, they will most certainly be applied to cultural remains and artifacts. It is encouraging to see the medical community contribute to the continued quest of understanding our species’ journey on earth through the application of powerful nondestructive imaging methodology and interpretational expertise.

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . X-ray in General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dual Energy Machines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Other Imaging Modalities and Their Appropriateness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ultrasound . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . MRI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Terahertz Imaging . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Medical Imaging for Archaeology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Radiation Damage to Ancient DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Benefits and Disadvantages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Future . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Before the discovery of X-rays, the only available method of scientifically investigating the structure of a mummified body was to dissect it. Since the discovery of X-rays by Roentgen in 1895, Egyptologists and others interested in paleopathology have been using medical imaging to aid research into ancient R. D. Loynes (*) KNH Centre for Biomedical Egyptology, University of Manchester, Manchester, UK e-mail: [email protected] R. Bianucci New Mexico Health Enhancement and Marathon Clinics Research Foundation, Albuquerque, NM, USA Warwick Medical School, Biomedical Sciences, University of Warwick, Coventry, UK Legal Medicine Section, Department of Public Health and Paediatric Sciences, University of Turin, Torino, Italy e-mail: [email protected]; [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_39

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biological artifacts. In this chapter, the history of medical imaging is briefly considered, particularly as it relates to the imaging of mummies. Various systems are discussed along with their suitability to mummy studies; and possible disadvantages of the use of ionizing radiation are also discussed as is the potential future direction of advances in this field. The subject of data security is introduced with reference to present day technology. Keywords

X-rays · Computerized axial tomography · Multidetector computed tomography · Magnetic resonance · Micro-CT scanning

Introduction Until the discovery of X-rays, the only available method of scientifically investigating the morphology of a mummified body was to dissect it. Such famous events include the public dissection of a mummy – Khnum-nakht – one of the occupants of the “Tomb of the two brothers” by Margaret Murray in Manchester in 1908 (David 2002: 31). Before this period, “unwrapping” mummies had been more of a public spectacle than a scientific exercise. However, whether or not the exercise was truly scientific, the end result was always complete destruction of a unique artifact. Since the discovery of X-rays by Roentgen in 1895, Egyptologists and others interested in paleopathology have been using medical imaging to aid research into ancient biological artifacts – both human and animal, one of the major benefits being the preservation of the artifact. The first person to use this technique was Koenig in 1896 who produced images of a human (child) and an animal (cat) mummy from Egypt (Adams 2016: 371–386). Another of the earliest examples was provided by Petrie in 1898, while the first royal mummy was imaged by Elliot Smith in 1903 (Ikram and Dodson 1998: 95). In the early days, this took the form of plain X-rays with the data produced on the only medium available at the time, that is, photographic film. Although a quantum leap forward at the time, these images are regarded now as somewhat primitive. Unfortunately, another phenomenon frequently experienced in the review of old, historic X-ray films is degradation of the image due to imperfect chemical fixation after development of the film. Over the subsequent 120 years, imaging has progressed and diversified to encompass several modalities of image production and systems of data capture and storage media. The topic of medical imaging is conveniently broken down into different groups of technologies – on the one hand the use of X-rays and all the subsequent developments involving their use and on the other the production of images using other parts of the electromagnetic spectrum. While this has improved medical diagnosis and management opportunities, not all modalities are appropriate for use in the field of paleopathology. This lack of appropriateness lies within the nature of the biological material itself.

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X-ray in General X-rays have a wavelength of approximately 1 nanometer (Fig. 1). At the time of discovery, the ionizing radiation produced by X-rays created a negative image on photographic film with bones shown as white or almost white images (having prevented the passage of most of the radiation) and soft tissues as grey forms – air being shown as black. The problem is that the complex anatomical structures of the human (or animal) body produce superimposed images (in fact shadows), which can make interpretation difficult. This difficulty is addressed in orthodox clinical radiography by taking images in two planes. Conventionally these are at right angles to each other. Unfortunately, this does not eliminate the issue completely and dilemmas of interpretation can still persist. As a result, researchers continued to address the problem. Bocage, Grossman, and Vallebona thus developed the idea first suggested by Mayer in 1914 and built their own Tomographic equipment (Seynaeve and Broos 1995: 284–288). Subsequently, Ziedses des Plantes published the most comprehensive study on tomography in 1931 (Seynaeve and Broos 1995: 284– 288). By swinging the X-ray tube back and forth around a specific fulcrum, it was possible to focus on one particular plane of tissue leaving the other planes out of

Fig. 1 The electro-magnetic spectrum

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Fig. 2 The principle of tomography

focus. By altering the focal distance, it was possible to produce a set of image slices through the body (Fig. 2). While tomograms were a significant advance, the development of the computerized axial tomography (CAT) scanner by Sir Godfrey Hounsfield in the late 1960s resulted in an enormous step forwards towards the current state of medical imaging. By rotating the X-ray tube through 3600 around the body, the image of a slice across the body is produced. The actual image cannot be recorded/captured on conventional X-ray film and has to be captured by electronic detectors (based on Cesium or Selenium) similar to those found in modern digital cameras and camcorders. These image components are then reconstructed using complex mathematical algorithms (Love et al. 2013). Therefore, the advent of powerful computers was essential to the development of CT scanning. Subsequently, the evolution of digital imaging and software to manipulate the image data permitted the creation of images in three standard (and an infinite number of nonstandard) planes in addition to 3D reconstruction (Fig. 3). As a result, it was possible to conduct Virtual Autopsies – or Virtopsies upon a body. Again, this depended on the change from an analogue to digital presentation of the image – a change which had huge implications for hospital diagnostic imaging departments. The continuous development of CT machines has enabled the initial image product of a slice separation or slice thickness of 1 or 2 cms to be reduced to a thickness in the region of 0.5 mms or less. In addition, the multiplication of detector rows from an initial 1 row (in itself a progression from the initial single detector, to a row of detectors) to a current array of up to 512 rows has resulted in impressive advances in image quality and definition. With the additional facility of image manipulation – for example, the deletion of part of the image data – it is possible to create pictures of selected parts of the subject without the distraction and masking

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Fig. 3 Standard body planes

of adjacent tissues. Again, these developments have only been made possible by a rapid increase in the computing “power” available.

Dual Energy Machines One further relatively recent development in the field of CT scans is the use of dual energy machines. These are scanners with (usually) two X-ray tubes, which can produce radiation at different energies. Although initially designed to enable double speed exposures (to permit the capture of images of fast-moving objects, for example, a beating heart), the tubes can also deliver different X-ray energy emissions so enabling imaging of two types of tissue with each exposure (for example allowing imaging of bone and soft tissues at the same time). The result is an increase in the definition of those images leading to easier demarcation of tissues of a similar but only slightly different radio-density. The slight disadvantage is that the files produced are usually much greater in size – therefore requiring Dicom reader software of greater sophistication and power (and, obviously more powerful hardware on which to run this software). Furthermore, dual energy machines are fewer in number and less accessible than single energy machines, usually only being situated in specialist referral centers. A further and more recent offering in the effort to produce images capable of differentiating tissues in this way is to produce one energy source and to capture the image using two distinctive detectors of different energy sensitivities.

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As has been alluded to above, CT scanning is essentially, more often than not a clinical phenomenon. It follows, therefore, that virtually all CT scanning machines are located in clinical facilities, that is, hospitals and clinics. This has the advantage that the machines are kept up to date, calibrated frequently and well maintained. Therefore, a well-developed and sustained relationship between museums and their local hospital/clinic is an essential factor in the on-going pursuit of the type of data that only CT scans can provide. The same comment can also be attributed to access to other modalities such as image intensifiers – see below. One other advance, which impacts on the use of medical imaging in the contemporary field, is the development and introduction of digital plain radiography and the use of digitized images and storage. Not only does this revolutionize the transfer of images it allows the development of computer software to manipulate the images. In many cases, it produces images of greater spatial resolution than CT but, on the other hand, no further reconstruction is available as the images are still in two planes and not the 360 format.

Other Imaging Modalities and Their Appropriateness Moving on now to other methods of image production, there are three further types of imaging currently available in the field of medical diagnosis. These are Ultrasound, MRI (magnetic resonance imaging), and Terahertz Imaging. The wavelengths in the electromagnetic spectrum that are used for these are indicated in Figs. 1 and 4. Ultrasound uses energy in the electromagnetic spectrum in the range 20 KHz to 2 MHz. The relationship of this range to 200 MHz is shown in Fig. 4 and this can be translated onto the more extensive range shown in Fig. 2 to obtain perspective.

low bass notes

animals and chemistry

20Hz

Infrasound

medical and destructive 2MHz

20KHz

Acoustic

Ultrasound

Fig. 4 Part of the electro-magnetic spectrum containing ultrasound

diagnostic and NDE 200MHz

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Ultrasound Ultrasound imaging relies upon the transmission of ultrasound waves through the object with the reflection of these waves at the various tissue interfaces thus producing the image. In order for this to be achieved, it is necessary to eliminate the interface at the surface between the probe and the object under investigation. In clinical practice, this is achieved by the use of a gel. However, the use of such a substance is obviously contraindicated in the case of ancient artifacts. Furthermore, the efficient transmission of the ultrasound waves requires the presence of fluid, clearly this is the very substance absent from desiccated tissues. Hence, ultrasound imaging is of little value in mummy studies.

MRI Clinical diagnostic MRI was developed in the 1970s by Lauterbur (1973) and Damadian (1971) and introduced into widespread clinical use in the 1980s and relies on the behavior of hydrogen atom nuclei when subjected to a magnetic field. When the protons are excited by the magnetic field, they flip their spins. When the magnetic field ceases, the protons return to a normal spin pattern and in doing so produce radio signals which are detected and measured. These signals are used to produce the image. The important feature is the presence of hydrogen in water (which forms 65–70% of the body in life). In mummies, this is one compound, which is absent (or almost absent) due to desiccation. Therefore, conventional MRI scans are of no use in most mummy studies. However, it is possible to alter the software to detect other elements and use these. In one instance sodium was used as the atom nucleus to be excited by the magnetic field and produced images in, for example, mummies of salt miners from Iran. However, this is an unusual example and to all intents and purposes MRI remains a rarely used tool in the investigation of mummies (Cockburn et al. 1998; Rühli et al. 2004). A further variation on the theme of MRI usage was demonstrated by Karlik et al. (2007: 105–110) when they optimized the software to detect very short relaxation times. This resulted in the ability to generate images of a desiccated brain. Clearly, the above comments apply to desiccated remains but not to remains containing water, for example Ice mummies and Bog mummies where these modalities do have a part to play in investigation and research. Shin et al. (2010: 329–334) added further to this concept when analyzing the body of a person from the sixteenth/ seventeenth century found in a Korean tomb. However, despite this, the use of MRI remains outside the mainstream of the majority of mummy research.

Terahertz Imaging Terahertz imaging is helpful in defining and imaging surface and immediately subsurface structures but not useful in exploring deeper structures (Ohrstrom et al.

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2010: 497–500). Its use in mummy studies and Paleopathology is, therefore, very limited and restricted to the investigation of such subjects as separating the images of wrappings from those of the body of the mummy to allow analysis.

Medical Imaging for Archaeology While not strictly the study of mummies, study of funerary artifacts from ancient Egypt is a natural corollary in this field of study. This can include the CT scanning of coffins to determine their construction – particularly in cases where various pieces of old coffins are reused and, in some cases, where the type of wood used can also be determined. The whole suite of funerary equipment also included canopic jars on numerous occasions. If sealed, the contents can be viewed by the noninvasive techniques of either conventional CT scans or micro-CT scans. Although the preceding sections cover the use of imaging as a diagnostic and analytical tool another function must also be mentioned. That is the use of medical imaging to aid in the collection of samples for other scientific analyses, for example, aDNA analysis. An original CT scan can be of great use in the location of targets for specimen collection. Subsequently, the use of an image intensifier with C arm can guide and direct an operator in the accurate collection of such samples while at the same time aiding the minimizing of damage to the artifact if instruments such as Bone Biopsy needles are used for the actual collection. An example of such a Bone Biopsy needle is shown in Fig. 5. In this particular case the external diameter of the needle is 3 mms and the internal diameter 2.4 mms. In the current age of the ability to successfully analyze very small samples chemically, such samples are quite adequate and produce only minimal damage to/intrusion into an artifact. This concept also fits in with the ethical dilemma of disturbing and taking tissue from human remains. In the UK the subject has been recognized by the Department for Culture, Media and Sport in Guidance for the Care of Human Remains in Museums which cites the Human Tissue Act (2004). Part 2: Section 9, stating that Fig. 5 Bone biopsy needle

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“the use of destructive sampling must have recorded scientific justification as a prerequisite. The smallest amount of tissue possible to complete the research should be taken by appropriately qualified staff. Also, all other nondestructive methods should be considered first.” However, a note of caution is also needed. The use of such a surgical instrument should be left to individuals trained, experienced, and skilled in such techniques as a great deal of damage can inadvertently be inflicted on the artifact by mishandling the device, while at the same time failing to actually obtain the desired specimen. Another ethical point to be taken into consideration is the subject of using clinical investigation devices for the investigation of mummies. However, the main objection here is the use of that machine on both live and dead bodies. It should be appreciated that this apparatus is regularly used to obtain images of dead bodies to produce forensic evidence for legal purposes. No objection to such use has been voiced and there is, in effect, no material difference between the two objectives. Also, after such investigations, it is common practice to perform a deep clean of the apparatus before use on live patients. Although not an immediately acknowledged role of academics, it does sometimes fall to them to contribute to museum exhibits and exhibitions. The product of medical imaging is a popular component of these experiences and can take the form of still images or video productions (such as rotating images of a body within a cartonnage, shroud, or coffin or alternatively progressive slices through a coffin/ mummy).

Radiation Damage to Ancient DNA The use of medical imaging techniques has one major advantage over other methods of investigation – it is noninvasive and, therefore, damage to unique and irreplaceable artifacts is avoided. This statement is made with one reservation – that relates to the potential of radiation damage to ancient DNA (aDNA). In living organisms, radiation damage is a well-accepted phenomenon. Mutations, DNA single-strand rupture and cell death [due to direct action of X-ray exposure and to indirect action of hydroxyl radicals (OH–) produced by water radiolysis (Roots and Osaka 1975)], base substitutions and deletions (Grosovsky et al. 1998) as well as structural changes (i.e., gene order rearrangements) have been described in biological contexts (Muller 1927; Wolff 1967; Liber et al. 1996). On another hand, cells have evolved mechanisms to detect and repair different types of damage that can occur to DNA, including moderate levels of radiationinduced damage (Clancy 2008); this control of DNA repair is tied to the regulation of the cell cycle through checkpoint mechanisms (Clancy 2008). When failures in these checkpoints occur, an accumulation of damage and mutations become manifest (Clancy 2008). For these reasons, all possible measures are used to minimize the exposure of patients to the inevitable radiation related to X-rays, CT scans, and other modalities using ionizing radiation (e.g., SPECT – Single Photon Emission CT and PET – positron emission tomography).

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Ancient DNA undergoes postmortem decay and, of course, no repair mechanisms exist. Oxidative and hydrolytic damage can cause structural modifications to the DNA (Lindahl 1993; Mitchell et al. 2005) which add to taphonomy. Hence, it has been hypothesized that, if ancient remains are exposed to significant levels of X-ray doses (i.e., repeated CT scans over time), their damaging effects could progressively add to one another and produce a reduction in the preservation of ancient DNA (Immel et al. 2016). Contradictory results in terms of radiation effects on DNA fragmentation have been obtained so far. Earlier studies by Grieshaber et al. (2008) and Paredes et al. (2012) attempted to assess the mechanisms of radiation-induced damage using Polymerase Chain Reaction (PCR) and electrophoresis-based analyses, respectively. Götherstrom et al. (1995) and Grieshaber et al. (2008) resorted to modern pig (Sus scrofa) foot bones as proxies for archaeological samples and target the degree of DNA fragmentation resorting to Polymerase Chain Reaction (PCR). Their results implied that multiple exposures to X-rays and CT scans produce an increased degree of DNA fragmentation. On the other hand, Paredes et al. (2012) extracted ancient DNA, before and after CT scanning, from desiccated bird skins. To infer the level of fragmentation induced by X-ray exposure, they used capillary electrophoresis-based DNA quantification. Pre- and post-CT fragmentation profiles did not show significant difference between the exposed tissues and the controls. More recently, the effects of X-radiation on fragmented DNA in dry, wet, and frozen states were simulated (Wanek and Rühli 2016) showing that the highest probability of radiation-induced DNA damage occurs in a wet state and, thus, supporting the notion that hydrolysis plays a critical role in aDNA preservation (Immel et al. 2016). Since the effects of X-ray irradiation had never been applied to authentic aDNA molecules from subfossil specimens, Immel et al. (2016) analyzed Late Pleistocene megafauna (i.e., cave bear, giant deer, bison, roe deer) using a polychromatic synchrotron beam covering a large range of dose level and energy relevant for both synchrotron and conventional X-ray sources (Immel et al. 2016). For doses below 200 Gy (Gray), no substantial degradation on ancient endogenous DNA was shown. Dosimetry confirmed that X-rays and conventional CT scans used in medical settings never reach a 200 Gy dose. An almost complete loss of ancient amplifiable DNA was recorded in sample aliquots exposed to a high X-ray dose of 170 kGy. A series of recommendations to perform conventional and synchrotron X-ray scanning of subfossils without preventing future aDNA investigation were propose by Immel et al. (2016) and are listed in Table 1.

Benefits and Disadvantages Having reviewed the currently available imaging options, the benefits and disadvantages should be considered. Clearly, the overarching benefit is the noninvasive nature of all imaging options. The next consideration is the quality of the product – that is, the image itself. There is little doubt that the greatest benefit lies with CT scans as they

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Table 1 General recommendations to apply to radiological investigations of ancient subfossilsa Recommendations for conventional CT of organic remains Scans should not be performed without a metallic filter, and the use of filters to remove the lower energies of the spectrum is advocated. The systematic use of at least 0.1 mm copper or brass filters is recommended for scanning such remains General recommendations for CT scans of organic remains Do not perform micro CT on wet artifacts, e.g., frozen specimens. Do not put the samples into water when performing a micro CT scan Do not perform scans at a higher resolution than absolutely necessary (dose increases by approximately the square of the increase in resolution) Do not perform multiple scans if comparable data already exists as the dose is cumulative. Data sharing (under the control of curators) and public databases should be made available Record all scanning parameters & c. and supply them to the owner of the artifact in order to preserve the whole scanning history of an artifact. Prior to the repeat scanning of an artifact, its scanning history should be reviewed to calculate the total dose accumulated to date (particularly when a synchrotron is used) After Immel et al. (2016)

avoid the problem of combination/superimposed images and allow virtually infinite options in the manipulation of the images. These include varying contrast, brightness, and magnification. As stated above, reconstructions can be performed in any of the three standard planes (sagittal, coronal, and axial) and an infinite variety of nonstandard planes. 3D reconstruction can be edited with reference to color, contrast, brightness, magnification, rotation and tissues can be removed to reveal a chosen view of other, selected tissues. This, in turn, allows comprehensive Virtopsies to be performed where appropriate. These investigations permit the development of interpretations relating to pathological conditions and, on occasions, causes of death as well as funerary techniques such as embalming where applicable. It also allows the identification of “enclosing” materials (such as linen wrappings or shrouds) and accompanying objects (such as amulets or ritual pouches and others). The major disadvantage being extreme difficulty in performing such investigations “in the field.” Although portable CT units are available, they invariably offer less sophistication than static units. Certainly, currently the option of dual energy machine usage is restricted to static units. Digital X-ray machines are available as portable units and are, therefore, suitable for use in the field and do produce high quality images. They also have the advantage of producing digital images that can be stored and transported on a computer or even a digital storage medium such as a memory USB stick. However, at present, they do not offer the option of significant post-production image manipulation and an extensive examination can be limited to multiple, overlapping X-ray “films/images” in two planes. Although early solutions to the subject of producing a CT scan with a portable digital X-ray machine are being developed, they are in an early stage of that development and seem to face a number of significant obstacles. While the older versions of X-ray machines are still available, the product is an analogue image on photographic film, which then requires chemical processing in a

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dark room. This can be a challenge in the field. A further consideration is that with the explosion in the routine use of digital imaging, the availability of the analogue Xray film and the chemicals necessary to process the film will, almost inevitably, become more and more of an increasingly tenuous supply issue. Having examined the subject of production of the image, the digital processing of that image needs some discussion. At this point it is important to recognize that the product of a CT scanner is in a Dicom format data file which is an agreed industry standard format that can be further processed by a variety of reader software programs. There are many options in terms of Dicom Reader software including those specifically designed for use in the medical diagnostic field and those designed for high level engineering design demands. The software also comes in formats which are specific to the manufacturer of the CT machine and those which are third-party and designed to handle files from many sources. Costs vary considerably and the platform upon which they are capable of running is usually a choice between Windows or Mac operating systems, although systems are emerging that will run on both platforms (there is at least one offering which will run on both of these and on Linux). Selection of the appropriate software is important as this can fundamentally influence the resulting images and their quality and, hence, the ability to differentiate between structures – particularly those of similar but slightly different densities and those of small size. File size handling is another important consideration when choosing software with only the more sophisticated programs (those devised for the design field) capable of handling the larger file sizes (such as those produced by micro-CT). With ever progressing advances in technology, the question arises as to how often consideration should be given to re-submitting artifacts for repeat imaging. Clearly this involves balancing the possible advantages which the new technology provides against the risks involved (e.g., transport damage and radiation damage). While it is the choice of the individual researcher, a CT scan with a slice thickness of more than 1 mm. might be seriously considered for re-imaging.

The Future The current position of medical imaging in mummy studies is, undoubtedly, likely to change in the future as more developments take place in the design of both CT scanners and the equipment delivering other modalities of imaging. Contemporary dialogue with developers and manufactures indicates that the present thrust of research and development is, on the one hand, more towards dual purpose machines and on the other, the refinement of image collection, transmission, and analysis. The dual-purpose machines being developed are designed to provide twofold modalities for speedier diagnosis in the clinical field – for example, a CT scanner which also provides PET or SPECT functionality. Clearly these secondary modalities are of no particular value in the field of Paleopathology as they depend upon the physiology of a living subject – by definition at variance with the subject under discussion.

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The developments in image collection, transmission, and analysis will lead to better definition and image manipulation. This would be of great benefit in the future. It is clear that this development will have to be accompanied by significant increases in the ability of Dicom Reader software to handle the ever-increasing size of image files. In other words, more complex Reader software running on ever more powerful hardware. It is implicit that the spatial resolution delivered by a Dicom Reader is related directly, not only to the sophistication of the software, but also to the sophistication of the hardware on which it is running. This includes not just the computer/server but the actual screen on which the image is delivered (in other words, Pixel/Voxel density is very important). Another factor to be remembered is the need for an ever-increasing capacity in the field of reliable, secure data storage. On this topic, it is important to note that whenever an artifact is imaged, the product, that is, the Dicom file in its totality should be held, not only by the hospital, but also by the museum possessing the artifact (preferably in, at least, duplicate). It can, in fact, become part of the provenance material of that object. Furthermore, it is important that the medium used for storage is kept up to date. Media do change from time to time and older media can, on occasions, be difficult or impossible to read years later when the appropriate hardware is no longer available. Flow charts of the journey to facilitate imaging and Dicom File storage can be found in Fig. 6. Currently in the early phase of development as a new medical imaging modality is Phase Contrast Imaging. This has the benefit of producing very detailed images of soft tissues – much enhanced on those produced by conventional X-rays and CT scans. Initial attempts at its development required massive equipment such as a Synchrotron (a type of particle accelerator) as the source of radiation. It is, currently, difficult to find references to the practice of this modality actual being in use in hospitals. Its usage, therefore, depends on transporting the subject to a dedicated Synchrotron site – usually a laboratory. However, the massively improved image quality should propel the advance of clinically appropriate machines (and their increased availability) in the near future. The development of Phase Contrast Imaging is certainly something to scrutinize over the next few years. While not strictly medical imaging, the use of micro-CT scanning must also be mentioned. The advantage of this modality is the enormous increase in image resolution. Unfortunately, the size of the examined object can be a limitation (with very few machines accommodating objects larger than 40–50 cms) and, therefore, it is usually limited to the examination of animals, birds, and fragments of human mummies. Another limitation is the large file size of data produced, which may require robust Dicom Reader software of a specialized nature. Finally, the subject of radiation does have to be mentioned. In micro-CT, this is much higher than in a conventional clinical scanner and may have to be a consideration when assessing the subject of aDNA damage. One of the most recent developments to be applied to scan analysis is the use of AI (Artificial Intelligence). Currently in a developmental phase and not usually available for routine clinical use, this function is said to be able to distinguish between tissues which are apparently the same to the naked eye. One potential

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Fig. 6 (a) Flow charts of “journey” to facilitate imaging and Dicom File storage (b) Fate of the file

problem in the field of mummy studies is that to develop it, AI needs to learn the subtle differences between tissues and their radiological densities and this requires exposure to many images. Clearly in the clinical field this does not presents a problem as thousands of new images are available each day. However, in the arena of mummy studies, the number is limited and this may delay its use in this field as AI learning will take so much longer.

Conclusion In 1895 the discovery of X-rays added a potent tool to the investigation of mummies. It was no longer necessary to perform intrusive and destructive processes in order to acquire details of the internal structure of those mummies. X-rays produced details of morphology which was previously only obtainable by autopsy/dissection. The initial residual problem was one of visualizing the subject in a 2-dimensional format when it was, in reality, a 3-dimensional object. Further research and development led to the production of the tomogram. However, it was not until the 1960s that the next and most important advance occurred. This was the development of computerised axial tomography by Godfrey Hounsfield. This, with the change to the format of digital imaging in the late 1990s, produced the current position of medical

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imaging with the ability to perform virtual autopsies. The result is data rich files for ongoing research but with NO damage to the invaluable and irreplaceable artifacts – the mummies. In this chapter, appropriate imaging modalities are discussed along with the subject of data security and storage (potentially future proofing that data). Although many medical imaging formats (modalities) have been developed and huge strides forwards have been achieved in the past two or three decades, not all are suitable for mummy research because of the nature on mummies themselves. Future advances in appropriate research techniques may propel medical radiology to being more useful in the fields of anthropology and paleopathology.

Cross-References ▶ Joseon Dynasty Mummies of Korea ▶ Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection ▶ Radiology Applications in Mummy Science

References Adams J (2016) Mummies, magic and medicine in ancient Egypt. Essays for Rosalie David. Manchester University Press, Manchester Clancy S (2008) DNA damage & repair: mechanisms for maintaining DNA integrity. Nature 1(1):103 Cockburn A, Cockburn E, Reyman TA (1998) Mummies, disease & ancient cultures. Cambridge University Press, Cambridge Damadian R (1971) Tumor detection by nuclear magnetic resonance. Science 171:1151–1153 David R (2002) Religion and magic in ancient Egypt. Penguin Books, London Götherstrom A, Fischer C, Linden K (1995) X-raying ancient bone: a destructive method in connection with DNA analysis. Laborativ Arkeologi 8:26–28 Grieshaber BM, Osborne DL, Doubleday AF et al (2008) A pilot study into the effects of X-ray and computed tomography exposure on the amplification of DNA from bone. J Archaeol Sci 35(3):681–687 Grosovsky AJ, de Boer JG, de Jong PJ et al (1998) Base substitutions, frameshifts, and small deletions constitute ionizing radiation-induced point mutations in mammalian cells. Proc Natl Acad Sci U S A 85:185–188 Ikram S, Dodson A (1998) The mummy in ancient Egypt. Equipping the dead for eternity. Thames & Hudson, London Immel A, Le Cabec A, Bonazzi M et al (2016) Effect of X-ray irradiation on ancient DNA in subfossil bones – guidelines for safe X-ray imaging. Sci Rep 6:32969. https://doi.org/10.1038/ srep32969 Karlik SJ, Bartha R, Kennedy K et al (2007) MRI and multinuclear MR spectroscopy of 3,200-yearold Egyptian mummy brain. Am J Roentgenol 189(2):W105–W110 Lauterbur PC (1973) Image formation by induced local interactions: examples employing nuclear magnetic resonance. Nature 242:190–191 Liber HL, Leong PM, Terry VH et al (1996) X-rays mutate human lymphoblast cells at genetic loci that should respond only to point mutagens. Mutat Res 163:91–97

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Lindahl T (1993) Instability and decay of the primary structure of DNA. Nature 362:709–715 Love A, Olsson M-L, Siemund R et al (2013) Six iterative reconstruction algorithms in brain CT: a phantom study on image quality at different radiation dose levels. Br J Radiol 86(1031):20130388 Mitchell D, Willerslev E, Hansen A (2005) Damage and repair of ancient DNA. Mutat Res 571:265–276 Muller J (1927) Artificial transmutation of the gene. Science 66:84–87 Ohrstrom LM, Bitzer A, Walther M et al (2010) Technical note: terahertz imaging of ancient mummies and bone. Am J Phys Anthropol 142(3):497–500 Paredes UM, Prys-Jones R, Adams M, Groombridge J, Kundu S, Agapow P-M, Abel RL (2012) Micro-CT X-rays do not fragment DNA in preserved bird skins. J Zool Syst Evol Res. https:// doi.org/10.1111/j.1439-0469.2012.00657.x Roots R, Okada S (1975) Estimation of life times and diffusion distances of radicals involved in x-ray-induced DNA strand breaks of killing of mammalian cells. Radiat Res 64:306–320 Rühli FJ, Chhem RK, Böni T (2004) Diagnostic paleoradiology of mummified tissue: interpretation and pitfalls. Can Assoc Radiol J 55(4):218–227 Seynaeve PC, Broos JI (1995) The history of tomography. J Belg Radiol 78(5):284–288 Shin DH, Lee IS, Kim MJ et al (2010) Magnetic resonance imaging performed on a hydrated mummy of medieval Korea. J Anat 216(3):329–334 Wanek J, Rühli FJ (2016) Risk to fragmented DNA in dry, wet, and frozen states from computed tomography: a comparative theoretical study. Radiat Environ Biophys 55(2):229–241 Wolff S (1967) Radiation genetics. Annu Rev Genet 1:221–224

Part III Ancient DNA Analysis and Mummy Research

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Jong Ha Hong, Chang Seok Oh, and Dong Hoon Shin

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analysis of Mitogenome . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . DNA Fingerprinting Techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tuberculosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Archaeoparasitology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chagas’ Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Hepatitis B Virus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Smallpox . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Helicobacter pylori . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Genetic Predispositions to Diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Breakthroughs in aDNA Analyses of Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Since the pioneering reports on successful extraction and analysis of ancient DNA (aDNA) in the mid-1980s, aDNA research has become a topic of great interest among academics and even the public as well. Inspired by the impressive preservation status of mummies, the earliest research on aDNA focused on mummies J. H. Hong (*) Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_56

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and mummified specimens across the world. Since then, mummy studies have produced a number of remarkable reports in parallel with the advances achieved in archaeogenetics. In this chapter, the brief history of aDNA analysis in mummy studies is summarized, and the prospects for this field will be discussed as well. Specific information on aDNA analysis in both its theoretical and practical aspects will be covered in detail in other chapters of this book. Keywords

Ancient DNA · Mummy · History · Prospect · Introduction

Introduction In the mid-1980s, one of the earliest successful analyses of ancient DNA (aDNA) was carried out on a nineteenth-century quagga, an extinct zebra species (Higuchi et al. 1984). By cloning and sequencing of mitochondrial DNA (mtDNA) amplicons, the researchers were able to obtain the first glimpse of an ancient genome (Frantz et al. 2020). Following this proof-of-concept study, aDNA research began to draw a great deal of attention from academics and the public as well. Looking back on the history of aDNA research, however, it was never a smooth advance, but rather, a series of crises and controversies. At the early stage of aDNA analysis, retrieval of aDNA molecules largely depended on polymerase chain reaction (PCR), a technique that was at that time instrumental to the analysis of small amounts of aDNA remnant in archaeological specimens. Due to technical difficulties and low throughput, however, genetic information retrieved by PCR was limited to a small number of loci (Frantz et al. 2020). Another major issue in the early days of aDNA study was the authenticity of PCR-amplified products. Most importantly, due to the poor preservation status of aDNA fragmented by oxidative or hydrolytic reactions, even a small amount of contamination from modern sources can dominate the PCR products. That is, PCR of targeted sequences might not exclusively amplify authentic aDNA fragments, but also modern exogenous, contaminant DNA (Marota and Rollo 2002; Willerslev and Cooper 2005). Therefore, as might be expected, there were academic disputes on whether aDNA amplified by PCR was actually authentic (Roberts and Ingham 2008). Many researchers agree that aDNA study is not free from controversy and thus has to be carried out with great care in order to provide authentic data (Donoghue et al. 2004). As a result of community-wide discussion (Pääbo et al. 2004), in the 2000s, a set of research criteria were suggested to reduce the likelihood of contamination in samples and to ensure the authenticity of aDNA analysis thereby (Willerslev and Cooper 2005). Nevertheless, those criteria can only indirectly determine the authenticity of aDNA data, and so the issue of potential contamination remained unresolved in PCR-based aDNA analysis. Technical advances during the last decade have provided a major breakthrough in terms of throughput and authenticity in aDNA study. First, next-generation

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sequencing (NGS) could provide the necessary throughput to enable shotgun sequencing of ancient genomes, even in cases where the amount of exogenous, contaminant DNA far exceeds that of authentic, endogenous aDNA (Frantz et al. 2020). Prior to the introduction of NGS techniques to aDNA analysis, only short stretches of mtDNA or a few other loci could be acquired from ancient specimens (Frantz et al. 2020). With the advancement of NGS, even ultra-short aDNA fragments (as short as 30–35 base pairs; bps) can now be analyzed by parallel sequencing. This cutting-edge technique thus rapidly replaced conventional PCR commonly used in archaeological science in the past. Next, we should note the novel techniques of DNA capturing that substantially enrich either the whole genome or a predefined set of genomic regions within the sequencing library. Indeed, by application of capturing techniques, extremely poorly preserved samples can be analyzed (Carpenter et al. 2013; Haak et al. 2015). Finally, it is now known that certain skeletal elements (e.g., the otic capsule protecting the inner ear bone) preserve endogenous DNA far better than others (Gamba et al. 2014). All of these recent advancements in aDNA analysis helped researchers to achieve much more authentic results that were previously unavailable with the conventional PCR technique (Frantz et al. 2020). From the earliest days of aDNA analysis, mummies were especially useful subjects due to their superb preservation status. The earliest report of mummy aDNA concerned a several-kilobase DNA stretch of HLA-DQA gene from an ancient Egyptian mummy (Pääbo 1985). In the case of mummies, aDNA analysis has been performed not only for bones but for various parts of soft tissues as well. Well-preserved parts of mummies have been used to conduct aDNA research at higher rates of success. In fact, certain parts of the mummified body (e.g., stomach) can yield genetic information (e.g., Helicobacter pylori) that cannot be obtained simply from bones. On this basis, many fascinating and ground-breaking reports have embroidered the history of mummy aDNA research that has been recognized for its unique academic significance. In this introductory chapter, we summarize a wide range of aDNA studies that have been conducted on mummified biological remains.

Analysis of Mitogenome Whereas there are only two copies of nuclear genomes per diploid cell, hundreds and even thousands of mitochondrial (mt) DNA copies exist in a single cell. Because of this high copy number, mtDNA analysis has been commonly performed in genetic investigations of mummies. Thanks to the protocols of forensic sciences, huge accumulations of relevant mtDNA haplotypes are available nowadays. Therefore, the mtDNA haplotype acquired from mummies can be easily analyzed and compared with other haplotypes reported from all over the world. Herein, we provide an overview of a few representative examples of mummy mtDNA studies. In 1991, the Tyrolean Iceman, a European Chalcolithic male, was found on the Tisenjoch Pass in the Ötztal Alps (Keller et al. 2012). The first genetic study on the

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Tyrolian Iceman (Ötzi) reported a hypervariable (HV) segment of mtDNA showing that his mitotype fell within the diverse range of contemporary central and northern European populations (Handt et al. 1994). In 2008, using PCR amplification and 454 sequencing, Ermini et al. (2008) retrieved the first complete sequence of the mitochondrial genome of the Iceman. They revealed that the Iceman belonged to a western Eurasian mtDNA haplogroup (K1), but a specific branch within it has not been found among modern European peoples. In the New World, mtDNA analyses on South American mummies continued. Monsalve et al. (1996) first revealed the mtDNA haplotypes of Colombian mummies (470–1849 AD). Luciani et al. (2006) also recovered aDNA from a pre-Columbian mummy (dated 980–1170 CE) found in Cuzco (Peru). Gómez-Carballa et al. (2015) studied a 7-year-old victim of Inca sacrifice who was discovered frozen on Aconcagua mountain (Argentina). The 500-year-old mummy’s complete mitogenome sequence belonged to haplogroup C1b, which thus far has not been reported for contemporary Native Americans (Gómez-Carballa et al. 2015). Besides South America, ancient mtDNA was also retrieved from 600-year-old North American mummies (Handt et al. 1996). In Egypt, mtDNA studies have targeted both human and nonhuman mummies. Genetic analyses were performed for the mtDNA control region of mummified cats (664–332 BCE). The sequences suggested that the cat mummies represented the mitotypes of modern domestic cats prevalent in the Middle East (Kurushima et al. 2012). Schuenemann et al. (2017) also presented mitochondrial and genome-wide genetic data from Egyptian human mummies (n ¼ 90). Their analyses revealed that ancient Egyptians shared an ancestry with the people of the Near East, and that Egyptian mummies could be considered as a repository of ancient genetic data revelatory of past human migration history.

DNA Fingerprinting Techniques Over the decades, aDNA analysis expanded its scope in archaeological as well as forensic sciences (Marota and Rollo 2002; Cipollaro et al. 2005). Many genetic analysis techniques in mummy studies were originally developed and first used in the field of forensic science. DNA analysis in archaeological science has been performed in a manner similar to that characteristic of crime scene investigation. In forensic science, autosomal short tandem repeat (STR) genotyping has been performed for human identification or paternity testing. Likewise, DNA analysis of archaeological samples has been carried out to obtain forensic DNA profiles of buried individuals. In order to analyze STR genotypes with small sample volumes, forensic and anthropological scientists have employed multiplex kits (Hummel et al. 1999; von Wurmb-Schwark et al. 2003; Iwamura et al. 2004; Ricaut et al. 2005, 2006; Amory et al. 2007; Di Nunno et al. 2007). Even so, complete autosomal STR profiles can be difficult to obtain from seriously degraded ancient specimens. Once secured, however, such data can provide us with substantial genetic information on long-buried individuals.

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DNA fingerprinting techniques have been used for the reconstruction of the family trees of historically important people (Gill et al. 1994; Clisson et al. 2002; Coble et al. 2009; Vanek et al. 2009; Baca et al. 2012). For instance, aDNA analysis was very useful in solving the mystery of the Romanov royal family (Gill et al. 1994; Coble et al. 2009). Amory et al. (2007) also reported STR genotyping on ancient hairs collected from sixteenth- to nineteenth-century Siberian mummies. Y-chromosomal DNA analysis was performed to infer the genetic sex of seventhcentury Taklamakan desert mummies whose external morphology was too incomplete to permit sex determination (Lin et al. 1995). DNA analysis was performed on the kindred of Tutankhamun, one of the most famous kings of all ancient civilizations. By a genetic fingerprinting technique, a multigeneration family tree of Tutankhamun could be successfully revealed (Hawass et al. 2010).

Tuberculosis Among the aDNA analyses of mummified tissues, the studies of ancient microbial DNA have opened a new horizon in paleopathology, as genetic data from ancient or medieval mummies thus obtained represent the evolutionary history of pathogenic microbes over time and place, thus answering historical questions unresolved so far. Mycobacterium tuberculosis, a pathogen commonly causing serious infections in human history, is particularly important in mummy research. A series of aDNA analyses provided invaluable scientific information on the evolutionary model of M. tuberculosis as well as the interaction between the ancient bacterial pathogen and its hosts: human beings and animals (Zink et al. 2002; Donoghue et al. 2004). In South America, there have been many reports of ancient tuberculosis in mummies. The report of Salo et al. (1994) was the first recovery of M. tuberculosis aDNA, specifically from lung tissue of a 1000-year-old female mummy found in Southern Peru. This aDNA report was significant, because it provided, for the first time ever, evidentiary proof of the presence of pre-Columbian human tuberculosis in the New World. In 1995, another case of pre-Columbian tuberculosis was genetically confirmed, that time for an eleventhcentury Chilean girl with Pott’s disease (Arriaza et al. 1995). In South America, the aDNA of M. tuberculosis and mycobacteria other than M. tuberculosis (MOTB) were successfully extracted from mummies (dated 140–1200 CE) of the Andean Mountain region and analyzed (Konomi et al. 2002). Recent whole-genome studies have confirmed pre-Columbian cases of tuberculosis and suggested the pinnipedinfecting M. tuberculosis lineage as the source (Bos et al. 2014; Nelson et al. 2020). Egypt is another place where important information on ancient tuberculosis has been acquired. Nerlich et al. (1997) managed to examine an Egyptian mummy of the New Kingdom found in the tombs of the nobles. They amplified M. tuberculosis aDNA and evidenced its presence in the ancient Egypt mummy’s lung. Zink et al. (2002) analyzed samples from ancient Egyptian mummies using spoligotyping for M. tuberculosis complex DNA. When compared with the international database of spoligotyping signatures, the Mycobacterium of the mummies was revealed to be of

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M. africanum or M. tuberculosis, but not of M. bovis. In particular, the Middle Kingdom samples showed a spoligotyping signature specific for M. africanum, whereas those of later periods expressed the M. tuberculosis patterns. These results did not support the conventional understanding that M. tuberculosis originated from M. bovis. Instead, human M. tuberculosis might have evolved from a precursor complex possibly related to M. africanum (Zink et al. 2002). A female mummy of ancient Egypt, nicknamed Dr. Granville’s mummy, was also subject to a scientific study of ancient tuberculosis. Morphological evidence showed a potential sign of fatal pulmonary exudates in the mummy. By nested PCR of IS6110, M. tuberculosis complex DNA was acquired from the mummy’s lung and gall bladder samples. It was confirmed that tuberculosis was likely to have been the Dr. Granville’s mummy’s major cause of death (Donoghue et al. 2010).

Archaeoparasitology DNA analyses on parasitological specimens (e.g., coprolites) from mummies provide information pivotal to a comprehensive understanding of changes in the genetic characteristics of many parasite species throughout history. The earliest studies on ancient parasite DNA were reported by Loreille et al. (2001) and Leles et al. (2008). After they successfully obtained Ascaris genes by PCR amplification performed on archaeological samples, aDNA analysis has been increasingly utilized for phylogenetic analysis of various parasite species.

Chagas’ Disease Trypanosoma cruzi, a pathogen causing Chagas’ disease, is a parasite commonly examined in archaeoparasitology. Many studies of ancient Chagas’ disease have been reported from the coastal region of northern Chile, where the extremely arid climate has preserved mummies very well (Guhl et al. 2014). DNA of ancient T. cruzi was first isolated from mummies (dated from 2000 BCE to 1400 CE) found in northern Chile. This molecular biological report identified Chagas’ disease in a case for which gross anatomical evidence was not available (Guhl et al. 1999). Ferreira et al. (2000) reported a conserved region of the minicircle molecule of T. cruzi using mummified specimens from the Museo Arqueologico de San Pedro de Atacama. Tissue specimens of as many as 283 human mummies (estimated date: 7050 BCE–1500 CE) from the low valley and coastal sites in northern Chile and southern Peru were also tested for a kinetoplast DNA segment of T. cruzi. Among the mummified specimens, 41% showed positivity for T. cruzi aDNA, revealing that Chagas’ disease was prevalent when the Chinchorro culture still prospered on the Andean coast (Aufderheide et al. 2004). Fernandes et al. (2008) evaluated T. cruzi DNA from a 560-year-old Brazilian mummy. Having found T. cruzi minicircle DNA in the mummy with a sign of megacolon, they argued that Chagas’ disease was prevalent in Brazilean society even before European

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colonization. A molecular study on the Atacama Desert mummies recently revealed the evolutionary history of T. cruzi and related Chagas’ disease (Guhl et al. 2014).

Hepatitis B Virus Hepatitis B virus (HBV) is a common viral pathogen that has caused large-scale morbidity and mortality around the world. However, as for its origin and evolution over the broad time-scale, there remains uncertainty (Patterson Ross et al. 2018). Information concerning HBV evolution has been provided by aDNA analyses on European and Asian mummies. A shotgun analysis of HBV for a mid-sixteenth-century Italian child mummy (NASD24), for example, was successful in completely reconstructing an HBV genome (Patterson Ross et al. 2018). The phylogenetic analyses showed a close relationship between the putative ancient HBV virus of the Italian mummy (NASD24SEQ) and modern D genotype sequences. However, the researchers also found that the HBV sequence of NASD24SEQ showed characteristics of an ancient origin (Patterson Ross et al. 2018). A similar pattern was observable in an HBV aDNA analysis of a pre-modern Korean mummy (Kahila Bar-Gal et al. 2012). These studies indicate that the genotypes of HBV diversified long before the sixteenth century; accordingly, potential similarities (loss of diversity) were found between modern and ancient HBV DNA in both the Italian and Korean mummies (Patterson Ross et al. 2018).

Smallpox Paleopathological claims of smallpox in ancient civilizations dates back millennia. Duggan et al. (2016) analyzed a genome of an ancient strain of variola virus from a Lithuanian child mummy of the seventeenth century. They revealed that smallpox evolution was more recent than commonly expected, with viral gene diversification occurring from the eighteenth to nineteenth centuries.

Helicobacter pylori H. pylori is one of the most prevalent bacterial pathogens involving human gastric mucosa. It is globally found in human hosts and shows a remarkable phylogeographic pattern reflecting human migrations in history. Maixner et al. (2016) screened for H. pylori aDNA remnant in gastrointestinal tract samples from Ötzi the Iceman and were able to retrieve a 5300-year-old H. pylori genome. Metagenomic analysis revealed that endogenous H. pylori DNA from the Iceman belonged to the Asian lineage. This means that the African H. pylori lineage might have been introduced into Europe after the time of the Iceman, admixed with the indigenous H. pylori, and formed the present-day hybrid lineage (Maixner et al.

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2016). Shin et al. (2018) also isolated ancient H. pylori DNA, this time from the stomach specimens of Korean mummies. However, full characterization of Joseon mummy H. pylori aDNA will have to await the application of cutting-edge techniques such as next-generation sequencing (NGS).

Genetic Predispositions to Diseases HLA class II histocompatibility antigen is a protein encoded by the HLA-DRB1 gene. DRB1 alleles are known to be closely associated with rheumatoid arthritis (RA). DNA analysis of the HLA-DRB1 gene to reveal genetic predisposition to RA has been performed on several mummies. In the case of Ötzi the Iceman, HLA class II alleles were analyzed; and the Ötzi’s type was revealed to be HLA-DRB1*1402, which is extremely rare among present-day Europeans, but common in Inuits and Native South Americans (Fisher et al. 2001). The genetic predisposition to RA was also evaluated for the sixteenth century partially mummified “Braids Lady” of Arezzo (Fontecchio et al. 2007), which study revealed genetic signs of RA features (DRB1*0101 allele). In oncogenetics, the mutations of the adenomatous polyposis coli (APC) gene are known to induce colorectal adenomas and carcinomas. Feldman et al. (2016) successfully sequenced E1317Q missense mutation in the APC genes of eighteenth-century Hungarian mummies. Their study served to demonstrate the need for historical cancer epidemiology by larger-scale analysis of specimens from different time periods and geographical locations. DNA studies of mummies also have proved that atherosclerotic cardiovascular disease (ASCVD) was much more common in ancient populations than expected. Keller et al. (2012) revealed, by whole-genome sequencing, that the 5000-year-old Tyrolean Iceman had a genetic predisposition to coronary heart disease. In a radiological examination, they identified major calcifications in arteries and the aorta as possible signs of atherosclerosis. They, therefore, searched for genetic risk factors of ASCVD by analysis of related SNPs in genome-wide association studies. In the study, they found multiple SNPs of a major risk locus for coronary heart disease, ischemic stroke, atherosclerosis, and sudden cardiac death (Keller et al. 2012). A similar genetic analysis was also performed on a seventeenth-century Joseon mummy with possible radiological signs of ASCVD (Shin et al. 2017).

Breakthroughs in aDNA Analyses of Mummies Often, ancient specimens contain extremely low levels of endogenous DNA, the remaining major portion being composed of contaminant DNA from the environment (Carpenter et al. 2013). As mentioned above, in recent years, technical breakthroughs for improved authenticity and throughput have been adopted in aDNA analyses. With the advancement of high-throughput sequencing, DNA capturing, and metagenomics, a revolution in aDNA research was inaugurated.

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NGS analysis guaranteed a more authentic and powerful investigation of ancient mummies’ genomes (Khairat et al. 2013). This technique has been used in archaeological science, especially in cases where the recovery of aDNA sequences was impossible due to extremely short endogenous DNA molecules. Keller et al. (2012) reported the complete genome sequence of Ötzi the Iceman, revealing indications for a recent, common ancestry of the Chalcolithic mummy and modern inhabitants of Sardinia. A novel NGS technology also has been applied to Graeco-Roman Egyptian mummies (806 BCE–124 CE) (Khairat et al. 2013). Loreille et al. (2018) conducted molecular sex estimation of an ancient Egyptian mummy by shotgun sequencing, demonstrating that nuclear genome data could be successfully recovered from the seriously damaged ancient specimens. To guarantee successful retrieval of endogenous DNA molecules from mummy samples, researchers have adopted capturing techniques to enrich the endogenous component of aDNA. Using RNA baits transcribed from human genomic DNA libraries, Carpenter et al. (2013) captured aDNA fragments from Peruvian mummies. Whole-genome capturing remarkably increased the sequencing yield, thereby showing its utility as an aDNA sequencing tool even in cases of very low levels of endogenous DNA remnant in mummy specimens. Metagenomics is another scope that mummy research promises to expand. Maixner et al. (2014) analyzed the metagenomics data from Ötzi the Iceman to screen bacterial ribosomal RNA (rRNA)-specific reads. In the analysis, they found metagenome sequences of T. denticola aDNA, revealing that disease-associated microorganisms could be detected by metagenomics for datasets from ancient human remains. The microbiome is another emerging subject in biomedical science, and certainly, the study of intestinal microorganisms is not rare in mummy studies either. The microbiome in the human intestine has a distinct set of microbial inhabitants such as bacteria, viruses, fungi, and others. Nowadays, the microbiome is known to be integrally linked to the health and disease of mankind. Many diseases (cancer, autoimmune diseases, etc.) are thought to be influenced by the gut microbiome, and indeed, to thwart the development of the disease, scientists seek to control the microbiome (Brody 2020). In the earliest days of aDNA research, scientists tried to investigate the normal flora of Ötzi the Iceman’s stomach and colon by PCR amplification (Cano et al. 2000). However, cutting-edge techniques (highthroughput sequencing and metagenomics) thereafter have been used for analysis of the microbiome. This trend in medical science, unsurprisingly, also has influenced recent mummy aDNA studies. Santiago-Rodriguez’s group has reported a series of microbiome studies using mummy specimens. First of all, by high-throughput sequencing and metagenomics, they characterized the paleofeces microbiome of an eleventh-century pre-Columbian Andean (Santiago-Rodriguez et al. 2015). The most abundant bacterial groups in the paleofeces were Clostridium and Turicibacter spp. aDNA sequences homologous to T. cruzi, C. botulinum, and human papillomaviruses (HPVs) were found there too. Unexpectedly, presumptive antibiotic-resistance genes were detected in the mummy paleofeces microbiome. This means that antibiotic resistance might not necessarily

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be caused by selective pressure otherwise commonly seen in a modern society (Santiago-Rodriguez et al. 2015). Another study of the same authors also focused on the gut microbiomes of two Andean mummies (tenth to fifteenth centuries CE) representative of the pre-European colonization period (Santiago-Rodriguez et al. 2016a). The mummies’ gut microbiomes mostly included anaerobes: Clostridiales or Bacillales. This finding is interesting because it corroborates the hypothesis that the Andean mummies were formed in a very-low-oxygen environment. The authors’ metagenomics exhibited T. cruzi and Leishmania donovani aDNA sequences as well, revealing that those parasites were prevalent among Andean individuals before colonization (Santiago-Rodriguez et al. 2016a). They also demonstrated ancient bacteriophage DNA in the naturally mummified cases also was found (Santiago-Rodriguez et al. 2016b). In the gut microbiomes of the pre-Columbian Andean mummies, a large number of viral sequences homologous to bacteriophages was found, thus presenting an excellent opportunity to reveal the ancient phageome. Finally, by high-throughput sequencing, Santiago-Rodriguez et al. (2017) characterized gut microbiomes and putative resistomes of Italian and Inca mummies, finding more antibiotics-resistance genes in the Inca mummies than in the Italian ones (Santiago-Rodriguez et al. 2017). Although most of the work on ancient nucleic acid has concentrated on DNA, Keller et al. (2017) showed evolutionary conservation of small noncoding microRNAs in Ötzi the Iceman. They were the first to demonstrate how specific ancient microRNAs in the Iceman’s various 5300-year-old tissue specimens could be obtained (Keller et al. 2017).

Conclusion Inspired by mummies’ impressive preservation status, the earliest aDNA studies chose them as the research focus. Since then, DNA research in mummy studies has advanced in parallel with developments in the general field of archaeogenetics. Especially, switching from targeted PCR-based data production to high-throughput NGS of whole metagenomic libraries marked a major qualitative transition in the field. In addition to far higher throughput, NGS has decisive advantages over PCR: it can access, without bias, the ends of aDNA molecules where most of the characteristic chemical damage accumulates, and it offers a statistical approach to the estimation of the level of contamination based only on sequence data. Therefore, we actively encourage researchers in the field to explore and adopt NGS techniques and genome-scale data analysis. Soft tissues, due to their scarcity relative to hard tissues in ancient skeletal remains, generally have comprised only minor fractions of ancient samples yielding genome-scale data in recent large-scale archaeogenetic studies. However, as vividly shown with respect to the diverse studies reviewed in this chapter, the genetic study of mummies has a potential to explore subjects that are impossible or extremely difficult to achieve by the genetic study of skeletal remains, such as microbiomes

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associated with soft tissues (e.g., gut, skin), genetic changes associated with tumors, and the epigenetic makeup of soft tissues, to list only a few. As the history of archaeogenetics to date suggests, laboratory and computational breakthroughs will continually expand the range of information stored in macro biomolecules. We firmly believe that future aDNA research on mummies will continue to evolve and overcome the current limitations.

Cross-References ▶ A Primer on the Population Genetic Analysis of Ancient Genomes ▶ Analysis of Low Copy Number DNA and Degraded DNA ▶ Ancient DNA and Paleoparasitology in Brazil ▶ Ancient DNA Study ▶ Application of SNP-Based DNA Phenotyping to Archaeological and Forensic Cases ▶ Current Trends in Ancient DNA Study Acknowledgments This work was supported by the Ministry of Education of the Republic of Korea and the National Research Foundation of Korea (NRF-2019S1A5C2A01083578). We specially thank to Dr. Choongwon Jeong’s comments on the conclusion of this chapter.

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Current Trends in Ancient DNA Study Beyond Human Migration in and Around Europe

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Population Genomics of Archaic Hominins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Archaeogenetics in Africa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Early Structure within Eastern Eurasians and the Coastal Migration Hypothesis . . . . . . . . The “Population Y” Hypothesis and Peopling of the Americas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Genetic History of Domesticated Animals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Past Genomic Diversity in Wildlife . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

The field of archaeogenetics, a study of DNA sequences from ancient biological remains, has experienced an explosive growth during the last decade. By exploiting genome-scale information from ancient DNA, archaeogenetics has been providing critical insights into the past evolution history of humans and other species. In this chapter, I summarize a few of the key research questions in human archaeogenetics outside of European population history, focusing on recent progresses and remaining issues on them. Also, I highlight two research trends in the archaeogenetics of nonhuman organisms, one on the domesticated species and the other on wildlife. In both cases, mummified soft tissues have substantially contributed as a source of ancient DNA. Finally, I conclude the chapter with a prospect for the research trend of the next decade, highlighting technical advances in investigating the past with ancient biomolecules.

C. Jeong (*) School of Biological Sciences, Seoul National University, Seoul, Republic of Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_10

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Keywords

Ancient genomes · Human genetic history · Animal domestication · Mummy · Wildlife genomics · Demography

Introduction The study of genome sequences from ancient biological remains, often referred to as archaeogenetics or paleogenomics, has bloomed for the last decade. Leveraging over the advances in laboratory and computational techniques, it has turned into a thriving and mature discipline. It provides a powerful tool to look into the genomic past of a population of interest, with a deep and broad implication on many disciplines, including but not limited to genetics, archaeology, anthropology, systematics, and conservation biology. Humans are by far the most studied species with regard to genome-scale ancient DNA (“aDNA”) data. In human study, much of the earlier genome-scale aDNA works focused on delineating the history of populations in Europe (Lazaridis et al. 2014; Allentoft et al. 2015; Haak et al. 2015; Mathieson et al. 2015). These aDNA studies cover a wide range of time period, from archaic hominins like Neandertals (Green et al. 2010; Meyer et al. 2013; Prüfer et al. 2017) to Pleistocene and Holocene hunter-gatherers (Seguin-Orlando et al. 2014; Jones et al. 2015; Fu et al. 2016; Sikora et al. 2017) to Neolithic, Bronze, Iron Ages, and to historical periods (e.g., Lazaridis et al. 2014; Allentoft et al. 2015; Haak et al. 2015; Schiffels et al. 2016; Lamnidis et al. 2018; Olalde et al. 2018, 2019). In Europe, demic diffusion is found to be strongly associated with two major cultural transformations, that is, transitions into Neolithic and Bronze Age (Lazaridis et al. 2014, 2016; Allentoft et al. 2015; Haak et al. 2015). These studies inspired many that followed to investigate presence and spatiotemporal dynamics of long-distance human migrations and subsequent admixture, a mixing of diverged gene pools into a single population. Migration and admixture are now among the key topics of interest in human aDNA studies. The study of human (pre-)history has no reason to be, and indeed has not been, limited to Europe and the surrounding regions. Nor is there a reason to limit the source of aDNA to skeletal elements. In fact, the very first genome-scale aDNA studies in anatomically modern human were conducted faraway from Europe and using soft tissues: preserved hair samples from an ~4000-year-old Paleo-Eskimo genome from Greenland (Rasmussen et al. 2010) and a historic Australian Aborigine genome (Rasmussen et al. 2011). Mummified soft tissues have also been used for extracting ancient genomes, such as the famous Ötzi the Iceman and the Egyptian mummies (Keller et al. 2012; Schuenemann et al. 2017). In parallel with active researches continued in Europe, aDNA studies are now investigating virtually every corner of the world: Middle East, the Americas, East Asia, Pacific islands, Siberia, and Africa (e.g., Jeong et al. 2016; Lazaridis et al. 2016; Skoglund et al. 2016; Posth et al. 2018; Sikora et al. 2019).

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Although humans are often considered as a model system in genomics, the barrier between a few genomically well-resourced species and the rest is becoming lower and lower due to technological advances, such as the introduction of next generation sequencing (NGS) and long-read sequencing (e.g., PacBio and Nanopore) (Rhoads and Au 2015; Jain et al. 2016). They make it more economical and feasible to build critical genomic resources like the draft/reference genome, variant/haplotype panels, and whole genome sequences of target individuals. aDNA studies in nonhuman species are quickly expanding its realm, now covering interesting topics such as ancient pathogen genomes (e.g. reviewed in Harkins and Stone 2015), early domesticated animals and their now-extinct wild relatives (Daly et al. 2018; Gaunitz et al. 2018; Verdugo et al. 2019), and lost biodiversity detectable from archaeological or museum specimens (Miller et al. 2008; Palkopoulou et al. 2015). With everadvancing genome-editing technologies, it is now even possible to experimentally evaluate functional implications of bygone genetic diversity (Lynch et al. 2015). In this chapter, I first highlight a few research questions in human aDNA studies outside of Europe that have been of great interest for the last few years (Fig. 1). These topics will likely to keep being investigated for the incoming decade. Then I move into a few recent landmark studies in nonhuman aDNA (Fig. 1). I conclude with a prospect that temporal aspect via aDNA will become a regular part of population genomic study of a large number of species in near future.

Population Genomics of Archaic Hominins It was arguably the most interesting early finding in archaeogenetics that archaic hominins left a legacy in the genomes of present-day human populations to a different degree (Green et al. 2010; Meyer et al. 2012). As a result of archaic admixture events, human populations outside of sub-Saharan Africa mostly derive ~2% of their genome from their Neandertal ancestors, and indigenous populations of Australasia (Australia, New Guinia and nearby regions) derive another ~3% from an archaic hominin group related to Denisovans in southern Siberia (Green et al. 2010; Meyer et al. 2012; Prüfer et al. 2014). East and South Asian populations also seem to have a trace of Denisovan-related ancestry, although to a much lesser degree around ~0.2% (Prüfer et al. 2014). The retrieval of archaic hominin genomes played a pivotal role in detecting and quantifying the archaic admixture signals in great detail (Sankararaman et al. 2014, 2016; Vernot and Akey 2014; Vernot et al. 2016), although eccentric patterns of genetic variation in present-day humans were used to narrow down candidate regions with archaic hominin ancestry (“archaic introgressed segments”) that were later confirmed to be of archaic origin by a comparison with the archaic genomes (Plagnol and Wall 2006). In addition to the archaic introgressed segments that preserve genetic materials from extinct archaic hominins, a growing number of whole or partial ancient genome sequences have been directly retrieved from skeletal elements of archaic hominins (Castellano et al. 2014; Prüfer et al. 2014, 2017; Meyer et al. 2016; Hajdinjak et al.

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Fig. 1 Geographic locations of representative ancient genome studies highlighted in this chapter. Square and diamond shapes represent studies using soft mummified tissues and skeletal elements, respectively. 1. Qeqertasussuk (Qt) – a Paleoeskimo genome of the Saqqaq culture derived from a hair sample. 2. Golden Ridge – an Aboriginal Australian genome derived from a historic hair sample. 3. Ötzi the Iceman – a Middle-Neolithic European genome from a mummified individual. 4. Abusir el-Meleq – the Egyptian Old Kingdom necropolis. 5. Denisova Cave – a cave site in southern Siberia that yielded hominin skeletal elements for the Denisovan and Altai Neandertal genomes. 6. Mota Cave – a cave site in highland Ethiopia from which a 4500-year-old individual’s genome was sequenced. 7. Grotte des Pigeons (Taforalt) – a site of the Iberomaurusian cultural context that yielded 15,000-year-old infant skeletons. 8. Tianyuan Cave – a Paleolithic cave site in northern China that yielded a 42,000-year-old skeleton. 9. Mal’ta – a Paleolithic site in the western Baikal region that yielded a 24,000-year-old boy’s skeleton. 10. Upward Sun River (Xaasaa Na0 ) – a terminal Pleistocene site in central Alaska. 11. Thistle Creek – a middle Pleistocene (560–780 kya) horse genome. 12. Taimyr – a late Pleistocene (35,000-year-old) wolf genome. The figure was produced using publicly available resources in rworldmap and mapdata packages in R v3.5.3

2018; Slon et al. 2018; Bokelmann et al. 2019). Inference on the relationship between these archaic genomes suggests that archaic hominins had a complex and dynamic population structure in spite of their small population size. First, archaic hominin populations held a complex population structure. A Neandertal genome from the far-east Denisova cave (“Altai Neandertal”) is more distantly related from all the other Neandertals found in Europe (Hajdinjak et al. 2018). Neandertals in Europe are also structured roughly mirroring geography, although a recent population turnover in Europe sourced from the Caucasus is suggested based on the phylogenetic tree of the European Neandertals nested by two Caucasus Neandertals from the Mezmaiskaya site (Hajdinjak et al. 2018). Likewise, Denisovan-related genetic segments in present-day populations are differentially related to the highcoverage Denisovan genome, thus implying the presence of diverged Denisovanrelated populations across Asia (Browning et al. 2018; Jacobs et al. 2019). Second, the high-coverage Denisovan genome may derive a small proportion of its ancestry from a deeply branching hominin lineage that is an outgroup to both modern humans

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and Neandertal/Denisovan clade (Prüfer et al. 2014). Together with the finding of an individual who has a Neandertal mother and Denisovan father (Slon et al. 2018), this shows a widespread admixture between different hominin lineages. Third, there seem to have been gene flows from a lineage leading to the anatomically modern humans to Neandertals prior to the main out-of-Africa event. Such a signal is shown in nuclear genome data of the Altai Neandertal, but not in the other European ones (Kuhlwilm et al. 2016), and also shown in the mitochondrial genome phylogeny of European Neandertals (Li et al. 2017). Future studies will further increase the number of available archaic hominin genomes, leading to the era of population genomics of archaic hominins. Archaeogenetic study of archaic hominins beyond Neandertals and Denisovans will also be critical to test hypothesized archaic introgressions (e.g., sub-Saharan Africa (Plagnol and Wall 2006)) or to characterize the nature of putative archaic hominin fossils, such as those in China (Li et al. 2017).

Archaeogenetics in Africa As the cradle of our species, Africa holds the largest fraction of human genetic diversity and the deepest branches of human populations (Cann et al. 1987). Population genomic studies of present-day humans clearly showed that the indigenous populations of southern Africa, often collectively called as Khoe-San, represent the first split within modern humans (Li et al. 2008; Schuster et al. 2010). However, extensive gene flows between sub-Saharan African populations complicated the inference of their population history using modern genomes only. Also, most of the deeply branching African lineages, such as Khoe-San, East African click speaker Hadza, Central African rainforest hunter-gatherers (often referred to as “pygmies”), still practice a hunter-gatherer life style and have been pushed off into smaller territory by the expanding populations with production economy (Clark and Brandt 1984). That is, the spatial distribution of the sub-Saharan African genetic diversity presumably has changed in a massive way for the last several millennia, heavily impacted by large-scale demographic events such as a Bantu expansion from West Africa (Patin et al. 2017). A few but growing number of archaeogenetic studies in Africa, finally overcoming climate hostile to biomolecule preservation, start to provide insights into the African genetic history. Early studies on ancient African genomes indeed support deep genetic roots of various hunter-gatherer populations in sub-Saharan Africa. First, ancient genomes from pre-pastoralist southern Africa have a great similarity with the present-day Khoe-San people, with clear pastoralists and European admixture signals in the latter post-dating the ancient genomes (Skoglund et al. 2017; Schlebusch et al. 2017). Using these pre-admixture genomes, more accurate estimates for the first population split within anatomically modern humans, that is, the age of their last common ancestor, are obtained to around 350–260 kya (thousands years ago) (Schlebusch et al. 2017). Second, ancient genomes from Eastern Africa (modern-day Ethiopia, Kenya, Malawi, Tanzania) form an ancient genetic cline that runs between a gene pool related to ancient southern Africans and one related to a 4500-year-old genome

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from Ethiopia (Llorente et al. 2015; Skoglund et al. 2017). Interestingly, Hadza, a click-speaking East African hunter-gatherer population, is the most closely related to the latter among the present-day populations (Skoglund et al. 2017). A near absence of Hadza-related ancestry among present-day East and South Africans supports a large-scale genetic turnover in these regions associated with the introduction of production economy. The genetic diversity among Africans so far observed may only provide an incomplete view on the past genetic diversity in Africa. The first Pleistocene genomes from the continent came from the 15,000-year-old site Taforalt with the Iberomaurusian technocomplex context, in modern-day Morocco (van de Loosdrecht et al. 2018). These genomes show a strong genetic affinity with present-day sub-Saharan Africans, to a degree much stronger than those of later north Africans or Near Easterners. Present-day west Africans, such as Yoruba (no ancient genome from west Africa has been made available so far), seem to act as the best proxy for their African-related ancestry, although there is a residual affinity to a more deeply branching lineages. Another study reports the asymmetric genetic relationship of two West African populations, Mende and Yoruba, in relation to worldwide populations (Skoglund et al. 2017). One possibility to explain this signal is to assume a presence of a lineage in West Africa, more deeply branching than the South African one, and its asymmetric contribution to present-day West African groups. To distinguish between these hypotheses that involve deep lineages and old admixture events, it is critical to make a direct observation of ancient gene pools that have not been sampled so far.

The Early Structure within Eastern Eurasians and the Coastal Migration Hypothesis The early history of anatomically modern humans in eastern Eurasia, including the dates and routes of peopling, still remain controversial. A central issue in this discussion is the so-called early coastal migration, which hypothesizes a two-phase peopling of eastern Eurasia (Lahr and Foley 1994). According to this hypothesis, the first phase of out-of-Africa migration followed the coastal route leading to southeast Asia and eventually to Australasia, while the later second phase mostly replaced the descendants of the first wave in mainland East Asia and became the ancestors of most present-day populations there. Present-day Australasians, as well as morphologically and ecologically somewhat distinct hunter-gatherer populations throughout southeast and south Asia, often called as “Negritos” are assumed to be the remaining descendants of the first wave (Hanihara 1992). Recent genomic studies are overall not in favor of the coastal migration scenario in its original form, that is, the early coastal population represents the first branch within Eurasians while ancestors of western and eastern Eurasians split more recently. Although the earlier study of a historic Australian Aborigine genome predating the European contact reported a signal that seemingly matched to a pattern expected by the coastal migration scenario (Rasmussen et al. 2011), the excess

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affinity of the Australian Aborigine with a lineage more basal to the other Eurasians can easily be explained by the archaic admixture from a Denisovan-related lineage. So far, genetic evidence for a contribution of a deeply branching Eurasian lineage into the Australasian ancestors remain negative to at best controversial (Malaspinas et al. 2016; Mallick et al. 2016; Pagani et al. 2016; Wall 2017). A major opinion in the field assumes Australasians and some other coastal groups, such as Onge from the Andaman Islands, to represent among the first splits within Eastern Eurasians (Lipson and Reich 2017). Specifically, these early branches or “deep” East Asian lineages branch off from the common ancestor of East Asians and Native Americans before their splits. Still, it remains an important question to identify the order of these early branching events and their spatiotemporal context. Due to the hostile climate conditions, ancient genomes from southeast Asia has only recently been made available (Lipson et al. 2018; McColl et al. 2018). Interestingly, these studies find a close genetic relationship between ancient huntergatherers associated with the Hoabinian culture and the present-day Onge people in the Andaman Islands. One of these studies co-analyzed Australasians (Papuans) together with Onge and the Hoabinian hunter-gatherers, and find a population graph including a deep shared ancestry between Papuan and the Onge-Hoabinian cluster most fitting (McColl et al. 2018). There are more East Asian genomes that are shown to harbor a “deep” East Asian ancestry in addition to the above-discussed ones. First, a 40 kya genome from the Tianyuan cave in northern China belongs to such a deep ancestry, but is different from the ones represented by Papuans or Onge (Yang et al. 2017; McColl et al. 2018). Second, sedentary hunter-gatherers from Neolithic Japan, referred to as “Jomon” following their pottery making tradition, also have a deep ancestry (McColl et al. 2018; Gakuhari et al. 2019; Kanzawa-Kiriyama et al. 2019). These studies agree that Tianyuan, Australasians, Onge-Hoabinian, and Jomon all derive, at least partially, from a deep East Asian ancestry, while providing conflicting opinions on the relationship between them. Future sampling of ancient genomes that represent key branches in the true but unknown population graph will be critical to understand the number and relationship of deep East Asian ancestries.

The “Population Y” Hypothesis and Peopling of the Americas Peopling of the Americas has been among the most intensely studied topics in archaeogenetics, and in human population genetics in general, outside of Europe. Setting aside much later Holocene migrations into North America by ancestors of Paleo- and Neo-Eskimos, it has been the most debated subject whether there was only a single ancestral population that migrated into the Americas during the Pleistocene, or there were multiple genetically distinct groups. This was initially driven by claims from some paleoanthropologists who observed morphologically distinct early individuals throughout the Americas (González-José et al. 2005). Although direct archaeogenetic investigations of these “Paleoamerican” specimen rejected their ancestry distinct from the other First Peoples (Raghavan et al. 2015), genomes of

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some present-day South American populations show unexpected signals against the single common ancestry of all First Peoples (Raghavan et al. 2015; Skoglund et al. 2015b). That is, some Amazonian populations show extra genetic affinity to populations outside of the Americans, especially Onge and Papuans, compared to the other Native Americans. The original studies which reported this ancestry, which is related to present-day Onge and Papuans and frequently referred to as “population Y,” did not provide a particular demographic scenario to explain the limited and scattered distribution of the population Y signal. Follow-up studies explored ancient genomes from South America and elsewhere to find individuals harboring the population Y signal to higher degree than the present-day populations, resulting in contrasting claims (Moreno-Mayar et al. 2018b; Posth et al. 2018; Scheib et al. 2018). Two of these studies primarily report population split and mixture within the clade of First Peoples as a sufficient explanation for the genetic diversity within ancient Native Americans (Posth et al. 2018; Scheib et al. 2018). In contrast, the other study finds the Ongerelated signal in early Holocene individuals from Lagoa Santa, Brazil (~10 kya) to a degree similar to present-day Surui people (~3% contribution) (Moreno-Mayar et al. 2018b). These individuals were also suggested to be of “Paleoamerican” type based on their morphology (Neves et al. 2007). A genomic study of the Pleistocene Tianyuan individual from northern China suggests that a mixture of Tianyuan- and Onge/Papuan-related lineages may serve as a good proxy for the hypothetical population Y (Yang et al. 2017), implying that the population Y signal may reflect genetic heterogeneity within the founding population in Beringia. Therefore, the population Y signal and its interpretation remain to be settled. It is a key objective for future studies to find ancient genomes that matches to the hypothetical population Y, or genomes that derive higher proportion of ancestry from this lineage. Archaeogenetic studies substantially improved our understanding on the Pleistocene origin of the ancestral First Peoples gene pool in Beringia for the last few years. First, Pleistocene genomes from archaeological sites in Siberia, such as Mal’ta, Afontova Gora, and the Yana Rhinoceros Horn site, revealed the presence of an ancient gene pool that contributed to the gene pool of the First Peoples (Raghavan et al. 2014b; Sikora et al. 2019). That is, First Peoples are admixed descendants of an eastern Eurasian and this Siberian population, often called “Ancient North Eurasians” (Raghavan et al. 2014b). Second, a terminal Pleistocene genome from the Upward Sun River site in Alaska (“USR1”) is shown to be the most basal lineage among the First Peoples (Moreno-Mayar et al. 2018b). Third, multiple studies provide new ancient genome data and models to explain later Siberian connections around the Bering strait, focusing on the dispersal of Paleo- and Neo-Eskimo-related populations around the Arctic (Raghavan et al. 2014b; Moreno-Mayar et al. 2018b; Flegontov et al. 2019; Sikora et al. 2019). Future studies are expected to reveal the Siberian part of the Native American population history to a detail by producing more ancient genomes. For example, the identity of the eastern Eurasian lineage that formed the ancestral First Peoples gene pool and its history in Eurasia remain to be explored. Also, the origin and dispersal of Paleo- and Neo-Eskimo populations

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outside of Beringia are poorly understood, especially with regard to the bigger context of Siberian population history.

The Genetic History of Domesticated Animals In parallel with human studies, the genetic history of domesticated animals has long been a key topic in population genetics (Larson and Fuller 2014). Earlier studies were based on genomes of modern animals, primarily focusing on “breeds” due to sample availability (The Bovine HapMap Consortium 2009; vonHoldt et al. 2010). These studies show a complicated evolutionary history of domesticated animals, highlighting pervasive introgression from local wild relatives during the early domestication process (Freedman et al. 2014; Frantz et al. 2015; Park et al. 2015). In addition, much of the genetic diversity in the wild source populations has been lost since the beginning of domestication: for example, wild horses and Aurochs are now extinct and the genetic diversity of wolves has been greatly reduced. These two issues, the effective multiple origins of domesticated animals due to local wild introgression and the loss of genetic diversity representing the local wild populations, have posited a major obstacle in reconstructing the genetic history of domesticated animals. Recently, surveys of ancient genomes began to appear for domesticated animals and their wild relatives. One line of studies traces well-preserved animal remains from permafrost, that is, natural mummies, much older than the onset of domestication process and thus looks into the genetic diversity of these animals in the distant past. For example, the oldest genome ever sequenced is from a horse lived in the Middle Pleistocene (560–780 kya) Yukon, Canada (Orlando et al. 2013). As expected from its old age, it falls outside of the present-day horse genetic diversity, represented by domesticated breeds and Przewalski’s horses, and provides a critical calibration point to infer the split time between horse and its sister species donkey (Orlando et al. 2013). Another example comes from Pleistocene wolves from the Taimyr Peninsula in northern Siberia (Skoglund et al. 2015b). This 35 kya wolf is largely outside of the present-day dog genetic diversity. However, interestingly, some dogs from the Siberian and New World Arctic, such as Siberian Husky and Greenland Sledge dog, share more alleles with the Taimyr wolf than other dog breeds do, suggesting introgression into these dog breeds from a lineage related to the Taimyr wolf. Human-made animal mummies have also provided invaluable information to explore the origins and nature of animals in the historic time period, especially in Egypt, including cats and sacred Ibis (Kurushima et al. 2012; Ottoni et al. 2017; Wasef et al. 2019). Extensive studies of ancient domesticated animals, now appearing in the literature, provide insights into the dynamic nature of their evolutionary history during prehistory. First, genomes from the earliest domesticated horses from the famous Eneolithic sites of Botai and Borly in northern Kazakhstan are again shown to be outside of the present-day and ancient domesticated horses (Gaunitz et al. 2018).

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Interestingly, Przewalski’s horses, a wild horse population considered to be the last descendants of pre-domestication horses by some, actually fall within the BotaiBorly cluster, suggesting that they descended from a domesticated population. Multiple lineages outside of the genetic diversity of the domesticated horses are found in ancient wild horses, suggesting a substantial loss of diversity (Schubert et al. 2014; Librado et al. 2017; Fages et al. 2019). Second, Neolithic and more recent cattle genomes from the Near East and Europe show an early genetic structure (Verdugo et al. 2019). Here the genetic structure of domesticated cattle substantially overlaps with that of wild Aurochs, suggesting strong local contribution from wild populations into the incoming cattle (Park et al. 2015; Verdugo et al. 2019). Still, the origin of zebu (Bos indicus) and African taurine cattle breeds remain obscure due to lack of Aurochs genomes that can explain their genetic profiles distinct from the Eurasian taurine cattle (Bos taurus). Third, ancient genomes even provide a new momentum to the studies of dynamic early history of the species with substantial present-day wild populations. Domesticated goat populations in the Near East and southeastern Europe were already clearly structured during the Neolithic, that is partially inherited into the present-day structure (Daly et al. 2018). This ancient structure is interpreted as a result of local mixture with wild caprine gene pool. In addition to a mixture with wild relatives, post-Neolithic goats from Anatolia and the Levant received a massive gene flow of goat populations from Iran and further to the east, pointing admixture as a pervasive process in the evolution of domesticated animals (Daly et al. 2018). A recent study of pigs (Sus scrofa) from the Near East and Europe also observes a large-scale genetic turnover: while the first domesticated pigs in Europe during the Neolithic were of a Near Eastern origin, Late Neolithic and later pigs mostly derive their ancestry from a source related to European wild boars (Frantz et al. 2019). Future archaeogenetic studies of domesticated animals are expected to expand its scope beyond Europe and the Near East and to contribute to understand their evolution as well as dynamics of human migration and changes in subsistence strategy.

Past Genomic Diversity in Wildlife A chance to directly observe ancient genomes have a broad implication beyond tracing the genetic history of humans and domesticated species. Systematics and population genetics of extinct species are among the most relevant applications of archaeogenetics. Genomes from extinct Elephantid species from Siberia and North America, such as woolly mammoth (Mammuthus primigenius) and American mastodon (Mammut americanum), reveal the genetic history of Asian Elephantids and their adaptations to high latitude (Miller et al. 2008; Rohland et al. 2010; Enk et al. 2014; Palkopoulou et al. 2015; Lynch et al. 2015). The taxonomic positions of other extinct megafauna species have also been explored using ancient genomes, including ground sloths (e.g., Milodon darwinii) (Delsuc et al. 2019), giant Gliptodont armadillos (Doedicurus sp.) (Delsuc et al. 2016; Mitchell et al. 2016), Moa

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(Anomalopteryx didiformis) (Baker et al. 2014), and elephant birds from Madagascar (Aepyornis hildebrandti and Mullerornis agilis) (Mitchell et al. 2014). Ancient genomes have also been used to trace the origin of biological materials leveraging over the known correlation between geography and genetic profile. It is a well-established approach in conservation biology using present-day genetic data, which have been used to trace illegally traded animals and animal remains, such as ivory (Wasser et al. 2015). A recent study traced the origin of walrus ivory in Medieval Europe and revealed that a good proportion of walrus ivory actually came from North America near Greenland, suggesting a key role of ivory trade in the economy of Norse settlements in Greenland (Star et al. 2018). Another study looked into remains of neotropical bird scarlet macaw (Ara macao cyanoptera) in the American Southwest and northern Mexico (George et al. 2018). By showing that all ancient samples from this region are genetically similar to each other and belong to one lineage, it was claimed that a long-standing breeding center had been maintained and functioned as a center for distributing this animal of symbolic importance across the American Southwest. Although a majority of archaeogenetic studies of wildlife species focus on a small fraction of genome, such as mitochondrial sequences, genome-scale studies will soon appear where higher genetic resolution helps.

Conclusion For the last decade or so, the study of ancient genomes has explosively expanded its range into all directions: space, time, and taxa. As the genomic studies of living species have been continuously advancing, archaeogenetics will follow its track and add a critical dimension of time into the genomic study of evolution. As the evolutionary study has a wide range of spatiotemporal focus, the quantity, quality, and type of data required to adequate resolve relevant questions also differ widely. For example, a study of recent evolutionary history within a single species, such as the history of human and domesticated animals, compares genetically similar individuals and groups and thus is benefitted most by high-resolution genomescale data and by large sample size. As reviewed in this chapter, population-scale genome-wide data are already commonly studied in this area, although it is still relatively infrequent to produce whole genome sequences with clearance of postmortem chemical damages (Rohland et al. 2015). Future studies will likely focus on continuing large-scale surveys as well as increasing sample and data coverage of specific regions and time periods for testing certain historical or archaeological hypotheses. Community-wide, these efforts aim at completing a map of the genetic diversity of humans and other species across space and time. On the opposite direction there is a study of deep time and deep phylogenetic relationship. Even a limited amount of sequence information provides a good insight into the relationship between phylogenetically distinct groups if retrievable. Therefore, the ultimate limiting factor here is preservation of biomolecules. Recent advances in the study of ancient proteins, paleoproteomics, leverage over superior preservation of

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some proteins like collagens, and are expected to provide a new breakthrough into deep time, including human evolution (Welker et al. 2015, 2016; Chen et al. 2019; Presslee et al. 2019).

Cross-References ▶ A Primer on the Population Genetic Analysis of Ancient Genomes ▶ Ancient DNA Study

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Ancient DNA Study Practical Aspects

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Removal of Sequencing Adapter and Barcode Sequences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mapping to the Reference Genome Sequence and Subsequent Read Filtering . . . . . . . . . . . . . . . . Assessment of Post-mortem Chemical Damages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Assessment of Contamination Level . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Genotype Calling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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The introduction of next generation sequencing into archaeogenetics transformed the field into a powerhouse of the genetic study of human history. Although the processing of ancient DNA sequencing data follows the same basic principle for that of high-quality modern genomes, there are properties specific to ancient DNA molecules that need to be taken into account during data processing and analysis. In this chapter, I summarize the up-to-date practice of key steps of ancient DNA sequence data, such as the removal of Illumina sequencing adapter, read mapping to the reference genome, tabulation of post-mortem chemical damages, estimation of contamination level, and genotype calling. For each step, I highlight the current practices and the underlying logic for them, as well as directions of future method development. Keywords

Ancient DNA · Next generation sequencing · Ancient genome data analysis · Chemical damage · Contamination estimation · Mummy C. Jeong (*) School of Biological Sciences, Seoul National University, Seoul, Republic of Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_11

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Introduction Genome-scale studies of ancient DNA (“aDNA”) are now frequently performed on many species in addition to humans, ranging from large-bodied domesticated and wild animals to microorganisms (Miller et al. 2008; Green et al. 2010; Rasmussen et al. 2010; Bos et al. 2011, 2014; Feldman et al. 2016; Daly et al. 2018; Gaunitz et al. 2018; Verdugo et al. 2019). Although a large fraction of ancient genome studies retrieved ancient DNA molecules from skeletal remains, mummified soft tissues of both human and nonhuman samples have also been utilized in several key studies of ancient genomes (Rasmussen et al. 2010; Rasmussen et al. 2011; Keller et al. 2012; Orlando et al. 2013; Skoglund et al. 2015; Ottoni et al. 2017; Schuenemann et al. 2017). The expansive trend of aDNA research closely resembles a boom of genome sequencing studies on present-day organisms with the introduction of the next generation sequencing (NGS) methods since mid-2000s (The 1000 Genomes Project Consortium 2012). Indeed, a great part of the bioinformatics tools to process and analyze aDNA sequence data are directly taken from those for modern sequence data, taking into account that sequencing data have essentially the same nature regardless of the age of samples. However, in practical aspects, computational and statistical analyses of aDNA sequence data require multiple tweaks from the standard analysis workflows of highquality modern sequencing data, reflecting unique characteristics of aDNA sequence data and constraints that they pose on the analysis. Due to these features, a naïve application of the standard workflows often results in biased or even false conclusions. Notably, biases and errors in the workflows may affect the analysis results in substantially different ways depending on the type of analysis and data. Therefore, it is of great importance for researchers planning genome-scale aDNA analysis to understand the unique nature of aDNA sequence data and to adjust their analysis workflows to take it into account. Importantly, the nature of aDNA sequence data does not vary between source materials, for example, soft mummified tissues vs. hard skeletal remains, and therefore the same workflow can be optimally applied to the mummy- and skeletal element-based aDNA studies. In this chapter, I outline a general workflow of aDNA sequencing data analysis, focusing on the human case. The same workflow can be applied to most non-human species with comparable genome size and ploidy, while smaller haploid genomes (e.g., those of bacteria) may be not need to consider some aspects of the issues raised in diploid genome analysis. For each step of the workflow, I highlight the general and aDNA-specific issues, widely used current approaches, and directions of future developments.

Removal of Sequencing Adapter and Barcode Sequences In contrast to high-quality, high-molecular-weight genomic DNA extracted from fresh tissue samples, DNA molecules extracted from ancient samples are fragmented into short pieces, often to the level of the median size less than 100 base pairs (bps)

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(Sawyer et al. 2012; Dabney et al. 2013). Therefore, a large fraction of molecules in aDNA sequencing libraries have an insert shorter than the read length, which typically ranges 100–250 bps in Illumina platforms or comparable NGS techniques provided by other companies. As a result, reads from such molecules with a short insert include common adapter sequences at the 30 end (Fig. 1). In some cases, extra sequences are also experimentally added between the common adapter and the insert in both 50 and 30 ends (Fig. 1). Like external barcode or index sequences located within the common adapter sequences, such “internal barcode” sequences are for allowing pooling of multiple libraries into a single sequencing lane while minimizing cross-contamination between libraries. However, unlike the external barcode sequences that are separately sequenced, these internal barcodes are sequenced as a part of the main reads. A naïve mapping of raw reads will critically reduce mapping efficiency and accuracy, due to the exogenous parts (i.e., internal barcode and common adapter sequences) included in many raw reads. Therefore, it is the first step of processing aDNA sequence data to trim the adapter sequences from raw reads prior to mapping.

Fig. 1 Five different scenarios for trimming Illumina adapter sequences from raw reads. (a–c) In the PE design, both the 50 and 30 ends of reads are directly observed. When the forward and reverse reads are (a) completely or (b) partially overlap, they can be merged into a single sequence that represents the whole insert. When the forward and reverse reads do not overlap (c), they are treated as standard PE sequences. (d–e) In the SE design, only the 50 end of reads is directly observed. If the read is longer than the insert (d), one can infer the end of the insert by detecting the Illumina adapter sequence at the 30 end of the read. When the insert is longer than the read, the 30 end of the insert is not observed (e)

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Removal of adapter sequences is a more straightforward task in the paired-end (PE) sequencing design than in the single-end (SE) one (Fig. 1). Specifically, both ends of the insert are clearly defined in the PE design, while only the 50 end is directly observed in the SE design. Depending on the insert and read lengths (IL and RL, respectively), the following five scenarios are possible for removing the adapter sequences: (i) PE-1 (IL  RL): both forward and reverse reads completely cover the insert; when the forward and reverse reads are aligned, unaligned parts are expected to match the adapter sequences. (ii) PE-2 (RL < IL 200 bases) and a synthetic target as an IPC (Gouveia et al. 2015). Likewise, many recently available kits can help analysts in determining the total amount of human DNA and human male DNA in a sample, as well as whether the sample is degraded or whether there are PCR inhibitors (Lee et al. 2014).

Detection of DNA Damage from MPS Data In nearly all cases, DNA extracted from ancient samples exists as a mixture of endogenous DNA and environmental DNA. Therefore, tremendous efforts have been made to control and identify contamination throughout the process of data generation and interpretation. During data interpretation, DNA damage patterns that cause nucleotide misincorporation have frequently been assumed to be the result of contamination. However, as the sequencing of millions of reads became possible using MPS, such damage patterns started to be used to identify ancient DNA sequence reads. In particular, the nucleotide misincorporation induced by the deamination of cytosines and 5-methyl cytosines thus has been proposed to be a powerful tool to distinguish between recent and ancient DNA sources (Briggs et al. 2007). Both in vivo and in vitro studies have shown that the rate of deamination of cytosine is 20 times more than that of adenine, thereby resulting in the predominance of the transition from C to T and complimentary transition of G to A in ancient DNA sequences (Lindahl and Nyberg 1972). Cytosine deamination rates are typically high at the 50 -overhangs, with an increased C to T transition rate toward sequencing starts and complementary increase in G to A transition rates toward read ends (Briggs et al. 2007). This phenomenon has been demonstrated in the comparative analysis of tissue remains that vary in age between 18 and 60,000 years, where three molecular features, i.e., DNA fragment length, base composition at strand breaks, and C to T transition, were investigated in their mitochondrial DNA sequences (Sawyer et al. 2012). DNA fragment length did not decrease consistently over time, but the frequency of C to T transitions was observed to increase toward the 50 -ends of DNA fragments over time. Therefore, a framework for calculating the likelihood of a fragment being authentic has been developed by measuring these postmortem degradation patterns (Briggs et al. 2007; Skoglund et al. 2014), and this has evolved into a statistical framework that provides estimates for degradation parameters such as the average length of

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overhangs, nick frequency, and cytosine deamination rates in both overhangs and double-stranded regions (Jónsson et al. 2013). The mapDamage 2.0 package has been defined as a Bayesian computational framework modeled with the simple assumption that mutations and postmortem DNA damage are independent within a fragment and damage events only depend on the relative position from the sequence ends (Jónsson et al. 2013). Ultimately, the program helps identify ancient DNA sequences by permitting quality scores for bases in a sequence read to be rescaled based on the probability of being damaged. However, this approach requires multiple read sequences to obtain scores, so it can only be applied to MPS data (Linderholm 2016). In a recent study on ancient Egyptian mummies (Schuenemann et al. 2017), DNA was extracted from 151 mummified individuals, and the authenticity of the ancient DNA retrieved from the MPS library was assessed using the mapDamage 2.0 program. The comparison of DNA preservation in ten individuals revealed that the yields of preserved DNA in soft tissues were 10 times lower than in the bone and teeth and the DNA damage pattern differed depending on the tissue type, with an average of 19% of damage in soft tissues and 10% of damage in the bone and teeth.

Analysis of Degraded DNA and LCN DNA Since the first retrieval of ancient human DNA (Pääbo 1985), technical advances have been frequently made to overcome the analysis challenges for LCN and degraded DNA. Early analysis of ancient DNA has focused on mitochondrial DNA, but recent advances in DNA analysis technologies such as MPS made it possible to routinely retrieve and analyze nuclear DNA from ancient samples.

Mitochondrial DNA Analysis Since mitochondrial DNA is maternally inherited, it allows us to study the ancestry and migration of human populations along the maternal lineage. Additionally, mitochondrial DNA has hundreds to thousands of copies per cell, and therefore, it can often be amplified for genetic investigation even when nuclear DNA analysis fails. Each cell has hundreds of mitochondria, and on average, each mitochondrion contains four to five copies of mitochondrial DNA (Satoh and Kuroiwa 1991). Therefore, the average copy number of mitochondrial DNA is calculated to be about 500 per cell, whereas only two nuclear DNA copies are present in a cell. This small, circular form of DNA is also less susceptible to exonucleases that degrade DNA than genomic DNA. Alternatively, as mitochondrial DNA is typically analyzed in samples with very little DNA and amplified with higher PCR cycle numbers (e.g., 36 cycles), it is more sensitive to contamination than nuclear DNA (Butler 2012). Therefore, mitochondrial DNA should be extracted with care and should be amplified in a clean laboratory with non-template controls.

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Even at high copy numbers, as mitochondrial DNA has a small size of 16,569 bp, it constitutes only approximately 0.25% of the cell’s total genome content. The mitochondrial DNA genome consists of a coding region, which encodes 37 RNAs and proteins, and non-coding region called the control region. Because the control region includes the replication origin but does not code for any gene products, it is prone to have more mutations and polymorphic sites compared with the coding region. Therefore, sequence analysis of the control region, which includes three hypervariable regions (HV1, HV2, and HV3), has long been used in the forensic field to identify individuals and families. To analyze mitochondrial DNA sequences, custom-made amplification is usually performed before Sanger sequencing, and primer pairs are selected depending on the level of sample degradation and target region (Edson et al. 2004; Berger and Parson 2009; Lee et al. 2008a). Therefore, the HV1 and HV2 regions, which encompass regions spanning 342 bp and 268 bp, respectively, can be amplified using several small PCR amplicons from highly degraded DNA samples. With recent advances in MPS, many amplification kits have become available for both control region and whole mitochondrial genome amplification for MPS analysis of degraded DNA. However, some quality-related issues have been raised when analyzing mitochondrial DNA sequences that were reconstructed using multiple fragments (Bandelt et al. 2001, 2004). For example, artificial recombination sequences can be created by combining the HV1 data of a sample with the HV2 sequence of another sample. In the forensic field, a methodology based on mitochondrial DNA haplogroup determination and comparison with existing mitochondrial DNA haplotypes was proposed to prevent errors during the reporting of mitochondrial DNA sequences (Lee et al. 2008b). The mitochondrial DNA phylogenetic tree shows evolutionary relationships among various mitochondrial haplogroups, which are based on similarities and differences in their sequences, and the phylogenetic approach to localize a sequence to a part of the phylogeny can be very useful for systematic error detection. This is because mitochondrial DNA haplogroups are defined by the presence of specific mutations that can cluster sequence groups. Since mitochondrial DNA sequences belonging to the same haplogroup share a common maternal ancestor, they are reported to carry the same or similar mutations. Thus, if the sequence from a fragment shows a typical mutation motif for a haplogroup while the sequence from another fragment of the same sample carries mutations specific for another haplogroup, this may imply contamination or artificial recombination caused by sample switching. When haplogroup determination fails, a neighborhood search for sequences in the database can provide information about potential closely related subsets of sequences, which will enable comparison and thereby allowing researchers to detect errors in the sequences (Lee et al. 2008b).

Use of Small-Sized PCR Products Degraded DNA and very low amounts of DNA can be more successfully analyzed using PCR primers that produce reduced sized amplicons (Wiegand and Kleiber

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2001). This was well demonstrated by an analysis of degraded samples from the remains of fire victims in 1994 (Lygo et al. 1994). In this analysis, where short tandem repeat (STR) markers were amplified to identify victims and determine their relatives, smaller STRs were found to be more effective than the larger loci. Since then, several mini-STR assays have been developed with most amplicon sizes below 200 bp (Hill et al. 2007), which aided in the identification of World Trade Center victims using burned or damaged bone samples. In addition, telogen hair with little nuclear DNA has been successfully analyzed using this mini-STR approach (Opel et al. 2008). To analyze ancient DNA, PCR amplification of overlapping fragments of small length has also been suggested (Pääbo et al. 2004). In a study concerning two ancient Korean mummies found in medieval tombs (Kim et al. 2011), mitochondrial DNA, Y chromosomal STR, and the ABO gene were successfully analyzed using a small size amplicon strategy. The mitochondrial DNA control region sequences were analyzed using a mini-primer set comprising fragments ranging in size from 146 to 170 bp (Lee et al. 2008a). Y-STRs were amplified using a commercial kit composed of 17 STRs and an in-house Y-miniplex plus system composed of 8 Y-STRs with an amplicon size ranging from 80 to 230 bp. Among the 17 loci of the commercial kit, it was the 8 Y-STR loci with amplicon sizes of less than 180 bp that showed reproducible results with replicate analysis. This demonstrates that using small size amplicon strategy can increase the likelihood of obtaining reliable DNA typing results from degraded DNA. To facilitate ABO genotyping, PCR primers were also designed to produce amplicons as small as 72–84 bp. Although these two mummies date back to approximately 500 years ago, DNA extracted from the ribs and soft tissues such as muscle and liver could be successfully analyzed by amplifying small PCR products. The identification and characterization of the skeletal remains of King Richard III was also performed using the small size amplicon strategy as well as MPS methods (King et al. 2014). The alleged remains of the king, who died in 1485, were excavated in 2012. His identity was confirmed by skeletal features and mitochondrial DNA matched with a living relative who was 19 generations removed from Richard III on the maternal line. The hypervariable regions (HV1, HV2, and HV3) of the mitochondrial DNA control region were successfully amplified and directly sequenced using multiple overlapping fragments ranging from 153 to 250 bp in size. For whole mitochondrial genome sequencing, 24 MPS sequencing libraries generated from 16 extracts were enriched using on-array hybridization. Eye and hair color typing was also conducted using enriched library sequencing with the probes designed specifically for pigmentation-associated single nucleotide polymorphisms. For any further confirmation, DNA fragments under 100 bp were PCR amplified and then sequenced on an MPS machine. Based on the results, King Richard III was predicted to have blue eyes and blond hair. The small size amplicon strategy has been successfully applied to many ancient DNA analyses. However, because only DNA fragments that are long enough to allow for the hybridization of two forward and reverse primers can be PCR amplified, direct PCR amplification, even with a small size amplicon, provides limited power to reconstruct sequences from short DNA fragments (Dabney et al. 2013).

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Improving DNA Recovery Since ancient DNA is usually found in trace amounts, it is crucial to maximize DNA recovery for subsequent ancient DNA analysis. Several comparative studies of DNA yields using various extraction techniques revealed that most methods could only recover a small portion of endogenous DNA (Mumy and Findlay 2004; Barta et al. 2014). Average DNA loss during extraction ranged from 45% to 99%, and this percentage was affected by DNA fragment length. Efforts have been made to improve DNA extraction methods, and the retrieval of ancient DNA molecules is also known to be seriously hindered by their small size. The amount of recovered DNA fragments and their lengths showed an inverse relationship. Therefore, a method for efficiently recovering DNA fragments of less than 50 bp has been developed (Dabney et al. 2013). Conventional methods used for DNA extraction from ancient bones and teeth revealed varying DNA recoveries depending on fragment size: 150 bp fragments showed 72% recovery, while 35 bp fragments showed only 22% recovery (Rohland and Hofreiter 2007; Dabney et al. 2013). Conversely, the newly suggested method allowed DNA fragments to be recovered equally at recovery rates of up to 90% for all fragments ranging in size from 35 bp to 150 bp (Dabney et al. 2013). The method has optimized the DNA extraction process by applying a larger volume of binding buffer consisting of guanine hydrochloride, sodium acetate, and isopropanol and replacing the silica suspension with a silica column to allow for increased loading of the binding buffer. As MPS became popular in ancient DNA analysis, it became more important to improve DNA extraction by increasing small fragment recovery. This was well demonstrated by comparing the fragment length distribution between two ancient DNA samples extracted using different extraction methods (Hofreiter et al. 2014; Dabney et al. 2013; Orlando et al. 2013). Using the new method with increased small fragment recovery, DNA fragments shorter than 50 bp could be successfully obtained from a 400,000-year-old cave bear (Dabney et al. 2013). However, the fragment range of DNA extracted from a 700,000-year-old horse using the conventional method was 40–140 bp, with an average fragment size of more than 70 bp (Orlando et al. 2013). The potential DNA recovery extrapolated from the cave bear fragment length distribution suggested that more DNA might have been recovered from the horse sample if a more efficient DNA extraction method had been applied (Hofreiter et al. 2014). Optimized DNA extraction methods are the basis for building a good MPS library from ancient DNA samples and are constantly being improved.

Massive Parallel Sequencing MPS has revolutionized ancient DNA research by overcoming nearly all limitations involved in the analyses of degraded DNA and LCN DNA. Unlike direct PCR sequencing, which only provides a small portion of informative sequence between two primer binding sites, MPS allows for full sequence analysis of short fragments by attaching priming sites for PCR amplification and sequencing using adapter

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sequences added at the fragment ends. In addition, because ancient DNA-specific damage patterns can now be measured directly from MPS reads, the procedures to detect contamination and authenticate ancient DNA sequences, such as replication, became unnecessary. More importantly, billions of reads generated from MPS enable high coverage nuclear and mitochondrial genomes to be reconstructed from short DNA fragments of ancient samples, while it is not possible to obtain complete nuclear genomes using PCR-based approaches. To analyze ancient DNA using MPS, DNA fragments should be converted into DNA libraries. DNA libraries are created by connecting artificial DNA segments called adapters to both ends of the DNA fragment, and this adapter sequence is later used to initiate sequencing reactions and to amplify the resulting library. Since the library molecule can be amplified by PCR whenever needed, the original DNA sequences are effectively immortalized while becoming embedded in the library (Gansauge and Meyer 2013). The first ancient DNA studies applying MPS methods used a standard library preparation protocol originally developed for high-throughput sequencing of modern DNA (Noonan et al. 2005). However, this method, which converts doublestranded DNA fragments into a library, resulted in a significant percentage of DNA strand loss and was found to be unsuitable for ancient DNA analyses. Therefore, a single-stranded DNA library preparation method has been developed specifically for MPS analysis of ancient or damaged DNA (Gansauge and Meyer 2013). A single-stranded library can be directly prepared from DNA extracts by ligating adaptors to each end via heat denaturation and a series of enzymatic reactions. Because ancient DNA is generally retrieved in very small amounts, DNA quantification is not usually attempted for library preparation. To convert ancient DNA fragments into a DNA library, an aliquot of DNA extract is first treated with phosphatase to remove phosphate groups from the 50 and 30 end of the DNA fragments. Then, a 50 phosphorylated biotinylated adaptor oligonucleotide is ligated to the 30 end of the heat-denatured single-stranded DNA fragment. The resulting adapter-ligated single-stranded DNA strands are immobilized on streptavidin-coated beads, and the complement strands are then synthesized using polymerase and 50 -tailed primer complementary to the adapter sequence. After removing the 30 adenine overhangs using T4 DNA polymerase, a second adapter is attached via blunt-end ligation. The resultant library molecules are released from the beads by heat denaturation and PCR amplified with tailed primers and full-length adapter sequences, including sample indices. Since the PCR amplification of library molecules is an essential process of MPS analysis, it is also important to select the appropriate DNA polymerase to minimize bias that may occur during PCR (Dabney and Meyer 2012). Using this method, 30%–70% of DNA fragments were estimated to be successfully converted to library molecules (Gansauge and Meyer 2013). Single-stranded library preparation has several advantages over double-stranded library preparation (Gansauge and Meyer 2013). Because this method uses biotinylated adapter molecules, it minimizes the loss of DNA fragments that can occur during the size-selective purification process of double-stranded library preparation. Additionally, unlike double-stranded library preparation, a single-stranded

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library can be prepared from damaged DNA strands, even DNA molecules with single-stranded breaks on both strands and DNA molecules with nucleotide modification. The single-stranded library approach provides an independent opportunity for each strand to be converted to the library, and adapter sequences are ligated to the DNA fragments without loss of the original ends of the DNA fragments since there is no need to perform blunt-end repair. As a result, DNA fragmentation patterns can also be investigated from this at high resolution. Alternatively, this single-stranded library preparation is optimized for sequencing short DNA fragments and is, therefore, not suitable for DNA fragments longer than 120 bp. MPS libraries can be directly sequenced, via a technique called shotgun sequencing. However, very low percentages of endogenous DNA in ancient DNA libraries often preclude shotgun sequencing due to the high cost for enough coverage. Ancient DNA exists as a mixture of endogenous DNA and environmental contaminants such as bacterial and fungal DNA, the endogenous DNA ratio of which is often less than 1% (Carpenter et al. 2013). Thus, with shotgun sequencing, most of the sequencing capacity is occupied by environmental DNA, and sequencing is costly to recover enough coverage for endogenous DNA of interest. To overcome these limitations, a capture or hybridization enrichment procedure was developed to increase the proportion of endogenous DNA in MPS sequencing libraries (Mamanova et al. 2010). This approach uses DNA or RNA bait molecules with sequence similarity to the target endogenous DNA (e.g., the exome or mitogenome). The library is hybridized to the immobilized bait probes (on-array capture) or hybridized to the bait probes and then immobilized (in-solution capture), while non-hybridized fragments are removed by washing. Therefore, eluted library molecules should contain more endogenous DNA than environmental DNA. Both capture methods have similar reproducibility, but in-solution hybridization capture has reported to have several advantages over on-array hybridization capture methods (Mamanova et al. 2010). In-solution hybridization capture uses excessive probes over library molecules, so a smaller amount of DNA library is required for enrichment than when using on-array hybridization capture method. In addition, unlike onarray hybridization capture, which works with microarray slides, no special equipment is required. Hybridization enrichment favors smaller fragments in which shorter fragments are captured with higher specificity than longer fragments. This hybridization enrichment method can increase sample preparation time and cost but can effectively lessen sequencing time and cost. In addition, pooling samples prior to hybridization can reduce target enrichment costs (Mamanova et al. 2010), although this procedure creates the potential for cross-contamination due to index switching between samples. In this way, together with hybridization enhancements, MPS has paved the way for recent major advances in ancient DNA analysis. The introduction of MPS has brought many advantages to ancient DNA analysis, but increased sequencing yield has also created new demands and challenges for effective analysis of MPS data (Hofreiter et al. 2014). Unlike Sanger sequencing, these methods do not check the sequence at every site; therefore, the analysis pipeline, which includes algorithms to reconstruct the original sequence of a sample

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from many short sequence reads, should be carefully configured. Thus, the importance of bioinformatics is also increasing. However, the entire genome cannot be fully sequenced using MPS methods because of the presence of long repetitive sequences, such as STRs, microsatellites, or transposable elements. Their characteristic sequence structure makes it difficult to identify their correct position in the genome, thereby precluding the reconstruction of the entire genome. Finally, even MPS cannot retrieve the sequence when the DNA is too degraded and fragments are too short to be analyzed.

Conclusion DNA analysis has been used to investigate the genetic background and population history of ancient humans. However, high DNA degradation and LCN of endogenous DNA frequently thwart ancient DNA analysis. Because DNA degradation occurs due to nonenzymatic damage as well as enzymatic damage, environmental conditions (e.g., temperature and the presence of water) can affect the quality and quantity of preserved DNA. Although desiccated tissues (i.e., mummified tissues) are likely to be protected from hydrolytic damage, DNA extracted from ancient human remains are usually highly degraded and present as a mixture of endogenous DNA and environmental contaminant DNA. Targeted amplification by PCR allowed for the efficient retrieval of DNA fragments of interest. Nuclear DNA as well as mitochondrial DNA could be successfully analyzed using multiple overlapping small-sized PCR amplicons. However, miscoding lesions caused by DNA damage and preferential amplification of exogenous modern DNA during PCR frequently raise issues associated with contamination. Robust implementation of replicate analysis and controls throughout the entire experimental process could help mitigate issues surrounding the PCR method. Advances in technology through the introduction of MPS have greatly improved the quality and standards of ancient DNA analysis and expanded research opportunities. Simultaneous sequencing of billions of sequence reads allowed for the analysis of whole genomes or mitogenomes from ancient DNA. By converting DNA fragments to library molecules, ancient DNA is successfully immortalized and can now be amplified for analysis when needed. Single-stranded library preparation requires more work than double-stranded library preparation, but it has been reported to have more advantages (e.g., providing independent opportunities for each strand to be converted to the library), thereby allowing us to get much a higher yield of ancient DNA molecules, especially from poorly preserved samples. In addition, high-throughput sequence reads enable the detection of damage patterns typically represented by C to T transition. Since C to T transition caused by cytosine deamination increases toward sequence ends, it is possible to differentiate between mutation and transition due to DNA damage. More interestingly, ancient DNA sequences can be authenticated based on the predicted probability of being damaged, which is calculated using cytosine deamination rates and the relative position of

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deamination from the sequence ends. As a result, some components of the original authentication criteria, such as replication, are now deemed obsolete. Moreover, library molecules created from a mixture of endogenous DNA and environmental DNA can be selectively enriched using hybridization enrichment or capture methods. Some ancient samples included less than 1% endogenous DNA, but hybridization enrichment of library molecules facilitated the analysis of these samples. As single-stranded library preparation and hybridization enrichment are both optimized for short DNA fragments, MPS can be more effective for ancient DNA analysis. As the sequence analysis of short DNA fragments became possible using MPS, efforts have been made to increase DNA recovery, particularly short DNA fragment recovery, from ancient samples, and a method that can recover DNA fragments of less than 50 bp is currently available. However, there is still much more room for technological development in the field of DNA recovery and hybridization enrichment (Rohland et al. 2018). Although many issues surrounding the analysis of degraded DNA and LCN DNA have been resolved by the introduction of MPS and improved DNA extraction methods, vast amounts of sequence reads have produced new challenges, such as the need to develop new computational methods. In addition, there is a need to increase the accuracy of available reference DNA sequence information to facilitate the alignment of short sequence reads to the reference. With continued efforts, ancient DNA analysis techniques will continue to evolve.

Cross-References ▶ Application of SNP-Based DNA Phenotyping to Archaeological and Forensic Cases

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A Primer on the Population Genetic Analysis of Ancient Genomes

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Choongwon Jeong

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Relatedness and Recent Consanguinity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Population Structure and Data Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Modeling of Population Relationship: F-Statistics and Their Extensions . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

The genetic analysis of ancient biological remains is increasingly relying on next generation sequencing and genome-wide data it produces. In addition to bioinformatics processing of high-throughput data produced by next generation sequencing, downstream population genetic analyses are critical to fully utilize rich evolutionary information stored in genomic variation data. In this chapter, I summarize representative population genetic methods that are applied to genome-wide data from ancient sources, such as mummies. They aim at detecting genetic relatives, overviewing the genetic profile of the analyzed individuals in the context of worldwide genetic diversity, and formally testing population relationship. For each method, I outline its general principle and provide practical guidance for conducting it. Keywords

Mummy genomics · Population genetics · Genetic relatedness · Principal component analysis · ADMIXTURE · F4 statistics · qpAdm

C. Jeong (*) School of Biological Sciences, Seoul National University, Seoul, Republic of Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_49

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Introduction The study of ancient genomes provides a window to directly observe genetic profiles of individuals and populations who lived in the past, stretching back to as far as hundreds of thousands of years (Krause and Pääbo 2016). Archaeogenetic studies of our own species are at the forefront of this trend, and findings from the study of ancient human genomes, especially those obtained from mummies or naturally mummified soft tissues, revolutionized our understanding of human history in various temporal and spatial scales (Rasmussen et al. 2010; Rasmussen et al. 2011; Schuenemann et al. 2017). For example, the genetic relationships between the anatomically modern humans and various archaic hominins, such as Neandertals, had been hotly debated for decades in the field of paleoanthropology (Stringer 2014), but archaic hominin genomes retrieved in the last decade now provide a solid background to reach a scientific consensus on the deep genetic history of hominins (Green et al. 2010; Meyer et al. 2016; Reich et al. 2010). Likewise, a direct comparison between the genomes of Mesolithic hunter-gatherers and early Neolithic farmers in Europe sealed a century-long debate on the mode of the Neolithic transition in Europe, supporting that human migrations from the Anatolian farming societies to Europe predominantly drove the diffusion of the Neolithic lifestyle across Europe (Allentoft et al. 2015; Haak et al. 2015; Lazaridis et al. 2014). A recent boom in the study of ancient DNA (“aDNA”) is driven by the access to genome-scale data, similar to the earlier transition into genome-wide data in the study of present-day genetic materials (Li et al. 2008; Mallick et al. 2016; The 1000 Genomes Project Consortium 2015). Microarray genotyping, and later next generation sequencing (NGS), have provided a high-throughput to routinely investigate millions of genetic variants across the genome for hundreds and thousands of individuals since the late 2000s (Bergström et al. 2020; McCarthy et al. 2016; Walter et al. 2015). Due to the highly fragmented nature of aDNA molecules (Sawyer et al. 2012), the microarray technology was not applicable to archaeogenetics. Therefore, NGS has been the main technological platform for archaeogenetics to achieve genome-scale data production. Genome-wide data have a statistical power to decode the past genetic history of individuals and populations to a level that far exceeds what can be provided by the analysis of a small number of markers (Li and Durbin 2011). This robustly holds true even if the markers used in the genome-wide analysis, i.e., mainly single nucleotide polymorphisms (SNPs), have marker-wise much smaller amount of information than the traditionally investigated loci, such as mitochondrial or Y chromosome haplotypes. This is because the relationship of sampled sequences from a single locus (referred to as the “gene tree”) often deviates from that of sampled individuals/ populations (referred to as the “population tree”), mainly due to two factors: gene flow and a phenomenon called the incomplete lineage sorting (Fig. 1). For example, while the species tree of (Gorilla, (Chimpanzee, Human)) robustly holds with a separation for two to four million years between the Gorilla lineage and the common ancestors of Chimpanzee and Human prior to their split, still only about 70% of gene trees across the genome match the species tree, while the remaining 30% finds

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Fig. 1 Population tree vs. gene tree. The population tree of three populations (P1, P2, P3) is shown as a tree of block segments, while the relationship of three sequences (a-c; the gene tree) is shown by the lines. (a) The gene tree (a, (b, c)) matches the underlying population tree (P1, (P2, P3)), where P2 and P3 are closer to each other than they are to P1. (b) The gene tree ((a, b), c) does not match the population tree, showing an example of the incomplete lineage sorting. Going back in time, sequences b and c do not converge into their common ancestor while (P2, P3) are separated from P1. Once all three populations merge into their common ancestral population, sequence b or c may coalesce (i.e., meeting their common ancestral sequence) with sequence a first before they coalesce each other, as in this case. (c) The population relationship in this case does not follow a simple tree: Instead, P2 received a recent gene flow from P1 (marked by a block outlined by dotted lines). The gene tree ((a, b), c) matches a population for the introgressed loci, but not the one without admixture

Gorilla closer to either Chimpanzee or Human than Chimpanzee and Human are to each other (Scally et al. 2012). Therefore, it is crucial to investigate the distribution of gene trees across the genome, even if the individual gene tree can only be inferred with substantial noise, rather than relying on a small number of gene trees, for accurately reconstructing the genetic relationship between individuals/populations. Genome-wide markers represent a large number of partially independent gene trees across the genome due to past recombination events between markers. In this chapter, I provide an overview of various population genetic analyses of genome-wide aDNA data, focusing on pseudohaploid genotype data (i.e., single base random sampling for each marker) on a preascertained set of genome-wide SNPs. Considering that this type of data is most abundant in the recent archaeogenetic studies (Haak et al. 2015; Mathieson et al. 2015), it has a practical importance to understand how these analyses work, how they are different from the analyses of high-quality diploid genotype data, and how we should interpret the results. It will provide a useful first-step tutorial for mummy researchers who are interested in the retrieval and analysis of ancient mummy genomes.

Relatedness and Recent Consanguinity Many population genetic analyses utilize allele frequency to infer the genetic history of a population and assume the independence between individuals within a group/population when calculating allele frequency from individual genotype data. Therefore, in

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large-scale genetic studies, quality control (QC) procedures typically include detection of close relatives within study participants. In addition, findings of genetic relatives or individuals from a recent consanguineous mating are of great interest within the archaeological and anthropological contexts (Mittnik et al. 2019). Among the most tantalizing archaeological example is the repeated consanguineous mating in the family of a famous Egyptian Pharaoh Tutankhamun (Hawass et al. 2010). Genetic relatedness means that two individuals share a common ancestor in a recent past (which is defined in a loose and arbitrary manner) so that large segments of their genomes are exact copies of each other, commonly inherited from their shared ancestor. These shared segments are called the identity-by-descent (IBD) segments. Geneticists characterize genetic relatedness in terms of the distribution pattern of the IBD segments: i) the coefficient of relationship (frequently referred to as “r”), which is the probability of an allele randomly sampled from an individual and has an IBD copy in the other, ii) IBD1 (commonly referred to as “k1”), which is the probability that only a single allele of an individual in a locus has an IBD copy in the other, iii) IBD2 (commonly referred to as “k2”), which is the probability that both alleles of an individual in a locus have their IBD copies in the other, and iv) the coefficient of inbreeding (frequently referred to as “f”), which is the probability that two alleles of an individual are IBD to each other. Various statistical methods have been developed for accurately inferring these parameters using dense genome-wide diploid genotype data (Manichaikul et al. 2010; Moltke and Albrechtsen 2014; Purcell et al. 2007). However, the standard statistical methods do not perform properly when accurate diploid genotype calls are not available. It is a common issue of studies based on a small number of low-coverage sequence data, including most aDNA studies. Various methods have been recently developed to overcome this issue, taking into account the nature of aDNA sequence data (Kennett et al. 2017; Korneliussen and Moltke 2015; Lipatov et al. 2015; Monroy Kuhn et al. 2018). The pairwise mismatch rate (PMR) is a simple method for inferring the coefficient of relationship (r) using pseudohaploid genotype data of a pair of individuals (Kennett et al. 2017). PMR is defined as the proportion of SNPs that two individuals have different alleles as their pseudohaploid genotype out of all SNPs in a panel that both individuals have genotypes. It can be easily adjusted to incorporate diploid genotype calls by taking a value of 0.5 when either individual has a heterozygous genotype. For a SNP i with population allele frequency pi, the expected PMR between a pair of individuals with the coefficient of relationship r can be written as below: ðExpected PMRÞ ¼



 r r r 0þ 1  2pi ð1  pi Þ ¼ 1   2pi ð1  pi Þ 2 2 2

Because r is invariant across SNPs, one can rewrite the genome-wide expected value as: ðExpected PMRÞ ¼

N
N 
 1 X r r 1 X  2pi ð1  pi Þ ¼ 1  1 2p ð1  pi Þ N i¼1 2 2 N i¼1 i

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Here r/2 corresponds to the probability that the two alleles sampled are IBD, and Hi ¼ 2pi(1-pi) corresponds to heterozygosity, which is the probability that two randomly sampled alleles in a population differ from each other. Therefore, one can expect that the PMR values between duplicated samples (e.g., two independent libraries from the same skeletal element) are around half of the values between unrelated individuals. One can set the baseline either by taking the mode of PMR values across individual pairs (assuming that there are many pairs of unrelated individuals) or by taking the PMR values from known duplicates. Either way, the expected PMR value linearly decreases with increasing r within the range [Hi, Hi/2]. Due to its reliance on pseudohaploid genotypes, PMR cannot distinguish between IBD1 (k1) and IBD2 (k2). For example, parent-offspring and full-sibling pairs both have r ¼ 0.5, but markedly different k1 and k2 values: (k1, k2) ¼ (1, 0) and (0.5, 0.25) for parent-offspring and full-sibling, respectively. Consanguineous mating, i.e., mating between relatives, also produces nonzero k2 values in the offspring from it. Multiple methods extend the standard probabilistic model for estimating k1 and k2 coefficients from diploid genotype data to incorporate uncertainty in genotypes in the form of genotype likelihood, such as lcMLkin and NGSRelate (Korneliussen and Moltke 2015; Lipatov et al. 2015). Genotype likelihood refers to the probability of obtaining the observed read data given a certain diploid genotype. k1 and k2 values for each pair of individuals in the data set are incorporated into the model as extra parameters that affect the probability distribution of individual genotypes per locus. Still, these methods are expected to perform well ideally in a genome-wide coverage of 2x or higher: although this level of coverage is considered “low” in genome sequencing of present-day individuals, a vast majority of ancient samples yields coverage much lower than ~1x, resulting in noisy estimates of k1 and k2. In many cases, noisy estimates of these parameters are still robust enough to distinguish between parent-offspring and full-sibling relationships, especially when combined with uniparental haplogroup, genetic sex, and age at death information. However, robust detection of more distant relatives with ultralow coverage aDNA data remains challenging. In addition to making individuals share more alleles on average, genetic relatedness also makes such allele sharing be locally distributed across the genome in the form of IBD segments. IBD segments tend to get shorter as the genetic relatedness between individuals becomes more distant, due to the recombination events accumulated over generations. That is, close relatives tend to have long stretches of IBD segments that reduce the genetic distance between two individuals (e.g., reducing PMR to ~3/4 and ~ 1/2 of the expected value without IBD for IBD1 and IBD2 regions, respectively), while more distant relatives have relatively shorter stretches of such IBD segments. Identifying IBD segments in low to ultralow coverage, ancient genomes remain a challenging task, but there are special cases that utilize such information with regard to the archaic hominin admixture. First, a study of a 40,000-year-old individual from Romania identified long genomic segments with high density of Neandertal-specific alleles as well as an overall high level of genome-wide Neandertal ancestry, allowing the authors to propose that she had a

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Neandertal ancestor as recent as four generations ago (Fu et al. 2015). Second, a study of an archaic hominin genome from southern Siberia revealed that this individual was heterozygous for most SNPs across the genome where known Neandertal and Denisovan genomes have different alleles, a phenomenon that can be explained only if she was a first-generation hybrid between a Denisovan father and a Neandertal mother (Slon et al. 2018). Going beyond these special cases of utilizing highly diversified lineages, a recent study provides a statistical method for identifying genomic segments where two chromosomes of an individual are IBD, often referred to as runs of homozygosity (ROH) (Ringbauer et al. 2020). Utilizing linkage disequilibrium (LD) information from a large number of modern genome sequences, it allows such an inference robust for ultralow coverage genomes as low as ~0.5x and for relatively small segments as short as 4 centiMorgans (cM). Future improvements in the probabilistic detection of IBD segments in low-coverage aDNA data will be a key to understanding the social organization of past human societies.

Population Structure and Data Overview Together with a search for close relatives, the first steps of genome-wide aDNA analysis include a summary of the genetic profile of ancient individuals in the context of worldwide genetic diversity among the present-day populations. Two methods are extremely popular and routinely used for this task: principal component analysis (PCA) and model-based genetic clustering based on the STRUCTURE model, such as ADMIXTURE (Alexander et al. 2009; Patterson et al. 2006; Pritchard et al. 2000). Both methods summarize a high dimensional genotype matrix (the number of SNPs  the number of individuals) to a much smaller number of variables per individual, such as PC coordinates or the proportion of each ancestry component (Fig. 2). PCA is a popular dimension reduction technique that projects each individual originally in an M-dimensional space of genotypes across M SNPs into a much smaller K-dimensional space of top K PCs. For this, one first calculates a covariance matrix of normalized genotype data and then diagonalizes the covariance matrix via eigen decomposition or singular value decomposition (Patterson et al. 2006; Yang et al. 2011). For example, among the most popular programs for running PCA of genotype data is the smartpca/eigenstrat program (Patterson et al. 2006), which standardizes genotype values (coded as 0, 1, or 2 for the number of “1” alleles) by subtracting the average genotype value across all individuals used for PCA and then by dividing it by the corresponding standard deviation of the genotype values assuming the Hardy-Weinberg equilibrium. The mean and standard deviation of the genotype values pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi ffi for a SNP where p is the frequency of the “1” allele should be 2p and 2pð1  pÞ. Most aDNA data are pseudohaploid genotypes due to low coverage and have missing data in a substantial fraction of SNPs. These features make it difficult to apply the standard PCA techniques to aDNA data. First, pseudo-haploidization by single allele random sampling creates an extreme level of artificial genetic drift

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Fig. 2 Dimension reduction of multilocus biallelic SNP genotype data by PCA and ADMIXTURE. The genotype matrix of M SNPs and N individuals places each individual in an M-dimensional space of genotypes. Typically, M is in an order of 104–106. Both PCA and ADMIXTURE map each individual into a much smaller dimension of top K PCs or K ancestry components, typically in an order of 101

(i.e., the ancient individual has zero heterozygosity) and results in high genotype error rates (same as per-base error rate which is in the order of 102 to 104). To exclude the impact of this artificial genetic drift and genotyping errors, in most cases, PCs are calculated for a set of high-quality diploid individuals (e.g., a large number of present-day individuals) and pseudohaploid ancient genomes are projected onto the top PCs (Lazaridis et al. 2014). By doing so, one can obtain more reliable and replicable results but misses a chance to directly look into the genetic history of ancient individuals not shared with those used for PC calculation. Second, the projection of ancient individuals onto the precalculated PCs is not a straightforward task when the projected individuals have missing data in a substantial fraction of SNPs. For example, the smartpca program assumes low missing rate and fill in missing genotypes with population mean values to streamline computational procedures (Patterson et al. 2006). This genotype fill-in procedure creates an artificial displacement of ancient individuals in the PC space toward the origin (i.e., toward zero in the PC coordinates). In earlier studies, this issue was dealt with a complex procedure involving the Procrustes transformation (Skoglund et al. 2012). For each ancient individual, one creates a data set composed of a reference set of present-day individuals (who will be used for calculating PCs) and the target ancient individual and for SNPs covered by the target ancient individual. By doing so, it removes the issue of high and varying missingness among ancient individuals. Then PCA is performed for each data set, using the same set of individuals but different sets of

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SNPs. Finally, PCA results are merged into one by the Procrustes transformation of PC coordinates of the reference present-day individuals. These days, such an approach is impractically inefficient when tens or even hundreds of ancient individuals are frequently coanalyzed. In the latest versions of the smartpca program, the missingness issue is resolved in a more efficient and accurate way: mainly, this method infers PC coordinates of projected samples by taking into account only the SNPs each projected individual is covered (for the formal description of the idea, please see https://github.com/chrchang/eigensoft/blob/master/POPGEN/ lsqproject.pdf). I recommend to apply this option “lsqproject: YES” to all cases when the sample projection onto the precalculated PCs is performed. The exclusion of ancient individuals from calculating PCs also creates a small amount of regression toward the origin, mainly because individual-specific genetic drift is not incorporated into PCs. That is, when a subset of individuals from a population is used to calculate PCs while the remaining individuals are projected, the projected ones tend to regress slightly toward the origin rather than being projected on top of the individuals included in PC calculation. This effect is more pronounced when the underlying genetic structure is subtler. The “shrinkmode: YES” option in the smartpca program is developed to resolve this issue, but it substantially increases the runtime. While I do not recommend a universal application of the shrinkage option due to an increase in the computational burden, I would like to highlight that the users should be aware of this issue and avoid a misinterpretation of data. Another set of popular methods share the same generative model for genotype matrix data that was first implemented in the STRUCTURE program (Pritchard et al. 2000). Since the initial publication of STRUCTURE, many programs implemented this model with different inference methods (e.g., maximum likelihood vs. Bayesian methods) and varying computational speeds, including the widely used ADMIXTURE program (Alexander et al. 2009; Novembre 2016). This model summarizes the genetic profile of each individual into a vector of K elements, corresponding to the proportional contribution from K distinct gene pools (Fig. 2). These K gene pools (frequently referred to as either ancestral populations or ancestry components) are characterized by their allele frequency for each of the M SNPs used in the analysis. Then, each individual is envisioned as a random sample from a population formed by a mixing of the K gene pools with the individual-specific mixture proportion. Integrating out local ancestry, the STRUCTURE model calculates genotype likelihood as below (Alexander et al. 2009):   Pr Gij ¼ g ¼

  2g 2 g pij 1  pij g

and pij ¼

K X k¼1

qik f kj

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Here, Gij is the genotype value of the i-th individual in the j-th SNP, which takes a value of 0, 1, or 2. fkj refers to the frequency of the “1” allele in the k-th gene pool, and qik refers to the proportion of ancestry that the i-th individual takes from the k-th gene pool. Then, pij becomes the allele frequency of the mixed population to which the i-th individual belongs. The likelihood of having a certain genotype is modeled as a random binomial sampling of two alleles assuming the Hardy-Weinberg equilibrium. The likelihood of data is obtained by multiplying the above per-SNP per-individual likelihood across SNPs and individuals, assuming independence of SNPs and individuals. That is, LðGjF, QÞ ¼

YY

2

i¼1 j¼1

gij

!

K X k¼1

!gij qik f kj

1

K X

!2gij qik f kj

k¼1

Because the STRUCTURE model simply excludes missing data from its genotype likelihood calculation, it does not show a substantial issue corresponding to the regression of high missingness samples toward the origin in PCA. Still, the inferred genetic profile (i.e., the ancestry proportion) of extremely low-coverage individuals should be considered noisy and interpreted with great caution. Also, high genotyping error rates in the pseudohaploid ancient individuals may create artificial allele sharing between them, especially in low-frequency alleles. Therefore, it is advised to filter out low-frequency SNPs (e.g., minor allele frequency < 1%) prior to running the program, e.g., by using “– maf 0.01” flag in PLINK (Chang et al. 2015). In both PCA and ADMIXTURESTRUCTURE-like clustering, the main goals of the analysis are i) to detect most distinct individuals/populations, and ii) to identify those showing intermediate genetic profiles: that is, those who are positioned between distinct clusters in PCA or those who are modeled as a mixture of multiple genetic components in ADMIXTURE (Fig. 3). Taken together, it is common to observe a genetic cline (or a gradient) in these analyses, either as a group of individuals scattered along a line in the PC space, or as those modeled as a combination of two or more ancestry components in varying proportions in ADMIXTURE. A naïve interpretation of this genetic cline would be to assume the observed genetic profile as a true representation of the recent evolutionary history: for example, an individual with an intermediate genetic profile may be interpreted as an admixed descendant of two or more ancestral populations (Fig. 3). However, a genetic cline in PCA or ADMIXTURE may be created by other reasons, including a serial founder event, strong genetic drift in a set of individuals, or unbalanced sampling, to name a few (Fig. 3) (Lawson et al. 2018; van Dorp et al. 2015). Therefore, it is important to consider PCA and ADMIXTURE mainly as a tool for the firsthand data exploration and hypothesis generation, not as a formal test of population relationship. Formal tests of population relationship are expected to be insensitive to private genetic drift and sample size per group.

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Fig. 3 Genetic clines in PCA or ADMIXTURE. In (a) PCA or (b) ADMIXTURE, a population (P2) may show a genetic profile intermediate of others (P1 and P3). (c) Such a profile may reflect a true admixture in the intermediate group (left), but other scenarios such as extreme genetic drift (middle and right) can also produce such a pattern

Modeling of Population Relationship: F-Statistics and Their Extensions A group of formal tests of population relationship, collectively called F-statistics, is widely used in genome-wide analysis (Patterson et al. 2012). These methods explore correlations in allele frequency differences between multiple groups to infer their genetic relationship: Especially, they focus on testing and quantifying admixture or gene flow, which are defined as a deviation from a tree-like relationship. F4 statistic and a nearly identical Patterson’s D statistic (which is frequently referred to as the ABBA/BABA test) are first developed to test a differential relationship of modern human populations with an archaic hominin group Neandertal (Green et al. 2010). If there was no recent gene flow between Neandertals and a subset of human populations nor a long-standing population structure involving Neandertals, it is expected that any two modern human populations (or individuals) should form a sister clade to the exclusion of Neandertals, making the following tree topology fit to genome-wide data: (Chimpanzee, (Neandertals, (Modern1, Modern2))). If there was a gene flow between Neandertals and Modern2 (in one direction or the other), this symmetry does not hold anymore: Instead, within the introgressed

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loci, Neandertals and Modern2 are closer to each other than they are to Modern1. Assuming that we observe a single allele per group in each locus, F4 statistic quantifies the difference in the number of sites where i) Neandertal and Modern2 have one allele and the others have another (thus “BABA” – Neandertal and Modern2 have “A” allele while Chimpanzee and Modern1 have “B” allele), and ii) Neandertal and Modern1 have one allele and the others have another (thus “ABBA”). If the two numbers are significantly different, the above tree-like relationship does not hold anymore, and a gene flow between some of the four populations should be postulated to adequately explain data. Indeed, this simple but powerful method provided the first firm proof for the Neandertal gene flow into the ancestors of present-day populations out of sub-Saharan Africa, and the Denisovan gene flow into the ancestors of present-day Australasians (Green et al. 2010; Meyer et al. 2012). Generalizing the above single allele per group case into the population frequency case, the F4 statistic of four populations (O, A, B, and C in this order) can be written as: F4 ðO, A; B, CÞ ¼

M   1 X  O p  pAi pBi  pCi M SNP i¼1 i

where pXi refers to the allele frequency of the i-th SNP in population X. The F4-ratio test is the first attempt to characterize details of the inferred admixture, here defined as a deviation from a strictly tree-like relationship, such as the admixture proportions, by utilizing multiple F4 statistics (Durand et al. 2011; Patterson et al. 2012). It involves five populations (here referred to as O, A, B, C, and T ) and two F4 statistics (Fig. 4). O refers to a distant outgroup, A and B represent proxies for the two major ancestral lineages, C is another group that falls on the A lineage, and finally, T refers to an admixed descendant of the A and B lineages. Let us assume a scenario that C is a sister clade of the true ancestral population of T on

Fig. 4 Schematic population graphs for F-statistic-based characterization of admixture. (a) The F4-ratio test aims to estimate admixture proportions (α) of the target T using two F4 statistics. (b) QpAdm both tests the admixture model and infers admixture proportions (αk) using references (Rk) and outgroups (Oi). Among the outgroups in the graph, O1 and O2 do not help because they are symmetrically related to the two sources, S1 and S2, and thus can be excluded from the test. The others, O3-O5, provide a statistical resolution to distinguish contributions from the sources because they are differentially related to the sources. Importantly, for all outgroups, each source-reference pair, S1-R1 and S2-R2, is symmetrically related

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the A side (A0), to the exclusion of a more distant proxy A: i.e., the tree (A, (C, A0)) holds. In this case, the ancestry proportion of T from the A lineage (α) is expected to be in the following form: α¼

F4 ðO, A; B, T Þ F4 ðO, A; B, CÞ

The above relationship holds because the expected allele frequency of an admixed population T is a linear combination of the sources A0 and B0. That is,   0 0 E pTi ¼ αpAi þ ð1  αÞpBi Then, using F4(O, A; B, C) ¼ F4(O, A; B, A0) and F4(O, A; B, B0) ¼ 0, it can be shown: F4 ðO, A; B, T Þ F ðO, A; B, A0 Þ F ðO, A; B, B0 Þ ¼α 4 þ ð1  αÞ 4 ¼α F4 ðO, A; B, CÞ F4 ðO, A; B, CÞ F4 ðO, A; B, CÞ Extending the idea of summarizing multiple F4 statistics, qpAdm analyzes a matrix of F4 statistics to test multiway admixture models (Lazaridis et al. 2016). The model fitting relies on principles of linear algebra, but the underlying idea is quite simple. Let us assume that a target population is an admixed descendant of K distinct source populations (S1, . . ., SK) with a nonnegligible mixture proportion (α1, . . ., αK), and there are K proxy reference populations (R1, . . ., RK) that are closely related to the sources: In the best scenario each Rk is a sister clade of Sk (Fig. 4). For a set of “outgroup” populations (O0, O1, . . ., ON) that are somewhat more distant but still differentially related to S’s, the following relationships hold:   F4 Oi , O j ; Sk , Rk ¼ 0 for all i, j, k   F4 Oi , O j ; T, Rk 6¼ 0 for all k for some ði, jÞ pairs K X

  αk F4 Oi , O j ; T, Rk ¼ 0

for all i, j, k

ðiÞ ðiiÞ ðiiiÞ

k¼1

That is, qpAdm minimizes the difference, measured by F4 statistics, between T and a weighted average of R’s. If it can find ðb α1 , . . . , b αK Þ that satisfies the symmetry condition (iii), one can interpret it as an adequate admixture model of T given the outgroups. If the best combination of R’s still deviates from T, the admixture model does not adequately mimic T: It may suggest that the current R’s are not a good representative of the true sources (S’s), in the sense that the relationship between R’s and S’s is not as simple as the above condition (i) suggests, or the current set of R’s is missing a representative for some S’s. The qpAdm program operates on the N  K matrix of F4 statistics (X), the elements of which are given as:

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Xij ¼ F4 ðOo , Oi ; T, Rk Þ When the above symmetry condition (iii) holds, not all columns of the matrix X are independent due to the following constraint: K X

αk Xik ¼ 0

k¼1

In other words, if the admixture model is adequate, there are only (K-1) independent columns in the matrix X instead of K columns. If the admixture model does not fit, all K columns are expected to be independent. Therefore, qpAdm infers (α1, . . ., αK) values assuming that only (K-1) columns are independent and quantifies the model fit in terms of the difference between T and a combination of R’s. Small p-values indicate that the model does not fit the target. The F4-ratio test can be seen as the simplest precursor of qpAdm, using T as the target, (A0 and B) as the references, and (O and A) as the outgroups. That is, the 1  2 F4 matrix becomes: X ¼ ðF4 ðO, A; T, BÞ F4 ðO, A; T, CÞÞ Although qpAdm does not have the statistical power to test the admixture model itself in this case, one can easily turn it into a valid qpAdm test by adding another outgroup D that is more closely related to B than to A (Fig. 4):  X¼

F4 ðO, A; T, BÞ F4 ðO, A; T, CÞ F4 ðO, D; T, BÞ F4 ðO, D; T, CÞ



QpAdm does not explicitly model all populations used in the analysis into a single population graph, which refers to a population tree corroborated by gene flow edges between branches. However, it provides a powerful statistical approach to reconstruct a part of this graph, mainly the relationship between the target and the references. Owing to its power and simplicity, it has been widely adopted as a key method to formally explore the nature of genetic clines identified by descriptive methods, such as PCA and ADMIXTURE.

Conclusion In this chapter, I reviewed three sets of population genetics methods that utilize genome-wide SNP data. These methods provide powerful tools to identify and characterize (i) genetic relatedness, (ii) a global overview of the genetic relationship between the target groups and worldwide populations, and (iii) a formal test of population relationship, respectively. These topics are at the basis of reconstructing the genetic history of any individual/population, both humans and nonhumans.

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These methods can be applied to genomic information of any source, including mummies, and therefore have been and will be widely utilized in the genomic study of mummies.

Cross-References ▶ Ancient DNA Study ▶ Current Trends in Ancient DNA Study

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Application of SNP-Based DNA Phenotyping to Archaeological and Forensic Cases

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . DNA-Based Prediction of Pigmentation Traits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Eye Color Prediction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Hair Color Prediction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Skin Color Prediction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pigmentation Prediction in Archaeological and Forensic Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . Prediction of Pigmentation Traits in Historical Figures and Forensic Identification Cases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Prediction of Pigmentation Traits in Ancient Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Perspectives on SNP-Based Phenotype Prediction in Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

388 388 389 395 397 399 399 402 404 405 405 405

Abstract

DNA phenotyping refers to the prediction of appearance traits of a sample donor via DNA analysis. In the forensic field, pigmentation traits, such as eye, hair, and skin color, have been most studied, and several validated DNA test systems are available for categorical color classification. Current DNA test systems provide considerable prediction accuracy, but using different classification schemes, different sets of markers, and different training and validation sets in model development it can generate different prediction results. Technological advances in DNA analysis have allowed many researchers to pursue SNP-based prediction of pigmentation traits in archaeological and forensic cases. This chapter introduces the scientific basis and H. Y. Lee (*) Department of Forensic Medicine, Seoul National University College of Medicine, Seoul, Korea Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_50

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analytical techniques of SNP-based pigmentation trait phenotyping, and presents several examples of their application. Keyword

Ancient DNA · Single nucleotide polymorphism · Pigmentation traits · Eye · Hair · Skin

Introduction Before genetic analyses were made available, anthropological studies of human population variation were largely dependent on physical characteristics, such as skin, eye, and hair color, body size and stature, limb proportions, and dry versus wet ear wax (Stoneking 2017). These studies not only documented variation in physical characteristics, but also investigated influences that could affect these variations. However, understanding the genetic background of these phenotypic variations, as well as the impact of climate and diet on them, can better explain the process of human adaptation and evolution related to these variations. Therefore, many researchers have tried to find a genetic basis that can explain these variations. Aided by methodological and theoretical advances in genome-wide association studies (GWASs), significant progress has been made in understanding some phenotypic traits, particularly those genes involved in hair, eye, and skin pigmentation. Compared to most physical characteristics that are complex polygenic traits, in which a large number of genes contribute to the phenotype, human pigmentation traits are considered to have a relatively simple genetic basis, and just a small number of genes provide most of this phenotypic information (Kayser 2015). Consequently, DNA-based pigmentation prediction has become possible with considerable accuracy, and has begun to be used in forensic evidence analysis to predict the appearance of unknown sample donors. In addition, thanks to advances in massive parallel sequencing (MPS) technology, the availability of partial and complete genome sequences from ancient samples has also increased, maximizing the possibility of obtaining information about the phenotypic traits of archaic humans (Neandertals and Denisovans) as well as of ancient modern humans. Moreover, development of targeted MPS methods based on PCR amplification or specialized hybridization capture methods that target a predefined set of phenotype-associated single nucleotide polymorphism (SNP) positions have facilitated high coverage data even from degraded DNA samples in a more cost-effective manner (Peltzer et al. 2018). In this chapter, current knowledge of DNA-based prediction of eye, hair, and skin pigmentation will be summarized, and its application to archaeological and forensic cases will be discussed.

DNA-Based Prediction of Pigmentation Traits Among various phenotypic traits, pigmentation traits such as eye, hair, and skin color variations can currently be predicted using genotype data on the level of common color categories. This is possible because genes and predictive SNP markers that are

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Fig. 1 Some of the important steps and genes involved in melanogenesis. Possible gene expression regulation by one gene to another is indicated by a dashed line arrow

either causal or associated with these characteristics have been identified (Schneider et al. 2019). Figure 1 shows some of the major steps and enzymes involved in melanin biosynthesis in humans. Melanin is a light-absorbing polymer synthesized by melanocyte in specialized lysosomes called melanosomes (Frudakis et al. 2003), and gives color to skin, hair, and eyes. Within the melanosomes, tyrosinase (TYR) catalyzes the hydroxylation of tyrosine to 3,4-dihydroxyphenylalanine (DOPA), which is then oxidized to dopaquinone to form the precursors of eumelanin (black or brown color) and pheomelanin (yellow or red color). Melanocortin-1 receptor (MC1R) is a major signaling molecule in melanocytes that responds to the α-melanocyte stimulating hormone by inducing the expression of enzymes responsible for eumelanin synthesis (Nasti and Timares 2015). The solute carrier family 24 member 4 (SLC24A5) and solute carrier family 45 member 2 (SLC45A2) are membrane transporter proteins that promote melanogenesis (Stoneking 2017). Polymorphisms within these genes or in regulatory regions near these genes can cause color variations in skin, hair, and eyes. Table 1 shows previously identified SNPs that can be applied to the prediction of pigmentation traits. Several statistical predictive models based on reference datasets consisting of SNP genotypes and associated phenotypes have been developed. Although many models demonstrate high prediction accuracy, different prediction results can be obtained based on the number and composition of SNP markers, and the differences in the underlying logic and supporting data for model construction.

Eye Color Prediction Unlike human hair and skin color, there are several major components that participate in determining the color of the eyes. Only a minimal correlation was observed among skin, hair, and eye colors within or between individuals of a given population, but good concordance was obtained between populations; populations with darker eye color tend to have darker skin and hair colors (Frudakis et al. 2003). These observations suggest that the genetic determinants for pigmentation can be different between tissues (Frudakis et al. 2003).

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Table 1 SNP for DNA-based prediction of pigmentation traits. (Redrawn with modification from Kayser (2015)) SNP rs3768056 rs13289 rs16891982 rs26722 rs28777 rs35395 rs4959270 rs12203592 rs1540771 rs10756819 rs1325127 rs1408799 rs2733832 rs683 rs35264875 rs3829241 rs1042602 rs1126809 rs1393350 rs10831496 rs10777129 rs12821256 rs12896399 rs17128291 rs2402130 rs1129038 rs1667394 rs12592730 rs12913832 rs2238289 rs3935591 rs6497292 rs7183877 rs916977 rs11636232 rs1724630 rs1004611 rs1037208 rs10852218 rs11855019 rs123439067

Gene LYST SLC45A2 SLC45A2 SLC45A2 SLC45A2 SLC45A2 EXOC2 IRF4 IRF4-EXOC2 BNC2 TYRP1 TYRP1 TYRP1 TYRP1 TPCN2 TPCN2 TYR TYR TYR GRM5/TYR KITLG KITLG SLC24A4 SLC24A4 SLC24A4 HERC2 HERC2 HERC2 HERC2 HERC2 HERC2 HERC2 HERC2 HERC2 HERC2 MYO5A OCA2 OCA2 OCA2 OCA2 OCA2

Chr. 1 5 5 5 5 5 6 6 6 9 9 9 9 9 11 11 11 11 11 11 12 12 14 14 14 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15

Predicted phenotypes Eye color Hair color ✓ ✓ ✓

✓ ✓

✓ ✓

✓ ✓ ✓

Skin color ✓ ✓ ✓ ✓ ✓ ✓

✓ ✓ ✓ ✓

✓ ✓ ✓

✓

✓

✓

✓

✓ ✓

✓ ✓

✓ ✓ ✓ ✓

✓ ✓ ✓ ✓

✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ (continued)

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Table 1 (continued) SNP rs12441727 rs1375170 rs1375164 rs1448484 rs1448485 rs1448490 rs1470608 rs1498519 rs1545397 rs1597196 rs1800401 rs1800404 rs1800407 rs1800414 rs1874835 rs1900758 rs2036213 rs2305252 rs2311470 rs2594935 rs2871886 rs3099645 rs3794606 rs4778137 rs4778138 rs4778177 rs4778190 rs4778232 rs4778241 rs6497268 rs7179994 rs728405 rs735066 rs7495174 rs749846 rs8023340 rs8024968 rs895828 rs895829 rs924312 rs924314 rs977588

Gene OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2 OCA2

Chr. 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15

Predicted phenotypes Eye color Hair color

Skin color ✓

✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

✓

✓

✓ ✓

✓

✓

✓

(continued)

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Table 1 (continued) SNP rs977589 rs1426654 rs2470102 rs4424881 rs3114908 rs8051733 rs12931267 179insC N29insA rs1110400 rs11547464 rs1805005 rs1805006 rs1805007 rs1805008 rs1805009 rs3212355 rs2228479 rs885479 Y152OCH rs9894429 rs1015362 rs2424984 rs4911414 rs6058017 rs6119471 rs2378249 rs6059655 rs7277820

Gene OCA2 SLC24A5 SLC24A5 APBA2 ANKRD11 DEF8 FANCA MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R MC1R NPLOC4 ASIP ASIP ASIP ASIP ASIP ASIP/PIGU RALY DSCR9

Chr. 15 15 15 15 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 16 17 20 20 20 20 20 20 20 21

Predicted phenotypes Eye color Hair color ✓ ✓ ✓ ✓

Skin color ✓ ✓ ✓ ✓

✓ ✓

✓ ✓ ✓ ✓

✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓ ✓

✓ ✓

✓ ✓ ✓

✓ ✓ ✓ ✓ ✓ ✓

✓

Variation in eye color is the result of differential deposition of melanin granules within a fixed number of stromal melanocytes in the iris (Imesch et al. 1997). This means that the number of melanocytes or the proportion of melanocytes in the iris stroma is not the major contributor to iris color, but rather the genetically determined density of melanin pigment granules in stromal melanocytes. Much of the initial knowledge on iris color variation has been derived from molecular genetic studies of rare pigmentation defects in humans and mouse and Drosophila model systems (Frudakis et al. 2003). Early studies of oculocutaneous albinism (OCA) in humans revealed that many pigmentation defects were due to lesions in the TYR gene (Oetting and King 1999; albinism database, http://www.ifpcs.org/albinism/). Subsequent studies of other TYR-positive OCA phenotypes revealed additional genes

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required for normal iris pigmentation; many of these genes are part of the TYR pigmentation pathway (Fig. 1) and include oculocutaneous albinism 2 (OCA2), tyrosinase-like protein 1 (TYRP1), MC1R, and adaptin 3B (AP3b) (Hamabe et al. 1991; Abbott et al. 1991; Robbins et al. 1993; Ooi et al. 1997). DNA-based eye (iris) color prediction was first attempted in 2007 in two separate studies (Frudakis et al. 2007; Sulem et al. 2007). Frudakis et al. used 33 SNPs in the OCA2 gene, and Sulem et al. used 9 SNPs in the MC1R, SLC24A4, TYR, KIT Ligand (KITLG), and oculocutaneous albinism 2-HECT and RLD domain containing E3 ubiquitin protein ligase 2 (OCA2-HERC2) genes, as well as the 6p25.3 region. In both studies, prediction of blue versus brown eye color was mainly dependent on variants of OCA2, and the other variants used by Sulem et al. added resolution to the discrimination between blue and green eye color. In 2008, three parallel studies reported the HERC2 gene, which regulates OCA2 expression, as the most important determinant of iris color (Sturm et al. 2008; Eiberg et al. 2008; Kayser et al. 2008). Sturm et al. found that a single SNP in the HERC2 gene, rs12913832, could predict eye color much better (R2 ¼ 0.68) than the previously reported OCA2 haplotype. Eiberg et al. also showed that the OCA2-HERC2 haplotype, including rs12913832, is strongly associated with blue eye color, and a specific OCA2-HERC2 haplotype was present in 97% of individuals with blue eyes. Kayser et al. proposed a model composed of three SNPs (HERC2 rs916977, OCA2 rs11855019, and OCA2 rs7495174), which provided a relatively high prediction accuracy expressed by an area under the receiver characteristic operating curve (AUC) value of about 0.8 for brown and blue eye color. At the molecular level, the conserved region around HERC2 rs12913832 represents a regulatory region controlling constitutive expression of OCA2, and the C allele at HERC2 rs12913832 leads to decreased expression of OCA2, ultimately resulting in blue eye color (Sturm et al. 2008). The first comprehensive DNA-based eye color prediction study investigated 37 SNPs in eight previously reported eye color–associated genes (HERC2, OCA2, SLC24A4, SLC45A2, TYR, interferon regulatory factor 4 (IRF4), TYRP1, and ASIP) in 6168 Dutch Europeans; 67.6% of individuals had blue eyes, 22.8% had brown eyes, and 9.6% had intermediate eye color (Liu et al. 2009). In this study, a prediction model based on multinomial logistic regression constructed using the data of 24 SNPs from 3804 individuals showed high prediction accuracy in the model validation set composed of 2364 individuals; this model demonstrated AUCs of 0.93 for brown eyes, 0.91 for blue eyes, and 0.73 for intermediate eyes. Based on these results, the same research group later proposed a robust and sensitive tool, termed IrisPlex, which includes a SNaPshot chemistry-based assay for multiplex genotyping of a minimal set of six SNPs (HERC2 rs12913832, OCA2 rs1800407, SLC24A4 rs12896399, SLC45A2 rs16891982, TYR rs1393350, and IRF4 rs12203592) and a prediction model implemented with an interactive website (https://hirisplex.erasmusmc.nl/). This system has been further improved by inclusion of 9466 individuals from various parts of Europe, and provides enhanced AUCs of 0.95 for brown eyes, 0.94 for blue eyes, and 0.74 for intermediate-colored eyes. However, because the majority of eye color prediction was achieved by HERC2 rs12913832, the IrisPlex model does not provide a prediction result for inputs

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lacking a genotype at this SNP site. Most incorrect predictions reportedly occur in individuals with heterozygous genotype at HERC2 rs12913832 (Kayser 2015). Similar results have also been reported in other studies (Valenzuela et al. 2010; Mengel-From et al. 2010). Valenzuela et al. investigated 75 SNPs of 24 genes that had been previously implicated in human and animal pigmentation studies using 789 European and non-European samples. They showed that a multiple linear regression model consisting of three SNPs, HERC2 rs12913832, SLC45A2 rs16891982, and SLC24A5 rs1426654 provided an R2 value of 74.8% for prediction of blue eyes solely based on HERC2 rs12913832, with the second and third SNPs SLC45A2 rs16891982 and SLC24A5 rs1426654 only marginally contributing to the increased R2 value of 76.4%. However, this cross-ethnicity study including Europeans and non-Europeans was unable to differentiate between true eye color and ancestral eye color effects, and the SNP SLC24A5 rs1426654 was found not to be directly involved in eye color at all (Kayser 2015). Mengel-From et al. investigated the predictive values of various combinations of SNP alleles in the HERC2, OCA2, and SLC45A2 genes in 395 Danes, and showed that four SNP-diplotype consisting of three HERC2 loci in strong linkage disequilibrium (LD) and one OCA2 locus (rs12913832, rs1129038, rs11636232, and rs1800407) provided a likelihood ratio of up to 29.3 for dark eye colors (brown and hazel) and up to 10.7 for light eye colors (light blue, gray, and green). In fact, prediction of intermediate eye colors has been the greatest limitation in eye color prediction, and the beneficial effect of including linked SNPs closely located to the lead SNP for intermediate eye color prediction has also been demonstrated in subsequent studies (Ruiz et al. 2013; Allwood and Harbison 2013). Ruiz et al. tested 37 SNPs located on pigmentation-associated genes in 416 individuals using two single base extension (SBE) assays to study SNP genotype-based prediction of skin, hair, and eye pigmentation (SHEP 1 & 2). They confirmed the presence of a genetic cluster corresponding to a complex and continuous range of intermediate eye color phenotypes that are distinct from blue and brown eye colors. The authors then reported improved intermediate (green-hazel) eye color prediction with an AUC value of 0.82 by employing Bayesian likelihood-based classification approaches, as well as a subset of 13 SNPs, including six IrisPlex SNPs and seven SNPs that are in LD with HERC2 or OCA2 SNPs of the IrisPlex (HERC2 rs1129038, rs11636232, rs7183877, and rs1667394 and OCA2 rs4778241, rs4778232, and rs8024968). In addition, the online classifier, i.e., Snipper, has the advantage of being able to submit incomplete SNP profiles that are commonly observed when analyzing challenging DNA samples. The difference in prediction results according to model algorithms used has also been demonstrated. In Yun et al. (2014), the prediction results obtained using the two different model algorithms, FROG-kb (http://forg.med.yale.edu/FrogKB) and Snipper (http://mathgene.usc.es/snipper), were compared with IrisPlex SNP genotype data of 905 individuals from European, genetically intermediate (Khanty, Uygur, and Yakut), and East Asian populations. The two model algorithms provided the same brown eye color predictions for East Asian populations but were inconsistent in the eye color prediction of almost 40% of the intermediate and European individuals. These results suggest that the difference in underlying logic and supporting data of the prediction model can lead to different predictive results.

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Recently, the interaction between SNPs, called epistasis, has begun to attract attention in the field of phenotype prediction, including pigmentation trait prediction. In epistasis, the expression of one gene is affected by the expression of one or more independently inherited genes. It has been reported that the interactions between HERC2 rs12913832, OCA2 rs1800407, and TYRP1 rs1408799 contributed to improving the prediction accuracy for green eyes (Pospiech et al. 2014). In addition, quantitative measurement of pigmentation traits is expected to improve future eye color prediction by reflecting detailed phenotype information. To compute quantitative measures of eye color, red, green, and blue (RGB) values can be taken from high-resolution digital images. In GWASs of eye and skin color in an African-European admixed population (Beleza et al. 2013), a principal curve was drawn to be fitted to the green versus blue scatter plot using RGB values taken from study samples and other reference samples, and the T-index, defined by the arc length of the data point from the end of the curve, which corresponds to the lightest eye color, was used for quantitative eye color measurement. Blue/green and brown eye colors roughly corresponded to the ranges of 0–0.15 and 0.15–0.4, respectively. Genotype and ancestry-based association analysis for eye color was carried out in 625 individuals; the result showed that the two major loci, HERC2 (OCA2) rs12913832 and SLC24A5 rs2470102, accounted for blue versus brown eye color and different shades of brown eye color. However, identification of additional genes and predictive SNPs will be needed to facilitate the quantitative approach to DNA-based eye color prediction.

Hair Color Prediction Melanocytes produce two different forms of melanin, i.e., eumelanin and pheomelanin (Fig. 1). People who produce eumelanin tend to have black or brown hair and dark skin, while those who produce pheomelanin tend to have red or blond hair and light-colored skin. In the biochemical pathways influencing melanin production, MC1R plays an important role controlling which type of melanin is produced by melanocytes. When MC1R is activated, it triggers a series of enzymatic reactions that lead to the production of eumelanin, but if inactivated or blocked, melanocytes make pheomelanin instead of eumelanin. Therefore, DNA-based hair color prediction was first attempted using 12 MC1R variants (Grimes et al. 2001), in which 98% of the individuals with homozygous genotype at one of these mutations, or with two different mutations, were red-haired. In 2007, the entire MC1R gene was sequenced in 184 individuals divided into two groups, i.e., red-haired donors and non-red-haired controls (Branicki et al. 2007), and five MC1R SNPs (rs11547474, rs1805006, rs1805007, rs1805008, and rs1805009) were indicated for prediction of red hair color. The prediction for all categorical hair colors was first reported as a part of a GWAS for pigmentation (Sulem et al. 2007). In this report, two MC1R SNPs (rs1805007 and rs1805008) were used to predict red-haired individuals with an accuracy of 70%, and SNPs from six additional genes (interferon regulatory factor 4-exocyst complex component

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2 (IRF4-EXOC2), TYR, KITLG, SLC24A4, HERC2, and OCA2) were used to predict other hairs colors with much less accuracy. In 2011, Branicki et al. suggested a model composed of 22 SNPs in 11 genes that can predict red, black, brown, and blond hair colors with AUC values of 0.93, 0.87, 0.82, and 0.81, respectively. Later, these SNPs and the prediction model were included in the HIrisPlex system, which enables categorical eye and hair color prediction. The HIrisPlex system is composed of a multiplex assay for 24 genetic variants and two models to predict eye and hair color (Walsh et al. 2013). Among 23 SNPs and one in del marker (Y152OCH, N29insA, rs1805006, rs11547464, rs1805007, rs1805008, rs1805009, rs1805005, rs2228479, rs1110400, rs885479, rs28777, rs16891982, rs2402130, rs12896399, rs1042602, rs1393350, rs12821256, rs4959270, rs12203592, rs1800407, rs12913832, rs2378249, and rs683), 11 were from the MC1R gene, two each from the SLC45A2, SLC24A4, and TYR genes, and one each from KITLG, EXOC2, IRF4, OCA2, HERC2, abstract agouti signaling protein/phosphatidylinositol glycan anchor biosynthesis class U (ASIP/PIGU), and TYRP1. During the construction of HrisPlex model, six SNPs from the IrisPlex were used for eye color prediction, and 22 SNPs, excluding TYR rs1393350 and SLC24A4 rs12896399 of the IrisPlex, were used for hair color prediction. This model was built based on 1243 training individuals, and produced accurate predictions in 87.5% of black, 80% of red, 78.5% of brown, and 69.5% of blond haired individuals from the 308 individual test set (Walsh et al. 2013). Further updates were integrated later, including the enlarged HIrisPlex genotype database (Walsh et al. 2014); the latest version of the HIrisPlex model based on 1878 individuals’ genotype data provides an AUC value of 0.92 for red, 0.83 for black, 0.80 for blond, and 0.72 for brown hair color. In addition, by utilizing the highest p-value approach combined with a stepwise model, this enhanced model provides additional shade prediction, i.e., light or dark, thereby, enabling hair color prediction with more detailed categorization, such as blond/dark blond, dark blond/brown, brown/dark brown, and dark brown/black. This model is available online (https://hirisplex.erasmusmc.nl/) and provides predictions for individual eye and hair color using complete and partial HIrisPlex profiles (Walsh et al. 2014). However, inputs missing all 11 MC1R SNPs and inputs missing a combination of HERC2-SLC45A2-IRF4 do not provide hair color prediction results due to a large decrease in the AUC value. Recently, Söchtig et al. tested 63 pigmentation-associated SNPs, including 24 hair color markers from HIrisPlex, in 605 European individuals using three SBE-based multiplex assays (SHEP 1, 2&4), and suggested 12 SNPs located in six genes strongly associated with hair color variation. They included rs28777 of the SLC45A2 gene, rs35264875 of the two pore segment channel 2 (TPCN2) gene, rs1129038 and rs12913832 of the HERC2 gene, rs4778138 and rs7495174 of the OCA2 gene, rs12931267 of the FA complementation group A (FANCA) gene, and rs11547464, rs1805006, rs1805007, rs1805008, and rs1805009 of the MC1R gene (Söchtig et al. 2015), eight of which are also present in the HIrisPlex system. By using Bayesian likelihood-based classification approaches, Snipper, the model based on these 12 SNPs, provides prediction accuracy similar to HIrisPlex model, with an

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AUC value of 0.94 for red, 0.84 for black, 0.86 for blond, and 0.64 for brown hair color. Overall, most inaccurate predictions have been observed in blond- and brownhaired individuals, which may be because current models were constructed using the genotype data from individuals including those who had been blond in childhood but have undergone hair color change with age. There is, however, another age-dependent hair color change phenomenon, i.e., hair graying or hair whitening, which has not been considered in the current model building process. Future research on the mechanism underlying hair color change with age will contribute to the improvement of hair color prediction models (Kayser 2015).

Skin Color Prediction Even in the time of the ancient Greeks, significant differences in human skin color across continents attracted attention, and many explanations and hypotheses have been proposed (Stoneking 2017). Skin color variations has long been thought to be the result of natural selection and highly correlated with UV exposure level by many researchers. However, that raised further questions, such as why was dark skin color preferred in areas with high UV exposure, and light skin color in areas with low UV exposure (Stoneking 2017). A hypothetical scenario is that dark skin pigmentation had occurred shortly after the loss of body hair, as in all other apes, the skin under the hair is light in color, and the skin in the parts of the body without hair is dark (Stoneking 2017). Such hair loss would have caused changes in skin properties of our ancestors. In fact, dark skin is highly resistant to parasites, and has low susceptibility to skin cancer when exposed to high UV light. However, the most plausible explanation for selective advantage for dark skin color is related to vitamin B9, also known as folic acid or folate (Branda and Eaton 1978; Jones et al. 2018a). Folate in skin and blood can be degraded by exposure to UV light, and folate deficiency can cause neural tube defects, one of the most common birth defects. On the other hand, melanin protects folate from UV radiation–induced degradation, and people with dark skin color have lower risk of developing neural tube defects. Another hypothesis relates lighter skin color farther from the equator to vitamin D; although vitamin D can be synthesized in our body upon exposure to sunlight, prolonged vitamin D deficiency can cause rickets, which manifest the softening and weakening of bones in childhood. Interestingly, larger amounts of melanin in darker skin inhibit vitamin D synthesis from sunlight, which is not typically an issue for people who live in sunny climes. However, in northern climes with less sunlight, there might have been selection for lighter skin pigmentation to facilitates vitamin D synthesis (Stoneking 2017), a popular evolutionary model for skin pigmentation referred to as the vitamin D–folate hypothesis (Jones et al. 2018a). Recently, new evidence in support of this hypothesis has been reported, including correlation between UV exposure and genetic variants in vitamin D and folate metabolism genes. For example, the incidence of the methylene tetrahydrofolate reductase

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(MTHFR)-C677T variant, which is closely related to aberrant folate metabolism, is lowest in populations with dark skin color (Jones et al. 2018b). However, there is much less genetic knowledge of skin color variation compared to eye and hair color variations. Since most skin color variation is expressed between populations on different continents, cross-ethnicity studies tend to fail to differentiate between true pigmentation effects and ancestry effects (Kayser 2015). Therefore, few GWASs have been reported that identify markers associated with skin color variations. In a previous study using both Europeans and non-European samples (Valenzuela et al. 2010), three pigmentation candidate SNPs (HERC2 rs12913832, SLC45A2 rs16891982, and SLC24A5rs1426654) could explain 45.7% of skin color variations, but as previously mentioned, it appeared that the effect of SLC24A5 rs1426654 was likely a European ancestry effect. In 2014, the first comprehensive skin color prediction study was reported using the genotype data of 59 pigmentation-associated SNPs from 285 Europeans and non-European individuals, including Americans, Africans, Eurasians, and Asians (Maronas et al. 2014). SNP scoring was performed using three SBE-based multiplex assays (modified SHEP 1, 2, and 4), and 10 SNPs in eight genes were finally selected for skin color prediction with an AUC value of 0.99 for white skin, 0.97 for black skin, and 0.80 for intermediate skin color in a test set of 118 samples. The ten SNPs include rs10777129 in the KITLG gene, rs13289 and rs16891982 in the SLC45A2 gene, rs1408799 in the TYRP1 gene, rs1426654 in the SLC24A5 gene, rs1448484 in the OCA2 gene, rs2402130 in the SLC24A4 gene, rs3829241 in the TPCN2 gene, and rs6058017 and rs6119471 in the ASIP gene. In 2017, a global skin color prediction study was published investigating 77 pigmentation-associated SNPs in 2025 individuals from 31 populations (Walsh et al. 2017). A total of 36 SNPs were selected and used to develop a statistical prediction model. Those 36 SNPs from 16 genes are as follows: SLC24A5 rs1426654, IRF4 rs12203592, MC1R rs1805007, rs1805008, rs11547464, rs885479, rs2228479, rs1805006, rs1110400, rs3212355, OCA2 rs1800414, rs1800407, rs12441727, rs1470608, rs1545397,SLC45A2 rs16891982, rs28777, HERC2 rs1667394, rs2238289, rs1129038, rs12913832, rs6497292, TYR rs1042602, rs1126809, rs1393350, RALY heterogeneous nuclear ribonucleoprotein (RALY) rs6059655, differentially expressed in FDCP 8 homolog (DEF8) rs8051733, PIGU rs2378249, ASIP rs6119471, SLC24A4 rs2402130, rs17128291, rs12896399, TYRP1 rs683, KITLG rs12821256, ankyrin repeat domain 11 (ANKRD11) rs3114908, and basonuclin 2(BNC2) rs10756819. An assay to genotype 41 SNPs, including these 36 skin SNPs and three statistical models to predict eye, hair, and skin color, were implemented as HIrisPlex-S, which provides high prediction accuracies with an AUC of 0.97 for light, 0.83 for dark, and 0.96 for dark black skin color. When using a five-category classification, AUC values of 0.74 were obtained for very pale, 0.72 for pale, 0.73 for intermediate, 0.87 for dark, and 0.97 for dark black skin color. This system enables intra- and intercontinental skin color prediction, and is currently available online along with the IrisPlex and HIrisPlex systems (https:// hirisplex.erasmusmc.nl/). Quantitative measurement of skin color may be able to improve DNA-based skin color prediction. In Beleza et al. (2013), the melanin index, which indicates melanin density estimated from melanin reflectance at 650 nm using spectrophotometry was

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used in a GWAS for skin color. Because the study population was African-European admixed, individual genomic ancestry was estimated based on SNP information, and genotype- and ancestry-based association analyses for skin color were carried out in 685 individuals. The four major loci, including SLC24A5 rs1426654, glutamate metabotropic receptor 5/tyrosinase (GRM5/TYR) rs10831496, amyloid beta precursor protein binding family A member 2 (APBA2) rs4424881, and SLC45A2 rs35395 were identified to explain 35% of total skin color variation. However, surprisingly, the genetic component with the largest effect was genetic ancestry, which accounted for 44% of skin color variation, and together with the four major loci accounted for 57% of total variance. Currently available DNA test systems that have been developed and validated along with suitable statistical models for prediction of eye, hair, and skin color are shown in Table 2.

Pigmentation Prediction in Archaeological and Forensic Samples DNA-based prediction of phenotypic traits such as eye, hair, and skin color was developed primarily to assist in crime scene investigations, narrowing the search for unknown suspects. However, this method is also useful to reconstruct the genetic legacy of historical individuals, as well as ancient humans. Although DNA extracted from archaeological and forensic samples is generally highly degraded, using short PCR fragments can improve genotyping success. In this regard, SNPs have been suggested as ideal candidate markers for analyzing degraded DNA due to their potential to be amplified in PCR products as small as 50 bp. Development of SBE-based multiplex assays, which combine multiplex PCR with SBE reactions, also facilitated retrieval of reliable data from multiple SNP loci. This method theoretically allows simultaneous amplification of up to 52 SNPs, and provides high sensitivity down to about 30 pg of input DNA (Fortes et al. 2013; Kayser 2015). In addition, advances in MPS technology and DNA recovery significantly increase the possibility of extracting genetic information from degraded DNA samples as partial or complete genome sequences. Moreover, development of targeted MPS methods based on PCR amplification or hybridization capture approaches that target a predefined set of phenotype-associated SNP loci can facilitate the acquisition of phenotypic information from highly degraded DNA. Therefore, many studies have already reported SNP-based prediction of pigmentation traits in archaeological and forensic cases.

Prediction of Pigmentation Traits in Historical Figures and Forensic Identification Cases Although predicting pigmentation traits from DNA at a crime scene can provide investigative leads to track unknown suspects, it can also be useful in mass disaster victim and missing person identification cases, especially when there is no reference

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Table 2 Currently available forensic DNA test systems for prediction of eye, hair, and skin color. (Redrawn with modification from Schneider et al.(2019)) Test system IrisPlex

Feature Eye color

Markers 6 SNPs

Technology Multiplex SBE

SHEP 1, 2

Eye color

13 SNPs

Multiplex SBE

HIrisPlex

Eye and hair color

24 SNPs

Multiplex SBE

SHEP 1, 2, 4

Hair color

12 SNPs

Multiplex SBE

SHEP 1, 2, 4 modified

Skin color

10 SNPs

Multiplex SBE

HIrisPlex-S

Eye, hair, and skin color

41 SNPs

Multiplex SBE and targeted MPS

Statistical model Multinomial logistic regression (N ¼ 9466) freely available at https:// hirisplex. erasmusmc.nl/ Snipper (N ¼ 416) freely available at http:// mathgene.usc. es/snipper/ Multinomial logistic regression (N ¼ 1878) freely available at https:// hirisplex. erasmusmc.nl/ Snipper (N ¼ 605) freely available at http:// mathgene.usc. es/snipper/ Snipper (N ¼ 118) freely available at http:// mathgene.usc. es/snipper/ Multinomial logistic regression (N ¼ 1423) freely available at https:// hirisplex. erasmusmc.nl/

Test accuracy AUC: blue 0.94, brown 0.95, intermediate 0.74

AUC: blue 0.999, brown 0.99, greenhazel 0.82

AUC (hair color, 22 markers): red 0.92, black 0.83, blond 0.80, brown 0.72 AUC (eye color, 6 markers): refer to IrisPlex AUC: red 0.94, blond 0.86, black 0.84, brown 0.64

AUC: white 0.99, black 0.97, intermediate 0.80

AUC (skin color, 36 markers): very light 0.74, light 0.72, intermediate 0.73, dark 0.88, dark to black 0.96. AUC (hair color): refer to HIrisPlex AUC (eye color): refer to IrisPlex (continued)

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Table 2 (continued) Test system ForenSeq™ DNA Signature Prep Kit (Verogen, USA)

Feature Eye and hair color

Markers 24 SNPs

Technology Targeted MPS

Statistical model Unknown Presumably initial versions of the IrisPlex and HIrisPlex models available via commercial ForenSeq™ Universal Analysis software (Verogen)

Test accuracy Initial version of IrisPlex model for eye color, AUC: blue 0.91, brown 0.93, intermediate 0.73 Initial version of HIrisPlex model for hair color, AUC: red 0.9, black 0.78, blond 0.75, brown 0.72

SNPs, single-nucleotide polymorphisms; targeted MPS, targeted massively parallel sequencing; SBE, single base extension

DNA profile for comparison. In addition, because pigmentation traits are not reflected in skeletal features, DNA-based prediction of pigmentation traits can be very helpful in reconstructing the appearance of famous historical figures using their remains. Many studies have reported pigmentation trait prediction results from human skeletal remains using sequencing, multiplex SBE, and targeted MPS methods. The first SNP-based pigmentation trait prediction applied to the identification of a famous historical individual was the prediction of the eye color of the Polish astronomer Nicolaus Copernicus (1473–1543) using monoplex PCR and sequencing (Bogdanowicz et al. 2009). His identification was based on the mitochondrial DNA result match between the putative skeletal remains and hair discovered in his calendar. To predict his appearance, the HERC2 rs12913832 allele was analyzed, and the resultant homozygous CC genotype implied that Corpernicus had blue eyes. Prediction of eye and hair color using the multiplex SBE-based HIrisPlex system was first performed on 26 teeth or bone samples including fourteenth-century Polish and modern human remains (Draus-Barini et al. 2013). Since in almost all samples analyzed the eye and hair color of the deceased individuals were unknown, actual predication accuracy could not be obtained from this study. However, accuracy estimates for the HIrisPlex system from previous reports were high (Walsh et al. 2013, 2014), and therefore, the prediction results were presumed to be accurate. In this study, a tooth from the Polish general Władysław Sikorski (1881–1943) was analyzed, and the HIrisPlex model predicted that he had blue eyes and blond hair, which was verified by reliable historical scripts. This same multiplex SBE-based HIrisPlex system was used to predict eye and hair color of 49 World War II victims excavated in Slovenia (Chaitanya et al. 2017).

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HIrisPlexSNP analysis has been also used along with a targeted MPS approach based on the hybridization capture method to predict the eye and hair color of King Richard III (King et al. 2014). For the identification of the remains, MPS libraries were built, and sequencing targets, i.e., mitochondrial DNA, Y-chromosome SNPs, and HIrisPlex SNPs, were enriched using the hybridization capture method. However, the on-array hybridization capture approach yielded insufficient coverage of HIrisPlex SNPs and Y-chromosome SNPs; therefore, MPS was carried out for multiplex PCR products containing HIrisPlex SNPs and Y-chromosome SNPs. The remains were identified as King Richard III by a perfect mitochondrial DNA match between the sequence of the remains and one living relative. The HIrisPlex SNP analysis results indicated that the king had blue eyes and blond hairs in childhood. In 2020, eye, hair, and skin color predictions were reported for 63 bone samples using PCR-based targeted MPS technology (Kukla-Bartoszek et al. 2020). In this study, 41 HIrisPlex-S SNPs were successfully analyzed in 35 samples, and complete genotypes were obtained using as little as 50 pg of input DNA. With this HIrisPlx-S system, separate models for eye, hair, and skin color prediction are available, and therefore, these three pigmentation traits could be predicted independently even in samples with partial profiles. The results of the pigmentation predictions showed typical European phenotypes, because the samples were all from regular forensic identification cases or had been excavated from cemeteries and a detention center for war victims in Poland. For example, eye color was predictable in 57 individuals (91%), of which 34 (54%) had blue eyes, 15 (24%) had brown eyes, and 8(13%) had inconclusive results. For the 16 bone samples identified through the STR profiles, the prediction results could be compared with antemortem data obtained from the families: eye color prediction results were consistent in 60% of samples, hair color in 86% of samples, and skin color in 75% of samples.

Prediction of Pigmentation Traits in Ancient Samples Analysis of DNA of ancient humans can provide insights into the timing and origin of selection events involving pigmentation alleles (Stoneking 2017); studies of Neanderthal and Denisovan genomes would provide further insights into this issue. Therefore, a fragment of the MC1R gene was amplified and sequenced in two Neanderthal remains to predict pigmentation traits (Lalueza-Fox et al. 2007). Analysis of the MC1R gene sequence was expected to provide the information about the appearance of Neanderthals, as MC1R variants that have reduced function are well known to be associated with red hair and pale skin color in modern humans. However, the two Neanderthal remains showed a nonsynonymous mutation that is not found in modern humans. Functional analysis suggested that this variant also reduced MC1R activity, so that Neanderthals likely had red hair and light skin color. It is interesting that different genetic variations in the MC1R gene commonly cause red hair and light skin color in modern humans and Neanderthals. This suggests that inactive MC1R variants may have evolved independently in both modern humans and Neanderthals (Lalueza-Fox et al. 2007).

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Advances in technology following the introduction of MPS have greatly improved ancient DNA analysis. By applying MPS technology to ancient human remains, whole genome sequencing (WGS) data from Neanderthals, Denisovans, and modern humans have been established, and can be explored to predict pigmentation (Green et al. 2010; Reich et al. 2010; Cerqueira et al. 2012). However, earlier ancient DNA WGS datasets often had the disadvantage of too low coverage for use in predicting traits based on certain SNP genotypes. In 2012, the allele states of 124 pigmentation-associated SNPs in 33 genic and seven intergenic regions were analyzed in the Denisovan genome at 1.9X coverage to predict pigmentation phenotypes (Cerqueira et al. 2012). The results showed that the Denisovan individual tested carried alleles associated with the dark skin, brown hair, and brown eyes of modern humans. Shortly after this study, high coverage genome sequence from the same Denisovan individual was generated using a single-strand library preparation method (Meyer et al. 2012). With this new approach, the high coverage Denisovan genome could be obtained at 30X coverage, and it was possible to determine the alleles for 11 SNPs that were previously undetermined, identifying three SNPs whose genotypes were different from the states observed in the 1.9X coverage genome (Meyer et al. 2012). The predicted pigmentation phenotypes were the same as reported in Cerqueira et al. (2012), but the possibility still remains that this Denisovan individual might have had other pigmentation-associated SNPs that do not exist in modern humans. In 2020, WGS data obtained from ancient and contemporary Native Americans were analyzed to predict pigmentation phenotypes using 61 SNPs from the HIrisPlex-S and Snipper SNP sets (Carratto et al. 2020). This study used low-tomedium coverage (0.5–20X) WGS data from 20 Late Pleistocene/Early Holocene ancient Native American samples and high coverage (>35X) WGS data from 22 contemporary Native Americans. To ensure reliable and accurate results, sequencing reads with a quality score lower than 30 were excluded, and predictions were reported only when 50% or more of the alleles were determined. Pigmentation traits were predictable in 29 samples, including 7 of 20 ancient samples and all 22 contemporary Amerindian samples. The seven ancient Native Americans were predicted to have black hair, intermediate/brown eyes, and intermediate/dark skin color. In all samples analyzed, the hair color prediction results, i.e., black hair, were consistent between HIrisPlex-S and Snipper. However, the prediction results for eye and skin color were not consistent in several contemporary Amerindian samples. HIrisPlex-S predicted that every individual would have brown eyes, but Snipper predicted that 12 contemporary individuals would have brown eyes, seven green/hazel eyes, and one blue eyes. HIrisPlex-S also predicted dark skin with dark black influence or dark black skin with dark influence for all individuals, but Snipper predicted intermediate skin color for all individuals except one, whose skin color was predicted to be white. Although the real phenotype of each sample was not known, the prediction results were consistent with the average phenotypes expected from the geographic region where the samples were originated (Carratto et al. 2020). However, Snipper was estimated to be more reliable than HIrisPlex-S in eye color prediction as IrisPlex has always had

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difficulty in predicting intermediate green/hazel eye color, and Snipper predicts blue color eyes when it fails to predict intermediate eye color. Therefore, the eight samples that showed inconsistent eye color prediction results may actually have had intermediate eye color. Skin color prediction results were not consistent between the two models, but this is mainly due to the different classification schemes used by the models, as dark skin defined by HIrisPlex-S classification looks like intermediate skin defined by Snipper classification (Carratto et al. 2020). These results demonstrate that the use of different classification schemes, different sets of markers, and different training and validation sets in model development can produce different prediction results. In addition, as an individual’s ancestry can account for much of the phenotypic inheritance, it is necessary to identify markers unique to regions outside Europe, and to train and validate models in more diverse population groups.

Perspectives on SNP-Based Phenotype Prediction in Mummy Studies SNP-based phenotype prediction has never been attempted in mummies, perhaps because mummified human bodies can retain well their antemortem physical characteristics, such as face structure, body height, hair morphology, etc. Besides hair color, the pigmentation traits, however, cannot be easily inferred from general anthropological analysis of the mummified individual, due to the possibility of changes in eye and skin color during mummification and thereafter. For example, ancient Egyptian mummies used during the mummification process a salt-like substance called natron, and additionally plant resins, which are known to darken the skin color. It is also well known that the eyes after death become dulled, and the cornea is readily deformed. In Egyptian mummies, eye color information might be obtained from rough painting of the iris on the linen pads over the eyelids, or from artificial eyes made of small black and white stones (Sandison 1957), but in most cases, it is not easy to obtain exact information about the eye color of mummified individuals. Therefore, if DNA is available, SNP-based prediction of eye, skin, and hair color may be helpful in obtaining information to reconstruct the detailed appearance of mummified individuals. When predicting pigmentation traits, it is also important to determine appropriate markers and predictive models. However, since most of the pigmentation-associated markers currently available have been identified from Europeans, and the models have been trained and validated using European samples, their prediction accuracy in other population groups may not be as accurate. For examples, Melanesian blond hair is known to be caused by an amino acid change of R93C in the TYRP1 gene product (Kenny et al. 2012), but the causal mutation is not involved in European blond hair, and is not included in current hair color prediction models. Thus, for better DNA-based phenotype prediction, ancestry analysis may be required prior to predicting pigmentation traits.

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Conclusion Advances in GWAS technology have brought significant progress to the understanding of pigmentation traits, such as eye, hair, and skin color, by facilitating the identification of associated genes and SNP markers. Accordingly, pigmentation traits have become predictable using the genotypes of a set of associated SNP markers and statistical predictive models. Since pigmentation trait information can be used to reconstruct the appearance of sample donors, it has begun to be used to obtain investigative leads for unknown suspects in forensic cases. Therefore, several forensic DNA test systems have been developed, which provide relatively high prediction accuracies for eye, hair, and skin color. These systems were designed using highly sensitive multiplex SBE assays or targeted MPS assays, which also facilitate the acquisition of phenotypic information from challenging samples such as ancient DNA. Therefore, SNP-based pigmentation trait prediction has been successfully applied to ancient human skeletal remains as well as to whole genome sequence data of archaic humans, e.g., Neanderthals and Denisovans. Prediction results were mostly consistent with the phenotypes expected from the geographic region where the samples had originated. However, as most of the predictive markers and models currently available are based on data obtained from Europeans, their predictive power may not be as high as expected in other populations. Therefore, for better prediction, it is necessary to identify and validate more markers from various populations. It is also notable that the two Neanderthal individuals tested were predicted to have had red hair and light skin color due to a nonsynonymous mutation in the MC1R gene. This mutation has not been found in modern humans, but through functional analysis it was found to be related to reduced MC1R activity. Therefore, when interpreting archaic human data, it should be noted that there may exist functional mutations not found in modern humans.

Cross-References ▶ Analysis of Low Copy Number DNA and Degraded DNA

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Part IV Archaeoparasitology

Mummies, Parasites, and Pathoecology in the Ancient Americas

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Contents Introduction: Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Avoiding Destructive Autopsy in Mummy Parasitology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Laboratory Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Methods of Choice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parasites Found in Mummies: Parasite Life Cycles and Nidi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Adenocephalus pacificus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Hookworms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Whipworms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ascaris . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pinworms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Echinostoma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Fleas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Trypanosomes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pathoecology in Mummy Populations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chinchorro Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chinchorro and Chagas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lessons of Pathoecology for the Chinchorro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chiribaya of Southern Perú . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Coastal Site – San Gerónimo . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Farming Village – Chiribaya Baja . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Administrative and Economic Center – Chiribaya Alta . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . El Yaral – Herding Village . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chiribaya Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ectoparasites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

412 413 413 414 414 414 416 417 417 418 418 419 419 420 420 421 421 422 423 423 424 424 425 425 426 426

A. Araújo Departamento de Endemias Samuel Pessoa, Escola Nacional de Saúde Pública, Oswaldo Cruz Foundation, Rio de Janiero, Brazil K. J. Reinhard (*) School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_13

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Intestinal Helminths . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chagas Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Summary of Chiribaya Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Isolated Mummy Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lapa do Boquete, Peruac¸u, Minas Gerais, Brazil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chagas Disease in the Texas-Coahuila Border . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Prehistoric Human Migrations – Mummies and the Big Picture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Serial Nature of Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummy Studies and The Stockholm Paradigm: Understanding How Parasites Colonize New Hosts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Mummy studies can make important contributions to the field of parasitology. Several theoretical approaches emerged in the field relevant to archeoparasitology and especially mummy studies. Three of these areas are summarized below. The ecological approach to reconstruction of infection patterns is pathoecology. A concept key to understanding prehistoric infection occurrence is nidality. A new evolutionary perspective has been defined as the Stockholm Paradigm. In this review, we propose an approach to mummy studies that provides outlooks relevant to current and future trends in parasitology.

Keywords

Parasitology · Pathoecology · Nidality · Methods · Stockholm paradigm

Introduction: Pathoecology The term “pathoecology” was coined by KR Reinhard in 1974 (Reinhard 1974a, b). Reinhard was an epidemiologist who worked in several areas of Native American public health. He applied pathoecology to the integrative study of historic climate and health records to define the impact of climate variability on health patterns before modernization of First Nations. Because statistically rigorous epidemiological data were not available for early historic periods, KR Reinhard (1974a) suggested the term “retrospective pathoecology” for application to reconstructing the ecology of past disease. KJ Reinhard began applying pathoecology to prehistory some 20 years after KR Reinhard coined the term. He began with the establishment of a link between the emergence of parasitic disease and Ancestral Pueblo cultural development (Reinhard 1988, 1990, 1992; Reinhard et al. 1987). Related to this was analysis of mummified remains (Reinhard and Hevly 1991). Later, this approach was applied to the bioarcheological problem of porotic hyperostosis (Reinhard 1992). Reinhard later shifted his research to Andean mummies with Jane Buikstra.

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Concurrently with the work presented above, archeological parasite research was established in Brazil, including analyses of mummies (Ferreira et al. 1979, 1980, 1983; Araújo et al. 1981; Novo and Ferreira 2016; Araujo and Reinhard 2015). A research consortium was established between Araújo-Ferreira laboratory and that of Reinhard. This collaboration was formalized in the mid-eighties (Reinhard et al. 1986) and continues up until this writing. One focus of investigations was on defining the distribution of parasites in the Americas (Araujo et al. 2013, 2015; Reinhard and Pucu 2014; Reinhard et al. 2016), with collaboration with colleagues in Peru and Chile to produce a series of papers (for review, see Araújo et al. 2011). The pathoecological approach was introduced into the literature by a collection of papers relating to Andean mummy collections (Martinson et al. 2003; Reinhard and Buikstra 2003; Reinhard and Urban 2003; Arriaza et al. 2010, 2013a). The approach was also applied to isolated mummies from other parts of the Americas (Reinhard and Araujo 2015, 2016).

Methods Avoiding Destructive Autopsy in Mummy Parasitology Mummies are a limited resource. All types of parasitological analysis can be conducted without extensive destruction of mummies. Tissue samples can be extracted from mummies using endoscopic techniques or through very small incisions made at points strategically selected to minimize the effect on mummy integrity. Thus, there is never a parasitological justification for extreme autopsy of mummies (Kiple et al. 2001). Indeed, beginning with the 1998 Mummy Congress in Arica, Chile, a new emphasis on mummy conservation dominates modern mummy studies. Until 20 years ago, there were justifications for destructive autopsy of mummies. Then, hundreds of mummies in the Andes were discovered every year and the museums were full of preserved bodies. Since 1990, workers in South American countries, especially Perú, Chile, Argentina, and Brazil, have spent considerable amounts of money building new mummy repositories specifically for the preservation of mummies. Therefore, there is no longer a justification based on the limited resources for preservation. Also until 20 years ago, some researchers justified destructive mummy autopsy based on the rationale that extant radiographic techniques could not produce images comparable to examining directly the gross pathology in mummies. Since that time, many of us have focused our attention specifically on the refinement of radiographic CT-scan methods for three-dimensional reconstruction of pathology. From the perspective of parasitology, we urge each mummy specialist to commence parasitological analysis with three-dimensional analysis of mummies using CT technology as recommended in this volume, ▶ Chap. 22, “Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice.” Using the images, endoscopic strategies for sampling tissue specific to each mummy

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can be devised. Also, gross pathology associated with infection can be visualized at this time. It is never advisable to autopsy a mummy. Preliminary planning of analysis focused on conservation and preservation should be employed in parasitological analysis. Effective, minimally invasive, endoscopic sampling can produce a great deal of parasitological data from any given mummy.

Laboratory Analysis The methods applied in the analysis of digestive organs and coprolites are reviewed by Camacho (Camacho et al. 2018). Methods for analysis of other internal structures are presented in case studies (Searcey et al. 2013; Kumm et al. 2010) and also reviews in this volume (▶ Chaps. 21, “Archaeoparasitology of Korean Mummies” and ▶ 26, “Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent”). Methods for processing and quantifying parasite eggs are identical to other microfossils. These methods are presented in this volume in the chapter ▶ Chap. 22, “Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice.”

Methods of Choice Three issues must be considered when conducting parasitological analysis of mummies: identification of parasites species, quantification of infection, and assessing if any disease was caused by the infection. For the latter consideration, visualizing disease evidenced by gross pathology is possible in some cases. Also, the number of parasites that infected the individual in life can be used to state whether disease was experienced. The value of quantification has been reviewed (Reinhard 2017; Camacho et al. 2017). These authors emphasized that infection and disease are two different things. Most parasite infections do not provoke disease. Parasite burden is a factor that affects pathology. Higher numbers of eggs signal pathology in mummy studies. Therefore, quantification is essential in estimating whether or not a prehistoric individual experienced disease. This is illustrated in Table 1.

Parasites Found in Mummies: Parasite Life Cycles and Nidi Each parasite has a distinct life cycle. Thus, when a parasite species is identified in a mummy, one knows immediately details of the ecology of that parasite’s transmission. These include the presence of an intermediate host, reservoir host, aspects of human behavior, and even details of the climate in which the parasite circulated.

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Table 1 Whipworm egg per gram counts from various sites. The egg per gram (epg) data are converted to worms per gram dividing epg by the average output of a female whipworm of 14,000 eggs per day (John and Petri 2006). A gram of coprolite is a small fraction of daily fecal output of 140–150 g for adult humans (Saito et al. 1991). Therefore, these data represent for the mummies from Nivelles and Piraino represent up to 375 and 555 female worms parasitizing the individuals at time of death. Depending on the numbers of male worms, the actual worm burden could have been twice as high. These levels of infection certainly caused symptoms. However, the low epg counts for the remaining cases were likely asymptomatic. It is noteworthy that New World infections represent very low worm burdens. This indicates that prehistoric infection was controlled by First Nation peoples as reviewed by Reinhard and Pucu (2014). This control is an important element of prehistoric American pathoecology Site Nivelles Interment, Belgium Piraino Mummy, Sicily Inca, Arica, Chile Chiribaya, Arica, Chile Vilnius Mummy, Lithuania Chiribaya Mummy, Ilo, Peru Chiribaya Mummy, Ilo, Peru Zape, Mexico Zweeloo, Netherlands

Epg 51,630 34,529 5,400 1,800 4,779 2,240 435 1,127 Traces

Worm(s) per gram 3.7 2.5 0.4 0.1 0.3 0.2 0.03 0.1

References Rácz et al. (2015) Kumm et al. (2010) Santoro et al. (2003) Morrow et al. (2014) Martinson et al. (2003) Jiménez et al. (2012) Searcey et al. (2013)

When several parasites are found in a mummy or a mummy population, many details concerning disease transmission can be inferred (Martinson et al. 2003). Each parasite species provides different information regarding pathoecology. Because mummies preserve information about endoparasitism and ectoparasitism and often demonstrate pathology of disease resulting from infection, mummies are the best resource for pathoecological study (Reinhard 1998). When combined with dietary data (White et al. 2006), a complete reconstruction of the interaction of parasitism and diet patterns can be achieved. Indeed, the methods of parasitological analysis are part of intestinal residue analysis as presented in ▶ Chap. 22, “Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice” (this volume). The association of parasites and host shows modes of infection and is the foundation for pathoecology. Another element of the concept is the locale of infection. For application to the pathoecology of infectious diseases, the nidality theory of Pavlovskiĭ (1966) is particularly useful for this purpose (see Slepchenko and Reinhard (2018) and Herbold (2005) for reviews). This theory holds that most vectors, hosts, and pathogens are united in the landscape by environmental determinants. These determinants control their distribution and abundance. The term “nidality” is derived from the word nidus (Pavlovskiĭ 1966). A nidus (plural nidi) is a breeding place, where pathogens or infectious agents are situated and reproduce. In this concept, the entire ecosystem defines the micro-scale transmission of pathogens in finite nidi. As reviewed by Herbold (2005), a disease agent occupies a specific ecologic niche. Within - niche variation and change will alter the

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established host–infectious agent–environment relationships. Typically, nidality is applied to the development of disease prevention strategies in the modern world. For those of us who work with remains from the ancient world, nidality is a particularly useful concept to define how and where people became infected with parasites (Reinhard and Bryant 2008). To apply nidality mummy studies, we must understand the life cycles of parasites discovered in the Americas. More general information about these parasites can be found in Roberts et al. (2004) and Rey (2001). A diversity of parasites can be found in mummies. Some of those found in American mummies are presented here.

Adenocephalus pacificus Adenocephalus pacificus (formerly Diphyllobothrium pacificum) eggs were recorded in South American mummies of the Pacific coast. This tapeworm (Order Pseudophyllidea) is a parasite of pinniped mammals including sea lions (Ottaria). Infection occurs when infected fish are ingested. The nidus of infection occurs where humans and pinnepeds share the same source of fish. Adults develop in mammal intestine reaching near 10 m of length. Ovoid eggs measuring 50–60 μm pass with feces, and they are characterized by a lid-like operculum in one end and a small knob on the other called the abopercular protuberance. The eggs embryonate in water and later, free-swimming coracidia exit the eggs, The coracidia are eaten by copepod first intermediate hosts. In the copepods, the procercoid stage develops. When infected copepod is eaten by marine fish, procercoid develops to another stage called plerocercoid in the muscle tissue. When the muscle is eaten by a mammal, an adult tapeworm develops in the intestine from the plerocercoid. Marine mammals are most commonly infected. Humans can be infected by ingesting raw or undercooked fish. Jean Baer, a Swiss parasitologist, recorded this infection in humans during an outbreak in Peru, in the 1960s (Baer 1969). He called attention to ancient archeological ceramic plates, similar to those used by modern people to serve ceviche, which is a source of infection in humans today. He hypothesized that diphyllobothriasis was a prehistoric problem. Callen and Cameron (1960) had already discovered Adenocephalus eggs in a prehistoric Peruvian burial. With regard to A. pacificus specifically, Patrucco et al. (1983) and Ferreira et al. (1984) confirmed Baer’s hypothesis by recovering A. pacificus eggs in coprolites. Human infection with A. pacificus increases during El Niño events because warmer water increases the diversity of intermediate hosts and amplifies the web of infection so that prey fishes of human populations are more likely to be infected. From the pathoecological perspective, warmer water increases the parasite reproduction in the coastal nidus. Sexual reproduction of the parasite takes place in sea mammals and warmer water optimized the chance the eggs will produce larvae that enter intermediate host tissues.

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Hookworms The clinical picture called ancylostomiasis or hookworm disease is associated with the presence of Ancylostoma duodenale or Necator americanus in the human intestine. Both can be found alone or associated in the same person. A. duodenale and N. americanus are considered specific to humans and close-related ape species. The larvae penetrate the human skin (A. duodenale larvae can also be swallowed), enter the blood system, molt in the lung tissue, and cause pulmonary irritation in the host. Some larvae are expelled, but most are swallowed and reach the small intestine as young adults. Males and females attach to the mucosa, sucking blood continuously. After mating, females begin to lay eggs that pass out with feces. The eggs hatch and the first stage larva leave the feces. In 2 or 3 weeks the third larva stage develops to infect a new host. The nidi for hookworms is soil close to fecal deposits. Areas near latrines are common nidi for hookworm infection (Schad et al. 1983). But fecal-contaminated agricultural fields could also have served as hookworm nidi (Behnke et al. 2000).

Whipworms Trichuris trichiura (whipworm) is a parasite species found in humans worldwide. However, people only become aware that they are infected after coprological examination. Adult worms live in the large intestine; they measure 30–50 mm, and the anterior part is thin as a thread, and the posterior end is thick, giving the worm the appearance of a whip. They bury the anterior end in the intestinal mucosa, and females lay 3,000–20,000 eggs per day. Eggs are very characteristic, with the shape of a tray or a barrel, with prominent opercular plugs in each polar extremity. Eggs need warm temperature (22  C), shade, and moisture to mature in the soil and became infective after 3 weeks. When swallowed with food or water, juveniles hatch in the stomach and young adults penetrate the large intestine mucosa. When development is completed, the posterior end protrudes into the intestine lumen while the slender portion remains embedded in the mucosa. The worms can live for several years. T. trichiura infection nidi are associated with fecal deposits. Embryonated eggs can become widely dispersed in human settlements. Jones (1985) found that on an average, there were 400 whipworm eggs per gram of sediment from nonfecal deposits in Medieval York England and as many as 20,000 per gram in fecal deposits. Fisher et al. (2007) report the same for Albany, New York, in historic times. Whipworms have been found in New World Mummies. From prehistoric contexts, trichuriasis has been diagnosed in Chile (Pizzi and Schenone 1954) and Gentio II Cave, Minas Gerais Brazil (Ferreira et al. 1980, 1983). Colonial mummies are also positive for whipworm including Itacambira, Minas Gerais Brazil (Araujo et al. 1984), and Peru (Fouant et al. 1982).

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Ascaris Ascaris lumbricoides is the largest nematode parasite found in humans. The worms are round, cylindrical, and the males have the posterior end curved ventrally, while the females have the end pointed straight. Females can reach 50 cm long, but males are smaller, measuring 15–30 cm long. Most infected people have 6–10 worms, but some people are parasitized by more than 500 worms. The infection is commonly asymptomatic, but sometimes only one worm can penetrate the appendix, gall, or pancreas duct, causing obstruction with severe consequences, which are sometimes fatal. The size of the worms, and their number in infected children, can cause intestinal obstruction with wall rupture causing death by septicemia. They are found in the jejunum moving permanently against peristalsis, but they can migrate to the stomach being expelled through coughing or by the nose. Females lay near 200,000 eggs per day. They pass continuously with feces and they are easily recognized by microscopic analysis. For the archeoparasitological diagnosis it is important to recognize taphonomic process that can affect morphologically the eggs. Eggs mature in the soil after 2 weeks under temperature ranging from 20  C to 30  C, and became infective after another week. Eggs can remain in the soil and infect people after months or even years. Eggs may be swallowed contaminating food or water, or inhaled with dust dispersed by the wind. Like whipworm, the nidi for Ascaris infection are fecal deposits. Ascaris infection was remarkably rare in the prehistoric Americas (Leles et al. 2010; Reinhard and Pucu 2014). Both Ascaris and whipworm were very common during all time periods in Europe. The control of these fecal-borne helminths is a hemispheric pathoecological feature of the Americas.

Pinworms The life cycle of Enterobius vermicularis is simple and very well adapted to allow transmission success. E. vermicularis is the biological agent of pinworm infection, a species found only in humans and close related primates. Females are cylindrical, with thin ends, measuring 1 cm. Males are smaller, with the posterior end strongly curved ventrally. They live in the caecum of the large intestine in humans, attached to the mucosa. Gravid females migrate to the perianal region where they deposit, dying soon after oviposition. The flat-sided eggs are very characteristic, measuring 50–60 μm of length by 20–30 μm of width, and inside a larva is already formed. To become infective they must be in contact with oxygen. When ingested they hatch in the small intestine of the new host (or in the same individual: that is called auto-infection), developing to adults while migrating to the caecum. There is no pulmonary cycle. Once in the large intestine the worms copulate and began to produce eggs. Eggs in feces are found in only 5–10% of infected people. However, the diagnosis is easy when adult worms are found in the perineum, underclothes or bed.

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Transmission is primarily airborne. The female worms lay large numbers of eggs on the perianal folds. These eggs are widely dispersed on air currents. Eggs can be aspirated or swallowed, infecting the same host or another individual. Pinworm transmission is independent of environment conditions, and human infection is found worldwide. Nidi for infection are houses, especially crowded houses, where eggs are disseminated on air currents. Although pinworm eggs are commonly found in coprolites from certain regions and time periods in the Americas (Camacho and Reinhard 2020; Reinhard et al. 2016), only one mummy has been diagnosed with enterobiasis (El-Najjar et al. 1980).

Echinostoma Species of Echinostoma (Digenea: Echinostomatidae) are flukes that infect wild animals, and some species are found in humans, commonly in Asia. Generally, the species are not very host-specific. Fresh water molluscs are the first intermediate host. Each egg hatches and a miracidium emerges to penetrate a snail. Many snail genera host miracidia including Physa, Lymnaea, Helisoma, Biomphalaria, and others. Metacercaria occur in molluscs, fish, and tadpoles. Ingesting intermediate host infects vertebrate definitive hosts, including reptiles, birds, or mammals, especially those related to aquatic environments. No mummy cases were reported in the Americas, except the finding of Echinostoma sp. eggs in a Brazilian mummy (Sianto et al. 2005). This illustrates the importance of mummy studies in expanding our knowledge of the range of parasites that infect humans.

Lice Most of the time lice live on human blood causing no problems to their hosts except itching and sometimes culturally-defined repugnance. They may transmit infectious diseases, such as typhus and rickettsial disease. Epidemics associated with lice were always associated with poverty, famine, war, economic exploitation, and conditions of inequality among people. The Order Anoplura comprises a number of small arthropods adapted to live only in a given host. The species are very host-specific, and they can exhibit a well-defined anatomical preference, as in the case of the genus Pediculus in Homo sapiens. Lice are small wingless insects, with thoracic segments united in a flat body. There are three species adapted to humans: they belong to the genera Pediculus and Pthirus. Pediculus humanus (or P. humanus corporis) lives in clothes and attaches eggs to them. Pediculus capitis (or P. humanus capitis) lives in head hair and attaches eggs to hair shafts. Pthirus pubis is smaller and preferentially lives in pubic hair. P. pubis can sometimes be found in the eyebrows, moustaches, and eyelashes. Changing behavior of the human species modified the distribution and abundance of these two species. Certain human habits of the last centuries almost eliminated

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P. humanus. These habits include showering, changing clothes to sleep, washing clothes, and sleeping alone or with just one other person. In contrast, Pediculus capitis is still commonly found. P. pubis is found mainly in pubic and axillary hairs, were they keep attached to the base of the hairs, sucking blood continuously. They are transmitted to one person to another during sexual intercourse, or by sharing the same clothes. Salivary glands emit a secretion introduced into the skin producing an inflammatory reaction. Scratching of the lesions may introduce bacterial infections causing adenopathies. Lesions can be found in the pelvic region, axillae, and the waist. Mummies are the main source for louse studies in the Americas. Numerous discoveries have been documented (Ewing 1924; El-Najjar et al. 1998; Rick et al. 2002; Martinson et al. 2003; Reinhard and Buikstra 2003; Arriaza et al. 2012, 2013a, b). As discussed in subsequent population-level pathoecology sections, these are a rich source for pathoecological studies.

Fleas Fleas (Insecta, Siphonaptera) are wingless insects. The body is flat, the head is distinct from the body and the hind legs are adapted to jump. Adults feed on blood of mammal hosts. They transmit bacteria and cestodes to humans. There are more than 2,500 known species, but only a few are important to humans. Pulex irritans, the house flea, may have been associated with humans even before the beginnings of civilization. For Peruvian mummies, P. irritans has been diagnosed (Dittmar et al. 2003). Xenopsylla cheopis is a rodent flea which is one of the most important vectors of plague. Ctenocephalides canis and C. felis are commonly found in domestic dogs and cats. P. irritans can also feed on dogs, but after feeding it abandons its host to lay eggs on the soil. Larvae hatch from the eggs and resemble maggots as they feed on organic debris and pupate. Because larvae live in the soil and can survive for long periods before molting, probably this species was already feeding on humans since the origin of humankind. Tunga penetrans has many popular names, such as sand flea, chigoe, “nigua” in Spanish and “bicho-do-pé” in Portuguese. Females are parasites that penetrate the host skin, usually pigs and other mammals, but also humans. Tungiasis causes extreme itching, inflammation, and pain. Secondary infection occurs frequently. Tetanus and gangrene are the most severe complications.

Trypanosomes Chagas disease, which is caused by the protozoan parasite Trypanosoma cruzi, has a multifaceted pathoecology. Because of this, it is very important in mummy studies. Its basic life cycle involves many species of reduviid bugs in the family Triatominae – winged insects called “kissing bugs” that act as vectors for the disease. The disease normally cycles among a wide variety of host animals including marsupials, edentates (such as the armadillo), carnivores, rodents, and bats. Humans most often

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become infected with T. cruzi when infected triatomines (assassin bugs) emerge nocturnally to feed on sleeping people (Reinhard et al. 2003; Roberts et al. 2004; Reinhard and Araújo 2012). The symptoms of Chagas disease are diverse. There are two stages of infection with distinct symptoms. In about 1% of cases, acute symptoms occur 1–2 weeks after infection. These include fever, fatigue, facial swelling around the bite site, and enlarged lymph glands. These symptoms last from 4 to 8 weeks and then disappear. Chronic disease develops 10–20 years after initial infection in about one-third of infected people. Cardiac problems such as cardiomegaly, arrhythmia, and cardiac arrest are common indicators of the chronic stage of this disease. Problems with the digestive system, including megaesophagus and megacolon, are also symptoms of the chronic stage, which in some cases causes death. Additionally, oral transmission of Chagas disease has been discovered (ShikanaiYasuda et al. 1991; Prata 2001). This occurs when humans eat infected food that is either contaminated with the feces of infected triatomines, or they accidentally eat the infected bugs. Either can occur inadvertently in contaminated products that are processed into consumable foods. Another potential method of transmission for Chagas disease is eating the meat of infected animals when the meat has not been fully cooked.

Pathoecology in Mummy Populations Chinchorro Pathoecology Bernardo Arriaza is an active sponsor of Chinchorro pathoecology. The Chinchorro are one of the best known mummifying cultures. They were a pre-agricultural culture with a diverse subsistence base. That subsistence base included fish and sea mammals from the marine environment and seeds, rhizomes, and fruits from freshwater wetland plants. The Chinchorro inhabited the coast, estuaries, and river valleys close to the coast. At this point, it is not obvious that they had permanent houses. There is abundant evidence, however, of small shelters of temporary nature. Most Chinchorro mummies were eviscerated (Arriaza 1995). However, in the later phase of the culture, they produced mummies without evisceration. These mummies have been extensively analyzed for intestinal helminths. These analyses reveal only A. pacificus. Interestingly, there is significant variation between the sites that have been analyzed. Sixteen mummies were examined from the cemetery of Morro 1–6. Five of them were positive for A. pacificus eggs. This is a 31% prevalence of infection. In contrast, none of eight mummies from the site of Morro 1 were positive for A. pacificus eggs. Morro 1 and Morro 1–6 samples can be dated between 5240 and 3560 years BP. Dietary analysis of the mummies indicates that they did not have equivalent subsistence strategies (▶ Chaps. 22, “Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice,” and ▶ 25, “Dietary Stress in Combat: Coprolite Analysis of a Korean War Marine Killed in Action”).

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We believe this was due to ecological variation between the times the cemeteries were established. We suggest that the ecological variation that resulted in different subsistence was connected to the cause of variation in A. pacificus infection. El Niño events could explain both (Arriaza et al. 2010). During modern El Niño, warm water moves to the Andean coast along with increased diversity of intermediate hosts for A. pacificus. As a consequence a greater number of human prey fish are infected and more human cases have been recorded during El Niño events. The site of MO 1, which has no infection with A. pacificus relied on small anchovy-sized fish. Indeed, 63% of the coprolites contained these bones of these small fish which are not normal hosts for A. pacificus. In contrast, at the heavily parasitized MO 1–6 site, only 13% if the coprolites contained anchovy sized bone. Plant remains in the mummies also reflect a difference in food diversity. MO 1–6 contains more tubers and seeds. In contrast, MO 1 is dominated by rhizomes. In all likelihood, the MO 1–6 Chinchorro were more reliant on the larger fish that are more common A. pacificus hosts. Bones of Sarda chilensis, Trachurus murphyi, Mugil cephalus peruanus, Scomber japonicus peruanus, Paralichtys ssp., Cilus monti, and Pimelotopon ssp. have been found in Chinchorro trash middens (Arriaza 1995; Arriaza et al. 2010). Of these, at least S. chilensis, T. murphyi, and M. cephalus are known intermediate hosts for A. pacificus. We suggest that the presence of A. pacificus infection is associated with the abundance of these fish during El Niño events. Thus, the pathoecology of infection is associated with a global environmental phenomenon.

Chinchorro and Chagas Aufderheide et al. (2004) discovered a high prevalence, 39–40%, of Chagas disease during Chinchorro times based on molecular analysis of mummies. They were somewhat confounded in explaining the high prevalence of Chagas for the Chinchorro culture. They suggested that “The most plausible explanation for this phenomenon suggests that, before human occupation of the studied region of South America, the sylvatic cycle of Chagas disease involved a vast reservoir among wild mammals and that the disease was transmitted by a large number of insect vectors that existed before human occupation of the studied region of South America. Upon settlement of this coastal segment 9,500 years ago, humans intruded upon and became participants in this sylvatic cycle, perhaps augmented by various forms of trypanosome ingestion, including consumption of infected food. The Chinchorros may have established a sedentary lifestyle elsewhere that included a domestic cycle before they moved to the northern Chile coast, bringing the vector and the disease with them. However, present archeological knowledge provides no evidence of links between the Chinchorros and any other postulated site. The proposed intrusion of the earliest Chinchorro people into a fully developed sylvatic cycle with gradual transition into a domestic cycle in subsequent generations or cultures appears to us to be the most plausible interpretation of our findings.” The pathoecology of Chagas infection for Chinchorro is explained. An indigenous triatomine, Triatoma spinolai, lives in beach environments on the coast of the

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Atacama Desert in the area of Arica, Chile. Today it feeds on dogs, rodents, sea birds, sea lions, and other marine mammals. T. spinolai is a natural vector of Chagas disease. Modern fishermen have been infected with T. cruzi by T. spinolai. In more recent work, aDNA analysis indicates that the original type of T. cruzi infecting Chinchorro was the Discrete Taxonomic Unit TcBat (Guhl et al. 2014). Thus, the origins of Chagas disease in the area have been revealed to be the result of transmission of a newly described DTU and a sylvatic triatomine vector. The largest Chinchorro domestic sites are in coastal environments. The trash mounds in which Chinchorro shelters were constructed were ideal nidi for Chagas disease transmission. The sites are now constructed of layers of shells, animal bones, and vegetal fibers. Such detritus would attract carion feeding birds and mammals. In these sites, infested with T. spinolai, the sylvatic cycle intuited by Aufderheide et al. (2004) was established. Thus, in prehistory, even the temporary use of these large and deep sites would have exposed Chinchorros to Chagas disease.

Lessons of Pathoecology for the Chinchorro The parasitism of hunter-gatherers is largely defined by their natural environment. This was certainly true for Chinchorros. The variation in fish-borne parasitism can be interpreted pathoecologically as a result of fluctuations currents which enhanced parasite life cycles in coastal environments where Chinchorros fished. They exploited the locally available fish species. When more species were infected as intermediate hosts, Chinchorros became infected as definitive hosts. The life cycle of Chagas disease has many variations. The Chinchorros were unlucky to make their homes in an environment where a good vector of T. cruzi existed. As a result, Chinchorros suffered a remarkably high level of Chagas disease for hunters and gatherers.

Chiribaya of Southern Perú The Chiribaya region extended from the Tambo Valley, Perú to the Azapa Valley, Chile. The excavations of Chiribaya sites in the Moquegua Valley, Perú directed by Jane Buikstra represent the largest archeological project specific to recovering Chiribaya pathoecological data. Martinson et al. (2003) present a literature review for the Moquegua excavations relative to parasitology. Radiocarbon dates suggests that the Chiribaya culture began in the Moquegua Valley as early as AD 700. The Chiribaya population increased in the lower valley during the succeeding centuries and reached their maximum population at approximately AD 1350. A “mega” El Niño event cut short the further development of the Chiribaya at approximately AD 1350. This El Niño event destroyed Chiribaya agricultural systems and reduced the population by as much as 80%.

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In the Moquegua Valley, Chiribaya territory extended from the coast to 3000 m above sea level in the Andes as reviewed by Martinson et al. (2003). Most sites are concentrated below 1000 m. As of this writing, 28 Chiribaya sites and 21 Chiribaya cemeteries are known to have existed or still exist in the area of the city of Ilo in the lower Moquegua Valley. Some of these sites have been destroyed by development and grave robbers. Six sites existed on the coast and the rest inland. There is a difference in subsistence between coastal and valley sites. Valley populations gathered lomas resources and cultivated crops. Coastal site subsistence activities included hunting and collecting marine mammals, fish, and invertebrates. Lomas formations are tightly defined plant communities, like islands of vegetation, separated by absolutely desolate desert, completely devoid of plant life. Important for pathoecology, Chiribaya architecture was organized in rectangular compounds on terraces defined by stone retaining walls (Martinson et al. 2003). The perimeter walls are made of cane that encloses rooms, armadas, storage areas, and halls. Houses themselves are made of cane walls plastered with adobe. These substantial compounds, and their building materials, are ideal habitats for triatomine disease vectors.

Coastal Site – San Gerónimo San Gerónimo is about 200 m from the mouth of the Moquegua (also Osmore) River (for literature review, see Martinson et al. 2003). The site was relatively small, about 12 ha. There are two published radiocarbon dates: AD 1020 and AD 1156. Archeologists excavated 90 Chiribaya burials before the site was destroyed by development. The cemeteries were not formal, and the tombs were made in trash middens. Normally, each tomb contained a single individual. The burials were flexed in seated position and wrapped in textiles. There was a diversity of offerings found with the burials including textiles, ceramic vessels, wooden keros, miniature boats, tools for use in textile manufacture, and fishing implements. A minority of tombs contained many finely made objects which show status differentiation at the site. For San Gerónimo hunting and gathering ocean resources was the economic basis for the site. Men are buried with fishing tools and the site middens are largely composed of fish and shellfish residue. But like other estuary Andean cultures, San Gerónimo inhabitants had estuarine and lomas resources, and possibly herded camelids. The location of this site near the river might have exposed its inhabitants to insectborne diseases. The fact that the site is down-stream from larger sites made it susceptible to fecal-borne parasites.

Farming Village – Chiribaya Baja Chiribaya Baja is the type-site for the Chiribaya culture and is located approximately 8 km from the coast. It is a small village of 14-hectares composed of residential

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terraces above the floodplain. The fact that the site is adjacent to the region’s largest tract of arable land reveals its farming past. It was the largest agricultural site in the valley. Typical for Chiribaya, houses are adobe-plastered rectangular cane and wood structures stone terraces. Many of the tombs at this site contained mummy bundles accompanied by quotidian, minimalist offerings. Relatively few decorated items were placed with the mummies. However, there was one section of the cemetery with more elaborate tomb construction and grave goods. Thus, there was a range in status within this site.

Administrative and Economic Center – Chiribaya Alta Chiribaya Alta was as the Chiribaya cultural center (Martinson et al. 2003). It is a 36-hectare site located approximately 5 km inland. It is a complex site with 9 cemeteries, a wall and ditch system, residential and agricultural terraces, and large cane-walled structures. The importance of Chiribaya Alta is indicated by its location. It was built 100 m above the river with a steep drop to the valley floor below. It has a commanding view of the valley and the best entries into the valley. Chiribaya Alta’s location would have allowed the inhabitants of the site to monitor movement in and out of the valley. It overlooks all main terrestrial food sources including pasture for camelids, lomas, and agricultural land. This site dominated all available resources.

El Yaral – Herding Village El Yaral was an agro-pastoral site (Martinson et al. 2003). This conclusion is based on its location, artifacts from burials and trash deposits, from stable isotope, and from its association with herding areas. However, marine fish was traded to the site. It is located 50 km inland and 1000 m above sea level. It is a relatively small site of about 13 ha, built on a terrace and hill slopes above the river. Interestingly, the Moquegua River goes underground downstream from El Yaral and emerges again in the lower valley upstream from Chiribaya Baja. The site gives the appearance of complexity with over 300 residential terraces, two identified cemeteries, and two large communal buildings. Radiocarbon dates show that the site was in use from AD 1027 to AD 1252. The burial pattern at El Yaral is very different from the other Chiribaya sites due to the use of oval tombs at El Yaral rather than rectangular tombs. Also, the burial offerings were very simple without obvious shows of wealth. There does not seem to be an elite represented at this site. Like other Chiribaya sites, houses were built of cane walls set into wall trenches. The cane walls were plastered. Two cemeteries were excavated, but as of this writing, the mummies have not been analyzed for endoparasites. However, the human mummies have been analyzed for ectoparasites. Llama mummies have been analyzed for endoparasites and ectoparasites.

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Chiribaya Pathoecology Several studies of Chiribaya parasitism have been published (Leguía and Casas 1999; Dittmar et al. 2003; Holiday et al. 2003; Martinson et al. 2003; Reinhard and Buikstra 2003). Evidence of diet, trade, status, occupation, and site formation from several villages were used to interpret the evidence of several parasitic diseases. Martinson et al. (2003) showed that parasitism was defined by occupation, trade, status, domestic animals, and site location relative to fresh water access.

Ectoparasites Lice of the species Pediculus humanus humanus (body lice) and Pediculus humanus humanus (head lice) were found in the clothing associated with mummies and in the hair of mummies. Reinhard and Buikstra (2003) analyzed the pathoecology of head lice and found significant variation between the Chiribaya sites and between ages and sexes. Interestingly, the pattern of lice was directly opposite of the modern pattern. Chiribaya men had the greatest infestation followed by women. Children were the least infested. This pathoecology was defined by male-specific behaviors. Men had very ornate hairstyles and wore hats. In desert environments, lice are killed by desiccation. By wearing hats, men maintained a more humid scalp environment. Therefore, the nidi for louse proliferation were the covered heads of men. The braided and plaited hairstyles of men facilitated the lice’s ability to hide during delousing. El Yaral and Chiribaya Alta had the best comparative data sets. From El Yaral, 60% of men, 0% of women, and 14% of children were positive for head lice. From Chiribaya Alta, 62% of men, 42% of women, and 29% of children were positive. For all sites, 56% of men, 38% of women, and 24% of children were positive. Lice proliferate in social groups. The big difference in the prevalence between men and women at El Yaral suggests that there was social distance between sexes, perhaps defined by sexual division of labor. This was less so at Chiribaya Alta were a larger proportion of women were infected. Dittmar reported body lice from mummy-associated clothing (personal communication). In 1997, she had the advantage of examining clothing before curatorial staff cleaned the clothing for storage. Reinhard found no body lice on Chiribaya clothing because he had access to the clothing after cleaning when he did his analyses. Therefore, analysis of mummy’s clothing for ectoparasites should be done before the clothing is cleaned for curation. At this stage of Chiribaya pathoecological analysis, we can only say the Chiribaya were positive for body lice, but we have no idea of the comparative prevalence between sites. Dittmar et al. (2003) found fleas of the genus Pulex on dog and guinea pig mummies. Fleas have had an important role in the transmission of disease to and among human populations over history. Plague and murine typhus are two diseases transferred by fleas. The dogs and guinea pigs analyzed by Dittmar were excavated

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from Chiribaya Baja. The nidi for infestation was undoubtedly the Chiribaya houses in which humans, dogs, and guinea pigs were domiciled and where flea eggs were laid and hatched. Dittmar (2000) studied 112 guinea pig mummies sacrificed at Yaral. Two genera of biting lice, Mallophaga, were found. Nineteen mummies were positive for Trimenopon hispidum and 9 for Gliricola porcelli. The life cycle occurs completely on the host. The eggs are cemented to hairs. They cannot transfer to humans. Four mummies had the blood-sucking mite unidentified species in the genus Ornithonyssus. These mites are active at night and take refuge in the day in structure. They can also attack humans and cause dermatitis. Microthoracius spp. were found Llama mummies El Yaral. This sucking louse reduces wool production in the modern world. The control of these lice must have been important to the Chiribaya who made their clothing of llama wool.

Intestinal Helminths The most common helminth was the tapeworm, A. pacificus. San Gerónimo inhabitants specialized in fishing. Trade in fish spread the disease to the other villages. Three separate analyses of coprolites and mummies were done. Martinson (2002) found that 20 of 29 samples (69%) from Chiribaya Baja, and 1 of 5 samples (20%) from Chiribaya Alta were positive for the eggs of this parasite. Holiday et al. (2003) found that 6 of 26 samples (23%) from Chiribaya Baja, and 1 of 3 samples (33%) from San Gerónimo positive for tapeworm eggs. As reviewed by Martinson et al. (2003), Reinhard found tapeworm eggs in 2 of 8 (25%) individuals from San Gerónimo and 7 of 11 (63%) mummies from Chiribaya Baja. These authors published quite different results. All authors used the same methods. Indeed, Holiday and Martinson were trained by Reinhard. The variation in results is minimal for San Gerónimo. The variation is threefold for Chiribaya Baja for which the authors had equivalent sample sizes. However, Holiday et al. (2003) analyzed different contexts from Chiribaya Baja. This variation reported by the authors reflects different prevalence between cemeteries. Like the Chinchorro, it could be due to temporal changes in El Niño effects. Thus, the variation could well be an aspect of El Niño pathoecology. One other helminth species, T. trichiura or whipworm, was found. Two of 9 (22%) San Gerónimo coprolites analyzed by Martinson et al. (2003) were positive for whipworm. No other mummies or coprolites analyzed from Chiribaya Alta or Chiribaya Baja were positive for whipworm. This indicates that fecal-borne parasitism was a problem only for San Gerónimo. The Osmore River that flows passed all of the villages emerges just a few kilometres inland of Chiribaya Alta and Chiribaya Baja. Therefore, the water at these sites was not as polluted by feces from inland sites. However, San Gerónimo, located at the river’s mouth, had only one source of polluted water after the river flowed past the other Chiribaya sites. The nidi for whipworm infection were the latrines of upstream villages.

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Several intestinal parasites were found in llama mummies. All are fecal borne. These include Lamanema chavezi, Nematodirus lamae, Capillaria spp., and Trichuris spp. The fact that fecal-borne worms infected llamas indicates that llamas had limited access to fresh pasture or they were herded into fecal-contaminated pens.

Chagas Disease The gross pathology of Chagas disease, megacolon, and megaesophagus, was observed in Chiribaya mummies (Martinson et al. 2003). Later, Aufderheide et al. (2004) tested 70 Chiribaya mummies from various sites using molecular biology. He found a prevalence of 47%. With nearly half of the mummies testing positive for Chagas disease, this was the most significant health crisis caused by parasites in the Chiribaya region. The pathoecology of Chiribaya Chagas disease focused on the houses which were friendly habitats to the triatomine vectors. Triatomines prefer structures with abundant tiny hollow spaces to hide in daylight. The plastered, cane walls of Chiribaya houses were perfect environments for triatomines. The domestic animals of the Chiribaya, dogs and guinea pigs, are excellent reservoir hosts for the disease. Thus, the primary nidus for trypanosome proliferation and transmission was inside of the houses where humans, reservoir hosts, vectors, and parasite were domiciled.

Summary of Chiribaya Pathoecology The analysis of Chiribaya pathoecology illustrates the integration of various parasitological diagnostic procedures with archeological analysis to reconstruct pathoecology. The diagnostic procedures included the examination of mummy gross pathology, microscopic identification of parasite eggs and cysts, entomological reconstitution of ectoparasites, and molecular biological diagnosis of infection. When the parasite data are combined with archeological evidence, the cultural influence on pathoecology can be discerned. For A. pacificus, the natural coastal nidus was amplified by trade which transferred infected fish meat inland. For T. trichiura, fecal contamination carried infective eggs downstream to San Gerônimo. The universal use of cane walled, plastered houses, built on masonry terraces created ideal nidi for the transfer of Chagas disease. The micro-nidus for louse proliferation was in the artificially-moist habitats under the hats of men. The male habit of head cover meant that men’s heads were highly parasitized. Domesticated animals all exhibited crowd-infection ectoparasites. The lice, mites, and fleas proliferated among the domiciled dogs and guinea pigs. The fecal-borne disease among llamas had its nidus in pens and pastures where the animals fed. The study of Chiribaya parasitism represents the goal of pathoecology.

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Isolated Mummy Pathoecology Lapa do Boquete, Peruaçu, Minas Gerais, Brazil When pathoecology is applied to individual mummies preserved in isolated contexts, much can be learned from disease ecology. This is demonstrated by the analysis of an adult male mummy from the site of Lapa do Boquete in the Peruac¸u river valley in the Brazilian state of Minas Gerais (Sianto et al. 2005; Reinhard and Araújo 2016). The site and mummy were excavated by the Brazilian archeologists Andre Prous and Renato Kipnis who sponsored the pathoecological analysis of the mummy they discovered. A large mass of feces that probably obstructed the descending colon at the time of death was observed in CT-scans of the mummy (Sianto et al. 2005; Reinhard and Araújo 2016). Dietary analysis of one fragment of the mass confirmed the fecal origin. Manioc starch and fiber (Manihot spp.), bean seed coats (Phaseolus vulgaris), probably Myrtaceae fruit epidermis, and fish bone were found. This shows that the individual had a mixed agriculture diet with wild, collected foods. Two species of helminth parasite, one common and one rare, were found in the fecal mass. Hookworm was present and has been recovered from other sites in North and South America. In addition, an unusual intestinal parasite, Echinostoma spp. was found. Echinostoma spp. is an intestinal fluke. Diagnosis of Chagas disease was based on gross pathology and molecular analysis. The molecular analysis of Chagas disease T. cruzi ancient DNA showed that the individual was infected by a common Brazilian strain of T. cruzi (Fernandes et al. 2008). Each of these parasites has a distinct life cycle and reveals different components of the Lapa do Boquete pathoecology which can be reconstructed from his parasites. In general, his environment was moist and warm to support wetland adapted intermediate and definitive mollusc hosts for Echinostoma spp. These included species in family Strophocheilidae the shells of which were commonly found in the site and may have been the involved in the Echinostoma infection (Sianto et al. 2005). Also, the man was active on shaded, warm, sandy-loamy deposits where hookworm larvae were active. While Triatoma infestans was originated from the Andean region, Triatoma brasiliensis may have been the main Chagas vector responsible for Trypanosoma cruzi transmission in caves and rock-shelters in the South American lowlands during prehistoric times. T. brasiliensis is well adapted to wild environment and also to human dwellings (Costa et al. 2003). Today, it is the main species transmitting T. cruzi in the Brazilian northeast. For Chagas disease in cave settings, the cave itself was the nidus. People generally deposited vegetal material and stone in their habitations which created a good environment for triatomine bugs. By working and sleeping in caves, humans were in close proximity to the infection vectors. Eating incompletely cooked meat from infected reservoir hosts including rodents and armadillos can cause infection

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(Reinhard and Araujo 2015). Kipnis (2002) showed that armadillo meat was commonly eaten in a poorly cooked state. Armadillos are a reservoir host for Chagas disease. Each infection of this mummy had a distinct nidality. Chagas had a restricted nidus. It was the enclosed area where the T. cruzi parasite, the triatomine vector, and humans were associated. The caves in the Peruac¸u Valley were the primary nidi for Chagas infection. Because we do not know which species infected humans, the nidus for Echinostoma infection is difficult to define. We suspect E. luisreyi was the species that infected this mummified individual (Sianto et al. 2005). Sadly, this is a newly discovered species with a poorly known natural life cycle. We can only say that the nidus for Echinostoma infection was the broad area of the Peruac¸u River and its tributaries where humans were associated with the parasites intermediate hosts.

Chagas Disease in the Texas-Coahuila Border The combination of gross pathology and molecular biology has been essential in diagnosing Chagas disease in South American mummies. It has been successfully applied in North America as well. In 2003, the gross pathology of a mummy discovered on the Lower Pecos region of Texas-Coahuila border near the town of Langtry was published. The gross pathology was impressive. Reinhard et al. (2003) described the extent of megacolon in this mummy which has no precedent in the clinical literature. They wrote: The large intestine is nearly complete. The appearance of this organ is consistent with megacolon. There are ten segments of the colon that are preserved today. Most segments exhibit a grossly enlarged diameter. The average diameter of the large intestine was 6 cm. There are 112.9 cm of the large intestine, compacted with partially digested food, which are “mummified.” The reason the intestine “mummified” is due to the fact that it was filled with compacted food remains that desiccated and retained the form of the colon. Of the intestine segments that could be weighed, 1,170 g of feces were present. The total volume of the preserved large intestine was 3,710 cm3. The SMM mummy contrasts with normal mummies. As noted above, normal fecal pellets are present ranging up to 3 cm in section. These are separated from each other in the haustra of the intestine. The diameter of a normal, mummified intestine is approximately 3 cm. Normally, the amount of feces in the large intestine weighs less than 30 g. In contrast, the SMM mummy contains feces of 6 cm in diameter and at least 3,710 cm3 of feces are within the mummy. Never has a large intestine been found that is completely filled with partially digested food. Analysis of the contents of one intestinal segment revealed bones of fish, four rodents, a bat, 250 grasshoppers, plant fibers, seeds, and grass pollen. The entire colon is filled with these sorts of foods. Therefore, the findings of this study demonstrate that this mummy suffered from megacolon in life (Reinhard et al. 2003).

Recent molecular analysis of tissues from the mummy recovered ancient DNA of a Columbian variant of T. cruzi. This is the northernmost confirmed case of Chagas disease in prehistory. The pathoecology of this confirmed case of Chagas disease can be reconstructed based on the archeology of the region.

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The most common, natural pathoecology of Chagas disease in this desert region involves the T. cruzi parasite, several species of triatomine bug vectors, and woodrat species hosts in the genus Neatoma. Four triatomine vectors are commensals in woodrat nests. These are Triatoma neatomae, T. rubida, T. sanguisuga, and T. gerstaekeri. These commensals live in with woodrats in the species N. mexicana and N. micropus. In the nests, the bugs transmit the T. cruzi infection to the woodrats (Reinhard et al. 2003; Reinhard and Araujo 2015). Humans interrupted this normal cycle by hunting woodrats and other hosts and also by creating mounds of burned rock left over from agave roasting activities. Analysis of coprolites shows that Neotoma spp. woodrats were a very common food for Archaic hunter-gatherers such as the SMM mummy (Reinhard et al. 2003; Reinhard and Araujo 2015). By eating woodrats and depleting their population and altering their environment, the triatomine vectors of Chagas disease were without their normal host and normal environment. At the same time, humans made ideal environments for triatomine bugs in habitation caves (Reinhard and Araujo 2015). Within caves and rock-shelters, humans cooked food, ground food, and slept. The cooking of agave especially resulted in the accumulation of rock and vegetation in caves. Such accumulation of rocks and vegetation is optimal for the four bug commensals as well as three other triatomine vectors of the disease, T. protracta, T. indictiva, and T. lecticularius. Eating food contaminated with infected triatomine bugs also transmits the disease. Bedrock grinding areas were used by Archaic people in these triatomine environments. Thus, by predation on woodrats, modifying cave environments, and developing subsistence methods that did not separate bug environments from food processing, humans made themselves susceptible to infection by T. cruzi. As seen in the SMM mummy, some infections resulted in Chagas disease.

Prehistoric Human Migrations – Mummies and the Big Picture Araújo and Ferreira have championed the idea that the modern and ancient distribution of human-specific parasites reflected prehistoric human migrations. Araújo et al. (2008) summarize this theory with evidence of hookworm and whipworm from mummies and wireworm and ascarids from coprolites. The importance of mummy studies in this debate was fundamental. Previous work had been disputed by anthropologists who doubted the accuracy of whipworm and hookworm diagnoses (Reinhard et al. 2001). Doubters argued that evidence from coprolites was invalid because human origin of coprolites from latrines and refuse was uncertain. However, the evidence from mummies was unequivocal because the human origin of coprolites from mummies is obvious. Armed with hookworm and whipworm data from mummies, Araújo et al. (2008) successfully argued that the prehistoric evidence of American human-specific parasitism showed that multiple migrations of humans occurred, at least one of which had a non-Beringean route.

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The Serial Nature of Mummy Studies As can be seen in this review of the pathoecology of American mummies, mummy studies of parasitic disease are sequential. By this, we mean that a particular parasite becomes the focus of intense study for a period of years and then is replaced subsequently by another species of interest. Clearly, in the first decade of the twenty-first century, T. cruzi and associated Chagas disease is a main organism of interest. In large part, this is due to the surprising nature of pathoecological studies. Such studies are surprising because so many discoveries are contrary to accepted scientific beliefs. Also, some parasites have diverse pathoecology, such a T. cruzi, that each mummy or mummy population provides a new and different view of the parasite. Like John Godfrey Saxe’s poem “The Blind Man and the Elephant” concerning six blind men identifying different parts of an elephant, each study provides a different perspective on the parasite and taken together a very complex image of diverse pathoecology emerges. With regard to the last few years of study, we can see that the earliest humans in the Andes intruded on a preexisting T. cruzi life cycle and became infected (Guhl et al. 2014). Later, by domesticating guinea pigs and dogs in cane-walled houses, Andean agriculturalists domiciled T. cruzi. On the east side of the Andes, ancient Brazilian agriculturalists intruded on yet another life cycle variant of T. cruzi and armadillos. Finally, far to the north, hunter-gatherers in Texas and Coahuila amplified the preexisting environment for Chagas disease by predation on the normal hosts. Other diseases of interest in the past included hookworm. Indeed, in the last two decades of the twentieth century, the evidence of hookworm from mummies and coprolites was the topic of lively debate in the parasitological and anthropological literature. This stemmed from the fact that few scientists believed that hookworm was present in the Americas before European invasion. Prior to our interest in archeological parasitology, the discoveries of Darling and Soper made in the early 1920s in Paraguayan South American natives, regarding hookworm infection, were very interesting (for review, see Araujo et al. 1988). They discovered that the indigenous people were infected mainly by A. duodenale, in a rate of 13:1 of the other species found, Necator americanus. At that time, both species were considered to have been imported, the first by Europeans, and the second by the slave trade from Africa. Native Americans were considered free of the most common intestinal helminth diseases. With the new data, as N. americanus infection predominated over A. duodenale in the colonial population of European and African origin, Darling and Soper hypothesized that N. americanus had, indeed, come from the Old World. But A. duodenale would have another origin, dated from pre-Columbian times. Based on Darling and Soper data, the American parasitologist Harold Manter, and also the Brazilian parasitologist Olimpio da Fonseca, in the 1970s, argued that the infection by A. duodenale would have come to the Americas with prehistoric migrants that used another way than the Bering Land Bridge (Araujo et al. 1988). They both argued that the parasite life cycle would have been interrupted under the low temperatures prevalent in Siberia

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and Alaska, as soon as the migrants started to move from the southern parts of Asia to the cold northern regions. Analysis of mummies confirms hookworm infection in pre-Columbian times. Parasite egg and larvae were found in coprolites collected from a mummified body found in an archaeological site in Minas Gerais, Brazil, dated 2,500 years old (Ferreira et al. 1983), and adult worms were found still attached in the intestine of a mummified Peruvian body, dated of 900 AD (Allison et al. 1974). These discoveries fed into a preexisting debate regarding hookworms in coprolites that was resolved in 2001 (for review, see Faulkner and Reinhard (2014)). Once hookworm and other parasites were accepted as prehistoric American parasites by the general scientific community, interdisciplinary research shows how very ancient coastal migrations during periods of climate change opened the Americas for tropical parasites. This is an impressive application of pathoecological interpretation in mummy studies.

Mummy Studies and The Stockholm Paradigm: Understanding How Parasites Colonize New Hosts In the world today, infectious agents are evolving, emerging, and changing their host distributions. In the modern world, the components of host switching have been defined by the Stockholm paradigm. Change in climate results in ecological perturbation that drives change of faunal assembly and structure (Brooks et al. 2014, 2019). Host switching by infections organisms is elucidated by the concept of “ecological fitting.” Ecological fitting refers to how parasite phenotypic/genotypic flexibility provides an opportunity for rapid host switching. Therefore, the parasite has traits that “fit” a new host, allowing the parasite to adapt to a new host. The paradigm derives its validity by examining evolutionary adaptations deep in time as well as emerging infectious diseases throughout human history and in modern times. The paradigm is based in conditions of changing climate. However, human adaptation as they colonized environments in prehistory resulted in exposure to endemic species of parasites. Thus, some of the paradigm components can be applied to understanding host shifts in prehistory. The molecular data from prehistoric South American mummies fit the Stockholm paradigm of parasite switching. T. cruzi genetic variant are referred to as discrete typing units (DTUs). In very early in prehistory, T. cruzi switched to human hosts as generalist DTUs known as TcBAT and TC IV. The Stockholm Paradigm specifies that over evolutionary time, parasites will evolve to adapt to the new host population. This is exhibited by DTU evidence over time in as evidenced by T. cruzi aDNA in mummies. Interestingly, domestic cycles evolved that exhibit some specificity to humans and human lifestyle. TcII emerged as a human infection as early as 3,000 years ago, with TcV and TcVI by emerging by 1600 years ago. The whole picture of T. cruzi evolution in mummies exhibits macroevolutionary and microevolutionary change. The earliest T. cruzi DTUs had a degree of adaptive plasticity such that the sylvatic strains switched to Homo sapiens when human occupation overlapped the sylvatic nidi of endemic

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transmission. Thus, in ancient times, T. cruzi became an ancient emergent disease. Later in archeological time, genotypic divergence resulted with the expansion of domestic cycles across south and central America as more T. cruzi DTUs evolved. In modern times, continuing expansion of T. cruzi makes Chagas disease an emergent infection exacerbated by human induced climate change (Cizauskas et al. 2017). In this example, we can see how parasitology and paleopathology define a continuum of emergent and reemergent infections from the remote past up to today. Data from the field of mummy studies revealed this evolutionary story of ancient host switching followed by centuries of emergence and reemergence.

Conclusion These examples illustrate the unique role mummy studies have to offer parasitology. As theoretical perspectives emerged in parasitology over the last decades, data from mummies were used define parasitological patterns. Specifically, the nidus concept of Pavlovskiĭ, the pathoecology concept of KR Reinhard, and the Stockholm Paradigm of Brooks and his colleagues can all be addressed with data from mummies. In this regard, population-level data, as well as single mummies, are unique sources of data to look at changing parasitological conditions over time.

Cross-References ▶ Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice ▶ Archaeoparasitology of Korean Mummies ▶ Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Archaeoparasitological Research on Korean Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Evidence of Trematode Infection in Korean Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parasitological Infection from the Joseon Period to the Twenty-First Century . . . . . . . . . . . . . . . . Ancient Parasite Species Rarely Discovered in Korean Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ancient DNA Analyses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Other East Asian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Archaeoparasitology is now a rapidly progressing research field, providing comprehensive context about parasitism of the past and the origins and evolution of each parasite species. As in other continents, historical parasitic infection patterns in East Asia have been successfully reconstructed by archaeoparasitological research. By interdisciplinary collaboration with archaeologists, parasitologists M. Seo (*) Department of Parasitology, Dankook University College of Medicine, Cheonan, South Korea e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea K. J. Reinhard School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE, USA D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_14

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in South Korea also have achieved meaningful academic progress in studies on various specimens uncovered at archaeological sites. Among them, coprolites of Korean mummies of the Joseon Dynasty period (1392–1910 CE) have been the most productive, due to their excellent preservation. Actually, Joseon mummies are now of central importance to parasitologists bringing to light the parasitic infection patterns of premodern Korean societies over broad temporal and geographical range. Coprolites, moreover, through ancient DNA (aDNA) analysis, further our understanding of each parasite’s paleogenetics on a global perspective. Keywords

Archaeoparasitology · Korea · Ancient DNA · Mummy

Introduction Ancient parasite eggs are commonly reported in studies of mummified remains around the world. Most frequently, coprolites inside mummy intestines or sediments associated with the pelvic bones prove positive for parasites (Araújo et al. 2000; Seo et al. 2014a; Reinhard et al. 2017). Parasitologists examine such specimens to reveal the parasitic infection patterns among ancient or medieval people and even to reconstruct the patterns of parasitism in historical societies (Ferreira et al. 2000; Horne 2002; Harter et al. 2003; Reinhard et al. 2003; Aufderheide et al. 2004; Fernandes et al. 2005; Seo et al. 2007). To date, archaeoparasitological reports based on mummy research show that parasitic infection in many regions has not remained constant over historical time, but rather has fluctuated (Ferreira et al. 2000; Reinhard and Araújo 2008; Shin et al. 2012a). Archaeoparasitologists have reconstructed the patterns of parasitism in, among other regions, East Asia (Seo et al. 2017). The data accumulated thus far, however, have not been sufficient for in-depth surveys that could uncover general patterns of parasitism through time and space. Until recently, archaeoparasitological knowledge has remained only incipient in East Asian countries. Happily, though, after collaboration between archaeologists and parasitologists began in East Asia, archaeoparasitology has achieved steady and meaningful progress (Seo et al. 2014a). Pioneering research in East Asia has been completed on various specimens from different archaeological sites. Mummies and their associated samples have been successfully collected and maintained in institutes or museums for future parasitological analysis (Seo et al. 2014b). Scholars proved that coprolites from ancient Korean mummies are among the most productive specimens due to their excellent preservation, far better than other kinds samples from archaeological sites in East Asia (Seo et al. 2017). Through full-scale parasitological analysis, clues to an understanding of the changing parasitic infection patterns through time are accumulating (Seo et al. 2014a, b, 2017).

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Archaeoparasitological Research on Korean Mummies In Korea, very well-preserved mummies have been discovered only in graves dating to the Joseon Dynasty (1392–1910 CE) (Kim et al. 2006). These graves in fact were built in large numbers throughout the Korean peninsula, mostly during the sixteenth to eighteenth century. Reviewing historical texts, Chu (1999) characterizes Joseon graves: “When lime, sand, and red clay (filled around the coffin) became hardened like a stone after a long while, the coffin could be protected from the possible intrusions of insects or grave robbers” (Shin et al. 2008). Actually, though the complete preservation mechanism was not scientifically understood at the time, various types of cultural artifacts as well as human remains including mummies have been abundantly retrieved in good condition from Joseon graves (Seo et al. 2014b). The preservation status of Joseon mummies has been so exceptional in fact, that specimens obtained from them proved very useful for archaeoparasitological studies. Over the last decade, Joseon mummies have been regarded as invaluable subjects for investigations into the exact parasitic infection patterns in Korean history (Seo et al. 2014a; Kim et al. 2016). Coprolites have been identified on computed tomography (CT) imaging as well-demarcated masses inside the mummy intestine. These images serve as guides to researchers using endoscopy in autopsies (Seo et al. 2014b). Various research techniques are then applied to the specimens to examine ancient parasite eggs recovered therefrom (Seo et al. 2014a, b, 2017). Briefly, in a wet lab, microscopic examinations are performed on ancient samples (Van Cleave and Ross 1947; Camacho et al. 2018). Mummy fecal samples are rehydrated by standard methods of analysis (Van Cleave and Ross 1947; Reinhard and Urban 2003; Camacho et al. 2018). After rehydration, the final precipitate in solution is pipetted onto microscopic slides and examined under light microscopy (Seo et al. 2007). In 2001, the first-ever archaeoparasitological results on a Korean mummy specimen were reported. Endoscopy revealed the presence of coprolites inside the abdominal cavity of a sixteenth to seventeenth century child mummy (Yangju) (Kim et al. 2006). When the samples were examined under microscopy, Trichuris trichiura, Ascaris lumbricoides, and Clonorchis sinensis eggs were found. In fact, this report clearly showed that ancient parasite eggs could be preserved intact in Korean mummy coprolites even after a long burial duration, and, in turn, that a mummy specimen could be an ideal research subject for future archaeoparasitological analyses (Seo et al. 2007; Shin et al. 2009). From that time onwards, a series of examinations on Korean mummy coprolites (Fig. 1) has revealed ancient parasite eggs and larvae representing T. trichiura, A. lumbricoides, C. sinensis, Gymnophalloides seoi, Paragonimus westermani, Strongyloides stercoralis, Metagonimus yokogawai, and Trichostrongylus spp. (Seo et al. 2007, 2008; Shin et al. 2009, 2012a, b). Under electron microscopy, eggs obtained from Korean mummies have exhibited ideal morphology that is very consistent with modern counterparts (Shin et al. 2009). In fact, the preservation of ancient samples found in Korean mummies has been so perfect that even larvae of Strongyloides stercoralis and Trichostrongylus spp. were found, specifically during

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Fig. 1 The location of Korean mummies for which archaeoparasitological studies were performed. (1) Yongin, (2) Jinju, (3) Sapgyo, (4) Hadong-2, (5) Hadong-1, (6) Sacheon, (7) Gangneung, (8) Dangjin, (9) Mungyeong, (10) Waegwan, (11) PJ SM, (12) Seocheon, (13) Yangju, (14) SN1-2, (15) SN3-7-1, (16) SN2-19-1, (17) SN2-192, (18) GJ1-2, (19) Hwasung, (20) Andong, (21) YG2-4, (22) YG2-6, (23) Dalsung, (24) Junggye (Credit: Jong Ha Hong)

the microscopic examination of a mummy (Gongju) unearthed from a seventeenth century Joseon grave (Shin et al. 2009). Indeed, good preservation quality is crucial to parasitologists’ capacity to answer many questions about parasitic infection patterns in Korean history. A series of examinations of coprolites found in ancient or medieval graves have shown a close relationship between the preservation of human and cultural remains and the presence of ancient parasite eggs inside of or on them (Shin et al. 2011a; Seo et al. 2014b). We discovered that tombs with better-preserved human remains and associated artifacts also had better preservation of eggs and larvae; conversely, those tombs with poorer preservation of human remains had either poorly preserved parasites or no parasite preservation whatsoever. This means that the existence of delicate human and cultural remains (e.g., hairs or textiles) can be used as indicators of tombs offering the best parasitological analysis potential (Seo et al. 2010).

Evidence of Trematode Infection in Korean Mummies Studies on each ancient trematode species from Korean mummies have provided detailed knowledge on parasitism in Joseon society. Among those species, C. sinensis still infects approximately 35 million people worldwide, causing clinical or subclinical signs known collectively as clonorchiasis (Keiser and Utzinger 2005;

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Sun et al. 2013). In Korea, C. sinensis was also one of the most commonly reported parasites until relatively recently (12.1% in 1970) (Shin et al. 2013). Ancient C. sinensis eggs also have been frequently reported from Korean mummies (Shin et al. 2013). One of the more impressive cases was a Korean male mummy (Waegwan) found in a seventeenth-century Joseon grave. According to the mummified man’s clan genealogy, he had lived from 1624 to 1685 CE. Both textiles and human remains were superbly preserved in this case. Under microscopic examination, C. sinensis and T. trichiura were observed in a coprolite specimen. Genetic analysis also showed that the ancient C. sinensis DNA sequence was not so different from those of C. sinensis still prevalent in East Asian countries (Shin et al. 2013). C. sinensis infection generally occurs when undercooked or raw freshwater fish harboring infectious metacercariae are ingested (Shin et al. 2013). The historical literature, moreover, provides descriptive examples of some of the raw seafood cuisine enjoyed by Koreans of the Joseon period. Certainly, both the historical and the archaeoparasitological data suggest, at the very least, that consumption of C. sinensis metacercariae-infecting fish probably was not rare among members of Joseon society. Another trematode frequently found in Joseon mummy coprolites is P. westermani. Its endemicity has been widely reported, and it continues to infect about 293.8 million people worldwide (Shin et al. 2012a). The archaeoparasitological data thus far obtained reveals that the P. westermani infection rate during the Joseon period was as high as 33.3% (Seo et al. 2014a, 2017). The most impressive finding on ancient paragonimiasis to date is the first-ever detection of an ectopic case, found in a seventeenth-century female mummy (Hadong-2) (Shin et al. 2012a). Paragonimus metacercariae generally involve the human lungs. However, in the case of ectopic paragonimiasis, the metacercariae accidentally migrate to other organs, causing many complications. In that parasitological study on the female mummy noted just above, numerous Paragonimus eggs were discovered in the liver, lung, intestine, and coprolite specimens (Shin et al. 2012a). By PCR, Paragonimus aDNA sequences were successfully amplified, showing 100% identity to those of contemporary P. westermani reported in Korea and Japan, which form a separate cluster distinct from those of P. westermani in South Asian countries (Philippines, Thailand, Malaysia, etc.). This study would provide the first report on hepatic paragonimiasis found in mummies around in the world. Korean mummy study has since uncovered another, more arresting case of ectopic paragonimiasis (Shin et al. 2017). On CT images of a male mummy found in Cheongdo, a radiopaque cyst-like liver mass showing a radiolucent pattern at the center was identified. To reveal the nature of the liver mass, we autopsied and bisected it. The cross-section showed two, outer-capsular and inner-trabecular segments. The gross pattern of the inner-trabecular segment well corresponded to the liver mass’s radiolucent area. In a subsequent parasitological examination of the obtained liver mass specimen, many ancient Paragonimus spp. eggs were detected, which indicated that the man had suffered from ectopic paragonimiasis involving the liver. We should note that such liver masses caused by hepatic paragonimiasis are very rare, even in clinical medicine (Shin et al. 2017).

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Paragonimus infection occurs as the result of the ingestion of an intermediate host by a definitive host (Liu et al. 2008). And significantly, according to the historical research, the Joseon people consumed raw crayfish medicinally for measles, and enjoyed raw crab prepared with soybean sauce as a seasonal delicacy. (Shin et al. 2012a, 2017). Considering these historical facts along with the repeated observation of hepatic paragonimiasis among Korean mummies, we presume that the P. westermani infection prevalence in Joseon societies was very high (Shin et al. 2017). G. seoi infection is another interesting finding of ancient trematode infection in Korean mummies. G. seoi is especially noteworthy in that it was discovered and reported, as late as 1993, as a new parasite species in Korea (Shin et al. 2012b). Interestingly, the ancient G. seoi eggs were found in the coprolite of a sixteenth to seventeenth century mummy (Hadong-1) who lived in a southern coastal county of South Korea (Seo et al. 2008). As the female mummy’s county was very close to both a sea and a river, marine or freshwater fish and all sorts of seafood would have been readily available. Actually, in the coprolite specimen from the female mummy, we found many ancient eggs of M. yokogawai, C. sinensis, and G. seoi, all of which are known to be contracted by ingestion of raw or undercooked fish (Seo et al. 2008). This was an unexpected finding, in that there had been no other discoveries of G. seoi eggs in that particular Korean county. Indeed, it was an especially noteworthy finding, as a 2001 nationwide survey of Korea showed G. seoi confined to southwestern counties of the Korean peninsula (Chai et al. 2001). In this light, it seems that, over the course of the intervening hundreds of years from the mummy’s time to the twenty-first century, the spatial distribution of G. seoi infection was reduced from a wider coastal span to a much more restricted locality (Seo et al. 2008, 2014b). This contraction hypothesis of G. seoi infection has been supported by an additional case of ancient G. seoi infection from a nonendemic region (Shin et al. 2012b). In this mummy specimen (Sapgyo) of the sixteenth century, G. seoi eggs were found along with the eggs of M. yokogawai, C. sinensis, and T. trichiura (Shin et al. 2012b; Seo et al. 2014b). This male individual might have been infected due to ingestion of metacercarie-contaminated oyster. Ongoing Korean mummy research no doubt will contribute to the resolution of the controversy generated by the contraction theory of G. seoi infection in history (Shin et al. 2012b).

Parasitological Infection from the Joseon Period to the TwentyFirst Century National survey data in Korea showed very high parasitic infection rates as late as the 1970s. This is evidence that rates of parasitic infection among Koreans fell only in very recent times, mostly due to the nation’s rapid industrialization in the late twentieth century. In the case of the pre-twentieth century dynastic eras of Korean history; however, parasitic infection patterns remain obscure (Seo et al. 2007). Recent and continuing archaeoparasitological research entailing examination of

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coprolite samples from Korean mummies is beginning to clarify the picture (Seo et al. 2017) (Tables 1 and 2). By analysis of the parasitological results for Joseon mummies (n ¼ 24) together with the national survey data for the late twentieth century (1971, 1981, 1992, 2004 and 2012), the parasitic infection rates among Koreans throughout history can be estimated (Seo et al. 2014a, 2017). The results to date are summarized in Table 2. One of the most remarkable findings from this data is that the Joseon period rate of infection by nematode parasites is not much different from the twentieth century rates (Seo et al. 2014b, 2017). Specifically, the relevant infection rates of the Joseon period were 58.3% (14/24) for A. lumbricoides and 83.3% (20/24) for T. trichiura, while the corresponding rates according to the 1971 survey data (Korea Association of Health Promotion 2012) were 54.9% and 65.4%, respectively. Those differences between the Joseon period and the 1971 survey data are not very remarkable. From 1971 on, however, the Ascaris- and Trichuris-infection rates in South Korea decreased dramatically, a radical change presumably caused by the country’s rapid modernization and industrialization, as noted above. As a result, the egg-positive rates of Ascaris and Trichuris in 2012 fell to 0.03% and 0.41%, respectively (Seo et al. 2017). We next compared the C. Sinensis and P. westermani rates of infection among Joseon mummies with the twentieth century national survey data (Seo et al. 2017). Whereas, the C. sinensis infected about 25% (6/24) of Joseon mummies, its infection rate in 1971 was as low as 4.6%. As for P. westermani while 33.3% (8/24) of Joseon mummies were infected, only 0.09% of South Koreans were so afflicted by 1971 (Korea Association of Health Promotion 2012; Seo et al. 2017). Reconstruction of pre-twentieth century parasitic infection rates is especially important for any clear picture of parasitological history in Korea, because the patterns of infection-rate decrease over time differ among parasite species (Seo et al. 2017). In comparing the trematode-infection rates between the Joseon period and 1971, sharp declines were observed for Clonorchis (from 25% to 4.6%) and for Paragonimus (from 33.3% to 0.09%). On the other hand, in the cases of Ascaris and Trichuris, the Joseon and twentieth century data did not significantly differ, as already noted. Rather, remarkable decreases in Ascaris and Trichuris infection rates were identified between the 1971 and 1992 survey data (from 54.9% to 0.3% for Ascaris; from 65.4% to 0.2% for Trichuris) (Seo et al. 2017). We remain unsure as to why the Ascaris and Trichuris infection rates fell later than did the trematode rate (Seo et al. 2017). As to the historical causes of infection, we note East Asian farmers’ practice, since the seventeenth century, of recycling parasite-egg-contaminated “night soil” (human excrement) for use as a fertilizer in vegetable farming. In this way, Ascaris and Trichuris eggs readily contaminated vegetables, whereupon consumption of such vegetables would have initiated a new infection cycle of soil-transmitted parasitism (Kim et al. 2014; Seo et al. 2017). Only once chemical fertilizers – products of modernization and industrialization – came to replace night soil, could Ascaris and Trichuris infection rates be significantly reduced. This explains, of course, why those rates of infection decreased so suddenly and dramatically after the 1970s (Kim et al. 2014; Seo et al. 2017).

Hadong-1 Sacheon Gangneung Dangjin Mungyeong Waegwan PJ SM Seocheon Yangju SN1-2

Sapgyo Hadong-2

• • •

•

• • • • • •

•

•

•

•

• •

•

•

Jinju

•

•

•

•

Yongin

•

Ancient parasite eggs discovered in mummy feces Ascaris Trichuris Enterobius Clonorchis Paragonimus lumbricoides trichiura Hookworm vermicularis sinensis westermani • • •

Table 1 Archaeoparasitological results of Korean mummiesa

• •

•

Metagonimus yokogawai

•

•

Gymnophalloides seoi

Strongyloides stercoralis

Trichostrongylus spp.

Estimated Taenia date 15th–16th century 15th–16th century 16th century 16th–17th century 17th century 1620–1630 1622 1633 1647 1685 1699 17th century 1640–1700 1605–1733

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a

• • •

•

•

•

•

•

•

•

•

Hwasung Andong YG2-4

YG2-6

Dalsung

Junggye

•

•

•

•

•

This table was originally published in Seo et al. (2017) and modified by addition of related information

• • •

• • •

•

SN2-19-1 SN2-19-2 GJ1-2

SN3-7-1

•

•

•

•

16th–17th century 1765  10 1755  10 17th–18th century 18th century Ca. 1571 15th–16th century 15th–16th century 16th–17th century 16th–17th century

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Table 2 Estimation of parasite infection prevalence in Korea from Joseon period to 2012a

Ascaris lumbricoides Trichuris trichiura Clonorchis sinensis Paragonimus westermani Total egg positive rate

Survey of (%) Joseon samplesb 58.3 83.3 25 33.3 100

1971c 54.9 65.4 4.6 0.09 84.3

1981c 13.0 23.4 2.6 0 41.1

1992c 0.3 0.2 2.2 0.0 3.8

2004c 0.05 0.27 2.4 0.002 3.7

2012c 0.03 0.41 1.9 0.0 2.6

a

This table was originally published in Seo et al. (2017) By archaeparasitological studies on mummies c National survey data by Korea Association of Health Promotion (2012) b

Ancient Parasite Species Rarely Discovered in Korean Mummies There have also been several unique findings of archaeoparasitological research on Korean mummies. The pinworm, Enterobius vermicularis, for example, currently infects nearly a billion people worldwide (Iñiguez et al. 2003), and accordingly is one of the parasite species commonly identified among the twenty-first century Korean population. In the historical perspective, E. vermicularis originated in Africa, and then spread to the other continents in the course of prehistoric human migration (Gonçalves et al. 2003; Iñiguez et al. 2006; Shin et al. 2011b). Many archaeoparasitologists have thus speculated that ancient pinworms crossed the Bering Land Bridge during their human host’s historic migration to the Americas (Gonçalves et al. 2003; Shin et al. 2011b). Ancient E. vermicularis eggs were observed in a 10,000-year-old human coprolite sample discovered in Utah (Fry and Moore 1969) and diagnosed in autopsies performed on mummies from Arizona (El-Najjar et al. 1980). These eggs also have been found in ancient coprolites unearthed in Peru, Chile, and Argentina (Reinhard et al. 2016). Old-World specimens of this species, by contrast, have been very scarce (Bouchet et al. 2003; Iñiguez et al. 2003; Camacho and Reinhard 2019). To date, only two such cases have been reported from archaeological samples: one for Roman latrines, and another for an Egyptian mummy (Herrmann 1985; Horne 2002; Seo et al. 2014b). It is noteworthy that there have been no reports of archeological E. vermicularis from East Asian countries despite its endemicity even in the twenty-first century. In this regard, E. vermicularis eggs in a seventeenth-century Joseon mummy (Dangjin) is a rare finding indeed (Shin et al. 2011b). Another rarity with respect to Korean-mummy coprolite-based archaeoparasitology is taeniasis. Ancient Taenia spp. eggs in a 3200-year-old Egyptian mummy, in embalming-reject jars from Egypt, in a Christian Necropolis mummy, and in Han-Dynasty Chinese mummies have been reported (Bouchet et al. 2003; Gonçalves et al. 2003; Harter et al. 2003; Le Bailly et al. 2010; Lee et al. 2011), as has cysticercosis from an Egyptian mummy of the Ptolemaic Dynasty (Bruschi et al. 2006). The sole case of ancient taeniid eggs in Korea thus far is that of a specimen

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collected from a seventeenth-century Joseon mummy (Gongju) (Lee et al. 2011). Such rarity is puzzling, especially considering that Korea is still an endemic area for taeniasis (Lee et al. 2011). Among 1920s Koreans, the infection prevalence of Taenia tapeworms was 7–16% (Lee et al. 2011). Therefore, the scarcity of taeniid eggs in archaeological specimens might be due to the fragile nature of Taenia egg shells, which makes them unsuitable for long-term preservation (Harter et al. 2003; Lee et al. 2011). Finally, we have not been able to find ancient hookworm eggs in any coprolite specimens obtained from Korean mummies. Ancient hookworm larvae and eggs have been reported by parasitological studies globally (Ferreira et al. 1987; Reinhard 1990; Šebela et al. 1990; Gonçalves et al. 2003), and hookworm currently infects approximately one billion people around the world (Gilles 1985). Considering that many Koreans (23.5%) suffered from hookworm infection in 1961 (Soh et al. 1961), the lack of hookworm eggs in Joseon mummy specimens, thus far, is unexpected (Seo et al. 2014a).

Ancient DNA Analyses Over the past decades, aDNA analysis has become a significant research field of interest within the discipline of archaeological science (Marota and Rollo 2002; Geigl and Grange 2018; Hong 2019). Since the first recovery of A. lumbricoides and A. suum, aDNA from coprolites unearthed at a fourteenth-century archaeological site in Belgium (Loreille et al. 2001), there have been further attempts to analyze parasite aDNA from archaeological or mummified specimens. Parasitologists have performed these analyses to facilitate the diagnosis of parasitic infection, the discrimination of regional differences in parasite genetic sequences, and the identification of migration routes of ancient populations, among other purposes (Iwagami et al. 2003; Doanh et al. 2009; Blair et al. 2016). Despite these achievements, detailed understanding of parasites’ genetic history requires still more DNA data, not only from modern samples but also from ancient specimens representing much wider geo-historical scopes. As of now, the number of aDNA analysis reports is insufficient for derivation of any detailed conclusions on ancient parasite genetics. Fortunately, the conditions associated with medieval Korean mummies are ideal for preservation of coprolites, which have proved to be integral to successful aDNA analysis (Seo et al. 2014b, 2017; Hong 2019). Utilizing such egg-containing specimens, we have phylogenetically analyzed multiple DNA regions of T. trichiura, A. lumbricoides, P. westermani, and C. sinensis aDNA to uncover the genetic characteristics of the parasites prevalent among pre-twentieth century Koreans. In this way, we have been able to expand the spatial-temporal scope of historical parasitism (Hong et al. 2017, 2019a, b, c; Hong 2019). In aDNA analysis, multiple DNA regions of parasite species are amplified and detected by electrophoresis. By BLAST searching with the consensus sequence, similar sequences are sought at, for example, the National Center for Biotechnology Information (NCBI). Phylogenetic analysis is performed by the Maximum

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Likelihood (ML) method. In order to secure more authentic aDNA analysis outcomes, the Criteria of Authentication categorized by Hofreiter et al. (2001) must be followed. In the course of aDNA analysis, researchers wear masks, head caps, gloves, and gowns. Every tool used in aDNA analysis must be sterilized before use, and aDNA work must be done in specialized facilities dedicated to aDNA analysis (Hofreiter et al. 2001; Ho and Gilbert 2010). Additionally, the Institutional Review Board (IRB) has reviewed and overseen the ethical aspects of aDNA analyses (Hong et al. 2017, 2019a, b, c; Hong 2019). The phylogenetic ML trees of the T. trichiura, A. lumbricoides, P. westermani, and C. sinensis aDNA and their modern counterparts with bootstrap values are summarized in the Figs. 1, 2, 3, and 4

Fig. 2 Maximum likelihood (ML) tree of Ascaris and other nematode species NAD1 gene. The percentages of replicate trees in which the associated taxa clustered together in the bootstrap test are marked next to the branches. The ancient Ascaris sequences are represented by red dots (Credit: Jong Ha Hong)

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Fig. 3 Maximum-likelihood tree of Trichuris SSU rRNA DNA region. The ancient Trichuris trichiura sequences are represented by red dots (Credit: Jong Ha Hong)

below. The DNA analyses on the specific ancient parasite eggs found in Korean mummies are described in the following paragraphs. Ascaris lumbricoides: Utilizing Korean mummy coprolites, analyses of A. lumbricoides cytochrome b, cytochrome c oxidase subunit 1 (CO1), internal transcribed spacer 1 (ITS1), and NADH dehydrogenase subunit 1 (NAD1), DNA regions were performed (Hong et al. 2017). According to the ML tree (Fig. 2), the taxa of the Ascaris spp. of the Joseon period are distinctly clustered from those of the Baylisascaris, Parascaris, and Toxascaris spp. Since A. suum and A. lumbricoides spp. do not make separate clusters in the trees, it can be reaffirmed that they are very difficult to differentiate with any genetic markers (Liu et al. 2012; Shao et al. 2014). This nonseparate-clustering aspect also is an indication that the genetic characteristics of the A. lumbricoides sp. of the Joseon period mummies were not uniform but somewhat diverse (Hong et al. 2017). Trichuris trichiura: This species currently infects 800 million people globally (Manz et al. 2017). The earliest aDNA analysis for T. trichiura was performed on an SSU rRNA sequence isolated in mid-eighteenth century human remains of the Joseon period (Oh et al. 2010). From that point onwards, additional

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Fig. 4 Maximum likelihood (ML) tree of Paragonimus ITS2 DNA region. The ancient P. westermani sequences are represented by the red dots (Credit: Jong Ha Hong)

investigations have been conducted on ancient samples from a number of countries. In brief, Myšková et al. (2014) were successful in obtaining the Trichuris SSU rRNA gene from eighteenth to nineteenth century specimens unearthed at a Czech Republic excavation site. Søe et al. (2015, 2018) reported sequences of T. trichiura mtDNA isolated in fifth to eighteenth century samples from Bahrain, Lithuania, Denmark, and the Netherlands (Hong et al. 2019a). Notwithstanding the comprehensiveness of the T. trichiura aDNA data reported to date, it is not sufficient for any definitive conclusions on T. trichiura genetics over a wide geohistorical range. Utilizing mummy coprolite samples representative of the fifteenth to eighteenth century Joseon period, multiple genes of SSU

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rRNA, ATP8, and ITS2 have been analyzed to determine the paleogenetics of T. trichiura (Hong et al. 2019a). According to those phylogenetic results, the Korean T. trichiura sequences belonged to a separate cluster distinct from those of the other genus Trichuris cases reported to date. The DNA markers were found to be useful in the diagnosis of T. trichiura, even in cases where the ancient parasite eggs examined were indeterminate for morphological identification of the species (Fig. 3) (Hong et al. 2019a). Paragonimus westermani: The genus Paragonimus comprises a species-rich group ranging across the tropical regions of Africa, America, Southeast and South Asia, as well as temperate zones such as East Asia and North America (Blair et al. 1999). Almost 50 species and subspecies have been reported for Paragonimus to date. Among them, approximately 16 species have been revealed to cause diseases in humans (Blair et al. 2016). Of the genus Paragonimus, the most commonly reported species is P. westermani, which is prevalent in Taiwan, Korea, China, Japan, and the Philippines (Blair et al. 2016). Scholars conjecture that this species currently infects over 20 million people in the world (Blair et al. 2016; Hong et al. 2019b). In genetic analyses, modern P. westermani have been clustered into at least two different groups (South/Southeast and East Asia) showing distinct geographical distributions (Iwagami et al. 2003; Doanh et al. 2009; Blair et al. 2016). However, as for Paragonimus spp. found in archaeological specimens, very few reports on the relevant paleogenetics have been made thus far. In studies on Paragonimus eggs microscopically detected in the coprolites of the fifteenth to eighteenth century Korean mummies (Seo et al. 2014a, 2017), multiple genetic markers (CO1 and ITS2) were tested for comprehensive understanding of the evolutionary history of P. westermani (Hong et al. 2019b). By NCBI/BLAST analysis, the sequences from Joseon mummies were very similar to P. westermani CO1 or ITS2 regions already reported. Every P. westermani taxon could be differentiated from the other genus Paragonimus spp.; and even the P. westermani sequences were distinctly clustered into several groups. In the case of CO1, P. westermani sequences were clustered into South and Southeast Asia (India, Philippines and Thailand), East Asia (Korea, Japan, and China), and a third group (India and Sri Lanka) (Hong et al. 2019b). P. westermani ITS2 sequences reportedly also have been separately clustered into Southeast Asia (Thailand), South Asia (India) and East Asia (Korea, Japan and China) groups, except for some P. westermani ITS2 sequences from South and Southeast Asia being clustered together with the East Asia group. Actually, P. westermani CO1 and ITS2 sequences of Korean mummies belong to the East Asia group (Fig. 4) (Hong et al. 2019b). Clonorchis sinensis: Parasitologists can diagnose C. sinensis through DNA testing on internal transcribed spacer (ITS: Xiao et al. 2013; Tatonova et al. 2017), nicotinamide adenine dinucleotide hydrogen dehydrogenase (NAD) (Sun et al. 2013; Xiao et al. 2013), and cytochrome c oxidase subunits (CO: Sun et al. 2013). Molecular tests reveal that C. sinensis is genetically distinct from other trematodes (Fig. 5) (Xiao et al. 2013; Tatonova et al. 2017; Hong et al. 2019c). Archaeoparasitologists also have tried to reveal the paleogenetics of C. sinensis by studies on mummy samples retrieved from archeological sites.

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Fig. 5 Phylogenetic analyses of ancient C. sinensis and the other trematodes ITS1 DNA region. The ancient C. sinensis sequences are represented by red dots (Credit: Jong Ha Hong)

Utilizing the coprolite specimens of Korean mummies, researchers have tested CO1, NAD2, ITS1, and NAD5 of C. sinensis aDNA, expanding the dataset to reveal the entire genetic history (Hong et al. 2019c). In the analyses, C. sinensis ITS1, NAD2, and NAD5 sequences were evidently distinct and clustered from those of other trematode species. The usefulness of molecular markers ITS1, NAD2, and NAD5 for differential diagnosis of C. sinensis from other trematode species could thus be reconfirmed (Hong et al. 2019c). In summary, aDNA analyses are based on new techniques developed over the past decades in the field of archaeological science. Genetic analyses on ancient parasite species using Korean mummy coprolites are a meaningful step, because they significantly expand the existing genetic pool of parasite species from Korean mummies we have examined. In our studies, each parasite DNA sequence of Ascaris spp., T. trichiura, P. westermani, and C. sinensis belonged to separate clusters that were evidently distinct from the other parasite species studied to date. The results also showed that the genetic characteristics of each parasite species were not uniform but rather were diverse to some degree. Thus

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too, the results also are successfully proving the usefulness of aDNA analysis in the genetic diagnosis of parasite species. However, it is also true that the number of aDNA cases reported to date, with the resultant data pools, remains insufficient to uncover detailed information on parasite genetics within a much wider geo-historical scope (Hong 2019). Final conclusions on ancient parasite genetics are still pending, awaiting the necessary additional aDNA analyses that will be reported in the coming years.

Other East Asian Mummies Other major achievements of archaeoparasitological study in East Asia have been reported from China, especially with respect to mummies of the Warring State, and the Han, Song, Ming, Yuan, and Qing dynasties. These reports are also remarkable for representing the actual parasitic infection patterns spanning different historical eras in China. The details will be presented in other chapters of this book, which is more specifically dedicated to mummy studies on the Chinese mainland. Note that in the case of Japanese mummies, scientific investigations were performed on them before archaeoparasitological approaches were developed.

Conclusion Archaeoparasitology is now rapidly progressing with the expanding cooperation of parasitologists and archaeologists. Most of the archaeoparasitological research completed in East Asia is owed to the discovery, maintenance, and research of wellpreserved mummies. As alluded to above, Joseon mummies are now of central importance to parasitologists seeking to uncover the parasitic infection patterns of premodern Korean societies. Also, aDNA research focusing on mummy coprolites is very significant to our understanding of each parasite’s paleogenetic patterns in the global parasitological perspective. Notwithstanding this wealth of significant archaeoparasitological data, more detailed research is still required in order to fully comprehend the reality of parasitism in East Asian history.

Cross-References ▶ Dietary Stress in Combat: Coprolite Analysis of a Korean War Marine Killed in Action ▶ Joseon Dynasty Mummies of Korea ▶ Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent Acknowledgments This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (NRF-2019R1H1A2080094).

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Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice

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Karl J. Reinhard, Julia Russ, Isabel Teixeira-Santos, You Zhou, and Vaughn M. Bryant

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Diet and Environment as Components of Pathoecology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Recovering Coprolites and Sectioning Intestines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analyzing Coprolite Residues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Reporting Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Comparing Coprolites from Mummies and Domestic Contexts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Case Studies of Hospice Revealed by Mummy Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analysis Illustrated – The Ventana Cave Mummy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dan Canyon Burial: Focus on Wild Grain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Skiles Mummy: A Debilitated Hunter-Gatherer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Coprolite analysis from mummies offers the opportunity to recover foods related to the last days of life. In some cases, the foods can be interpreted in paleopathological context. This is especially true of the America’s where mummies and

K. J. Reinhard (*) · I. Teixeira-Santos School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE, USA e-mail: [email protected] J. Russ · Y. Zhou Morrison Microscopy Core Research Facility, Center for Biotechnology, University of NebraskaLincoln, Lincoln, NE, USA e-mail: [email protected]; [email protected] V. M. Bryant Department of Anthropology, Texas A&M University, College Station, TX, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_15

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coprolites are most common. We review the data from three mummies. The first is an ancestral Pueblo Mummy from Utah, USA. The remains are of a child. Radiology shows that the individual experienced several periods of growth arrest. The foods provided to the child for several days before death consisted of wild grass. This species of grass has a low nutritional content, especially related to the energy expended to collect the seeds. However, it is available at times when there is a limited variety of foods available. Therefore, the diet can be interpreted as famine diet. A second mummy from the Lower Pecos Canyonlands on the Texas/ Coahuila border shows foods collected for an incapacitated individual. The paleopathological diagnosis for this individual is Chagas disease. This individual was starving for some weeks before death as indicated by stable isotope analysis. However, he was not starving because of the lack of food. He was starving as a result of megacolon and compaction of food in his distended abdomen. The food remains show that his community collected nutritious plants and animals for him to eat. The most noteworthy foods provided were derived from harvesting insects. Analysis shows that these insects were among the most nutritious foods available in this desert. The final case is of a mummified child from southern Arizona. Two separate coprolites show a reliance on a diversity of desert foods. These included mesquite pods, cactus fruits, cactus pads, and traces of other foods. We detail the methods employed for dietary analysis to provide future researchers with a guide as to sampling and analysis. Keywords

Diet · Final foods · Methods · Hospice

Introduction A paper on coprolite sampling by Wood and Wilmshurst (2016) focused on subsampling specimens for multiple lab analyses. These authors present a detailed sampling methodology for coprolites. In some respects, their recommendations provide a system for sampling that acknowledges the continuing advancement of coprolite analysis. However, in comparison to traditional methods, their sampling strategy divides coprolite samples into an unnecessary number of subsamples, at least six. This is especially destructive for small samples, such as ones obtained from mummies. Also, the recommendation to cut away at least 2 mm from the outer circumference of the coprolites and discard it will unnecessarily eliminate potential data. For samples from culturally secure contexts, such as mummies, coprolite samples are simply too small for discard or for making many subsamples. Generally, mummy samples are less contaminated with ambient microfossils. We are taking this opportunity to present an alternative strategy, illustrated with cases of mummies, based on modifications of a standard sequential analysis of coprolites that were first established decades ago but improved over time. This approach maximizes data recovery while preserving coprolites for future research.

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Analysis of Mummy Digestive Tract Contents with Examples of Relevance to. . .

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Especially with regard to mummy studies, researchers must adapt to a variety of preservation conditions. Samples from mummies can be consolidated or unconsolidated, mucilaginous or dry, extremely hard or friable, and miscible in strong bases or weak acids. Therefore, processing can involve a variety of nuanced approaches. Over the years, experiences have accumulated and therefore, we emphasize that it is important to build on those methods of the past and add newer molecular and immunological approaches as they evolve. Gut content analysis has been part of mummy studies for decades and in all regions of the world (Cockburn et al. 1998; Aufderheide 2003). P.V. Glob (1977) describes the results of intestinal tract analysis for some of the first bog mummies recovered. Sakurai et al. (1998) describe the same type of analysis for early research in China. Callen and Cameron (1960) pioneered coprolite and intestinal analysis in South America, and Harrington (1960) presented the first analysis of gut contents for North America. Lindow Man’s intestinal contents were the focus on intense analysis (Hillman 1986; Holden 1986; Scaife 1986). Hillman (1986) summarized the amazing potential of intestinal content analysis in addressing many significant questions. Holden (1994) summarized all aspects of the digestive process in the context of what dietary remains could be expected in mummy intestine analysis. Historical details of coprolite analysis have been published by Bryant (Bryant 1974a, b) and Bryant and Dean (1975, 2006). “Gut-level” data can address many types of data including, but not limited to: the person’s health, the season of death, culinary practices, food storage, geographic origin of a mummy, the social role of the person, medicinal plant use, and sometimes poisonous plant use. There has been a decline in microscopic studies recently. Most post-1995 mummy dietary analysis articles present stable isotopic or trace elemental methods applied on hard tissue, soft tissue, and hair. We contend that the visual analysis of gut contents should be a necessary companion to molecular and chemical reconstruction of ancient diets. We believe that stable isotopic or trace elemental data provide interesting information concerning long-term diet of mummies at a general level, whereas intestinal residue analysis provides detailed information about the amounts and types of foods eaten in the days before death. Importantly, the analysis of gut contents can prevent inaccurate conclusions derived from molecular and chemical analyses (Dickson et al. 2000; Reinhard et al. 2008). Molecular biology has generated increasing interest in the twenty-first century (Hofreiter et al. 2001; Mulligan 2006; Speller et al. 2010; Thomas et al. 2008). Although rarely applied to mummy gut contents, this topic has burgeoned into paleontological applications and archaeological studies. Among archaeological papers, analysis of diet focuses on food source DNA and viral sequences (Poinar et al. 2001; Rivera-Perez et al. 2015). Other papers address verification of human origin (Gilbert et al. 2008; Poinar et al. 2009; Jenkins et al. 2013). Relating to the evolution of diet and effects on human health, gut microbiome studies have come into full analysis (Tito et al. 2008, 2012; Cano et al. 2014; Santiago-Rodriguez et al. 2015; Warinner et al. 2015). Methods are now available in diverse labs so that microscopic and molecular analyses can be applied to the same samples revealing a richer reconstruction of diet (Battillo 2017).

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Diet and Environment as Components of Pathoecology Pathoecology as an interpretive construct is presented by Reinhard and Araújo (2014). Usually, coprolites are analyzed from mummy intestines. Coprolite analysis provides the environmental and dietary context of pathoecology. For example, Arriaza et al. (2010) defined the environmental effects of El Niño events that affected parasite prevalence in intermediate hosts and human diet. They showed how environmental variation defined available food resources, which in turn affected variation in parasite prevalence in Chinchorro mummies from different cemeteries. This was also confirmed by the studies of Jarzen and Bryant (2008) noting the present of helminth eggs in a Chinchorro age (7500–1500 BC) mummy from Peru. Although the term pathoecology was coined in the Americas (Reinhard and Bryant 2008; Reinhard and Araújo 2014), the analysis of mummies in Europe has inculded this holistic pathoecology approach for decades (Holden 1994). The analysis of the Tyrolean ice mummy, Ötzi, represents a particularly wonderful pathoecological approach to the understanding of a single mummy (Oeggl 1996). Among the dozens of published works concerning Ötzi, the analysis of coprolites was noteworthy for the reconstruction of Ötzi’s movement as well as his diet. Ötzi represents a discrepancy between chemistry and intestinal residue study (Oeggl 1996; Oeggl et al. 2007; Macko et al. 1999a, b; Dickson et al. 2000; Rollo et al. 2002). Macko et al. (1999a, b) analyzed the stable carbon and nitrogen isotope rations presence in Ötzi’s hair. They assert, “We have recognized a primary vegetarian component in the diet of the Neolithic Ice Man . . . the δ15N value of 7.0% for the Ice Man clearly points to diet which is essentially vegan” (Macko et al. 1999a:72). Dickson et al. (2000) commented on Macko et al. (1999a) by pointing out that they did not integrate the hair chemistry with previously published literature regarding intestinal residues (Oeggl 1996, 2000; Dickson 1997). They disputed the vegan interpretation based on the discovery of meat residue found in the colon of Ötzi. Dickson et al. (2000:1846) concluded, “When using isotopic analyses to reconstruct diet, the results should not be interpreted in isolation from their wider context. The reconstruction of an individual’s diet must be based on the isotopic values of both the individual of interest and possible food sources.” This is good advice, but it should go both ways (i.e., coprolite analysts should place their data in context of stable isotopic data as well). Only by confronting the contrasts between the diet reconstructions, can we develop an integrated reconstruction of diet (Reinhard et al. 2010, 2012). The analysis of bog mummies in Northern Europe included dietary reconstructions as well as environmental analysis. These have typically been incorporated with pathological and cultural evidence to explain the ecology of ritual sacrifice. Cumulatively, these represent a pathoecological approach to mummies over a broad geographic region. Thus, although pathoecology is not a term used in Europe, the pathoecological approach has been followed by European mummy researchers for many decades.

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Recovering Coprolites and Sectioning Intestines Coprolites, or fragments of coprolites, can be recovered via endoscopic techniques aided by radiography. The mummy should be examined for existing cracks or orifices through which coprolites can be extracted. Standard x-rays can be taken of the mummy to locate coprolites. Coprolites are relatively radiopaque and appear as internal objects of various sizes (Fig. 1). Optimally, three-dimensional computed tomography can be used to image the coprolites in a manner that shows the anatomical location of the coprolites relative to the previously identified cracks or orifices in the mummy (Arguelles et al. 2014). Then it is a matter of using a flexible endoscope to remove the coprolites without damaging the mummy. Ideally, several grams of coprolite should be removed. The amount of information recoverable from a coprolite is related to the size of the sample. Frequently, mummies do not contain coprolites, but do contain loops of their bowels with food contents. These can be sampled after radiographic study defines their presence. These sections can be removed, rehydrated and analyzed (Searcey et al. 2013; Reinhard et al. 2017; Camacho et al. 2018) (Fig. 2). If there are no cracks or expanded orifices in the mummy, it is possible to make an incision near the bowel loops or coprolites. Then the coprolites can be removed. This should be done very carefully with a goal of conserving the mummy. In modern mummy science, conservation must always be the primary concern. If coprolites cannot be removed without compromising the mummy’s internal and external structure, it is best to leave the coprolites in situ and unanalyzed. When attempting aDNA analysis, samples must be secured as quickly as possible after recovery. Molecular biology samples must be recovered by using standard aDNA recovery techniques (lab gloves, face mask, sterile utensils), before other recovery procedures are undertaken, due to the risk of contamination, which potentially could confound aDNA analyses (Malmstrӧm et al. 2015)

Analyzing Coprolite Residues As we mentioned earlier, Wood and Wilmshurst (2016) present a sampling method developed for Quaternary period coprolites. Although their methods acknowledge the needs for molecular analysis, their recommendations do not address diagnostic needs such as enzyme-linked immunosorbent assay. They recommend subsampling coprolites into at least six subsamples. In our experience, mummy samples sent to our labs are quite small, sometimes less than a gram. We advocate the application of several analyses to a single subsample so that the maximum data return can be gained from one sample (Fig. 3). This approach dates back to the beginning of modern coprolite analysis (Bryant and Williams-Dean 1974). It has since been applied to mummies (Reinhard and Hevly 1991).

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Fig. 1 For recovering coprolites from mummies, methods have changed drastically during the past two decades. Two decades ago, coprolites were recovered by autopsy. Images (A–C) show coprolites exposed in Peruvian mummies in 1990 autopsies. Obviously, this method is destructive and is inconsistent with the goal of conservation. Today, radiography can be employed to locate coprolites, if any are present, while preserving the mummy (Kiple et al. 2001). (D, E) show abdominal CT-scan sections of a Chilean mummy in the collection of the University of Nebraska State Museum. These show loops of intact bowel. Adobe Photoshop ® was used to enhance the contrast of D to detail the remains of the intestine. (F) shows coprolites visible in a CT-scan of a Peruvian mummy. Coprolites can be imaged in three dimensions as shown by image G of a Brazilian mummy from the state of Minas Gerais. With such images, analysts can plan recovery procedures for the coprolites that will not damage the structural integrity of the mummies (Image credits, A–C, Karl J. Reinhard; D, E, David Kiple, Advanced Medical Imaging, Lincoln, Nebraska; F, Zanier J.F.C. and B. Tessarollo (Universidade Estadual do Rio de Janeiro); G, Setor de Radiologia do Hospital das Clínicas da UFMG, published in Sianto et al. (2005)

Recovering residues from coprolites can be quite simple or somewhat complicated, depending on where a mummy is found (Holden 1990a, 1994). The simplest methods are associated with the analysis of bog bodies. Water is used to wash out the contents of the viscera, which are separated into macroscopic and microscopic remains. For the bog mummy called the Tolland Man, Glob (1977:32) wrote with regard to the digestive organs, “The organs were carefully rinsed, to remove contamination from the surrounding peat. Their contents were then washed out and proved to consist of a blend of finely reduced plant particles and seeds.”

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Fig. 2 The Searcey method of recovering material from intestinal sections (Searcey et al. 2013; Camacho et al. 2018). The process of rehydrating and processing of the El Chorrillo Mummy Guanche mummy is shown. (A) intestine section removed and ready for rehydration; (B) section 1 h after rehydration; (C) section after 24 h of rehydration showing expanded size and dark coloration of fluid; (D) washing fluid through a 250 μ m mesh, microscopic remains pass into beaker; (E) cleaning exterior of section and washing remains through mesh; (F) cleaned exterior of section, all macro and micro remains from the exterior have been saved for analysis; (G) opening of section to recover internal residues; (H) rinsing of interior of section; (I) gently scraping the interior with a flexible, blunt, plastic spatula to clean but not tear the delicate intestine tissue; (J) recovered macroscopic remains on the surface of a 250 μm mesh; (K) labeled centrifuge tube ready to receive microscopic residues for concentration through centrifugation; (L) process of concentrating microremains from separate beakers of internal and external microfossils (Images by Andrew D. Reinhard)

The simple method that is applicable to bog mummies is impossible to use on desiccated mummies from the Americas where actual coprolites are usually preserved in the large intestine. These will not rehydrate in water alone. Instead, a 0.5% solution of trisodium phosphate (Na3PO4) must be employed to rehydrate the

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Fig. 3 The sequential analysis of coprolites follows this general flow chart. Not all types of macrofossil and microfossil residues are presented here. However, these do represent the most common dietary elements an analyst is likely to encounter in many parts of the world

coprolites before they can be separated into microscopic and macroscopic residues for subsequent analysis (Reinhard and Hevly 1991; Reinhard 1993). However, South American mummies, especially those that are permeated with oily residue from decomposition, pose special problems (Holden 1989, 1990b, 1991a, 1994; Holden and Núñez 1993; Reinhard and Bryant 1995). Oily residues that permeate the coprolites often make them insoluble in trisodium phosphate. In such cases, a brief treatment (1 min) of the coprolites in 1–4% potassium hydroxide (KOH) disperses the oily residue and then the coprolites will respond to traditional trisodium phosphate rehydration. For quantification of microremains, Lycopodium spores are often used as tracers, but these must be added at the beginning of chemical processing (Warnock and Reinhard 1992; Reinhard et al. 2006). Important microremains can include diatoms, phytoliths, starch grains, parasites, pollen, and a variety of other rare remains (Fig. 3). Currently, most Lycopodium tracer spores can be purchased in tablets, each containing a known numbers of spores. Thus, a known number of spores can be added to a known quantity of coprolite while it is in the rehydration solution. Lycopodium sp. is a high-latitude and moisture-loving clubmoss that is not endemic to most areas where desiccated mummies are found. However, in northern latitudes where bog bodies and frozen mummies occur, Lycopodium is endemic and therefore not recommended as a tracer. For those regions, tablets of tropical species of Eucalyptus pollen are often used in the same manner. The rehydrated coprolite can be disaggregated with a magnetic stirrer until the microfossils, macrofossils, and added spores were thoroughly mixed. Once

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disaggregated, the coprolite solution is then poured through a mesh screen with openings of 250–300 μm into a clean beaker. The residue on top of the mesh must be rinsed thoroughly with a jet of distilled water or alcohol to force any remaining microfossils to pass through the screen and into the beaker. The microfossils in the beaker are then concentrated by centrifugation. The numbers of microfossils (pollen, phytoliths, starch granules, parasite eggs/larvae, diatoms, etc.) per gram of coprolite can be calculated using the pollen concentration formula discussed by Maher (1981), that is, microfossils/g will equal the microfossils counted times the total number of Lycopodium spores added divided by the number of tracer spores counted times the sediment weight or volume. Macroremains are larger than 250–300 μm and are transferred to filter papers from the screen where they are dried and identified. Plant fibers, seeds, fruit exocarp, leaves, plant epidermis, insect chitin, animal bones and hair, mollusk and eggshells, and many other types of macroremains can be identified, often to species, from such dried preparations (Fig. 4).

Fig. 4 Coprolite analysts must often be skeptical of their pollen identifications. We developed this flow chart to aid palynologists in critically evaluating their preliminary pollen identifications (Reinhard and Bryant 2007). It shows the sequence of questions the coprolite analyst must pose to each pollen identification

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Microremains are generally those items smaller than 250–300 μm. Analysis of microremains must be done in stages. Slides of microscopic residue must be examined for parasite eggs, phytoliths, starch grains, and any other types of microfossils of interest. These samples must be collected and prepared before the rest of the sample is processed for pollen because the acetolysis solution used to prepare pollen will destroy cellulose, alter starch, and some types of parasite eggs. In addition, other chemicals often used, such as hydrochloric and hydrofluoric acids, will destroy phytoliths and diatoms. For these reasons, we recommend retention of a subsample of the original microremains before continuing with the pollen processing phase (Reinhard and Hevly 1991; Reinhard and Bryant 1992, 1995; Reinhard 1993). When the analysis of microremains and macroremains is completed, then the interpretation phase concerning diet and origin can begin. Interpretation is often more complicated with microscopic remains than with macroscopic remains but integration of both sets of data is essential for complete reconstruction. Microscopic remains can come from a variety of sources. Pollen and spores can come from the foods eaten, inhalation, drinking water, or from the digestive tracts of animals eaten whole (Bryant 1975; Bryant and Holloway 1983; Bryant and Hall 1993). Phytoliths and starch granules most often come from dietary components, but because ancient people also ate small animals, including their digestive tracts, some phytoliths, and starch could arrive as contaminants from animals eaten as food. Diatoms can come from a variety of sources, including drinking water, epidermis of aquatic plants, and from the digestive tracts of small fish. Therefore, interpretation of microfossils must often rely on quantification of the numbers of microfossils as well as comparisons with complementary macroscopic data. Pollen interpretation is often complicated. Several important features related to pollen transit through the human intestinal tract have been demonstrated through experiments related to pollen consumption in modern feces (Dean 2006; Kelso and Solomon 2006). Dean used an experiment in which subjects consumed specific foods containing natural pollen (i.e., broccoli, mesquite flowers, strawberries). The resulting fecal samples were then collected over a period of weeks and each was examined. Kelso and Solomon conducted a similar study except they added specific quantities of pure pollen from selected species to the diets of their subjects. The results of both experiments show that essentially all pollen consumed by humans passes through the intestinal tract undamaged by digestion. Although the contents of cytoplasm in pollen grains are digested, the pollen walls, called exine, are unaltered. One important discovery from these experiments was that after ingestion, pollen grains of different sizes and shapes will travel through the intestines at different rates; some take only days while other types of pollen can take weeks to completely leave the digestive system. Therefore, pollen found in mummies might potentially be consumed days or even weeks before death. In actual studies of mummies, however, a different picture emerges (Reinhard 1993; Oeggl 1996; Rollo et al. 2002; Oeggl et al. 2007). Both Oeggl and Reinhard found evidence that pollen sampled from different regions of mummified intestinal tracts provide different clues about the environment and diet. Reinhard (1993) was able to show that pollen frequencies and pollen concentration values varied

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significantly in different regions of the colon in Peruvian mummies. Oeggl studied the Ötzi mummy and reported, “Sequential sampling of the food residues in the digestive tract of the 5,200-year-old-glacier mummy has made possible the analyses of a series of meals and, from the pollen content, the deduction of the environments in which the last meals were eaten. During his last 33 or so hours, Ötzi crossed different habitats in the Ötztal mountains over considerable distances from high near the timber line (at about 2,500 m), to the zone of warmth-loving trees (about 1,200 m or less), and finally very high in the zone of perennial ice (above 3,000 m).” Quantifying environmental pollen types can be complicated by the prehistoric food habits of ancient peoples. In some parts of the Americas, people ate foods that carried millions of dietary pollen grains (Reinhard and Hevly 1991; Dean 2006; Reinhard et al. 2006). Therefore, the pollen spectrum of many individual coprolite samples could easily be dominated by dietary types in such proportions that it would make finding environmental pollen types very difficult. Typically, archaeopalynologists often count 200 pollen grains per sample. Past research has shown that 200 grain counts are reliable for the statistical representation of the pollen spectrum in a sample (Jones and Bryant 1996; Traverse 2007). However, when one or more dietary types dominate the coprolite pollen spectrum, it might be necessary to count 1,000 or more pollen grains per sample in order to find uncommon pollen types that could offer clues about the environment. An emerging debate among coprolite analysts focuses on determining which pollen types reflect diet, which are medicinal, and which are background pollen types that reflect only the local environment. Recent papers and debates have defined the theory of pollen interpretation, as it applied to coprolites, and these papers are relevant to mummy studies. Chavez and Reinhard (2006) opened the debate by reporting the presence of ten pollen genera that included taxa from medicinal species. The coprolites ranged in age from 8450  80 BP and 7230  80 BP and were excavated from Piauí, Brazil. These same authors presented a theoretical faulttree analysis to determine which plants could have been used medicinally. They recommended that with regard to medicinal plants, the palynologist must ask several key questions of pollen evidence. These questions include: (1) is there evidence of prehistoric pathology that would have required treatment; (2) does the therapeutic property of the plant match paleopathological medicinal needs; (3) is it likely that pollen from the therapeutic plant will be carried by the part used ethnographically for medicine; (4) is it likely that pollen will persist in the prepared use of the medicine; (5) is the pollination strategy of the plant in question (insect vs. airborne dispersal) prone to be underrepresented in the normal pollen rain; (6) can pollen be used to make a precise identification of a medicinal genus or species; and (7) is the amount of pollen present in the coprolite consistent with a medicinal use? Palynologists should not interpret pollen taxa as reflecting medicinal use unless the analyst can address each of these questions in the affirmative. This intellectual construct was modified by us to address purported dietary, environmental, and poison use from the Moche Giant Burials in Perú (Reinhard and Bryant 2007). Geyer and his colleagues (2003) reported the discovery of 34 pollen types in the Moche Giants of poison, dietary, and environmental taxa. The Reinhard/Bryant research group (2007) detected interpretive errors in 28 of

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these pollen identifications (Reinhard and Bryant 2007). From this, they formalized a list of eight common error classes (Fig. 3). To the list of questions that Chaves and Reinhard (2006) posed of their pollen identifications, they added two more. First, is the pollen type of a food taxon endemic to the study area or of a food product that could have been traded to the site? Second, were the laboratory and microscopy facilities contamination-free and adequate for studies of pollen analysis? With regard to other types of microfossil remains, we believe that it would be appropriate to ask these same questions of phytoliths, starch, and other microfossils found in coprolites. Like pollen, these types of microfossils could have their origin in inhaled air, drinking water, earth attached to plant foods, nonplant foods processed with teeth, food residue in animal prey, and/or other sources. Reinhard et al. (2001) identify phytoliths in dental calculus that originated in strips of plant leaves used for textile weaving that were pulled through the teeth as part of preparing the leaves for use. Because similar types of phytoliths were found in coprolites from the region, the data from the dental calculus helped explain the phytolith source as being nondietary. When possible, the study of dental calculus on the teeth of mummies provides a valuable type of analysis. Microfossils from diet and also from plant processing are often trapped in dental calculus (Reinhard et al. 2001; Boyadjian et al. 2007; Wesolowski et al. 2010). By analyzing dental wear and microfossils in dental calculus, and comparing them with microfossils found in coprolites, the behavior patterns that introduce nondietary microfossils into the intestinal tract can be identified. For example, dental wear patterns consistent with coca chewing or plant fiber preparation, which leaves distinct dental wear patterns and sometimes trapped phytoliths in dental calculus, can be identified. The presence of nondietary phytoliths and fibers in coprolites can be better understood with complementary dental analysis. Macroscopic remains can sometimes be introduced into the human digestive system independently of dietary behavior. For example, sphagnum moss leaves most probably ingested accidentally with drinking water were found in the Tollund mummy’s intestine (Fischer 1998). Glob (1977) notes that some of the weed seeds found in the Grauballe bog mummy from Denmark may have come from seeds unintentionally harvested with domestic grains and then were eaten (Asingh and Lynnerup 2007). For peoples who ate animal intestinal contents, Bresciani et al. (1991) warn that large amounts of contamination could be introduced into the mummy from eating animal prey. Generally macroscopic remains are less likely to come from contamination than those found in microscopic remains. Quantification and identification of macroscopic remains are essential (Holden 1994). In general, quantification is based on the numbers of macrofossils found per gram or milliliter of residue. Pearsall (2001) summarized various methods used for quantifying and identifying macroremains. Often, the identification of macroremains can be quite complex while at other times it can be quite easy, depending on the diversity and type of macrofossils encountered and the degree of preservation of the remains. For example, Helbaek (1958) found 63 types of seeds in the Grauballe mummy. Such a wide diversity of seeds is an identification challenge for even the most expert botanists. His previous analysis of the stomach contents of the Tollund

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mummy (Helbaek 1950) was more complicated because many of the seeds had been fragmented by grinding and then degraded by digestion. Holden (1994) presented a method of quantification with a goal of estimating the relative amounts of different foods originally eaten in life. Because of the potential for finding a high diversity of materials with some of them poorly preserved, macrofossil identification should involve a number of steps or stages. First, it is advisable to sort the macrofossils into broad categories such as fibers, bones, seeds, charcoal, leaves, etc. Second, the items in each category must be quantified by counting, by volume, and/or by weight. Third, each macrofossil category should be identified as to its biological origin. For this task it is important to have access to reference examples of seeds, wood, charcoal, fiber, and other such materials. Whenever possible, it is also advisable to have access to reference collections of small animal skeletons that can be used to identify bone fragments found in coprolites. Regardless of the amount of time and effort spent, it is generally inevitable that a portion, and sometimes a large portion, of macroscopic remains will remain unidentifiable. This is regrettable, but this has been a problem since the beginning of coprolite analysis, as noted decades ago by Fry (1977). Molecular biology will make an impact on coprolite analysis in the coming decades (Poinar et al. 2001; Rollo et al. 2002; Thomas et al. 2008). Through this type of study, aDNA sequences of plants and animals can be recovered. Reinhard et al. (2008) recommend that molecular biologists carefully consider their evidence. Chloroplast DNA can come from ambient sources, even pollen. If insects or other small animals are eaten, then the plant sequences might be their digestive system. Rollo and colleagues do an excellent job of this. They report that pine and fern sequences are most likely contaminants from ambient pollen. Sequences of cereal grains indicate a dietary origin. Sequences from ibex and red deer show that Ötzi ate meat from these animals. At this point, the study by Rollo et al. (2002) stands as the model of molecular analysis. They interpreted their sequences in the light of the ecology of Ötzi’s region and in perspective of coprolite analysis.

Reporting Results The final coprolite report should address the basic types of macroscopic and microscopic residues. Typically, in the microfossil section one should present data on the parasites, pollen grains, phytoliths, starch grains, and animal hairs found in the coprolites studies. Occasionally, other types of microfossil items in coprolites would include tiny pieces of plant epidermis, fragments of plant fibers, diatoms, fungal spores, moss residue, and tiny pieces of insects. The macrofossil report should address at a minimum the identification of seeds, leaves, fruit exocarp, stem fragments, charcoal, insect fragments, reptile and fish scales, shell, and tiny stone fragments. Ectoparasites such as ticks, lice, and fleas were eaten in some cultures (Johnson et al. 2008). Even though these types of insects are rare in mummy coprolites, in some areas the remains of those insects would provide insights about the health of the individual under study.

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Once the analysis of coprolites has been completed, the researcher must then interpret what the data mean in a pathoecological sense. Thus, the pathoecology synthesis section should focus on using the macrofossil and microfossil data as keys in an interpretation of the diet, medicine, potential disease, nutritional adequacy, and when possible, clues about the environment. The health of the individuals who produced the coprolite specimens can be determined from the presence or absence of endoparasites, ectoparasites, and dental abrasives. The environmental synthesis of the area where the coprolite was formed can sometimes be derived from the coprolite pollen record revealing the “pollen print” of a certain region. This can be done because each region contains a spectrum of mostly wind-pollinated environmental pollen that is deposited on ground and lake surfaces throughout the year (Bryant et al. 1990). Comparative reference sediment samples collected from the locale where the mummy was found will reveal the “pollen rain or print” for that region. Sometimes, the coprolite analyst is able to infer the potential season of death for a mummy by examining the pollen in the digestive system. If the environment and/or seasonality cannot be determined from the coprolite pollen evidence, then hopefully an analysis of the macrofossil remains might reveal this type of information. Fruits and nuts mature and are most available during certain seasons. The reconstruction of diets comes from the integration of all types of macroscopic and microscopic residues present. Some microfossils and macrofossils are mutually supportive. Ground up fragments of grass seeds and grass pollen in a coprolite could suggest a diet of grass seeds. Likewise, if a coprolite contains manioc (Manihot esculenta) starch grains and also fragments of manioc tuber fiber, then those complement each other and strongly suggest a diet of manioc. Sometimes, dietary use is indicated by one type of residue but not the other. For example, fish bones and fish scales are common macroscopic components of coastal mummies from Peru, but no evidence of fish meat has yet been found in the residues from the same mummies. Alternatively, microfossils such as willow pollen might suggest the use of willow bark and flowers for medicinal use, even though willow leaves or wood are not represented. In the case of medicinal teas, the pollen is often present in the tea if leaves or flowers were used, but often the macrofossils from the same plant sources are rarely present in coprolites (Arguelles et al. 2014). In the final synthesis of coprolite information, all of the various types of data must be considered and explained (Fig. 4). If done correctly, each component becomes an important clue about diet, medicine, health, nutrition, the environment, and even the time of year when the individual died (Kumm et al. 2010; Piombino-Mascali et al. 2013).

Comparing Coprolites from Mummies and Domestic Contexts When possible, the analysis of coprolites from mummies should be compared to other data derived from independent coprolite studies from the same general region. One such study was conducted by Dominguez et al. (1992) to evaluate the suspected starvation diet represented by an Ancestral Pueblo mummy found in the American Southwest. Holden (1991b) compared Chilean mummy gut contents with coprolites

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and flotation samples from domestic contexts. This type of comparative coprolite/ mummy analysis can be very useful for determining if special foods were fed to sick individuals, who then died. Coprolites recovered from a Mimbres skeleton (Shafer et al. 1989) showed that the individual had been chronically ill and was fed a finely ground mixture of maize gruel mixed with mustard leaves and also fed some type of willow tea. Compared to the components found in other coprolites from the Southwestern region, the mummy coprolite was unique. For these types of reasons, the coprolite analyst should always consider that the diet represented in the coprolites recovered from mummies may or may not be consistent with foods ingested by the general population. For this reason, it is important to recover and analyze coprolites found separately from mummies and then use those data as control information against which the diet of the mummy can be measured.

Case Studies of Hospice Revealed by Mummy Analysis The final foods obtained by prehistoric communities and prepared for ill individuals are evidence by dietary study of mummies. Since the ill cannot gather and prepared foods for themselves, the intestinal contents of mummies reveal societal efforts to cure and/or comfort the dying. In some cases, the attempt at cure is hinted in the recovered data. However, the attempt to provide a level of comfort is represented in food, especially foods of unusually high quality evident compared to known diets for the respective culture. We address this aspect of prehistoric hospice with the following case studies.

Analysis Illustrated – The Ventana Cave Mummy The Basin and Range province of the Sonoran Desert in Southern Arizona was home to a long sequence of cultures beginning with Paleoamericans and continuing today with the Tohono O’odham Nation of Southern Arizona. The ranges in this area brought significant seasonal rains, which nourished a wide variety of edible wild plants. Maize farming was introduced relatively early in the region, over four millennia ago. However, the desert’s natural flora provided a wealth of desert succulents, grains, and legumes. Native cultures relied on wild plants for a large part of their subsistence. Therefore, the subsistence pathoecology was especially complex due to the diversity of wild and cultivated plants (Weber et al. 1996). Ventana Cave Burial 5 was of a partially mummified child recovered from the Tohono O’odham Nation of Southern Arizona. The burial was recovered in 1941 and 1942 (Haury 1950) and was curated at the Human Identification Laboratory, Arizona State Museum until it was repatriated. The burial is ascribed to the Hohokam culture and dates between A.D. 1000 and 1450 (Haury 1950). The child was 5–6 years of age at death. It was partially mummified, consisting of a nearly complete skeleton, some soft tissue, and hair. Coprolites were recovered in the 1950s when the body was defleshed so the bones could be examined. The

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coprolites were transferred to Reinhard’s care in 1983 and the results of a first analysis were published by Reinhard and Hevly (1991). In 1995, Reinhard visited with the Tohono O’Odham council to discuss the analysis results. The dietary residues from this analysis are now archived in the University of Nebraska Lincoln Laboratory of Palynology and Pathoecology. Four desiccated coprolites were found. Two were dark colored and granular and two were light brown and fibrous. In this case, two meals were represented in a single mummy. One-gram fragments of each type were submitted for analysis. The macroscopic remains of the dark, granular coprolite consisted of saguaro cactus seed (Carnegiea gigantea) and to a lesser amount the fibers of mesquite pods (Prosopis sp.). The seeds were milled as evidenced by the extreme fragmentation of the seeds. Whole seeds were rare and were imaged when present. When ground, the hilum of each seed tends to break off and preserve in recognizable form. Based on the count of hila and whole seeds, at least 439 saguaro seeds per gram of coprolite are present. Miscellaneous remains including 5 unidentified seeds, 11 insect fragments, 13 mesquite pod fragments, and one grass leaf were encountered. The microfossils from the dark, granular coprolite included very tiny saguaro seed coat fragments, supporting evidence that the seeds were finely ground (Fig. 5). The seeds of the Ventana Cave 5 are morphologically similar to those from C. gigantea and organ pipe cactus (Stenocereus thurberi). Crucially, the seed surface differs between the species. Organ pipe cactus exhibits a ruminate surface, covered with small lozenge-shaped cells with raised edges as illustrated in Figure 2H by Arroyo-Cosultchi et al. (2006). In contrast, saguaro seed cells do not exhibit the raised edges. Cactus pollen was very common. In the dark, granular coprolite, 500,000 pollen grains per gram of coprolite were present. Of these, about 11,000 were from prickly pear (Platyopuntia sp.) and 1,500 were from nondietary, wind pollinated plants. The remaining pollen grains were from a ceroid-type cactus, consistent with C. gigantean (Figs. 6 and 7). Traces of maize pollen (Zea mays) were present. Ceroid refers to cacti with columnar growth forms. Phytoliths of both types of cacti were recovered (Fig. 6). The identifiable cactus pollen and phytoliths were abundant as shown in Fig. 8. This image shows a preparation made directly from the material screened from the macroscopic fragments of the dark, granular sample. Pollen grains and phytoliths were especially abundant and obvious. However, fine fibers and animal hair were also visible. Both cacti are, and were, used as food by Sonoran Desert peoples. The buds develop and the flowers appear in April and June. Seeds from the saguaro fruit are prolific with as many as 2,000 produced in a single fruit. In June, fruits form. Of the plants gathered by southern Arizona peoples in the Sonoran Desert, the saguaro is the most important. The Tohono O’odham collect using long poles to pull the fruits down from the high arms. The fruits are eaten raw and also dried. The fruit pulp was crushed and spread as a jam on other foods. Some fruit was fermented and consumed during ceremonies. Saguaro seeds were parched in the sun and then milled to make meal cakes. Organ pipe fruits were used in the same ways.

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Fig. 5 The cactus seeds found in the mummy are most consistent with saguaro. The two upper images show views of modern seeds. The modern images and seeds are a match for the archaeological seeds in shape and surface microstructure. The lowermost images illustrate the fact that the seeds were finely ground (Credit: Karl Reinhard)

Plant stem tissue was also common, represented by tracheids from xylem vessel tube elements. These spiral structures, composed of lignin, are distinct evidence of plant stem consumption (8). The fibrous coprolite contained only 60,000 pollen grains per gram. Of these, 35,000 are from the ceroid-type cactus noted above and about 6,000 are from prickly pear. The remaining pollen is from nondietary, wind-pollinated plants. Microfossils from the fibrous coprolite included calcium oxalate phytoliths from the two types of

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Fig. 6 In the desert west on North America, pollen and phytoliths are commonly found in microscopic remains. Both reflect plant consumption. For the Ventana Cave mummy, two broad cactus categories are represented. The images on the left show a pollen grain and phytolith typical of ceroid, or columnar, plants such as organ pipe, saguaro, and barrel cactus. The pollen grain and phytolith on the right are typical of opuntioid cacti, including prickly pear. Prickly pear and cholla cacti produce mace-like crystals with long points. Ceroid cactus (those that have a columnar growth form) produce phytoliths composed of aggregates of tabular crystals (Jones and Bryant 1992)

cacti. Spiked phytoliths from prickly pear or cholla were evident as well as plated phytoliths of ceroid cactus. One fragment of plant epidermis, comparable with agave, was observed. Identification was based on comparative examination of modem epidermis sections of species of Agave, Yucca, and Dasylirion. Mesquite fiber was the most common macroscopic component of the fibrous coprolite fragments. Per gram, 46 mesquite pod fragments, two mesquite pod stems, and a mesquite leaf were present (Reinhard and Hevly 1991). Remains of 105 saguaro seeds were also discovered.

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Fig. 7 To identify the source of the ceroid pollen, the pollen from the mummy was compared to pollen collected from modern cacti. The archaeological pollen was shrunken and most grains were distorted. However, the surface morphology of the archaeological pollen compares well to saguaro, Carnegiea gigantea. The upper images show modern saguaro pollen, polar view (left) and equatorial view (right). The middle images show archaeological pollen, polar view (left) and equatorial view (right). The lower images show a mass of archaeological pollen (left) and modern pollen from a flower (right). The clustering of archaeological pollen suggests that pollen was eaten from a floral source (Credit: Karl Reinhard)

The Tohono O’odahm people, formerly known as the Papago, made dietary use of all of the plants found in the Ventana Cave mummy (Hrdlička 1908; Castetter and Underhill 1935; Bell and Castetter 1937; Castetter and Bell 1937; Bruhn 1971; Crosswhite 1980; Hodgson 2001). In the Ventana Cave region, mesquite (Prosopis juliflora) has been a dietary staple for millennia. One of us, Reinhard, commonly ate

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Fig. 8 These low magnification images of microscopic remains from the Ventana Cave mummy show components that are typical of desert North American coprolites including phytoliths, pollen, animal remains, and xylem. Comparing these images to Figs. 6 and 7, ceroid and opuntioid calcium oxalate phytoliths are identifiable. Shrunken and distorted pollen grains are evident. The arrows point to lignin spiral thickenings, tracheids, that are found in the vessel elements of xylem fibers in plants. These are long fibers that conduct water and nutrients. In the lower image, a mammal hair is visible (Credit: Karl Reinhard)

the fresh pods on desert treks. Reinhard occasionally consumed a thick drink made of the ground pods following O’odham recipes. Desert people have long eaten mesquite in this way and undoubtedly the evidence in the Ventana mummy shows that ground mesquite was eaten. In addition, Hodgson (2001:189) summarizes even more native dietary uses of mesquite including eating flowers and sucking nectar. The flowers were also soaked in water to make a sweet beverage. Some tribes chewed on mesquite resin. The trees produce fruit every year and are a consistent source of food. Mesquite pods mature in summer although exactly when depends on elevation but less so on abundance of precipitation. The trees have very long taproots that grow into ground water sources. This accounts for harvest reliability. The main food source is the fleshy interior of the pod that surrounds the seeds. Seeds were and are less commonly eaten. The dried pods were shattered and ground in mortars to make flour. The flour was prepared as an uncooked cake. Bell and Castetter (1937) note bruchid beetle larvae were often in the pods and were ground into the flour and eaten.

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The flour could also be mixed with water and imbibed as a sweet drink or gruel. Gasser (1982:226) reports that “8% of the mesquite pod is crude protein, 52% is carbohydrate, 25–30% is sugar, and slightly over 2% is fat.” He also emphasizes the productivity of mesquite harvest in which a single person can gather up to 175 pounds of dried pods per day and two people grinding can produce 88 pounds of flour daily. Undoubtedly the remains of mesquite in the Ventana Cave child represent the ingestion of mesquite flour. We cannot say whether this was ingested as a drink, stew, or cake. The saguaro had special significance to the Tohono O’odahm Nation and the specific relationship between the cactus and the culture contributed to a long history of research (Hrdlička 1908; Castetter and Underhill 1935; Castetter and Bell 1937; Bruhn 1971; Raab 1973; Crosswhite, 1980; Fontana 1980; Gasser 1982; Rea 1991; Hodgson 2001). Folklore and ceremony are integral to the saguaro harvest from collecting fruit to consuming the wine which is a final product related to the harvest. The wine ceremony is the key event that brings the rain allowing agriculture (Crosswhite 1980:5; Hodgson 2001:103). Tohono O’odham believed that the plants were spiritual beings equivalent to humans in soul (Nabhan 1982). The wine was one of several food products produced from saguaro. Saguaro fruits produced syrup, jam, dehydrated pulp, and seed flour. The harvest season was of special importance to the culture. Only the husk of the fruit was not eaten. The husks were put fresh side up at the harvest site to encourage rain. The fruit juice, pulp, and masses of seeds were collected and processed. The pulp and seeds, when dried, could be stored for months. The Ventana Cave mummy presents one finding that is inconsistent with the ethnography of plant use. The find of pollen and phytoliths that could come from saguaro are enigmatic. The flowers dry and fall off of the fruit when the fruit ripens and splits. The splitting of the fruits signals that it is time to harvest. Also, the husks that contain phytoliths are thrown away. Therefore, if the Ventana cave population were behaving as historic Tohono O’odham, we would not have found the pollen and phytoliths. The source of the inconsistency might be in identification errors on our part. The ceroid phytoliths are shared with many columnar cacti and could have conceivably originated with a different cactus. However, the pollen is consistent with saguaro and we do not think that an error in pollen identification is possible. Thus, we think that it is likely that the community prepared the buds, flowers, or pollen for the child. Buds from cholla were baked and eaten in the region, so a method of cactus bud preparation has been devised in prehistory. The succulent petals and pollen from the flowers could have been eaten directly. We believe that the evidence of 500,000 pollen grains per gram and the presence of aggregates of pollen are strong evidence that polleniferous flowers or buds were eaten. The challenge of harvesting flowers is significant. While collecting flowers for a pollen reference collection, one of us (Reinhard) was impressed at the difficulty of prying flowers off of needle covered cactus while precariously perched on a high ladder. However, to obtain the flowers, the Ventana Cave community must have accomplished this task. Compared to other cacti, saguaro flowers produce an abundance of pollen. The amount of pollen produced per flower was documented by Schmidt and Buchmann (1986). Each flower produces about 2,174 pollen grains. Therefore, the pollen concentrations of

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the Ventana Cave mummy represent the pollen production of about 230 flowers. The collection of polleniferous food for this individual was a remarkable demonstration of care for the child. Since fresh pollen was present in the mummy, it is obvious that the individual died in the saguaro flowering season of mid-May to early July. The peak flowering season is in mid-June to Late-July (Schmidt and Buchmann 1986). An exhaustive review of ceremony associated with saguaro is presented by Crosswhite (1980). For the Ventana Cave child, the fact that saguaro flowers were provided, combined with saguaro’s special folk importance to the culture, suggests that the flowers had ritualistic implications in this case. The analysis of this burial illustrates some key points of dietary reconstruction from mummies. First, more than a single meal may be represented in a mummy. Second, the presence of more than one type of micro- and macrofossil from a single source plant allows for definitive identification of that plant. Quantification of the numbers of micro- and macrofossils provides an idea of how much of the foods were eaten. In this case, the saguaro seed cake and mesquite flour were the major sources of food. When the nutritional values of the foods are referenced, then nutritional data can be inferred (Weber et al. 1996, Winkler 1982). Diverse food sources represent nutritionally rich diet. In this case, the combination of flowers, saguaro seed cake, and mesquite flour show that protein, sugar, fat, and micronutrients were available in the diet. Finally, the season of death can be inferred. In this case, the child died during the summer saguaro harvest.

Dan Canyon Burial: Focus on Wild Grain Ethnographic records show that some foraging groups focus most of their diet, even for weeks, on a single food source when seasonally available. By inference, this is true for prehistoric peoples. Therefore, the intestinal tracts of some mummies might be completely saturated with residue of the same food in all digestive regions, had the individual focused only on one or a few primary food sources. One of us (Reinhard) participated in the 1977 archaeological survey and archaeology conducted close to Dan Canyon. Dan Canyon is a small, south-flowing tributary of Moqui Canyon, which was the focus of our research. In turn, Moqui Canyon was formed by a tributary of the Colorado River. The canyons are cut through a red rock plateau and provided rock shelters and ecological diversity necessary for the mixed subsistence of sedentary Ancestral Pueblo people. The region was once a place of dramatic canyon land natural beauty. Much of this was destroyed by the establishment of the Glen Canyon Dam and Lake Powell, which inundated 108,335 square miles of canyon country including valuable archaeological resources. Besides this direct destruction, the Glen Canyon National Recreation Area, centered around Lake Powell, opened up remote areas that could be vandalized and looted by pot hunters using boats to reach exposed archaeological resources. Indeed, the 1977 archaeological work was designed to salvage archaeological sites from vandals and pothunters.

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The dam was the focus of Edward Abbey’s (1975) The Monkey Wrench Gang, focusing on environmental activists resisting the industrial destruction of the natural beauty of the Colorado Plateau. The disturbance of the Dan Canyon burial was one additional desecration of the cultural environment enabled by the creation of Lake Powell. The declining water level in Lake Powell and specifically in Dan Canyon resulted in the exposure of a partially mummified burial. Wave action of the lowering water level revealed the burial, which was recovered and analyzed with guidance from affiliated Native American tribes. Researchers from The University of Nebraska-Lincoln and the National Park Service conducted the analysis of a partially mummified individual (Dominguez et al. 1992). The burial was that of a child about 3.5 years of age. On March 6, 1991, the research results were presented to the Hopi Tribe and the human remains and artifacts were interred. Research completed by Dominguez’s research group (1992) included to the following observations. The burial was made near a small granary but isolated from any known habitation site. Offerings were present and included a spoon, a datable ceramic canteen, a stone pendent, sandal fragments, and basket fragments. These items and the burial were dated to between A.D. 1210 to 1260. The artifacts signal that the individual was associated with average socioeconomic status and the burial occurred during the final Ancestral Pueblo occupation of Glen Canyon. This was a period of local environmental degradation. Harris lines signaled episodic stress, and were the only pathological condition present. Seven coprolite aggregations, composed of 20 samples in total, were found with the burial. The fact that they do represent intestinal contents is verified by the presence of mummified intestinal wall skin partly covering one of the coprolites. All coprolites are dark colored and granular. Macroscopic analysis showed that all 20 coprolites found in areas of the intestinal tract were composed of caryopses, the grain seeds of grasses. Morphologically, these were consistent with rice grass, Achnatherum hymenoides (previously Oryzopsis hymenoides). The seeds are dense and heavy which is characteristic of A. hymenoides. Also typical for this species, the edible caryopses themselves are surrounded by bracts (scale-like leaves) which are adhering to the seeds. Therefore, bracts were ground with the seeds and can be seen and identified in the macroscopic remains. A. hymenoides caryopses were a common food among the prehistoric people of the American Southwest (Doebley 1984; Huckell and Toll 2004). The caryopses are especially easy to harvest from the plant (Doebley 1984). The plant was very important for Archaic huntergatherers and continued to be a dietary component of later agriculturalists. Analysis of coprolites from across the Southwest shows that A. hymenoides was more commonly eaten by the Ancestral Pueblo who lived in the area of Glen Canyon than in other regions of the Colorado Plateau (Reinhard 1992). Previous coprolite analysis shows that A. hymenoides was never eaten as a sole dietary component (Reinhard 1992). It was, however, often eaten with other plant and animal foods. Therefore, the Dan Canyon child is of interest because only A. hymenoides was present in the coprolites. Microscopic analysis, beyond palynology, was also carried out. No phytoliths or other dietary residues were observed. However, at the time of this analysis, methods for recovering starch grains and then identifying them had not been developed.

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Palynologically, three coprolites were analyzed from different regions of the colon. Pollen analysis of coprolites has been a wonderful source of dietary data for decades (Bryant and Williams-Dean 1975; Reinhard and Bryant 1992, 2008; Bryant and Reinhard 2012). Dietary and medicinal plants not represented by macrofossils are often suggested by pollen. In the case of the Dan Canyon burial, the vast majority of the pollen was from noncultivated grass. The concentration values of grass pollen from these three coprolites, each from a separate area of the colon, varied from 19,900 to 49,600 grass pollen grains per gram of coprolite. Geib and Smith (2008) note that A. hymenoides pollen diameter ranges between 30 and 50 μm. The general monoporate and spherical morphology of the pollen as well as the size is similar to reed grass, little barley grass, and panic grass. Therefore, association of identified seeds is essential to infer the pollen’s botanical source. In this case, the association of A. hymenoides seeds with the pollen is circumstantial evidence that pollen originated with the seeds. This inference is supported by experimental archaeology (Geib and Smith 2008). These researchers collected A. hymenoides grains in June at various places on the Colorado Plateau. They then processed pollen from raw seeds, chaff, and parched seeds. They noted that field burning was effective in harvesting seeds “by igniting piles of cut grass, which produced a shower of small, dark seeds” (Geib and Smith 2008:2087). The resulting chaff and seeds were variable with regard to pollen content. There were 12,180 pollen grains per gram of chaff and 160 pollen grains per gram of seed. For unprocessed, intact seeds, there were 8,490 pollen grains per gram. From their experiments, Geib and Smith conclude, “that grinding crushes the bracts thereby freeing pollen from under these floret structures. Some Paiute groups, for whom rice grass was a food staple, used specialized milling stones in order to crack the bracts free and thereby remove them in a second winnowing (Catherine Fowler, personal communication 1996), which would doubtless greatly reduce the concentration of grass pollen” (2008:2087). This experimental information indicates that the crushed seeds in the Dan Canyon child were processed though one milling and winnowing which left a significant quantity of pollen to be ingested. This suggests that millions of grass pollen grains derived from A. hymenoides were present in the intestine of this child. Thus, it seems that “binging” on grass seeds by this prehistoric individual had resulted in the saturation of the entire intestinal tract with pollen and macrofossils from this one food source. A. hymenoides is a food source and was used by Navajo, Hopi, and the Paiute. The individuals that buried the Dan Canyon child included baskets and leather pouches of A. hymenoides as part of the mortuary ritual, a practice that highlights the value placed on this food source. Low food diversity, in this case, does not mean poor nutrition. Rice grass is one wild grain under consideration for domestication today. Already A. hymenoides flour is produced and is sold as Montina and has a nutritional profile generally similar to wheat flour (Sands et al. 2014). It has 17% protein and for a 100 g serving has 70 g of carbohydrate, 24 g of fiber, and 3 g of fat. There are 380 calories per 100 g serving. Of the protein component of rice grass, 42% is composed of essential amino acids. These values are for modern processed A. hymenoides grain (Sands et al. 2014). However, traditional gathering of the grain in prehistory explored through experimental archaeology shows that the grain was less than ideal (Simms 1985). The

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harvest of ricegrass has a low caloric profitability of less than 400 calories per hour. However, it can be harvested in June before preferable plant resources are available. Therefore, although A. hymenoides grain was a nutritious food for the ailing child, the collection cost for his/her community was substantial. This is a lesson on the commitment communities made to care for members. Dominguez (1992:72) and his colleagues compare the burial assemblage with those from other sites in the region. They conclude that this is a conventional burial for a normal status individual. “The grave goods indicate extensive efforts to provide the child with equipment and food necessary for afterlife, despite nutritional shortages, and imply that this individual was well cared for.” Considering the high nutritional value of rice grass and the high value this plant had to the subsistence of Glen Canyon Ancestral Puebloans, the dietary evidence supports the archaeological conclusion of high expenditure by the community towards guardianship, nurture, and provision of this child.

The Skiles Mummy: A Debilitated Hunter-Gatherer In the a high rimrock overlooking the Rio Grande, Val Verde County, Texas, a 35- to 45-year-old male was interred between AD 636 and 1030 (Turpin et al. 1986). His body was initially laid on its side long enough for the fluid contents of the stomach and intestine to pool in the left side of the thorax (Fig. 9). The abdomen, distended with compacted feces, projected beyond the anterior iliac crests. Internally, the mass pressed up against the spine. All segments of the colon and rectum were swollen to six times the normal diameter as shown in Fig. 10 (Reinhard et al. 2003; Verostick et al. 2018). The mummy was recovered by ranchers in the early twentieth century and preserved in a private museum. It was described in the 1980s by archaeologists who applied the designation of SMM to this individual. The SMM mummy suffered the most extraordinary and excruciating case of Chagas disease found either in archaeological or clinical settings (Matsuda et al. 2009; Reinhard et al. 2003; Barth and Kundrotas 2011; Reinhard and Araujo 2014; Verostick et al. 2018). Chagas disease is a complication of Trypanosoma cruzi infection. In Latin America, an estimated ten million people are infected with a fatality rate of one in ten (Maguire 2006). The SMM case is extraordinary because gastrointestinal symptoms are rare today among populations living north of the equator (Maguire 2006). Also, T. cruzi infection is rare along the Coahuila-Texas frontier. The SMM mummy pathology stirred research into modern infection levels in Coahuila with the recognition that it signaled “an as yet unraveled transmission cycle of T. cruzi in the region” (Martínez-Tovar et al. 2014:173). Reinhard and Araujo (2014) hypothesized an archaeological transmission cycle based on Archaic alteration of the desert landscape that disrupted the cycle of transmission in woodrat nests that resulted in human infections. Heart disease is the more common complication of infection. Both diseases of the heart and digestive system appear 10–15 years after infection. Thus, the pathology shown in SMM reflects a long-term development of disease after initial infection.

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Fig. 9 The Skiles Mummy as presented in 2016. The dark gray objects in the abdomen, wrapped by deer hide straps, are the colon segments completely filled with digesta (Credit: Karl Reinhard)

Fig. 10 This colon segment was selected for analysis as presented in this chapter. It shows that a four-gram sample of the contents was removed for dietary analysis while leaving the intact shape of the colon segment for continued display. The size of the distended colon is evident in this image. Safe handling of the samples was facilitated by a shellac application to all coprolites applied in the 1930s (Credit: Karl Reinhard)

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The presence of megacolon in the region suggests that Chagasic heart disease may have also taken a toll among Archaic hunter-gatherers. In endemic regions, mummies show the pathological conditions of megaesophagus and megacolon. The latter condition is represented by the SMM mummy. The feeding habits of triatomine bugs transmit T. cruzi (Rassi et al. 2010; Martínez-Tovar et al. 2014; Reinhard and Araujo 2014). The insects blood-feed on sleeping hosts. Some species defecate at the site of feeding and the human host rubs the feces into the wound or into the eyes. If the insect feces are contaminated with the protozoa parasite, then the parasites move into the bloodstream. The infection can provoke a reduction of smooth muscle activity with resulting chronic constipation and, eventually, the condition of megacolon. The symptoms in the modern world are not necessarily fatal (Rassi et al. 2010). In early stages, the effected patient can be placed on a high fiber and high fluid diet. Laxatives and enemas are also prescribed. When fecal impaction occurs, manual extraction of the rectal contents is required. Especially in the rectum and sigmoid colon, coprostasis results in fecalomas which are masses of feces that are much harder than normal feces. Comparison of agave phytoliths between the SMM mummy and Hinds Cave coprolites shows that there was sufficient compaction pressure within the SMM colon to fracture phytoliths. This has not been reported in analysis of rhomboid Agavaceae phytoliths from coprolites (Danielson and Reinhard 1998; Riley 2012). Figure 11 illustrates the fracturing of phytoliths in SMM. Obviously, the compaction and blockage suffered by SMM was extreme. Patients with severe conditions can be treated by surgical resection. This option was, obviously, not available to Archaic hunter-gatherers in the area. The earliest published details of symptoms addressed by surgery were presented nearly five decades ago (Todd et al. 1969). In this case, an English woman was diagnosed with Chagasic megacolon after visiting Argentina. Her symptoms expressed over a six-year period were constipation, abdominal distension, and increasing difficulty in swallowing. She was able to defecate only after using large Fig. 11 This image of microscopic remains from the SMM mummy shows phytoliths from Agave spp. The majority of the microfossils in the mummy were fractured calcium oxalate phytoliths. It is likely that this mass of agave accumulated and compressed in the colon some weeks before death which resulted in the fracture of the calcium oxalate crystals (Credit: Karl Reinhard)

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doses of paraffin with disproportionate straining. The interruption in normal digestion resulted in a 14-pound weight loss over the last three years of her symptom history. Medical examination revealed abdominal distension and palpable fecal masses. Endoscopy showed a minimum of 25 cm of extremely expanded rectum and colon. Surgeons removed the sigmoid colon and rectum and the patient recovered. The removed colon section was distended, but not to the extreme distension exhibited by the SMM colon sections (Reinhard et al. 2003; Verostick et al. 2018). The case suggests that SMM may have suffered years of symptoms before the fatal blockage. A second case showing the extent to which a person can endure megacolon is illustrated by a case from Columbia in a man who suffered both heart and colon pathology (Flórez et al. 2010). The cause of death was a stroke. Autopsy revealed Chagasic megacolon and Chagasic alteration of the heart. The relevance of this case to the SMM mummy pathology is that the effected organs are identical. In both cases, ascendant, transverse, and descendant segments of the colon were all dilated with a maximum perimeter of 18 cm and a wall thickness of 1 mm. Although the SMM mummy intestinal wall thickness cannot be measured, the average circumference of the intestinal sections was 18.8 cm (Reinhard et al. 2003). This is very similar to the Columbian case and shows that extreme Chagasic colon is not necessarily fatal by itself. This also shows that SMM may not have died from septicemia due to intestinal bacteria leaking from the enlarged colon. He may have succumbed to associated circulatory conditions related to Chagasic heart disease. At the end of his life, SMM survived at least two, and possibly three, months with totally impaired intestinal function. Analysis of hair sections from the mummy reveals 13 months of diet before death (Verostick et al. 2018). During the last 2-3 months of life, SMM was starving. The previous 10 months showed a normal seasonal variation in isotope rations. This shows that there was a relatively rapid onset of the fatal Chagasic colon episode over 3 months. However, as indicated by the modern cases summarized above, he could have experienced more minor symptoms years before the fatal megacolon event. Archaic Chihuahuan diet has been analyzed through table isotopes (Bousman and Quigg 2006; Verostick et al. 2018). In the Lower Pecos Canyonlands, there was a high reliance on CAM plants. Riley (2008, 2012) verified through coprolite analysis that CAM succulents, especially agave, were essential in the regional diet. The CAM pathway is an adaptation in plants that is beneficial because it allows certain species to minimize photorespiration and thus save water in hot and arid environments. The SMMs colon contents were unusual, relative to the Lower Pecos Canyonlands Archaic diet because the main identifiable dietary items were grasshopper fragments with 33 grasshopper fragments per gram. These were rarely part of the Archaic diet and indicate that SMM was probably provided with special foods near the end of his life. Grasshoppers have very high protein content, exceeding 70% (Ramos-Elorduy et al. 1997). Chagas disease is an emerging infectious threat in Texas today. The SMM mummy shows that it is actually a reemerging infectious disease that provoked pathology in prehistory.

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Conclusions The case studies presented above illustrate the value of interpreting intestinal tract data based on newer perspectives developed in the last few decades. All three cases were analyzed around three decades ago. Reanalysis confirmed the initial findings. The value of the reanalysis of these case studies relates to additional insights derived from new papers in isotope analysis, botany, paleoethnobotany, ethnobotany, palynology, nutrition, and experimental archaeology. When these new data are derived from these sources, the new interpretations are more detailed. The merging of these lines of study resolves a true pathoecological picture of illness and death in an environmental context. The value of reanalysis of mummies has been emphasized by Lynerup (2015) and Asingh and Lynnerup (2007). For the Grauballe Man, which was the focus of this reanalysis, Harild (2007) found that weed seed, with some grain and meat, was eaten before death. This stands in contrast to Helbaek (1958) who reported 63 types of seed in the Grauballe mummy. The more sensational past interpretation of ergot poisoning from previous work was not supported by new studies. We suggest that continued studies of coprolites and mummies include reanalysis of older reports as well as applying as many of the new techniques as possible in analyses of new coprolite and mummy discoveries.

Cross-References ▶ Bog Bodies ▶ Dietary Stress in Combat: Coprolite Analysis of a Korean War Marine Killed in Action ▶ Endoscopy in Mummy Studies ▶ History of Mummy Studies ▶ Mummies, Parasites, and Pathoecology in the Ancient Americas ▶ Radiology Applications in Mummy Science ▶ Radiological and Ethical Considerations of Autopsy in Mummy Study ▶ Stable Isotope Analysis in Archaeological Science and Mummy Studies

References Abbey E (1975) 2000 the monkey wrench gang. Perennial, New York Arguelles P, Reinhard KJ, Shin DH (2014) Forensic palynological analysis of intestinal contents of a Korean mummy. Anat Rec 298:1182–1189 Arriaza BT, Reinhard KJ, Araújo A (2010) Possible influence of ENSO phenomenon on the pathoecology of diphyllobothriasis and anisakiasis in ancient Chinchorro populations. Mem Inst Oswaldo Cruz 105:66–72 Arroyo-Cosultchi G, Terrazas T, Arias S (2006) The systematic significance of seed morphology in Stenocereus (Cactaceae). Taxon 55:983–992

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Jessica Smith, Lauren Gipson, Dario Piombino-Mascali, and Rimantas Jankauskas

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mite Affiliations with Mummies in Forensic Contexts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mite Affiliations with Mummies in Archaeological Contexts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Current Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preliminary Findings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Arthropods have long been an important component of the study of mummified human remains due to their ability to resist natural taphonomic and decompositional processes. However, despite the large amount of research that has been produced regarding arthropod associates of human decomposition, one important taxon of arthropod has largely remained overlooked by the literature: that of the acari, or mites. The purpose of this chapter is to provide a brief overview of published cases in which mites have been found in association with mummified human remains in forensic and archaeological contexts. This chapter will begin J. Smith (*) Department of Anthropology, University of Nebraska-Lincoln, Lincoln, NE, USA e-mail: [email protected] L. Gipson Department of Forensic Science, University of Nebraska-Lincoln, Lincoln, NE, USA D. Piombino-Mascali Department of Anatomy, Histology and Anthropology, Faculty of Medicine, Vilnius University, Vilnius, Lithuania R. Jankauskas Institute of Biomedical Sciences, Vilnius University, Vilnius, Lithuania © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_40

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with a discussion of contemporary forensic cases and historical archaeological cases that feature mites recovered from mummies, as well as a review of other arthropod associates of decay that have been located alongside the mites recovered from these contexts. The chapter will conclude with a section discussing current research being conducted in the field of archaeoacarology with mites recovered from mummies found in a crypt beneath the Dominican Church of the Holy Spirit in Vilnius, Lithuania. Keywords

Acarology · Archaeology · Arthropods · Forensic Science · Mites · Mummies

Introduction Arthropods have long been an important component of the study of mummified human remains. Unlike many other invertebrates, arthropod associates of decay have bodies that are largely comprised of chitin, which allows them to better resist the taphonomic and natural processes of decomposition (Morrow et al. 2016). The corpocenosis (community of organisms found in association with corpses) is comprised of a wide variety of active arthropod decomposers, including various taxonomic families in the order Diptera and various taxonomic families in the order Coleoptera (Morrow et al. 2016). However, despite the large body of research that has been produced regarding this corpocenosis, there is one important taxon of arthropod that remains largely overlooked by the literature, and that is the Arachnida class known as Acari, or mites. Mites are morphologically tiny, but diverse joint-legged invertebrate arthropods of the class Arachnida and are related to spiders and scorpions. Roughly 55,000 species, 5,500 genera, and 1,200 subgenera representing approximately 540 families in 124 superfamilies of mites have been identified and described based on distinct morphological and behavioral characteristics (Krantz and Walter 2009). Many acarologists, however, believe that the estimated number of mite species only accounts for a small fraction of the extant mite species that inhabit earth. Some estimates suggest that the actual number of extant mite species could range from anywhere between 500,000 and 1,000,000 (Krantz and Walter 2009). Due to their noteworthy abundance, evolutionary plasticity, and relatively diminutive size, mites have been remarkably successful at colonizing a wide range of terrestrial, aquatic, and marine environments. Mites are a cosmopolitan taxon adept at flourishing in virtually every habitat capable of sustaining life. They can be found in freshwater, saltwater, oceanic trenches, tropical and temperate forests, grasslands, soil, arctic tundras, deserts, and human domiciles. Similarly, mites are quite adaptable and exploit an extensive variety of microhabitats, or small specialized habitats contained within larger surrounding habitats or ecosystems. The types of microhabitats mites can utilize include, but are not limited to, leaf litter, humus, decaying logs, puddles, lichen and mosses, hydrophytes, algae, beds and bed linens, clothing, skin,

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and fresh and mummified animal carcasses and human corpses. Accordingly, mites occupy many different ecological niches in these microhabitats and are adept at adapting to the ecosystems they inhabit. Because mites can exploit a wide variety of microhabitats, they must practice a broad range of feeding strategies. Mites are known to be predators, parasites, scavengers, carnivores, herbivores, and fungivores and can subsist on anything from plants, bacteria, and fungi to other mites and invertebrates, to the flesh of both human and nonhuman animals. The ubiquitous nature of mites and their propensity for colonizing and feeding on human cadavers have generated a substantial amount of interest from the disciplines of forensic science and archaeology. So much so that, in recent years, forensic acarology has become one of the fastest-growing branches within the field of forensic science. Forensic acarology takes the scientific study of mites one step further by using “information provided by the acari...in the investigations of crime scenes” (Rasmy 2011). Mites from forensic contexts are recovered from corpses in active decay, as well as corpses that have been partially or fully mummified. Previous research conducted within the field of forensic acarology has suggested that mites have the potential to make considerable contributions to modern criminal investigations and can serve as trace evidence and indicators of crime due to their close associations with corpses (Russell et al. 2004; Braig and Perotti 2009; Perotti et al. 2009; Perotti and Braig 2009; Rasmy 2011; Medina et al. 2013; Saloña-Bordas and Perotti 2014). More specifically, this research proposes that mites may be useful in the estimation of postmortem interval, as the life and death cycles of the mites can sometimes provide valuable information on how long a body has been in a specific location; to help determine whether a corpse has been moved or relocated, as mites are often microhabitat-specific (Medina et al. 2013); provide or reinforce information about the types of arthropod decomposers that have visited the body since deposition (Saloña-Bordas and Perotti 2014); and in establishing cases of abuse and neglect. Unfortunately, despite growing interest in forensic acarology, it remains in its infancy as far as research and practical applications go. Archaeoacarology is the study of mites that have been recovered from archaeological contexts. These mites are generally recovered from mummified human remains and associated grave goods. Much like forensic acarology, archaeoacarology is still very new and remains relatively unexplored save for a few isolated papers. Despite the paucity of literature surrounding archaeoacarology, some limited observations have been made about how it might be useful for the study of mummies in antiquity. Prior research has suggested that mites, when analyzed in conjunction with other arthropod associates of decay, could be valuable in providing insights into the taphonomic conditions associated with mummies, the environmental and depositional contexts surrounding the mummies after death, postmortem burial rituals, and environmental conditions, and could also assist curators in assessing the curatorial needs of mummies in their care (Morrow et al. 2015). The purpose of this chapter is to provide a brief overview of published cases in which mites have been found in association with mummified human remains in forensic and archaeological contexts. This chapter will begin with a discussion of contemporary forensic cases and historical archaeological cases that feature mites

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recovered from mummies. These sections will also include a review of other arthropod associates of decay that have been located alongside mites recovered from these contexts. The chapter will conclude with a section discussing current research that is being conducted in the field of archaeoacarology with mites recovered from mummies found in a crypt beneath the Dominican Church of the Holy Spirit in Vilnius, Lithuania.

Mite Affiliations with Mummies in Forensic Contexts The roots of forensic acarology can be traced back to the winter of 1878 when the corpse of a newborn infant was found on the wasteground of the rue Rochebrune in Paris. The corpse was described as having been “covered in some kind of linen cloth soaked with moisture and rotten at the points that made contact with the soil” (Perotti et al. 2009). An autopsy was performed on the body of the infant on January 15, 1878, by the famed French pathologist Paul C. H. Brouardel. Brouardel reported that the body had been completely mummified and was inhabited by various invertebrate arthropods, including mites and butterfly larvae (Benecke 2008). The substantial amount of colonization by arthropods prompted Brouardel to request the assistance of Edmond Perrier, a professor at the Muséum National d’Histoire Naturelle, and Jean Pierre Mégnin, a veterinary surgeon and entomologist. Monsieur Perrier was tasked with the examination of the butterfly larvae, while Jean Pierre Mégnin was asked to examine the mites. During his examination, Mégnin found that the outside of the corpse was covered in “a layer of fine brown powder that is exclusively composed of [casings/shed layers] of the mites and their feces” (Mégnin 1894). Additionally, he noted that the mites found on the outside of the corpse were not alive at the time of discovery but that a “large, writhing, active colony” (Mégnin 1894) of live mites was still present in the interior of the infant’s cranium. Mégnin found that all the mites recovered from the cranium and body belonged to a single species Tyroglyphus longior (Gervais), also known as Tyrophagus longior (Gervais). T. longior belongs to the family Acaridae (Astigmata) and was originally described by the French acarologists Paul Gervais in 1844 (Perotti 2009). T. longior was reported as a mite that is closely associated with dry meats and cheeses (Perotti 2009) but was also known to consume the fatty acids and soapy materials comprised of ammonia that are produced during dry decomposition (Brouardel 1879; Perotti et al. 2009). Mégnin theorized that the colony of mites discovered on both the body and within the cranium must have initially originated from a small number of phoretic hypopial nymphs that were deposited on the corpse by other arthropods (Benecke 2008), such as dipterans, coleopteran, or myriapods. Overall, Mégnin calculated that a population of approximately 2.4 million living or dead mites was present on the body of the infant (Benecke 2008). He also determined that after 15 days the first generation which was assumed to contain 10 females and 5 males matured and developed; after 30 days, 100 females and 50 males were present; after 45 days, 1,000 females and

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500 males were present; and after 90 days, one million females and 500,000 males were present on the body (Benecke 2008). Based on Mégnin’s discovery, as well as the discovery and classification of the butterfly larvae by Monsieur Perrier, the infant was estimated to have died approximately 5–8 months before her discovery. In the following years, Mégnin continued to publish articles about mites (Mégnin 1887, 1889, 1894, 1896) and began developing “his theory of predictable, ecological waves of insect life on corpses” (Benecke 2008), which culminated in the publication of his pioneering work La faune des cadavres: Application de l’entomologie à la médecine légale. In this book, Mégnin expanded and advanced his theory that corpses are exposed to eight successive waves of insects (discussed in more detail below), provided a description of the larval and adult stages for numerous families of insects, and provided drawings of the overall anatomy of the insects as well as drawings that focused on wing venation and posterior spiracles (Benecke 2008). The book also featured 19 case reports describing insects found in association with cadavers. The publications produced by Jean Pierre Mégnin were instrumental in advancing and popularizing forensic entomology, and to a lesser extent forensic acarology, as they contributed a substantial amount of information about the types of arthropod fauna that can be associated with graves and mummified or decomposing corpses. In Mégnin’s system, the first wave of arthropod succession coincides with the fresh stage of decomposition (as described in Damann and Carter 2014) and is characterized by the arrival of species belonging to the families Muscidae, Phoridae, and Calliphoridae. The second wave, which most closely correlates to the bloat or active decay stage of decomposition (Damann and Carter 2014), is characterized by the presence of Calliphoridae and Sarcophagidae. Wave three, which most closely corresponds to the active or advanced stages of decomposition (Damann and Carter 2014), is marked by the arrival of species from the Dermestidae and Pyralidae families. Wave four, which corresponds with the advanced stage of decomposition (Damann and Carter 2014), is characterized by the arrival of species from the Piophilidae, Anthomyiidae, Cleridae, and Fannidae families. Wave five also corresponds to the advanced stage of decay and is marked by the presence of Piophilidae, Lonchaeidae, Muscidae, Phoridae, Silphidae, and Histeridae. The final waves (six, seven, and eight) can be correlated to the skeletal remains stage of decomposition (Damann and Carter 2014). Wave six is marked by the arrival of various species of mites, including those from the families Dinychidae (Uropodidae), Trachytidae, Acaridae, Histiostomatidae, and Glycyphagidae. Wave seven is characterized by the presence of insect families Pyralidae, Tineidae, and Dermestidae. Finally, wave eight is marked by the arrival of species from the families Tenebrionidae and Anobiidae. Over time, the arthropod waves of succession created by Mégnin have been expanded and altered to fully encapsulate the often-transitory nature of arthropod colonization of corpses. The mummified infant case was especially influential because it represented the second case in France’s history in which entomology was used in a forensic context and the first time mites were used to estimate postmortem interval (PMI), thus opening the field of entomology and acarology to forensic investigators.

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After Mégnin’s pioneering work, forensic acarology experienced a considerable lull in its use in forensic casework. For nearly 100 years, forensic acarology fell by the wayside and was overshadowed by forensic entomology. This is because mites are microscopic decomposers that do not share the obvious nature of proliferating insects. It was not until the 1980s that mites were reintroduced as a viable source of evidence for forensic casework and the late 2000s when case reports were published that featured mites recovered from mummified human remains. Pimsler et al. (2016) described a fly-mite association discovered during routine indoor medicolegal death investigations in Texas, USA. The first case described by the research team involved an individual that was found deceased in his bedroom. At the time of discovery, the decedent was reported to be in an advanced state of decomposition and most of the remaining soft tissue was observed to be desiccated and mummified (Pimsler et al. 2016). During the autopsy, the Medical Examiner recovered dipteran (true fly) larvae and pupae from the body. Dipteran pupae were also recovered from the hair and scalp of the decedent. The pupae recovered from the hair and scalp were stored in tangled hair in a honeycomb arrangement to be reared to the adult stage so that they could be identified, while the larvae and pupae recovered from the body were divided into two samples, with half of the sample being preserved and the other half being kept alive (Pimsler et al. 2016). All adults reared from the hair sample were identified as Synthesiomyia nudiseta (Wulp) and displayed a substantial population of mites. Adults reared from the pupal samples collected from the body were identified as S. nudiseta (Wulp) and Sarcophagidae and did not exhibit any populations of mites. Initially, the mites were identified by Pimsler and her colleagues as an unknown species of the genus Myianoetus (Acariformes: Histiostomatidae). Myianoetus spp. includes species that specialize on ephemeral resources, such as dung and vertebrate carrion, and whose appearance in the environment is oftentimes erratic (Russell et al. 2004; O’Connor 2009; Pimsler et al. 2016). This genus of mite is known to be phoretic on flies that belong to several dipteran families, including Sphaeroceridae (Fain et al. 1980), Ceratopogonidae (Fain and Domrow 1980), Muscidae (Negm and Alatawi 2011), and Calliphoridae (Miranda and Bermúdez 2008). Phoretic mites are those mites that use other organisms for transportation purposes, without becoming parasitic to the transporting organism. Phoresy is a common and convenient strategy for mites, as they do not have wings and are often less mobile than their insect counterparts. As with other free-living astigmatid mites, Myianoetus spp. shows a morphologically and behaviorally specialized deutonymph instar which assists in dispersal by allowing the instar to persistently affix to suitable hosts (Krantz and Walter 2009; Pimsler et al. 2016). Later comparisons of the specimens collected in this case with comparative voucher specimens of deutonymphs and adult mites permitted Pimsler and colleagues to identify the mite population associated with S. nudiseta (Wulp) on the samples of hair as Myianoetus muscarum. Reinhard et al. (2017) discovered 2,357 mites and 1,425 mite eggs per ml of sample in tissue associated with the sacrum of a mummified homicide victim in Nebraska, USA. The mummified body examined was that of an adult female who expired abruptly from blunt force trauma to the skull. The decedent was discovered 6 years after her death in her Nebraska residence in the town of Chester, which is

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located on the border of Nebraska and Kansas. The body of the decedent was found unclothed and laying in the prone position on an unsecured section of carpet. Dirt that had accumulated on the carpet had been displaced onto the surface of the decedent’s skin. When found, the decedent was in a near-perfect state of mummification. At the time of autopsy, 100% of the body was covered by preserved skin and the body hair, head hair, and fingernails remained preserved and in place on the body. However, minimal preservation of the internal organs was observed. Reinhard et al. (2017) noted that insect activity was limited on the body, with only 37 dermestidtype burrows being recorded. Unfortunately, because the original goal of the analysis was to determine whether the mummified decedent was killed in or near her Nebraska residence, or if she had been killed at another place and transported back to her residence based on an evaluation of pollen and botanical evidence, the mites were not evaluated for the purpose of identification (Fig. 1). Kamaruzaman et al. (2018) described the biology of a species of Macrochelidae mite found on the corpse of a homeless man. In this case, the decedent was found outdoors on a patch of land close to a popular beach in “Solar Vistahermosa,” Alicante, Southeast Spain. The body was discovered in a faceup position lying on the ground under an umbrella. At the time of discovery, the decedent was fully clothed and was covered by a blanket up to his neck, with only his head exposed to the elements. The cadaver was noted to be in an advanced state of decay, with some partial mummification of the body. During the autopsy, the pathologist collected empty puparia and blowfly adults from the family Calliphoridae, including Calliphora vicina, Chrysomia albiceps, and Lucilia sericata; the larvae, pupae, and puparia of Hydrotaea capensis; the empty puparia, pupae, larvae, and adults of Synthesiomyia nudiseta; the larvae from Fannia scalaris; and the pupae of Conicera tibialis and Puliciphora rufipes

Fig. 1 Microscope image of a mite recovered from tissue associated with the sacrum of a mummified homicide victim in Chester, Nebraska, USA. (Image captured by Reinhard et al. (2017))

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(Kamaruzaman et al. 2018). Mites were also recovered from the body and were sent to the Acarology Lab at the University of Reading for preparation and identification. Two female mites belonging to Macrocheles muscaedomesticae were recovered from the body. M. muscaedomesticae is a cosmopolitan species commonly found in domestic, urban, and semirural habitats (Kamaruzaman et al. 2018; Durkin et al. 2019). This species of mite specializes in rotting organic matter, such as dung and compost (Krantz and Walter 2009). However, M. muscaedomesticae is also known to be a highly synanthropic species of mite and will often inhabit poultry farms (Perotti 1998; Kamaruzaman et al. 2018). Additionally, M. muscaedomesticae is phoretic on other synanthropic animals, with a preference for flies of the Muscidae and Fanniidae families that closely associate with humans. M. muscaedomesticae adults frequently feed on the eggs of Muscidae and Fanniidae, as well as acarid mites (Kamaruzaman et al. 2018). M. muscaedomesticae has only been reported on human corpses in two other cases in Europe. In the first case, M. muscaedomesticae were excised from the brain of a boy following a failed operation in France (Kamaruzaman et al. 2018). In the second case, one female mite was recovered from a human corpse found in a small wooded area on the North Downs in Southeast England (Kamaruzaman et al. 2018).

Mite Affiliations with Mummies in Archaeological Contexts Mites have long been utilized by archaeologists and paleopathologists to help study past populations and archaeological assemblages. Research employing mite remains from archaeological contexts began in the 1960s with coprolites recovered from archaic mummified human remains. Mites retrieved from these coprolites allowed researchers to reconstruct conditions that existed at the time of defecation, identify available food and dietary conditions, and understand the dynamics and ecology of past diseases (Radovsky 1970; Guerra et al. 2003; Baker 2009; Chaves da Rocha and Maués da Serra-Freire 2014). By the 1970s, archaeologists had begun to retrieve and identify mites from human remains recovered during archaeological excavations. However, these early studies did not systematically incorporate the mites into their site analyses but merely mentioned that mites had been recovered (Radovsky 1970). It was not until the 1990s and 2000s that this oversight was corrected, and mites began receiving a more thorough evaluation when uncovered in association with ancient mummified human remains. Baker (1990) was one of the first researchers to rigorously investigate mites found in association with a human mummy excavated from Tarapaca, Northern Chile. The recovered mites were extracted from the gut contents of a mummified individual by archaeologists at the Institute of Archaeology, University of London, and were referred to Anne Baker for identification purposes. Baker was able to identify the specimens as a new species, Lardoglyphus robustisetosus. The mite family Lardoglyphidae includes two genera and nine species (O’Connor 2009). These mites are known to infest stored food products, especially those food products that contain processed animal materials. Due to the high number of adults, nymphs, and hypopi contained within the intestines, it was hypothesized that L. robustisetosus

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was ingested with food infested by the mites and that this food possibly consisted of dried meat or pemmican (Holden and Núñez 1993; Baker 2009). Hidalgo-Argüello et al. (2003) reported on mites recovered from 13 tombs housing the mummified remains of Leonese royalty discovered at the CollegiateBasilica of St. Isidoro in Leon, Spain. Samples taken from the bottom of the tomb, which included bones, hairs, clothing, dust, and teeth, were found to contain both mite eggs and the remnants of adult mites. The researchers were able to identify nonparasitic mites belonging to the suborder Oribatida (described as Oribatei) and the family Aphelacaridae (Aphelacarus acarinus). Mites in the suborder Oribatida frequently occur in the top layer of soil and litter debris, and can also be found on plants, lichens, and mosses. These mites consume a variety of organic materials, including fungi, algae, decomposing plant material, dead collembolans, and occasionally nematodes. A. acarinus mites are a semicosmopolitan species that have been recorded in a wide variety of habitats, including mattress dust, stored grains, soil, bat guano, and the nests of subterranean termites (Baker and Craven 2003; Baker 2009). Other mites recovered from the Leonese samples include those from the superfamily Tarsonemoidea and the subfamily Podapolipidae. Mites of the subfamily Podapolipidae are known to be parasites of arthropods. Their hosts include cockroaches (Dictyoptera, Blattodea), hide beetles (Dermestes ater), and carnivorous ground beetles (Coleoptera, Carabidae) (Baker 2009). Mites from the Cheyletidae family were also found. Cheyletid mites can frequently be found in synanthropic habitats, such as grain stores, nests, farm detritus, and house dust. These mites often predate other mites, as well as the early instars of storage moths (Lepidoptera) and beetles (Baker 2009). While Hidalgo-Argüello et al. (2003) did not elaborate on the possible origins of the mites found in the sample, Anne Baker (2009) hypothesized that the prevalence of mite fragments and the absence of whole, live mites indicate that the infestation of mites was most likely not recent. Rather, Anne Baker (2009) believed that the mites gained access to the mummies and tombs when Napoleon’s troops occupied the basilica in the nineteenth century. During this time, the tombs were broken into and looted, and parts of the basilica were converted into barns and stables for horses. Morrow et al. (2016) reported the occurrence of a variety of mites, and other arthropods, in embalming jars containing the visceral remains of at least two prominent members of the Medici family and the materials used to prepare their bodies after death, such as sponges and cloths. Fragmentary mite remains were found in eight (Samples 2–7, 9, and 10) of the ten samples processed for the study. However, due to poor preservation of the recovered mite fragments and an absence of whole specimens, the researchers were unable to identify the mites taxonomically. In lieu of identification, Morrow and colleagues opted for the quantification of mite densities via counts of mite capitula and eggs observed in the samples. The mites were quantified using the Lycopodium quantification technique (Camacho et al. 2018) frequently employed in palynological and archaeoparasitological analyses. The use of this quantification technique allowed the researchers to demonstrate large mite densities in seven (Samples 3–7, 9, and 10) of the ten samples. Morrow et al. (2016) were also able to establish the presence of a host of other invertebrate

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arthropods that were present alongside the mites in the embalming jars. A total of six out of the ten samples processed contained the remains of other nonmite arthropods. Samples taken from jars 4, 6, and 9, which contained viscera and textiles, included puparia from Hydrotaea capensis (Diptera: Muscidae). Samples taken from jars 5, 6, 7, and 10, which contained body tissue, vegetal remains, and fibers, included phorid puparia. The researchers were also able to recover spider beetle (Coleoptera: Ptinidae) remains from jars 4 and 5. Morrow et al. (2017) recovered the fragmentary remains of mites and pseudoscorpions from grave deposits extracted from burials located in the Ychsma Polity of Pachacamac on the Central Coast of Peru. Sediment samples were taken from 21 burials and rehydrated in 0.5% trisodium phosphate for 48 h. After rehydration, the samples were screened through a 250-micron mesh. Any material larger than 250 microns was subsequently examined for the presence of arthropods. In total, Morrow et al. were able to recover pseudoscorpion fragments from two of the samples, as well as a host of fragmentary mite remains. Based on their findings, the researchers were able to conclude that the decomposition process resulted in a proliferation of both insects and mites. The mites, however, were too badly preserved to allow taxonomic identification. Morrow and her colleagues hypothesize that the poor preservation of the mites can be attributed, in part, to predation by the pseudoscorpions that infested the Peruvian mummies at Pachacamac.

Current Research The Dominican Church of the Holy Spirit sits atop several subterranean crypts containing the remains of clergymen and laypeople, which date back to the eighteenth/nineteenth century AD (Piombino-Mascali et al. 2014). When the crypts were first constructed, they consisted of a principle subterranean area established under the central nave (Piombino-Mascali et al. 2015) with a series of side chambers erected under the sanctuary aisles. During the last three centuries, the crypts have endured several disturbances (Piombino-Mascali et al. 2015). The first of these events occurred when Napoleon’s army established a military hospital in the church (Piombino-Mascali et al. 2015). The crypts were used as makeshift sepulchers, and the corpses of soldiers were deposited within them. By 1844, the Russian Empire had regained control of the church and began chronicling the crypt and its contents. Shortly thereafter, in 1854, a description of the crypts and their history was published by the Dominican Friar Wojciech Wincenty Bagiński (Piombino-Mascali et al. 2015). This publication provided a list of the aristocrats entombed within the crypts. It is also known that during the mid-1800s the human remains contained within the crypts were moved to two side chambers which were subsequently walled up. Additionally, the main accessway to the crypts was sealed. In 1858, the archaeologist Eustachy Tyszkiewicz was granted permission to examine the crypts to determine whether the body of Grand Duke Alexander had been entombed there (Piombino-Mascali et al. 2015). In his report, the archaeologist

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noted that columns of coffins had been created by stacking the coffins on top of one another and that a number of bodies, including those of unburied French soldiers from 1812, lay piled up within the crypt. Hundreds of preserved bodies were documented in the church cellars, with multiple rows of bodies leaned against walls. In 1901, students of the Nazarene Gymnasium were successfully able to reorganize two of the crypts and reinter some of the scattered bones in coffins. In 1906, noted historian Władysław Zahorski provided in-depth descriptions of the bodies, textiles, and coffins located in the crypts. However, at this time, Zahorski also reported that the corpses of the Napoleonic soldiers were no longer present. In the 1930s, during Lithuania’s occupation by Poland, more extensive efforts were made to organize the subterranean vaults by students at Stefan Batory University (Piombino-Mascali et al. 2015). During this time, debris was removed from the chambers. Coffins were moved from the side rooms to the main crypt and historical documents were recovered, which included the dates for coffins from the late seventeenth and eighteenth centuries. However, work on the project was terminated before its completion. By 1941, National Socialist Germany had assumed control over Lithuania during World War II (Piombino-Mascali et al. 2015), and in 1944, the crypts were designated as bomb shelters. In 1962, during Soviet occupation in Lithuania, there was renewed interest in cataloging and preserving the contents of the crypts by government officials (Piombino-Mascali et al. 2015). In late 1962, it was reported that the bodies contained within the crypts were in bad condition due to contact with air. The infiltration of air was linked to the presence of a storeroom for vegetables. At the same time, concerns over contamination of the crypts due to the presence of mice and flies were expressed (Piombino-Mascali et al. 2015). The committee of government officials overseeing the project decided that closing the chambers and sealing the mummies behind a glass window would be advisable. Thus, the mummies were sealed. Beginning in 1963, artifact information was collected, and coffin hardware and associated textiles were documented by a team of historians, ethnographers, and forensic scientists. Some articles of clothing and religious artifacts found in the crypts were also collected for exhibition. At this time, forensic scientist Juozas Markulis registered the remains of 500 individuals present within the crypts, with 200 of these individuals found to have undergone mummification. Analysis of these individuals showed that mummification occurred spontaneously due to low constant temperatures and environmental ventilation (Piombino-Mascali et al. 2015). Some bodies were mummified in the early stages of decomposition and exhibited bloating or adiopocere formation. By 1967, conditions in the crypt had significantly deteriorated. Markulis noted at the time that it felt as if humidity levels within the crypt had drastically increased. It was also reported that fungal-induced decomposition of the soft tissues on some of the bodies had initiated (Piombino-Mascali et al. 2015). Appeals were made to the ministry of culture to reestablish the initial environmental conditions by restoring airflow to the crypts. However, no actions were taken to save the mummies, and the site was eventually closed.

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Finally, in 2004, an anthropological survey of the chamber containing the greatest number of corpses was conducted (Piombino-Mascali et al. 2015). This study revealed that all mummies contained within the crypts were decomposing. Sealing the mummies behind glass had been ill-advised. In 2008, efforts to conserve the remaining mummies were made. In 2011, officials from the Dominican Church of the Holy Spirit requested research be performed that enabled the analysis, documentation, and preservation of 23 mummies and their associated body parts (Piombino-Mascali et al. 2015), thus leading to the creation of the Lithuanian Mummy Project. This project was designed to allow researchers to analyze and document various aspects of the lives of those individuals interred in the crypt, as well as provide analyses and documentation of the environment surrounding them in death. The investigations performed by the Lithuanian Mummy Project have become instrumental in uncovering information about the decomposition process of the mummies and provide a framework for surveying the organisms that can potentially operate as active decomposers to the mummies housed within the crypts. The University of Nebraska’s Pathoecology Laboratory is currently researching mummified tissue samples that were recovered and submitted by the Lithuanian Mummy Project for analysis. Previous research indicated that the mummified tissue samples were infested with a wide diversity of mites. This finding was confirmed by student volunteers during the fall of 2018 when the superficial layers of the mummified tissue samples were dusted for adherent detritus. The detritus collected from the tissue samples was then microscopically examined for the presence of mites. Both whole and fragmentary mites were discovered during this preliminary examination. This research project constitutes a pioneering study on mites that are associated with corpses. As such, four primary goals were derived for this project. The first goal seeks to ascertain whether different tissue samples will exhibit a differential diversity and abundance of mite assemblages related to coffin microhabitats. The second goal is to evaluate whether the mummies were thoroughly infested with mites to determine whether the mites colonized the mummies early in the history of the crypt or later when the mummies were disturbed and damaged. The third goal seeks to establish whether a brown powdery substance found on some of the mummies is mold, as it was previously identified, or actually layers of mites. Finally, the fourth goal of the study will be to attempt to identify whether predatory mite species and grazers both appear in the samples, which will indicate that a community of mites infested the corpses for a substantial amount of time. Moreover, a survey of other arthropod remains found in association with mites will be conducted. The novelty of this research project has also required the researchers to experiment with multiple recovery techniques to find the ideal method for extracting mites from mummified tissue. Early attempts at extracting the mites involved micropipetting individual mites from processed samples and placing them onto microscope slides. However, this method proved too time-consuming and was abandoned. Additionally, kerosene-floatation was attempted (Proctor 2001) but was found ineffective due to the abundance of lipids present in the samples. The current method being applied is soaking the mummified tissue in 0.5% trisodium phosphate and then screening the samples through a 50, 160, and 250-micron meshes. The resulting

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mites are concentrated by microcentrifugation into size classes based on mesh size. Microscope mounts are made with either glycerin or Hoyer’s solution for slide preparation. Slides are then viewed using a Nikon Eclipse microscope with an Infinity Image Capture system. A further attempt to mount mite specimens for scanning electron microscopy was made to facilitate more accurate identification of the recovered mite species.

Preliminary Findings The recovery technique described above has allowed researchers to successfully recover a wide array of both whole and fragmentary mite remains, as well as mite eggs, from the mummified tissue samples. So far, only two of the recovered specimen types have been positively identified to the family level. However, with the use of SEM, the researchers anticipate more identifications in the near future. The first specimen recovered belonged to the family Glycyphagidae (Fig. 2). Glycyphagidae is the largest in the superfamily Glycyphagoidea with 41 genera and 192 species (Krantz and Walter 2009). Mites in this family are known to occupy the nests of vertebrates, with specific species preferring the nests of rodents, insectivores, and didelphimorph marsupials that inhabit the New World. Glycyphagidae often exhibit a variety of body morphologies and setal form. However, virtually all mites within the family Glycyphagidae will display a pattern of external ridges on the venter surface of the subcapitulum (Krantz and Walter 2009). Other characteristics that are generally shared among Glycyphagidae include a rounded, soft body; extended and barbed idiosomatic setae; a cuticle that is covered with pointed microtrichia or lobed protrusions; elongated tarsi; and a shield-like, reduced, or absent prodorsal sclerotization (Krantz and Walter 2009). Some species in this family have also been found to colonize human domiciles, where they make up a significant part of the acarofauna of both house dust and stored food products (Krantz and Walter 2009). In recent decades, members of the Glycyphagidae family have been implicated as a source of clinically significant allergens, as well as occupational allergens among agricultural workers, bakers and pastry chefs, shopkeepers, cheesemakers, horse riders, millers, etc. The second specimen recovered belonged to the family Tarsonemidae (Fig. 3). The Tarsonemidae is a family of mite which was established in 1877 (Baker and Wharton 1952). Tarsonemidae has since been reexamined and reassigned into a complex system of super- and subfamilies. These families are based upon morphological and biological factors; for example, the subfamily of Podapolipidae is classified by its parasitic relationship with insects. Other subfamilies are classified on parasitism with plant species or necrophagous species. These classifications were established in 1939 by Henry E. Ewing. In 1953, Robert E. Beer rearranged genera to different families. He then defined Tarsonemidae with the addition of the subfamily Tarsonemidae which were comprised of two of Ewing’s subfamilies; Beer also included three other genera: Steneotarsonemus, Xenotarsonemus, and Rhynchotarsonemus.

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Fig. 2 Microscope image of a Glycyphagidae mite recovered from the Lithuanian mummy samples. Image captured by Jessica Smith

Tarsonemidae mites have been affiliated with agriculture since 1877 when the Steneotarsonemus bancrofti was associated with sugarcane plants. Many other Tarsonemidae mites, as well as the S. bancrofti, have been implicated in the agricultural world as pests. Those produce often associated with mite infestation include strawberries, pineapples, mushrooms, and other fungi (Ewing 1922; Baker and Wharton 1952; Jeppson et al. 1975). This family of mites can also be parasitic to insects, small and large animals, or even humans (Jeppson et al. 1975). Tarsonemid mites are generally very small creatures ranging anywhere from 100 to 300 micrometers, with males being much smaller than the females. Females of the Tarsonemidae family tend to be more oval in body shape, while the males tend to be rounder. The bodies of these mites are called the idosoma which is transected by a suture. This suture divides the anterior two pairs of legs and the posterior two pairs of legs. The anterior section of the body is called the propodosoma, while the posterior is known as the hysterosoma; the latter section is further divided into the metapososoma and the opisthosoma. Males of the tarsonemid mites have genital papilla or plates which contain aedeagus. Females of the species have special organs located between coxae 1 and 2. These organs are referred to as clavate sense organs, or pseudostigmatic organs and are most likely specialized sense organs (Jeppson et al. 1975). The mites also have setae which, on the females, connect to the fourth set of legs with a whip-like shape. The mouthparts of a tarsonemid mite are

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Fig. 3 Microscope image of a Tarsonemidae mite recovered from the Lithuanian mummy samples. Image captured by Karl Reinhard and Jessica Smith

located in the capsular head known as the capitulum (Jeppson et al. 1975). The capitulum contains a pair of chelicerae which, in some species, are inserted into the plant cells for feeding purposes, as well as paired palpi (Jeppson et al. 1975; Denmark 2000). The classification of mites in the Tarsonemidae family for the males is based primarily on the characteristics of the posterior legs in the males due to the variability of these features. The fourth pair of legs in this species are considered accessory copulatory appendages due to their use in the mite’s mating and premating behaviors. These legs are also generally separated into four segments, although a fusion of the tibia and tarsus can occur giving the appearance of three segmentations. Claws at the end of these legs can also vary from large, pronounced claws to almost nonexistent. The femur may also vary with the species as spurs and other modifications may be present to help with mating as the fourth pair of legs has little to do with movement. Setae may also be used to classify species within the family as the morphology and location of these appendages may range from normal characteristics to modified characteristics. Normal is classified as narrow at the base to tapered at the ends, while modified may include various other shapings (Jeppson et al. 1975). The life cycle of the tarsonemid mites has four separate stages (Baker and Wharton 1952; Jeppson et al. 1975). These stages include the egg, larva, quiescent

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nymphal stage, and the adult (Baker and Wharton 1952). However, it was originally thought that these stages were separated into egg, larva, immature female, adult male, and adult female (Ewing 1922). The latter stages were combined into a single category, and the immature female was renamed as the nymph stage (Ewing 1922). Female mites can only lay one egg per reproductive event, and the egg tends to be relatively large in comparison to the female with a white opaque appearance (Baker and Wharton 1952; Jeppson et al. 1975). Once the mite is ready to emerge from the egg, it hatches as a white opaque six-legged larva with the males being much smaller than the females (Jeppson et al. 1975). After this stage is complete, the mites enter a pupal or nymph stage where the mites transform into adults with the development of the genitalia and the fourth pair of legs (Jeppson et al. 1975). Once these features have developed, the mite is classified as an adult and may range in color depending upon its food sources (Jeppson et al. 1975).

Conclusion Like insects, mites have been discovered in a wide variety of forensic and archaeological contexts featuring mummified human remains. Their accurate identification has aided forensic scientists in estimating postmortem interval, determining whether a body has been moved or relocated, and reinforced information about the types of arthropod decomposers that have visited the body since deposition. Likewise, mites have also assisted archaeologists by providing valuable insights into the taphonomic conditions associated with mummies, the environmental and depositional contexts surrounding the mummies after death, postmortem burial rituals and environmental conditions, and the curatorial needs of mummies housed in museum collections (Morrow et al. 2015). Despite their ubiquity and the potential value they hold for analyzing crime scenes and archaeological assemblages, mites have rarely been utilized for these purposes. This is due in large part to two factors. The first factor is the difficulty in extracting mite specimens from recovered human remains. The second factor is the lack of experience among archaeologists and forensic scientists with mite taxon. Current research being conducted in the fields of forensic acarology and archaeoacarology is helping to remedy the first factor. The second factor, however, might prove more difficult to resolve. One suggestion for future researchers in the fields of forensic acarology and archaeoacarology is to focus their endeavors on constructing user-friendly dichotomous keys that make it easier for those who are unfamiliar with mites, and their associated anatomy and taxonomy to identify them when they are discovered during crime scene or site analyses.

Cross-References ▶ Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice ▶ Archaeoparasitology of Korean Mummies ▶ Bog Bodies and Natural Mummification of Siberia

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Mégnin JP (1889) Entomologie appliquée à la médecine légale à propos de la thèse de M. Georges Yovanovitch. Soc de Méd Lég de France Bull 21:249–251 Mégnin JP (1894) La faune des cadavres, application de l’entomologie à la médecine légale. G. Masson, Gauthier-Villars et fils: Paris Mégnin JP (1896) Note sur une collection d’insectes des cadavres, intéressants à connaître au point de vue médico-légal, offerte au Muséum. Bull Mus Natl His Nat 2:187–190 Miranda R, Bermúdez S (2008) Mites (Arachnida: Acari) associated with Calliphoridae flies (Diptera: Oestroidea) at three localities in Panama. Rev Colomb Entomol 34:192–196 Morrow JJ, Baldwin DA, Higley L, Piombino-Mascali D, Reinhard KJ (2015) Curatorial implications of Ophyra capensis (Order Diptera, Family Muscidae) puparia recovered from the body of the Blessed Antonio Patrizi, Monticiano, Italy (Middle Ages). J For Leg Med 36:81–83. https:// doi.org/10.1016/j.jflm.2015.09.005 Morrow JJ, Myhra A, Piombino-Mascali D, Lippi D, Roe A, Higley L, Reinhard KJ (2016) Archaeoentomological and archaeoacarological investigations of embalming jar contents from the San Lorenzo Basilica in Florence, Italy. J Arch Sci Rep 10:166–171 Morrow JJ, Taylor L, Peck L, Elowsky C, Owens LS, Eeckhout P, Reinhard KJ (2017) Pseudoscorpions of the family Cheiridiidae (Arachnida: Pseudoscorpiones) recovered from burial sediments at Pachacamac (500–1,500 CE), Perú. J Arachnol 45:370–375 Negm MW, Alatawi FJ (2011) Four new records of mites (Acari: Astigmata) phoretic on insects in Riyadh, Saudi Arabia. J Saudi Soc Agric Sci 10:95–99 O’Connor BM (2009) Astigmatid mites (Acari: Sarcoptiformes) of forensic interest. Exp Appl Acarol 49:125–133. https://doi.org/10.1007/s10493-009-9270-2 Perotti MA (1998) Moscas sinantrópicas (Diptera: Muscidae y Fanniidae) en producciones avícolas del sudeste bonaerense. Nat Neotrop 29:145–154 Perotti MA (2009) Mégnin re-analysed: the case of the newborn baby girl, Paris, 1878. Exp Appl Acarol 49:37–44. https://doi.org/10.1007/s10493-009-9279-6 Perotti MA, Braig HR (2009) Phoretic mites associated with animal and human decomposition. Exp Appl Acarol 49:85–124. https://doi.org/10.1007/s10493-009-9280-0 Perotti MA, Goff ML, Baker AS, Turner BD, Braig HR (2009) Forensic acarology: an introduction. Exp Appl Acarol 49:3–13. https://doi.org/10.1007/s10493-009-9285-8 Pimsler ML, Owings CG, Sanford MR, O’Connor BM, Teel PD et al (2016) Association of Myianoetus muscarum (Acari: Histiostomatidae) with Synthesiomyia nudiseta (Wulp) (Diptera: Muscidae) on human remains. J Med Ent 53(2):290–295. https://doi.org/10.1093/jme/tjv203 Piombino-Mascali D, Kozakaitė J, Tamošiūnas A, Valančius R, Panzer S et al (2014) Skeletal pathological conditions of Lithuanian mummies. Pap Anthropol 23(1):118–126. https://doi.org/ 10.12697/poa.2014.23.1.10 Piombino-Mascali D, Urbanavičius A, Daubaras M, Kozakaitė J, Miliauskienė Z, Jankauskas R (2015) The Lithuanian mummy project: a historical introduction. Lietuvos archeologija 41:131–142 Proctor HC (2001) Extracting aquatic mites from stream substrates: a comparison of three methods. Exp Appl Acarol 25:1–11. https://doi.org/10.1023/a:1010677700404 Radovsky FJ (1970) Mites associated with coprolites and mummified human remains in Nevada. Contr Univ Calif Archaeol Res Facil 10:186–190 Rasmy AH (2011) The humans lie but the spiders do not lie: an overview on forensic acarology. Egypt J Forensic Sci 1(3–4):109–110. https://doi.org/10.1016/j.ejfs.2011.07.001 Reinhard KJ, Milanello do Amaral M, Wall N (2017) Palynological investigation of mummified human remains. J For Sci 63(1):244–250. https://doi.org/10.1111/1556-4029.13463 Russell DJ, Schulz MM, O’Connor BM (2004) Mass occurrence of astigmatid mites on human remains. Abh Ber Naturk-mus Gorlitz 76:51–56 Saloña-Bordas MI, Perotti MA (2014) First contribution of mites (Acari) to the forensic analysis of hanged corpses: a case study from Spain. Forensic Sci Int 244:e6–e11. https://doi.org/10.1016/j. forsciint.2014.08.005

Ancient Parasites Seen in the Archaeology and Medical Contexts in the Han Dynasty, China

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Xiaoya Zhan, Wuyun Qi, and Hui-Yuan Yeh

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Backgrounds of the Han Dynasty (202 BCE to CE 220) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Parasites in the Han Dynasty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Roundworm (Ascaris lumbricoides) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tapeworm (Taenia sp.) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pinworm (Enterobius vermicularis) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Whipworm (Trichuris trichiura) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Flukes in the Han Dynasty: Oriental Schistosomiasis (Schistosoma japonicum) and Chinese Liver Fluke (Clonorchis sinensis) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

The Han Dynasty (202 BC–CE 220) is regarded as the golden age of ancient China for its significant contributions and deep imprintson history. The health condition during the era of the Han Dynasty has been one of academia’s focuses. With the emphasis on the Han Dynasty, this chapter integrates the knowledge of both archaeological contexts and traditional Chinese medical texts to discuss parasitic loads and the mechanisms behind these infections. By integrating the findings of paleoparasitology with historic textual sources, we present the six species that were identified by archaeological reports, namely, blood fluke (Schistosoma japonicum), Chinese liver fluke (Clonorchis sinensis), pinworm (Enterobius vermicularis), whipworm (Trichuris trichiura), tapeworm (Taenia sp.), and X. Zhan · H.-Y. Yeh (*) School of Humanities, Nanyang Technological University, Singapore, Singapore e-mail: [email protected] W. Qi Institute of Archaeology, Chinese Academy of Social Sciences, Beijing, China © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_41

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roundworm (Ascaris lumbricoides). Meanwhile, this chapter aims to examine the possible factors that had contributed to the parasitic infections during the Han Dynasty through different aspects, including dietary habits, daily behaviors, and economic activities. Possible factors such as the consumption of raw meat, poor hygiene, the application of night soil as fertilizer, and the physical proximity of pigpens and human latrineswould all contribute to parasitic infections during the Han Dynasty. Lastly, this chapter addresses the medical knowledge used to target parasitic diseases in the Han Dynasty. Keywords

Paleoparasitology · Han Dynasty · Early China

Introduction The Han Dynasty (202 BC–CE 220) is regarded as the golden age of ancient China for its political, economical, and cultural contributions. The deep imprints left behind by the Han Dynasty are significant until this very day. The ethnic Chinese still refer to themselves as “Han people” as an echo of the golden age, and “Han character” is a synonym for Chinese characters (Smith 2008). Significant progress and development of production activities, as well as myths and faiths, characterized the Han Dynasty as a highlight of China’s history. Several aspects, including the widespread application of iron agricultural tools, usage of cows to plow the fields, irrigation systems, and a nationwide shift in focus to agriculture as instituted by the government, contributed to the formation of this mighty empire. These factors built a solid foundation for the Han Dynasty and thus allowed this empire to achieve significant progress and development in production activities (Xu 2005). The founding of the Silk Road by Zhang Qian is another symbol of the Han Dynasty’s status as a growing empire. The interaction between the West and the East resulted in the introduction of new agricultural productions within China, while the West began to import products such as tea, silk, and porcelain from the East. However, such transnational interactions, despite being beneficial for the economy, heightened the risk of parasites and other related infectious diseases being introduced to the country. With an increase in population and rapid development in sedentary agriculture, the risk of catching infectious diseases among groups was exacerbated. Moreover, some infectious diseasesoriginated from animals which were in close contact with people, such as smallpox, tuberculosis, and hives from cattle; pertussis from pigs and dogs; and influenza from pigs and ducks (Diamond 2017).In addition, human activities such as migration accelerated the spread of infectious diseases. Studies have suggested that human activities via both land and sea trading routes along the ancient Silk Road were responsible for the wide spread of infectious diseases, such as the plague known as Black Death, that took place between Asia and Europe (Schmid et al. 2015).

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Among the diseases that were transmitted between groups were parasitic diseases, a unique kind of infectious disease. Parasites reside within humans and accompany them throughout the evolution of history. To understand the health of the people in the past and channels used in the spread of infectious diseases, it is useful to observe and analyze the eggs of parasites (Mitchell 2013). This is because the eggs of parasites are easily identifiable through a microscope and are often well-preserved in archaeological contexts. Meanwhile, multidisciplinary researches, including paleoparasitology, zooarchaeology, archaeobotany, archaeopalynology, starch analysis, and phytolith analysis, could reconstruct the dietary habits of ancient residents and offer a paleoecological perspective of the past (Reinhard and Bryant 2008). The Chinese have had a long history of struggling with parasites. The earliest finding of parasitic eggs from archaeological contexts dates back to the Neolithic Age (Zhang et al. 2006). Though paleoparasitological studies in China are limited compared to that of the Americas or Europe, current reports have shed light on the spread of infectious diseases and health conditions in ancient populations. This chapter focuses on parasites in the Han Dynasty, especially intestinal parasites, through ancient medical books and archaeological findings which depict the past health conditions of people and human activities. At the same time, this chapter hopes to provide a better understanding of the mechanisms behind the infection of parasites.

Backgrounds of the Han Dynasty (202 BCE to CE 220) The Han Dynasty was the second imperial dynasty in Chinese history and characterized by two phases, Western Han (202 BCE to CE 8) and Eastern Han (CE 25 to CE 220). The transition from the Western Han to the Eastern Han was marked by Wang Mang’s usurpation (CE 9 to CE 23) and Gengshi Emperor’s Period (CE 23 to CE 25). The Han Dynasty solidified and maintained the unification founded by the Qin Dynasty (221 BCE to 207 BCE), and the Han Dynasty achieved significant progress in different aspects such as agriculture, handicraft, military, and culture with the benefits of unification. One of the main achievements achieved by the Han Dynasty was the formation and development of the agrarian economy. The Warring States Period (fifth century BCE to 221 BCE) had established the basis for this tremendous agricultural development in the Han Dynasty in several aspects, including the wide application of iron farming tools, irrigation systems, and the emphasis on agriculture from Confucianism. The Han Dynasty inherited these advantages and continued with intensive farming (Xu 2005). Historical records suggest that multiple grains were cultivated and consumed during the Han Dynasty, including millet, wheat, barley, rice, beans, and hemp. Preservation of such grains in pottery barn models from Han tombs proves the existence of these grains (Xu 2005). Moreover, isotopic analysis indicates the dominance of C4 plants (millets) in the northern area and the increase of C3 (wheat and rice) plants in daily diets moving into the Han Dynasty; meanwhile, ancient residents in Southern China might have started to cultivate and consume

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wheat earlier than in Northern China (e.g., Hou et al. 2012; Zhang et al. 2013; Guo et al. 2016). Historical records also suggest the development of animal husbandry during the Han Dynasty. Many domestic animals such as pigs, sheep, and poultry like chickens were raised. The main specialized animals that were raised are pigs. The people of the Han Dynasty regarded life after death as a continuation of their mundane lives; hence, the basic human needs would also have to be fulfilled in the “other world” (Loewe 2005). Therefore, grave goods would be indicative of their lifestyle. Animals’ skeletal remains were commonly unearthed in the Han Dynasty tombs, regardless of the social status of the deceased. This is due to the traditional concept of animals where they are viewed as food, guardians, entertainment, and companions in “the other world” (Deng 2015). Low-ranking nobles and commoners would have a simple and common animal burial as compared to higher-ranking nobles. The burials also differ in geographical locations where low-ranking officials and commoners in the North had more chickens, pigs, and dogs buried with them. In contrast, fish and seafood were commonly seen in tombs in Southern areas (Deng 2015). Notably, only high-ranking nobles have access to seafood consumption. Thus, it is not surprising that they appear in the contents of the grave goods in the Northern regions. Isotopic analysis indicates that commoners in the Han Dynasty would consume pigs and dogs as their primary sources of meat (Guo et al. 2016; Zhou et al. 2017). The consumption of fish and seafood may differ from region to region: common people living in the south would consume more fish, while in the north, only upperclass people had easy access to seafood (Zhou et al. 2017). The well-known channel between the East and the West – the Silk Road – was officially founded during the Han Dynasty. The founding of the Silk Road, consequently, promoted interactions between the East and the West, bringing new crops, plants, and species to the Central Plains, including grapes, carrots, sesame, and fennel (An et al. 2004; Qiu et al. 2012).

Parasites in the Han Dynasty The health conditions of the people from the past were always an insightful topic for research, especially with the role of parasites in infections. Various parasites have been spotted in China, in both archaeological and medical contexts. Parasite eggs ranging from the Neolithic Period to the Qing Dynasty have been reported to be found in pelvic soil samples, mummies, and hygiene sticks (Yeh and Mitchell 2016; Yeh et al. 2016). Moreover, ancient medical books, such as Jin Gui Yao Lue (金匮要 略, known as Essential Prescriptions from the Golden Cabinet written by Zhang Zhongjing in the Eastern Han; also a part of Shang Han Za Bing Lun 伤寒杂病论), Zhu Bing Yuan Hou Lun (诸病源候论, known as Chao Shi Bing Yuan written by Chao Yuanfang in the Sui Dynasty), and Qian Jin Fang (千金方, written by Sun Simiao in the Tang Dynasty), recorded the existence of parasites and treatments for

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parasitic diseases. Many of the recorded parasites were intestinal parasites, including roundworms, tapeworms, and whipworms (Yeh et al. 2019). The Han Dynasty, which enjoyed a reputation equivalent to that of the Roman Empire,attracted particular attention from academia and the public. Pathological studies on human remains indicate that people in the Han Dynasty suffered from several diseases, including leprosy and syphilis (Zhang 1994).Moreover, anatomical studies on mummies from the Han Dynasty suggest the existence of arteriosclerosis (Peng 2009). Ancient medical texts have also recorded cirrhosis, gallstone, and gastrohelcosis in the Han Dynasty (Peng 2009). According to the historical records in the Han Dynasty, parasites and related diseases, especially intestinal parasites, have been observed and treated. There are also ancient medicinal books and official historical books such as Shang Han Za Bing Lun (伤寒杂病论, known as Shanghan Lun written byphysician Zhang Zhongjing in Eastern Han), Lunheng (论衡, written by Wang Chong in Eastern Han), and Hou Han Shu (后汉书, known as Book of Later Han written by Fan Ye, recording the history of the Eastern Han); “chang chong” (长虫), “chi chong” (赤虫), and “nao chong” (蛲虫) were frequently recorded in these books, and “chang chong” was identified as roundworm, and “nao chong” was identified as pinworm (Peng 2009). Other than the records in historical contexts, parasitological reports from archaeological sites also indicate parasitic infections in the Han Dynasty. Table 1 lists out parasitic findings from the archaeological contexts in mainland China, and three of them date back to the Han Dynasty. Details will be discussed in the following sections.

Roundworm (Ascaris lumbricoides) Roundworm, also known as Ascaris lumbricoides, might be one of the most common foodborne species that have infected humans (Torgerson et al. 2015). Humans get Ascaris infections via the fecal-oral route. Several factors, including intake of food or water that is contaminated by roundworms’ eggs, poor hygiene, and usage of night soil as fertilizer, promote the risk of being infected by roundworms (Dold and Holland 2011; Kim et al. 2014). The embryonated eggs of roundworms will hatch and develop into juveniles in the duodenum after ingestion. These juveniles will then move freely among the organs, including the liver, heart, and lungs. Through such movement, the juveniles will grow into adults and finally return to the small intestine where the adult females release eggs (Roberts et al. 2013). Characteristics of the roundworm egg, such as its oval shape and mamillated coat, make it easy to identify under the microscope. Usually, no symptoms would show if the infection by A. lumbricoides infection is minor. However, heavy infections of roundworms can cause vomiting, signs of malnutrition, and intestinal blockages. There are several paleoparasitological reports on roundworms in China from archaeological contexts, dating from the Neolithic period to the Ming Dynasty (CE 1368–CE 1644) (see Table 1). However, although there is a great variety of

Eastern Zhou Warring States Han Dynasty Han Dynasty Han Dynasty Song Dynasty Song Dynasty Ming Dynasty Ming Dynasty

Longhu, Henan

Hengyang, Hunan Province Guangzhou, Guangdong Province Guangzhou, Guangdong Province

Phoenix Hill, Jiangling, Hubei Province Changsha Mawangdui, Hunan Province Xuanquanzhi, Gansu Province Jintan, Jiangsu Province

Jiangling, Hubei

Dates Neolithic

Location Jiahu, Henan

M M F

Mummy

Mummy

28– 40 50

M

Mummy

NA

NA

Hygiene sticks Mummy

80

84

50

F

Mummy

55

NA

NA

Age NA

M

F

NA

Sex NA

Mummy

Context Pelvic soil Pelvic soil Mummy

Wealthy

Wealthy

NA

NA

Commoner

Wealthy

Wealthy

NA

NA

Social status NA

T. trichiura, C. sinensis, Fasciolopsis buski

A. lumbricoides

C. sinensis

Clonorchis sinensis, Trichuris trichiura, Ascaris lumbricoides Schistosoma japonicum, Clonorchis sinensis, Taenia sp., Trichuris trichiura Schistosoma japonicum, Trichuris trichiura, Enterobius vermicularis Clonorchis sinensis, Taenia sp., Ascaris lumbricoides, Trichuris trichiura A. lumbricoides, T. trichiura

Parasites spotted Ascaris lumbricoides, Trichuris trichiura, Taenia sp. Hookworm

Table 1 The summary of parasitic findings in China from archaeological contexts. (Adapted from Yeh and Mitchell (2016) and Li (1984))

Li 1984

Li 1984

Li 1984

Li 1984

Yeh et al. 2016

Li 1984

Lei and Hu 1984 Wei et al. 1980

References Zhang et al. 2006 Lan et al. 2011

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Zhongshan, Guangdong

Dongguan, Guangdong

Dongguan, Guangdong

Fuzhou, Fujian Province

Longyan, Fujian Province

Shaowu, Fujian Province

Yangzhou, Jiangsu Province Yangzhou, Jiangsu Province Fuqing, Fujian Province

Ming Dynasty Ming Dynasty Ming Dynasty Ming Dynasty Ming Dynasty Ming Dynasty Ming Dynasty Ming Dynasty Qing Dynasty M

Mummy

Mummy

Mummy

Mummy

Mummy

Mummy

M

M

F

F

F

M

F

Mummy

Mummy

M

Mummy

NA

NA

50– 60 NA

74

41

50

60

NA

Wealthy

NA

Wealthy

Wealthy

Wealthy

Wealthy

NA

Wealthy

Wealthy

C. sinensis

A. lumbricoides, T. trichiura, C. sinensis, Fasciolopsis buski A. lumbricoides, T. trichiura, C. sinensis

C. sinensis, T. trichiura

A. lumbricoides, T. trichiura

A. lumbricoides

C. sinensis, F. buski, A. lumbricoides, T. trichiura

A. lumbricoides, T. trichiura,

A. lumbricoides, T. trichiura,

Huang and Zhang 1995 Huang and Zhang 1995 Huang and Zhang 1995

Li 1984

Li 1984

Li 1984

Li 1984

Li 1984

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roundworms (Ascaris lumbricoides) found in ancient China, only one case is from the Han Dynasty. It is found on personal hygiene sticks at Xuanquanzhi, Gansu Province. Cloth-covered sticks from the Xuanquanzhi site functioned as personal hygiene sticks for wiping the anus after defecation. Dried feces still remained on some ancient hygiene sticks. The eggs of A. lumbricoides have been observed to be on the fecal samples that remained on the sticks (Yeh et al. 2016). Though there is a lack of reports regarding archaeological contexts of the Han Dynasty, roundworms may still be one of the most prevalent parasites in ancient China. There are records of roundworms along with its symptoms and treatments on various ancient records. Roundworms were often recorded in historical contexts as hui (蛔/蛕), jiao hui (蛟蚘), and chang chong (长虫). The earliest record related to roundworms is from Suwen, Huangdi Neijing, known as The Inner Canon of the Yellow Emperor,written by unknown authors between the Warring States Period and the Han Dynasty (fifth century BCE to CE first century): The symptom of stomach-caused coughing is vomiting after coughing, and severe coughing would lead to vomiting long worms out (胃咳之状, 咳而呕, 呕甚则长虫出, 胃咳之状, 咳 而呕, 呕甚则长虫出). (Wang 2009; Yeh et al. 2019)

This can be interpreted as vomit induced by ascariasis, an infection caused by roundworms (Sun and Zhao 2007). Another medical book in the Han Dynasty written by Zhang Zhongjing, Shang Han Za Bing Lun, also records the vomiting of roundworms: Jue Yin (厥阴) is a disease that would cause the patient to feel thirsty despite drinkinga great amount of water. The patient would also experience an uncomfortable sensation in the thoracic region and burning inside the stomach. The patient, despite being hungry, would not have the appetite to eat; after eating, the patient would suffer from vomiting roundworms (厥阴之为病, 消渴, 气上撞心, 心中疼热, 饥而不欲食, 食则吐蚘 (蛔), 下之利不止). (Zhang 1956; Yeh et al. 2019)

Other medical records dated after the Han Dynasty also noted the presence of roundworms. Some examples include Zhu Bing Yuan Hou Lun (诸病源候论), Qian Jin Fang (千金方), and Jing Yue Quan Shu (景岳全书).

Tapeworm (Taenia sp.) Cestoda is a class of worms under the phylum Platyhelminthes. The subclass, Eucestoda, is commonly referred to as tapeworms. Various species under the class of Cestoda are endoparasites, living in the digestive systems of vertebrates (Roberts et al. 2013). Though many species of Cestoda are able to infect humans, we only focus on Taenia sp. in this chapter. Three species of Taenia can infect humans: T. solium, T. saginata, and T. asiatica (Sato et al. 2018). The infection routes of these species involve the consumption of raw or uncooked meat and/or the internal organs of the prior hosts.T. solium and T. asiatica parasitize pigs, while T. saginata is

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commonly found in cattle. Hence, communities with the habit of consuming raw or uncooked pork and beef often have a higher infection rate of Taenia sp. Humans infected with tapeworms often show no symptoms. Occasionally, weight loss, nausea, diarrhea, and abdominal pain would occur, and segments of the worms can be spotted in the feces (WHO 2019b). Though the eggs of Taenia sp. are easily distinguishable from other parasites in the archaeological contexts with its characteristic morphology, it is hard to distinguish the three species individually under a low power light microscope (Zeibig 2014). T. asiatica presents similar morphologies to T. saginata while the life cycle is similar to T. solium. Hence, specific and precise identification of these three species cannot be done solely based upon microscopic observations. Sato et al. (2018) reported that the three species of Taenia could be distinguished through mtDNA techniques. This indicates a promising future for identifying Taenia sp. in paleoparasitology. However, molecular approaches are not widely practiced in archaeological contexts. In order to avoid potential misdiagnoses, the three species of Taenia are simply referred to as Taenia sp. There are only two paleoparasitological reports of tapeworms in China thus far, andboth date back to the Han Dynasty (see Table 1). One is from Xuanquanzhi, as mentioned above, where the eggs of roundworms have also been found. Tapeworms were also found in a mummy buried in Tomb No. 168 located in Phoenix Hill, Hubei Province. Wei et al. (1980) conducted an autopsy on the mummy and spotted the eggs of Taenia sp. in the intestinal canal. Although there are limited findings of tapeworms from archaeological contexts, there are several records of tapeworms in written sources. The practice of consuming raw or uncooked meat in China has a long history. Feng and Feng (2006) interpreted the record in Zhou Li (known as The Rites of Zhou, completed in the Warring States Period), which described the infected pork of Taenia sp.: Pao Ren (the position title for those who are in charge of food) would take charge of recognizing whether the meat was still edible based on the smell. If the pig is blind and pork smells bad, then the meat is not edible (庖人:辨腥臊香(臭)之不可食者。(猪)盲而交 睫, 腥(星)).

They believed the blindness of pigs was caused by a parasite that livedin their eyes, and the putrid smell was a symptom of pork taeniasis. This may be the earliest record of tapeworms in China. White worm (cun bai chong寸白虫 or bai chong 白虫) in ancient Chinese records has now been regarded as tapeworm through its color and size. Zhang Zhongjing recorded in his book Jin Gui Yao Lue (known as Essential Prescriptions from the Golden Cabinet,completed in the Eastern Han Dynasty): Eating beef and pork together would definitely cause white worm (牛肉共猪肉食之, 必作 寸白虫). (Zhang 1956)

Chao Yuanfang described the size and length of a tapeworm in Zhu Bing Yuan Hou Lun (known as Treatise on the Causes and Manifestations of Diseases, finished in Sui Dynasty):

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Cui Bai, one of the nine worms, is one-inch long, white in color and small in size. It would start to breed and hurt the host when the host’s intestinal organs are weak (寸白者, 九虫内 之一虫也, 长一寸而色白, 形小蝙, 因府藏虚弱而能发动). (Chao 1982; Yeh et al. 2019)

Sun Simiao, another ancient medical practitioner in the Tang Dynasty, wrote in Qian Ji Fang that a tapeworm could grow over 10 m long and cause the death of the host: White worms mate and lay considerable amounts of eggs. The female worms can grow to as long as four or five zhang. (Zhang (丈) is a traditional measuring unit in East Asia. One zhang is roughly equal to 3.33 meters today. In the Han Dynasty, one zhang ranged from 2.13 to 2.37 meters.) The worms could cause death too (白虫相生, 子孙转多, 其母转大, 长 至四、五丈, 亦能杀人). (Sun 1955)

These records suggest that ancient medicine practitioners in China had substantial knowledge of tapeworms regarding its appearance and paroxysm. Notably, in the Han Dynasty, Zhang Zhongjing already discovered that consuming pork and beef could cause the infection of tapeworm. Paleoparasitology has developed in other East Asian countries such as Japan and South Korea, and various reports have revealed the existence of different endoparasites on the mummies and ancient remains. Similarly, not many paleoparasitological reports of tapeworms have been recorded(Yeh et al. 2019). A low original infection rate of tapeworms and poor preservation would explain the lack of findings of Taenia sp. in archaeological contexts (Lee et al. 2011). However, according to historical records, the consumption of meat, especially pork, was quite popular in ancient China and was not limited to the Han Dynasty. Hence, it is likely that medical practitioners have built up their knowledge of parasites through experience. They must have witnessed an abundance of tapeworm-related cases, allowing them to deduce that the consumption of beef and pork could have caused the infection of tapeworms. Therefore, the low infection rate of tapeworms may not be sufficient to explain the lack of information on Taenia sp. in archaeological contexts in China. With increasing awareness and interest in paleoparasitology, we hope to gain more insights regarding Taenia sp. in the future.

Pinworm (Enterobius vermicularis) Pinworm (Enterobius vermicularis) is an intestinal parasite that is distributed widely across the world. It is especially prevalent among school-age children, regardless of socioeconomic levels. Humans are the only natural host of pinworms. The infection of pinworms often involves accidental ingestion of eggs, as a result of nail-biting, insufficient washing of hands, poor hygiene, and even inhalation (Shoup 2001). As many patients with enterobiasis show no clinical symptoms, a standard characteristic would be perianal itches that result from the production of pinworms. The adult worms mainly live in the cavities of the cecum, colon, and ileum. The female worms would lay eggs outside the anus of the host when the host falls asleep. This would cause perianal itches. When the host suffers from the itches and tries to

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relieve the itch by scratching, it is possible to contaminate one’s fingers and nails. A route of anus-finger-oral infection facilitates the repetitions of pinworm infections. Hence, a heavy load of parasites could live within a single host (Shoup 2001). Though the pinworm may be the most prevalent parasite that infects humans nowadays, it is not a commonly researched parasite among paleoparasitological studies in China. There is only one report on the finding of the eggs of E. vermicularis from archaeological contexts thus far. Wei (1973) reported the finding of eggs of pinworm (E. vermicularis), whipworm (Trichuris trichiura), and oriental schistosomiasis (Schistosoma japoniucm) from a female corpse dating back to the Han Dynasty, which is located at Mawangdui, Changsha, Hunan Province. As such, the condition of enterobiasis was already recorded in ancient books as early as in the Han Dynasty. Pinworm was recorded as “nao chong” (蛲虫) in medical and historical records such as Zhu Bing Yuan Hou Lun and Qian Jin Fang. One volume in Shi Ji (史记, known as Records of the Great Historian) called Biographies of Bianque and Canggong (扁鹊仓公列传)recorded that an ancient medical practitioner called Chunyu Yi in the Western Han once treated a female patient who suffered from “nao jia” (蛲瘕), which, based on other records, nao refers to pinworm: Chunyu Yi once treated a woman who was desperately ill. Other physicians diagnosed that this woman would die from the disease. However, Chunyu diagnosed that this woman suffered from nao jia, based on the swelling of the woman’s belly and howthe skin of the belly was yellow and rough (意治一女, 病甚, 众医皆以为寒热笃, 当死不治; 公诊其脉 日: 蛲瘕, 蛲瘕为病, 腹大, 上皮黄粗. . .). (Sima 1982)

However, the debate remains as to whether nao jia here is referring to enterobiasis as the description does not match the features of enterobiasis (Song 1980). On the other hand, Zhu Bing Yuan Hou Lun (written by Chao Yuanfang in the Sui Dynasty) stated that pinworms lived in the intestinal canal and were active near the region of the anus. Also, the host would suffer from itches caused by pinworm: Nao chong lives in the intestines (蛲虫居胴肠). Nao chong is one of the nine worms living in the intestines. If the digestive system is strong, the worms would not be active. If not, then nao chong would take advantage of it. The worms would cause itches or come out from the anus (蛲虫者, 九虫之一也, 在于肠间, 若府藏实, 则虫不妄动, 胃肠虚弱, 则蛲虫乘之; 轻则或痒、或虫从谷道溢出). (Chao 1982)

Although this observation is recorded only in the latter part of the Han Dynasty, Chao Yuanfang precisely described the habitus of pinworm that accurately matches our current modern knowledge of a pinworm.

Whipworm (Trichuris trichiura) Whipworm (Trichuris trichiura) is a common helminth, and the infection of whipworm (T. trichiura) would lead to trichuriasis. Humans can be infected with

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whipworms by consuming contaminated water and food. The eggs of whipworms are frequently seen in the feces of the human hosts. As a result, the infection rate would be higher among the communities that apply human feces as fertilizers. People infected with whipworms often have co-infections of roundworms and hookworms at the same time, owing to the similar infection routes of these parasites (WHO 2019a). Heavy loads of whipworms would lead to several symptoms, such as fatigue, abdominal pains, and diarrhea (WHO 2019a). The discovery of whipworms in the Han Dynasty was from a mummy unearthed at Phoenix Hill, Hubei Province (see Table 1). Most reports of whipworms (T. trichiura) are from the Ming Dynasty (see Table 1). Whipworms are also quite common in paleoparasitological research in other East Asian countries, such as South Korea (Zhan et al. 2019). Even though scholars encounter whipworms very often in the paleoparasitological contexts in East Asia, it is hard to determine if the ancient records refer specifically to whipworms.

Flukes in the Han Dynasty: Oriental Schistosomiasis (Schistosoma japonicum) and Chinese Liver Fluke (Clonorchis sinensis) Schistosomiasis is a parasitic disease caused by the infection of Schistosoma, which is commonly known as blood flukes. Of all the Schistosoma species that infect humans, Schistosoma japonicum is found mainly in Southern China and the Philippines (Colley et al. 2014). The life cycle of schistosomes is complicated due to the morphological transformations and the involvement of intermediate hosts of freshwater snails. S. japonicum uses the snails of Oncomelania sp. as the intermediate hosts (Colley et al. 2014). Snails infected with S. japonicum would produce cercariae into the freshwater. Once humans come in contact with the contaminated freshwater, it is easy to contract the parasite. The cercariae would penetrate the skin and release larvae awaiting maturation in the organs. A range of symptoms would appear after the infection of schistosomes, including fever, exhaustion, diarrhea, and abdominal pains lasting for weeks (Colley et al. 2014). Clonorchis sinensis, known as Chinese liver fluke, is another fluke that is found in China, Korea, Vietnam, and Russia (Wu et al. 2012). Similar to blood flukes, C. sinensis uses freshwater snails as the first intermediate host and freshwater fish and shrimp as the second intermediate host. Different from blood flukes, the infection route of C. sinensis is foodborne, as human body contract liver flukes by consuming raw or uncooked fish. C. sinensis would cause severe damage to the liver and biliary system. In worse cases, chronic infections of C. sinensis could cause cancers, including hepatocarcinoma and cholangiocarcinoma (Fried and Abruzzi 2010). The archaeological site, Mangwangdui in Hunan Province, is famous for its excavation of a female mummy. Other than the eggs of T. trichiura and E. vermicularis mentioned in the previous sections, the autopsy also revealed the existence of oriental schistosomiasis (Schistosoma japonicum) (Wei 1973). Another site dating back to the Han Dynasty, which led to the discovery of S. japonicum, is from Phoenix Hill in the Hubei Province. The mummy unearthed at this site also contained preserved eggs of C. sinensis (Wei et al. 1980). Xuanquanzhi is also

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another site in the Han Dynasty that contributed to the discovery of C. sinensis. Moreover, the eggs of C. sinensis have been found on mummies dating back to the Ming Dynasty, while there has been no other report on S. japonicum in ancient China thus far. Interestingly, Yeh et al. (2019) summarized that both Japan and South Korea lack the parasitological findings of S. japonicum from archaeological contexts. Similar to whipworms, there is a lack of key inputs from traditional medicine records on Chinese liver flukes. On the contrary, scholars have interpreted the word gu (蠱, a written character that refers to a worm inside the body) as oriental schistosomiasis in historical records (Jiang 1993). None of the records dating back to the Han Dynasty explicitly states the presence of blood flukes. Jiang (1993) explained that the records in Zhu Bing Yuan Hou Lun (amedical bookwritten by Chao Yuanfang, an imperial physician, around CE 610), including shui du bing (water-caused disease, 水毒病), along the Yangtze River, were referring to schistosomiasis. Sha shi (沙虱) in Zhu Bing Yuan Hou Lun was depicted: The mountains and rivers are infested with sha shi (沙虱) so fine that they can hardly be seen. They would attach onto people that enter infected rivers to bathe or walk through grass on rainy days. The people will then suffer from the infections. (山水间多有沙虱, 其细略不 可见, 人入水浴及水澡浴, 此虫在水中, 著人身, 及阴天雨行草中, 亦著人, 便钻入皮里). (Chao 1982; Yeh et al. 2019)

These records seem plausible to be about blood flukes. However, as Xiao (2006) pointed out, schistosomiasis was not fully understood by traditional Chinese practitioners back then.

Discussion As mentioned above, parasitological reports have revealed the presence of the eggs of whipworms (T. trichiura), roundworms (A. lumbricoides), tapeworms (Taenia sp.), pinworms (E. vermicularis), blood flukes (S. japonicum), and Chinese liver flukes (C. sinensis) at archaeological sites dating back to the Han Dynasty. Considering theinfection routes of these parasites, the diet and cooking methods in the Han Dynasty would be discussed with great attention. Cooking methods, such as steaming, frying, simmering, and roasting, already existed in the Han Dynasty (Yang 2009). However, eating raw meat was also popular then. Raw meat was cut into thin slices and served as kuai (脍). Li Ji (礼记, known as The Book of Rites,a record of the social forms, ceremonial rites, and administration of the Zhou Dynasty) stated that raw meat was commonly eaten with sauces. Moreover, the sauces would change along with the seasons: Consuming kuai in spring is suggested to be accompanied with scallions as the sauce, while in autumn, it is better to have mustard seeds (脍:春用葱, 秋用芥). (Ruan 1980)

Notably, kuai was served as a treat and reward during the Pre-Qin Period of China (Wu 2017). The tradition of consuming raw meat carried on into the Han Dynasty and soon became a cultural delicacy.

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Several books recorded the popularity of kuai during the Han Dynasty, as well as the collective knowledge of people regarding the risk of parasitic infections through the consumption of raw meat. Ying Shao, the author of Fengsu Tongyi (风俗通义, completed around 195 CE, recording the strange and exotic matters of interest during the Eastern Han), was surprised that people in Zhu’e County (an ancient county in Shandong) did not consume raw meat (“祝阿不食鱼生”) (Wang 1981). Hou Han Shu (known as Book of the Later Han, written by historian Fan Ye, in the fifth century CE, about the history of the Han from 6 to 189 CE) recorded an instance whereby Hua Tuo (华佗, a famous physician in the Eastern Han) treated a patient who had been infected with parasites due to the consumption of kuai: Chen Deng, an officer at Guangling, suffereddiscomfort and thus lost his appetite. Hua Tuo diagnosed his pulse and said, ‘You have worms in your stomach, which would gradually swell. This may be a result of eating raw fish.’ Hua Tuo immediately wrote a prescription and let Chen Deng take medicine. Soon after, Chen Deng puked several worms with red heads that still moved, and half of the worms’ bodies were located in the kuai. Chen Deng soon recovered from the initial discomfort. (广陵太守陈登忽患匈(胸) 中烦懑, 面赤, 不食。佗 脉之, 曰: ‘府君胃中有虫, 欲成内疽, 腥物所为也。’即作汤二升, 再服, 须臾, 吐出三升 许虫, 头赤而动, 半身犹是生鱼脍, 所苦便愈). (Fan 1973)

Medical books such as Jin Gui Yao Lue noted that eating kuai with dairy products would lead to parasitic infections: Eating sliced raw meat with dairy would lead to the growth of worms in one’s intestinal system, which causes acute infectious disease (食脍, 饮奶酪, 令人腹中生虫, 为疟). (Zhang 1956)

Kuai refers specifically to a dish of raw fish, raw pork, and raw beef that was consumed commonly in ancient China. However, modern clinical studies indicate that the consumption of raw or uncooked meat, including fish, beef, and pork, exposes consumers to a higher risk of parasites such as tapeworms and Chinese liver flukes (Schantz 1996; Lun et al. 2005). The ancient physicians in the Han Dynasty were very much aware that consuming raw or uncooked meat, especially beef, would lead to parasitic infections (Yeh et al. 2019). However, even with this knowledge, eating raw meat remained popular, and this practice continued throughout the generations. Another interesting possible factor that contributed to parasitic infection during the Han Dynasty would be the preparation process of animals for consumption (including killing and flaying) in the kitchen. Ren (1981) reported a stone slab that was unearthed from a Han tomb at Shandong, and this stone slab was analyzed, which was recorded by Huang (1990). This stone slab provided insight into how animals were prepared for consumption by depicting a scene in a kitchen. Steer, sheep, and pig (or boar) were slaughtered in the kitchen. Their remains and blood would have likely came into contact with surrounding people and tools; thus, this potentially increased the risk of a parasitic infection if the animals did carry parasites. Meanwhile, pictures of dogs were recorded on the stone slab, and a wandering dog was watching a live chicken. This implies that the kitchen is an open area, allowing

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animals like stray dogs to enter easily, thereby increasing the risk of spreading infectious diseases and parasites. The application of feces as fertilization was another risk factor for parasitic infections. The night soil of human and animals might have been used for farming as early as the Shang Dynasty (seventeenth century BCE–eleventh century BCE) (Xu 2005). Han Fei, a famous political philosopher in the Pre-Qin Period, wrote in Jie Lao (解老) that fertilizing the soil with excreta was of great importance for agriculture (积力于田畴, 必且粪灌) (Chen 1984). Thereafter, in the Han Dynasty, excreta from humans, animals, and green manure were applied to farming. The importance of night soil and other organic waste was noticed by ancient farmers and further emphasized in books such as Fan Shengzhi Shu (氾胜之书, an agricultural book written in the Western Han) (Ju et al. 2005). Moreover, it was common to build pigpens next to human latrines during the Qin and Han Dynasties. This construction allowed easier collection of night soil (Li 2016).This construction allowed pigs to eat human feces easily and, as a result, get infected with parasites such as tapeworms and roundworms. It was recorded in Shi Ji that the imperial concubine Lady Jia of Emperor Jing of Han (188 BC–141 BC) once encountered a wild pig in the latrine (“贾姬如厕, 野彘卒入厕”) (Sima 1982). This record suggested that the latrines were open enough for wild animals to enter easily. If the people of the higher class used loosely constructed latrines, the condition of the latrines used by the masses would likely be in an even more derelict condition. It will be tempting to suggest that people of different social statuses might suffer from different rates of parasitic infections. However, several factors suggest that parasitic infection might be prevalent throughout all social strata. First of all, the habitual consumption of raw meat was widespread, and the amount or type of meat did not differ among classes. An isotopic study revealed that people of all classes had a relative abundance of meat to consume; the only differentiating factor between social classes in the Han Dynasty would be the types of crops available for consumption (Zhou et al. 2017). Meanwhile, the development of husbandry during the Han Dynasty increased the number of farm animals, and redundant animals were slaughtered for consumption. Consequently, it resulted in an abundant supply of raw meat for consumption. Second, the application of night soil and the unique construction of latrines near pigpens exposed everyone to parasites, regardless of class. These lifestyles would even lead to repeated infections and/or cross-infection of parasites. Third, a lack of understanding about hygiene exacerbated the situation. As indicated by the kitchen scene on the stone slab, the killing and cooking of animals were carried out in the same location. Pets were free to enter kitchens too. Such open environments were hard to manage and do not ensure food safety. Meanwhile, it is hard to find out whether ancient people washed their hands or followed other hygiene protocols during the process of preparing food or after using toilets. Fourth, current reports indicate that people from high and low statuses were all infected with parasites. Table 1 shows that two out of the three reports on parasites in Han China were from wealthy people. The samples from Xuanquanzhi indicate that people who were traveling along the route took a break in this relay station. Little

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information has been reported on common people regarding paleoparasitology, probably due to the poor preservation of tombs of the common folks; thus, it would be less possible to spot parasites under the microscope. Furthermore, a limited understanding of paleoparasitology in fieldwork would impede the understanding of related works. We hope that more findings will be discovered, from the increased awareness of paleoparasitology and the growing development of techniques.

Conclusion In conclusion, the Han Chinese contracted parasitic diseases due to their lifestyles and diets, especially through the consumption of raw meat. These factors increased their susceptibility to parasitic diseases, and more than one parasite could infect the host at the same time. Six species of parasites have been identified in the three Han Dynasty remains till this very day. Being made aware of parasites, ancient medical physicians attempted to comprehend and treat such parasitic infections. Although they might not have the full comprehension of the infection routes and species, their works are still nonetheless remarkable. Much credit has to be given to them for noticing the ill effects of eating raw meat. Regarding the scarcity of reported cases of parasites in Han remains, this work combines the written materials and archaeological findings to explore the health condition of the Han people. With the increased awareness of paleoparasitology, further research on the parasites in the Han Dynasty is expected.

Cross-References ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies of Song-Ming Dynasty in China Acknowledgments This research was fully supported by NAP Start-Up Grant from Nanyang Technological University, Singapore.

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Dietary Stress in Combat: Coprolite Analysis of a Korean War Marine Killed in Action

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Karl J. Reinhard, Marina Milanello do Amaral, Gregory E. Berg, and Brianna Neu

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

We present our microbotanical and macrobotanical analysis of gut contents (coprolite) from the partial mummy of an American casualty from the Korean War known as Private First Class (PFC) Smith (PFC Smith’s name is anonymized. All other details are accurate.). Microscopic and macroscopic remains reveal a high fiber diet. Macrobotanical remains consisted of leaf and stem fiber and seeds from Potentilla (cinquefoil) and Sinapis alba Syn. Brassica alba (mustard). In general morphology, the mustard seeds from the coprolite are small compared to modern cultivated yellow mustard. Pollen analysis suggests a food source that included the buds of a Rosaceae family plant. Several Rosaceae genera produce pollen consistent with the recovered pollen including Potentilla. The coprolite K. J. Reinhard (*) School of Natural Resources, University of Nebraska- Lincoln, Lincoln, NE, USA e-mail: [email protected] M. M. do Amaral Superintendência da Polícia Técnico-Científica, São Paulo, Brazil G. E. Berg Defense POW/MIA Accounting Agency (DPAA), Joint Base Pearl Harbor-Hickam, HI, USA e-mail: [email protected] B. Neu Department of Anthropology, University of Nebraska, Lincoln, NE, USA © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_42

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matrix is composed of dark brown material that is inert. We suggest that the inert component is derived from a preservative/consolidant that was applied to the corpse at some time. This preservative/consolidant caused microscopic materials to adhere to each other which complicated pollen analysis. Given the analysis and subsequent findings, we conclude that the diet of this individual consisted of lownutrition raw mustard seeds and Potentilla foliage, which is a famine food. This is indicative of a starvation or scavenging-based dietary regimen. Keywords

Mummy · POW · Korean War · Palynology · Archaeobotany · Diet · Coprolite

Introduction The analysis of soldiers killed in conflict has significant emotional and, sometimes, political implications. Therefore, results of forensic study of killed in action (KIA) soldiers must be presented in an especially sensitive manner. In this case study, we examine the nature of sustenance that could be available to service personnel that are in a dramatic and dynamic retreat from a pursing enemy. The case focuses on a gut content sample from PFC Smith, who was an American Marine assigned to the first Battalion, seventh Regiment, first Marine Division, and deployed to the Chosin Reservoir in North Korea in 1950. In the first year of the Korean War, UN forces pushed back against the North Korean Army, driving them from the southern portion of the Korean Peninsula toward the Yalu River and the northern (Korean) border with China. Part of this push northward involved actions around the Chosin (Changjin) reservoir. The events of these actions are described in a variety of military texts (see Appleman 1987, 1989; Hammel 1994, Russ 1999 and others) and are often associated with this famous quote from Colonel Lewis Burwell Puller (aka “Chesty” Puller): “We’ve been looking for the enemy for some time now. We’ve finally found him. We’re surrounded. That simplifies things.” On the night of Nov 27, the Communist Chinese forces launched a massive counterattack at UN forces on both sides of the Chosin reservoir. Over the next several weeks, over 17,000 UN troops were listed as casualties, at the cost of some estimated 28,000 Chinese troops. With supply lines fragmented and/or disrupted, ammunition, medical aid, and even food were difficult to come by across the region. During this time, PFC Smith fought for 12 days, from November 27 until being injured near Kot’o-ri and dying on December 8, 1950. PFC Smith was killed due to a ballistic wound to his head; his remains were interred in a temporary cemetery in Hungnam, North Korea by UN forces, but were not recovered during the eventual evacuation. His remains were turned over to UN Forces during Operation Homecoming in 1954, but could not be positively identified, and he was buried in National Memorial Cemetery of the Pacific (NMCP) as an unknown. Based on the results of identification efforts by DPAA, he was finally returned to his family in the early 2000s.

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As noted, in 2006, the Joint POW/MIA Accounting Command, now DPAA, disinterred an unknown soldier buried in the NMCP based on new evidence of potential identity. The exhumed remains were partially mummified, particularly the abdominal region, and a preserved coprolite or possible gut contents were discovered. At the request of DPAA, we undertook the analysis of the coprolite/gut content sample. Gut content analysis has been part of mummy studies for decades and provided a wide array of surprising discoveries. The history of study is presented in a chapter in this volume, Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health and Hospice. Multidisciplinary work was established in the twenty-first century. Analysis of the Piraino 1 mummy from Sicily showed details of diet, medicine, and parasite infection involved in the death of a high-status figure (Kumm et al. 2010; Piombino-Mascali et al. 2013). Arguelles and her colleague (2015) revealed the reason of death, tea and honey consumption of a Korean Joseon dynasty general. On a population level, Reinhard and his colleagues (2011) showed the surprising reliance of ancient Chinchorro peoples in Chile on wild estuary plants.

Materials and Methods A coprolite was submitted for analysis to the Palynology Laboratory, School of Natural Resources at the University of Nebraska-Lincoln. We have been developing methods for application to coprolites from mummies, including Korean mummies, since 2008 (Arguelles et al. 2015; Kumm et al. 2010; Piombino-Mascali et al. 2013; Searcey et al. 2013; Verostick et al. 2019). We attempted to process this sample in the same way as we process Korean mummies of the Joseon Dynasty (Arguelles et al. 2015) and Chinchorro mummies from Chile (Reinhard et al. 2011). The sample was sent via FedEx in a sealed box. The sample inside the box was double sealed in plastic bags and labeled “CIL 2006-040, coprolite, 17.4 grams, 05 June 06.” The sample measured 3 cm in maximum diameter. The specimen was sectioned using a scalpel, and 0.5 g were submitted for palynological and botanical analysis (Fig. 1). Unlike other coprolites from mummies, this specimen was very hard, almost rock-like. One scalpel blade broke in the process of sectioning the sample for analysis. We have only encountered this consistency in colon remains that were solidified with shellac (Verostick et al. 2019). Further analysis confirmed that this sample had been solidified by an application of a preservative/consolidant, such as shellac. Standard processing for coprolites involved rehydration in 0.5% trisodium phosphate. This method, devised by Eric Callen, has been extensively reviewed (Bryant and Dean 2006; Bryant and Williams-Dean 1975). Normally, coprolites rehydrate in 48 h. In this case, the coprolite did not fully rehydrate in the expected 48 h, possibly in part, due to the solidifying agent applied to the mummy. In our experience, an alternative method is to rehydrate the coprolite in dilute HCl (Racz et al. 2015). We removed the coprolite from the rehydration solution, transferred to another clean beaker, and added 2% HCl to the sample. The sample successfully separated, and we

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Fig. 1 The coprolite sample measured 3 cm in maximum diameter. We used scalpels to section the sample. For palynological and microbotanical analysis, 0.5 g were submitted. For macobotanical and SEM analysis, 3.0 g were used. This specimen was rock-like comparable to mineralized coprolites. In this section, seed and fine fibers can be seen in a dark matrix with white specks. The composition of the dark matrix remains unknown (Credit: Karl Reinhard)

were able to continue processing the specimen via disaggregation with a stir bar on a stir plate. The sample was disaggregated in this way until the macroscopic remains were in suspension. However, unlike other studies, the microscopic remains did not fully separate into individual items and many pollen grains adhered to other microfossils due to the adhesive quality of the preservative/consolidant. At this time, one tablet of Lycopodium (batch 124,961, each tablet containing 12,489 spores plus or minus 491, University of Lund, Sweden) was added to the sample. The disaggregated remains were poured through a 250-μm mesh screen in a 600-ml beaker. Using distilled water jets from wash bottles, the macroscopic samples were washed while being separated with plastic laboratory mini spatulas. In this way, the microscopic remains are thoroughly removed from the macroscopic remains. Following screening, macroscopic remains were transferred from the mesh screens onto sterile filter paper, labeled, and left to dry. The dried macroremains are later examined using a binocular dissection microscope. The water and microscopic residues that passed through the screen were collected in the beaker and concentrated by centrifugation. The microscopic remains were immediately analyzed for starch, phytoliths, and other microscopic remains (Reinhard et al. 2012). In addition, the samples were scanned for parasite eggs. All remains were counted along with the added Lycopodium spores. After this analysis, pollen concentration was done following previously described methods (Arguelles et al. 2015; Reinhard et al. 2006, 2017). An acetolysis procedure was used to dissolve potential cellulose, hemicellulose, and chitin (Reinhard et al. 2006). The sample was washed with glacial acetic acid, to replace water; acetolysis reacts exothermically and violently with water. After the acid was

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decanted, acetolysis solution (9 pts acidic anhydride to 1 pt sulfuric acid) was mixed into the sediment. Then microscopic residue was heated at 99 °C for 10 min. After one glacial acetic acid wash, the sample was then washed repeatedly in distilled water until the supernatant was clear. The sediment sample was mounted on microscope slides with an additional drop of glycerin. A coverslip was then placed on the preparation and sealed with commercial nail polish. Following palynological procedures, a minimum of 200 pollen grains were counted and identified. Pollen counts were made with the Jeneval compound microscope at 403 and 1003 objective with10
oculars using DIC and bright field settings. A Ziessaxiomat compound microscope was also used for analysis. Imaging was done with the Jeneval system. Final quantitative analysis was accomplished using standard pollen concentration calculation based on the following formula: ððm  lÞ
tÞÞ  x, where m ¼ microfossils counted; l ¼ Lycopodium spores counted; t ¼ number of Lycopodium spores added to the sample; and x ¼ mass of sample. Pollen grains are classified as “pristine” “good,” “degraded,” or “unidentifiable.” Pristine grains are those that are perfectly preserved. Good grains are intact in shape, ornamentation, and aperturation but show some slight deformation. Degraded grains are exhibited one or a combination of eroded surface structure, folding, rupture of the wall, or fragmentation. Degraded grains are still identifiable to a taxon. Unidentifiable grains are so eroded, folded, torn, and/or fragmented that they could not be identified. A second 3.0 g sample was processed to recover and quantify remains. These were rehydrated and processed through screening as described above. However, in this case, the separated microscopic remains were processed for scanning electron microscope study (Morrison Microscopy Core Research Facility). The macroscopic contents were screened using four different sizes: a large screen of greater than 1 mm, a mid screen of less than 1 mm but greater than 0.5 mm, a fine screen of less than 0.5 mm but greater than 0.25 mm, and a superfine screen of less than 0.25 mm. This method was used to further distinguish the contents of the sample following published methods (Reinhard et al. 2012). Samples from the unique contents from each size range were placed on slides with a drop of glycerin for further microscopic analysis. The macroscopic remains were sorted using dissecting tools into types such as fiber, seed, and epidermis. Seeds were initially identified using text and web-based resources. Once provisional genus and species identifications were made, seeds were ordered from seed warehouses for comparison to the seeds found in the coprolite sample. Seeds and images of seeds were shared with a specialist at the National Research Institute of Cultural Heritage in Korea.

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Results The appearance of the coprolite was abnormal. Coprolites are usually light in weight, but this coprolite specimen was abnormally heavy. Most coprolites are friable and can be easily sectioned with a scalpel. The PFC Smith sample was so hard that the first scalpel blade broke when sectioning was attempted. After the coprolite was separated, we observed that an outer layer of preservative/consolidant had permeated the sample. Also, white deposits were noted within the specimen (Fig. 1). These are likely not simply calcium, but rather a mixture of chemicals, to include embalming compounds such as formaldehyde, methanol, and other solvents. The hardening compound also likely contains lye and plaster. It was made in a variety of ways across multiple theaters and times. The fact that the sample responded to HCl treatment confirms that it was mineralized with calcium derivatives. The combination of mineralization and solidification resulted in an abnormally heavy and hard sample. SEM analysis confirmed that the microscopic materials were dominated by granular debris (Fig. 2). No parasite eggs were found. Macroscopic remains consisted of intact seeds mixed with fibers which were encased in a granular, dark brown to black matrix. The fibers in the macroscopic remains are consistent with macerated stems and leaves (Fig. 3). The fibers were consistent with general leaf and stem morphology. The large screened materials, greater than 1 mm, were primarily composed of consolidated residue and leaf material and weighed 1.6 g. Around 3% of these large aggregates included white semi-transparent crystalline structures. The mid-screened material, less than 1 mm and greater than 0.5 mm, was composed of around 80% large aggregates of consolidated residue, 10% leaf and stem parts, and 10% circular white aggregates and trichomes. The mid screen weighed 0.19 g and featured sheets of plant cells, originally arranged in tiny tubes in about 20% of the residue clumps. The fine screen, between 0.5 mm and 0.25 mm, weighed 0.13 g and was composed Fig. 2 SEM analysis confirmed that the microscopic materials were dominated by granular debris. In this image, a plant epidermis fragment with defined cell walls is visible. Surrounding it are very fine granular residues that made up over 90% of the SEM preparation (Credit: Karl Reinhard)

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of about 90% leaf and stem material, 5% broken seeds, and 5% of the white aggregates. The superfine screened material, less than 0.25 mm, weighed 0.17 g and was composed of about 95% ground plant epidermis, 4% leaf fibers, and 1% white aggregates. One of the white aggregates was crushed and placed on a slide with glycerin. This aggregate was composed of linear crystals inconsistent with starch. The crystals are not unlike naphthalene which was used to repel insects. The crystalline material could be derived from formaldehyde and other embalming compounds and added as a preservative when PFC Smith was repatriated. The two types of seeds were consistent with yellow mustard (S. alba Syn. B. alba) (Fig. 4) and Potentilla (Fig. 5). The mustard seeds are completely consistent with the general morphology of comparative material in the lab. The mustard seeds were Fig. 3 Three fibers are laid over scales marked in 1 mm divisions. The fibers are consistent with macerated stems and leaves (Credit: Karl Reinhard)

Fig. 4 The mustard seeds are completely consistent with the general morphology of comparative material in the lab. From the published guides, mustard seeds are nearly globular in shape and finely pitted (Thomas et al. 2012). The color is orange for yellow mustard, S. alba (Syn. B. alba) (Credit: Karl Reinhard)

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Fig. 5 Four Potentilla seeds are laid over scales marked in 1 mm divisions. The size of these is consistent with Potentilla nutlet size (Credit: Karl Reinhard)

bright yellow with a very smooth outer surface. There were 0.1 g of mustard seeds in the sample. The brown material is unknown, but is very hard and inert. The identification of Potentilla was more difficult. According to the Korean botanist affiliated in the National Research Institute of Cultural Heritage in Korea, the seeds are most likely from a species of genus Potentilla. The species cannot be readily identified by mere inspection on the seed. Thus, accurate identification on the species is similarly difficult in this case. At a genus level, the seeds are light brown or reddish brown colored. Low veins project in diagonal and vertical directions on the whole seed surface. The seeds are about 1 mm in length and 1.3 mm in width. There were 0.4 g of Potentilla seeds in the sample. The microscopic remains are presented in Table 1. Trichomes were the most abundant plant microfossil. These are distinct cells that develop on the surface of many plant structures such as stems, leaves, and fruits. Over 200,000 trichomes were found per gram of sample. Trichomes occur on many plants, including mustard family (Brassicaceae) foods such as mustard greens and cabbage. They are also very common on species of Potentilla (Heo et al. 2013). Structures associated with xylem and phloem in vascular bundles, tracheids, vessel elements, and fibers were common. These totaled 65,000 combined per gram of material. The taxonomic source of the plant material could not be determined. However, the fragmentation of the leaves and stems, as seen also in the macroscopic remains, suggests that the leaves and stems were macerated before consumption. We compared the fibrous material to commercial Korean kimchi. The coprolite fibers were courser than those from kimchi. Thus, we do not think that the fibers were from a traditional Korean source. Root hairs were present and approximately 68,000 were found per gram of sample. As with the other microscopic remains, we cannot identify a taxonomic origin of the hairs; they only indicate that roots of some type were consumed. Very little starch was found. Starch tends to be abundant in intestinal residue from archaeological sites for cultures that are dependent on starch as primary sources of sustenance (Reinhard et al. 2012; Vinton et al. 2009). Recent specimens studied from a Korean mummy (Arguelles et al. 2015) and a Sicilian mummy (Piombino-Mascali et al. 2013) had very little starch even though other evidence indicated that starch was consumed in the form of noodles and bread. The starch in these two mummies was gelatinized by cooking before consumption and was fully digested. It may be that PFC Smith was provided starch in some form and that it was digested, therefore

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Table 1 The counts and concentrations of nonpollen microfossils made before pollen processing through acetolysis. The phytolith category refers to silicified plant trichomes Category Generalized cells Phytoliths Raphides Druses Total Stem/Leaf cells Epidermis Trichomes Total Root Cells Root hair Vascular stem cells Tracheids Vessel elements Total Miscellaneous Starch Fungal spores For quantification Lycopodium spores

Counts

# per gram

232 16 85

116,000 2000 42,500 160,500

35 410

17,500 205,000 222,500

136

68,000

96 34

48,000 17,000 65,000

1 8

125 1000

50

undetectable in coprolite analysis. However, the abundance of plant vegetative tissue elements suggests that he ate a high-fiber, low-starch diet around the time of death. No pollen grains were pristine (Table 2). However, the majority of Rosaceae pollen was in good condition with some deformation of the pollen grains (Fig. 6). It is noteworthy that many of the Rosaceae pollen grains were present in aggregates. The aggregates indicate that the pollen was consumed in food stuffs made from floral parts. Pollen of the rose family is often striate and can be used to make approximate genus determination (Chwil 2015; Hebda and Chinnappa 1994). The taxonomic source of the Rosaceae pollen from the coprolite could match a variety of genera in the family, including Potentilla. Oryza pollen comes from rice. After ingestion, traces of dietary pollen can persist in the digestive tract for days or weeks as explained in Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health and Hospice in this volume. It is likely that the rice pollen trace reflects food eaten some time before death. We are dubious about three pollen categories because the pollen was folded and abraded (Table 2). Therefore, we present the identifications of Citrus, Ranunculus, and Solanaceae as our best attempts at identifying poorly preserved pollen and we present them in the interests of full disclosure. Nine poorly preserved pollen grains exhibited morphology completely new to us and are classified as unknown. Fourteen pollen grains were crushed or fractured beyond the point of any classification. These

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Table 2 Korean MIA Pollen counts and concentration values. Rosaceae is an insect pollinated type and is not ingested with water and is not inhaled. Therefore, the pollen must have been ingested with food or drink Taxon Rosaceae Rosaceae clumps Solanaceae (?) Quercus Oryza Pinus Poaceae Ranunculaceae (?) Citrus (?) Typha-type Unknown Unindentifiable Total pollen Lycopodium

Common name Rose family Aggregates of rose family Nightshade family c.f. Oak Rice Pine Wild grass Buttercup family c.f. Citrus genus c.f. Cattail

Added exotic tracer spore

# counted 31 6 1 5 2 5 4 1 1 1 9 14 80 563

Calculated concentration 1377 266 44 222 88 222 176 44 44 44 400 622

Source diet diet envir. envir. envir. envir. envir. envir. envir. envir.

Fig. 6 Aggregates of pollen were recovered from the coprolite. In this case, the striate morphology of two grains in center left is consistent with the rose family and perhaps the genus Prunus. The third pollen grain on the right does not show the striations because the focus is through the center of the grain to show the pores in the colpi (grooves) of the grain. These are striate tricolporate grains. It is noteworthy that the pollen grain to the left is flattened (Credit: Karl Reinhard)

were also trapped in an adhesive material that made it impossible to view all sides of the grains. Three pollen types, Quercus (oak), Pinus (pine), and Poaceae (uncultivated grass), are wind-borne pollen types from the environment where

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PFC Smith fought. These were found in small amounts throughout the sample. Typha/Sparganum pollen comes from water plants and can be ingested through drinking water.

Conclusion The analysis of the coprolite allows us to address a reality of war: the adequacy of nutrition. Clearly, military/government issued foods were absent in the waning days of PFC Smith’s life. This is likely due to the ferocity and relentless nature of the Chinese counterattack and subsequent withdrawal of UN Forces from the Chosin reservoir area. The food that was eaten by PFC Smith was a low-calorie mix of fibrous plant material and seeds. In actuality, this was a starvation diet; it could easily have been composed of items that could be scavenged from the surrounding countryside or procured in local villages/houses. The cultivated foods evident were Asian. Pollen showed 88 grains per gram of rice pollen were found. However, this is not convincing evidence that rice was a major staple of PFC Smith’s diet at or around the time of death. The overwhelming abundance of plant structures from stems, roots, and leaves indicates that PFC Smith ate mostly fiber. In PFC Smith’s case, the main source of food was Potentilla. The seeds and pollen grains found in the sample were both consistent with this genus. The microscopic plant evidence is consistent with leaves, stems, and roots and an abundance of trichomes. These findings are consistent with consuming mats of Potentilla. Potentilla is a starvation food (Boesi 2014; Kim 2015; Ried 1977). Rhizomes, young shoots, and leaves were eaten in Tibet, China, and Korea in times of famine. The find of seeds indicate that PFC Smith ate fruits of Potentilla. These are not reported as edible. It is likely that the pollen grains were adherent to the fruits when they were consumed. Mustard seeds are part of indigenous Korean and Chinese cuisine. However, the consumption of raw seeds in large quantities is not part of the traditional cuisine. The seeds are generally ground and added to food. PFC Smith ate raw, unprepared seed, or the seeds were mixed into a hastily prepared food/seasoning with improper preparation. It is clear that the coprolite represents a high-fiber, low-calorie diet. The fact that famine foods were the main identifiable culinary elements indicates that the supply lines and other means of accessing food could become untenable during times of retreat and constant fighting. It is likely that PFC Smith supplemented his diet with items either scavenged, procured, or purchased from the local inhabitants or from the surrounding countryside.

Cross-References ▶ Analysis of Mummy Digestive Tract Contents with Examples of Relevance to Diet, Health, and Hospice

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▶ Radiological and Ethical Considerations of Autopsy in Mummy Study ▶ Showing Respect to the Dead: The Ethics of Studying, Displaying, and Repatriating Mummified Human Remains

References Appleman R (1987) East of Chosin: entrapment and breakout in Korea, 1950, vol 2. Texas A&M University Military History Series, College Station. ISBN 978-0-89096-283-1 Appleman R (1989) Disaster in Korea: the Chinese confront MacArthur, vol 11. Texas A&M University Military History Series, College Station. ISBN 978-1-60344-128-5 Arguelles P, Reinhard K, Shin DH (2015) Forensic palynological analysis of intestinal contents of a Korean mummy. Anat Rec 298:1182–1190 Boesi A (2014) Traditional knowledge of wild food plants in a few Tibetan communities. J Ethnobiol Ethnomed 10:75 Bryant VM, Dean GW (2006) Archaeological coprolite science: the legacy of Eric O. Callen (1912– 1970). Palaeogeograph Palaeoclimatol Palaeoecol 237:51–66 Bryant VM, Williams-Dean GW (1975) The coprolites of man. Sci Am 232:100–109 Chwil M (2015) Micromorphology of pollen grains of fruit trees of the genus Prunus. Acta Sci Pol Hortoru 14:115–129 Hammel E (1994) Chosin: heroic ordeal of the Korean war. Presidio Press, Novato. ISBN 978-089141-527-5 Hebda RJ, Chinnappa CC (1994) Studies on pollen morphology of Rosaceae. Acta Bot Gallica 141:183–193 Heo KI, Lee S, Yoo M, Lee S, Kwon Y, Lim SY, Kim S, Kim SC (2013) The taxonomic implication of trichome and epicuticular waxes in tribe Potentilleae (Rosaceae) in Korea. Korean J Plant Taxon 43:106–117 Kim MH (2015) Contemplation on the emergency foods in Korea under the Japanese occupation. J East Asian Soc Dietary Life 25:721–738 Kumm K, Araújo A, Piombino-Mascali D et al (2010) Archaeoparasitological investigations of a mummy from Sicily (18th–19th century AD). Anthropologie 48:177–184 Lech RB (2011) Tortured into fake confession: the dishonoring of Korean War Prisoner Col. Frank H. Schwable, USMC. McFarland, Jefferson Piombino-Mascali D, Zink AR, Reinhard KJ et al (2013) Dietary analysis of Piraino 1, Sicily, Italy: the role of archaeopalynology in forensic science. J Archaeol Sci 40:1935–1945 Rácz SE, Pucu de Araújo E, Jensen E, Mostek C, Morrow JJ, Van Hove ML, Bianucci R, Willems D, Heller F, Araújo A, Reinhard KJ (2015) Parasitology in an archaeological context: analysis of medieval burials in Nivelles, Belgium. J Archaeol Sci 53:304–315 Reid BE (1977) Famine foods of the Chiu-Huang Pen-ts’ ao. Southern Materials Centre, Taipei Reinhard KJ, Edwards SK, Meier DK (2006) Pollen concentration analysis of ancestral Pueblo dietary variation. Palaeogeograp Palaeoclimatol Palaeoecol 237:92–109 Reinhard KJ, LeRoy-Toren S, Arriaza B (2011) Where have all the plant foods gone? The search for refined dietary reconstruction from Chinchorro mummies. Yearb Mummy Stud 1:139–151 Reinhard KJ, Johnson KL, LeRoy-Toren S et al (2012) Understanding the Pathoecological relationship between ancient diet and modern diabetes through coprolite analysis: a case example from Antelope cave, Mojave County, Arizona. Curr Anthropol 53:506–512 Reinhard KJ, Milanello do Amaral M et al (2017) Palynological investigation of mummified human remains. J Forensic Sci 63:244–250 Russ M (1999) Breakout: the Chosin reservoir campaign, Korea 1950. Penguin Books, New York. ISBN 978-0-14-029259-6 Searcey N, Reinhard KJ, Gardner SL et al (2013) Parasitism of the Zweeloo woman bog body. Int J Paleopathol 3:224–228

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Thomas J, Kuruvilla KM, Hrideek TK (2012) Chapter 21 – Mustard. In: Peter KV (ed) Handbook of herbs and spices, vol 2. Elsevier, Amsterdam Verostick KA, Teixeira-Santos I, Bryant VM et al (2019) The Skiles mummy: care of a debilitated hunter-gatherer evidenced by coprolite studies and stable isotopic analysis of hair. Int J Paleopathol 25:82–90 Vinton SD, Perry L, Reinhard KJ et al (2009) Impact of empire expansion on household diet: the Inka in northern Chile’s Atacama Desert. PLoS One 4:e8069. https://doi.org/10.1371/journal. pone.0008069

Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent

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Sergey Mikhailovich Slepchenko, Min Seo, Jong Ha Hong, Chang Seok Oh, and Dong Hoon Shin

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Siberian Peoples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Joseon-Korean Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Chinese Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion: Old World Evidence Supporting Reinhard and Araujo’s Hypothesis . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

There was a pioneering speculation that the parasitism of mankind has changed continuously and sometimes dramatically in association with the social changes

S. M. Slepchenko (*) Institute of the Problems of Northern Development, Tyumen Scientific Center of the Siberian Branch of the Russian Academy of Sciences, Tyumen, Russia Surgut State University, Surgut, Russian Federation e-mail: [email protected] M. Seo Department of Parasitology, Dankook University College of Medicine, Cheonan, South Korea J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_45

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in history such as the emergence of complex societies or evolution of subsistence strategies. Though this assumption was based on the outcomes of archaeoparasitological studies at the Colorado Plateau of North America, very few supporting reports were available from the Eurasian continent to date. To corroborate this idea more convincingly, we aim to compare the parasitological results of different Eurasian archaeological sites where human populations of varying social complexity depended on different subsistence strategies throughout history. In this chapter, we thus reviewed archaeoparasitological reports on the Siberian native peoples as nomads, fishermen, and hunter-gatherers, Russian migrant-descendants, and Korean and Chinese pre-modern mummies as the people of agriculture-based society. The current report reveals that Reinhard and Araujo’s hypothesis (2008) about the relationship between serious parasitism and complex societies could be also applicable to the Old World archaeoparasitological cases. Keywords

Archaeoparasitology · Russia · Siberia · South Korea · China · Mummy

Introduction Human societies have experienced remarkable transitions in lifestyles according to the different stages of social evolution in history. Likewise, in case of archaeoparasitology, the pattern of ancient parasitism was not always the same at different spatial-temporal situations. Actually, the parasite infection pattern in history cannot be explained solely by simple host-pathogen interaction. Rather, in order to understand the actual ancient parasitism more comprehensively, parasite infection should be considered from socioeconomic as well as cultural perspectives. Therefore, not only to confirm the presence of ancient eggs or larvae of certain parasite species, by archaeoparasitology, we should also define ancient parasitism associated with social changes in history, especially the emergence of complex society or evolution associated with subsistence strategies. Reinhard and Araujo (2008, 2012) once proposed that the parasitism became serious in history when social complexity outstripped the sanitation and hygiene. A shift from hunter-gatherer’s band to agricultural society in history will be the case to show close relationship of parasitism and social complexity. This Reinhard and Araujo’s original assumption could be proven by researches on archaic huntergatherers and more recent agricultural ancestral Puebloans on the Colorado Plateau in North America (Reinhard and Araujo 2008). According to them, hunter-gatherer’s parasitism was not likely so serious as expected because of small band size, low population density, high mobility, and their dietary patterns (Reinhard and Araujo 2008). On the other hand, when farming began in history, the agricultural communities’ parasitism became serious compared to the counterpart hunter-gatherers. The heavy parasite infection in agricultural societies could have been caused by the contamination of drinking water, increase in population density, sedentary life in an

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apartment-style living, and the irrigation centered on agriculture, etc. (Reinhard and Araujo 2008). The parasitism of the mankind history has thus changed continuously and sometimes dramatically, especially around the time when people’s survival strategy changed. Despite these pioneering speculations on the parasite endemicity affected by social factors in history, in case of Eastern hemisphere or the Old World, archaeoparasitological evidence as to the transition of parasitism, especially associated with the emergence of complex society or the change in subsistence strategy, are still rare to date. To prove this historical event more extensively in spatial-temporal perspectives, archaeological reports of different countries or continents need to be thoughtfully examined. In this chapter, we therefore compare the archaeoparasitological studies so far from different regions of Eurasian continent. Using parasitological data of mummies or skeletonized specimens from Russia, Korea, and China, we substantiate the original assumption of Reinhard and Araujo (2008, 2012), corroborating the difference in parasitism between nomads, fishermen, hunter-gatherers, and agriculturalists according to their own subsistence strategies or social complexity stages.

The Siberian Peoples We used available parasitological, historical, and ethnographical data to reveal different parasitism patterns of various societies that have existed historically in the Old World countries such as Russia (Siberia), China, and South Korea. The cultural/ subsistence background of the societies addressed in this chapter is summarized in Table 1. The geographical locations of the archaeological sites are indicated in Fig. 1. Among them, native peoples in Siberia were mostly nomads, semi-nomads, fishermen, or hunter-gatherers (Slepchenko et al. 2015, 2016). Human populations of Siberia have overcome harsh environmental conditions by effective survival strategies. Until the migration of Russians, very few arctic or subarctic Siberian natives were influenced by the agriculturalists’ culture or lifestyle (Figs. 2 and 3). In Siberia, parasitological specimens were acquired from the burial grounds of Vesakoyakha, Kikki-Akki, and Nyamboyto sites discovered at Tazovsky and Krasnoselkupsky districts in the Yamalo-Nenets Autonomous Okrug. They represent the parasitism among the Siberian nomads (reindeer herders), fishermen, or hunter-gatherers (Slepchenko and Ivanov 2015; Slepchenko et al. 2016; Slepchenko and Reinhard 2018; Poshekhonova et al. 2019). The estimated date of their burials ranges from the eighteenth to the early twentieth century (Table 1). Archaeologists conjectured that Vesakoyakha burial grounds were created by the Siberian native Nenets in the nineteenth century. They were likely nomadic reindeer herders within the Vesakoyakha River Basin. The Kikki-Akki was the eighteenth-century burial ground constructed by Taz Selkups, the Siberian native hunter, gatherer, and fishermen (Slepchenko and Ivanov 2015; Slepchenko et al. 2016; Poshekhonova et al. 2018, 2019). Another burial ground at the Nyamboyto I was also formed during the early twentieth century, by the Nenet hunter-fishermen (Kvashnin and Tkachev 2014; Slepchenko et al. 2016) (Table 1).

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Table 1 Archaeological reports of this chapter

Lifestyle Sedentary

Migratory

a

Siberian native people After Before contact contact with with Russiansa Russiansb c Niamboito Khantyk Kikki-Akkid Vesakoiakha II–IVe BuchtaNakhodkaf Zeleny Yarg Nadym Gorodokh Doge-Bary IIi Nefteprovod IIj

Mansik None

Russian immigrants Russian old-timersl Yeniseiskm None

Koreans Korean mummiesn

Chinese Chinese mummies and skeletonized individualso

None

None

For Niamboito, Kikki-Akki, and Vesakoiakha II-IV3, it does not mean that they never contacted the Russians. Their life routines (purchase of goods or payment of tax, etc.) enforced only episodic and exceedingly rare contact with the Russians, at best b The peoples were living together with Russians. Their children went to kindergarten, studying together with Russians c Hunter-fishermen d Hunter-gatherer-fishermen e Nomad, reindeer herders f Hunter-fishermen migrating in the summer and settled in the winter g Hunter-fishermen possibly migrated in the summer and settled in the winter, but not confirmed yet h Hunter-fishermen, semi-nomadic population i A nomadic Iron Age community, geographically close to the neighboring agricultural areas j The middle Bronze Age hunter-gatherer (2000 to 1000 BCE) possibly wandered around Kan River k The populations of villages such as Vanzetur, Shaitanka, Nizhnie Narykary, Ustrem, Tegi, Vanzevat, Aneevo, Lombovozh, Sosva, and Sartynia located in Berezovskii District of the Khanty-Mansi Autonomous Okrug. Hunter-gatherer-fishermen living in small villages. Their villages began to increase in population with the advent of the Russians. They bought goods in a store l Russian immigrant descendants: teachers, doctors, nurses, kindergarten teachers, drivers, sellers, workers, etc. They were not engaged in agriculture; they bought everything in the store. They also went fishing and hunting m Farmers in the seventeenth to the eighteenth century n Korean mummies of the sixteenth to the eighteenth century. People living in agriculture-based society. Those mummies (n¼24) include reported cases from Yongin, Jinju, Sapgyo, Hadong-2, Hadong-1, Sacheon, Gangneung, Dangjin, Mungyeong, Waegwan, PJ SM, Seocheon, Yangju, SN1-2, SN3-7-1, SN2-19-1, SN2-19-2, GJ1-2, Hwasung, Andong, YG2-4, YG2-6, Dalsung, Junggye o Chinese mummies (n¼13) include those of Jiangling (Hubei), Changsha (Huibei), Phoenix Hill (Huibei), Jintan (Jiangsu), Hengyang (Hunan), Guangzhou (Guangdong, n ¼ 2), Yangzhou (Jiangsu, n ¼ 2), Fuqing (Fujian), Shaowu (Fujian), Longyan (Fujian), Fuzhou (Fujian). They were living in agriculture-based society

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Fig. 1 Geographical locations of the archaeological studies cited in this chapter. 1, Zeleny Yar burial ground; 2, Buchta-Nakhodka burial ground; 3, Nadym Gorodok settlement; 4, Vesakoiakha II–IV burial grounds; 5, Niamboito burial ground; 6, Khanty, Mansi and Russian old-timers living in villages (twentieth century); 7, Kikki-Akki burial ground; 8, Yeniseisk; 9, Nefteprovod II burial ground; 10, Doge-Bary II burial ground. Specimens were also obtained from the villages such as Shaitanka, Nizhnie Narykary, Ustrem, Tegi, Vanzevat, Aneevo, Lombovozh, Sosva, and Sartynia located in Berezovskii District of the Khanty-Mansi Autonomous Okrug (Credit: Sergey Slepchenko)

The Buchta Nakhodka 2 was possibly the oldest and the northernmost burial ground in the tundra zone of Western Siberia (Yamal Peninsula). The burial ground was located on the shore of a lake in the Yamalsky District, Yamalo-Nenets Autonomous Okrug (Slepchenko et al. 2019a). According to the archaeologists, the burial sites were composed of two different groups: those of the sixth to the seventh century and the twelfth to the thirteenth century CE. The burial ground was likely to be formed by native hunter-fishermen who were nomadic during summer but settled in winter season (Slepchenko et al. 2019a) (Table 1). Zeleniy (Zeleny) Yar grave site in the northwestern part of Siberia was constructed by another native hunter-fishermen who were settled or dispersed according to different seasons. At the Zeleny Yar site, the archaeologists found the twelfth- to thirteenth-century child mummy and a 1-year-old infant remain for whom archaeoparasitological studies were done (Slepchenko et al. 2015, 2019b) (Table 1). For the former child mummy, samples were collected from the abdominal region. After skin incision was made in the suprapubic region, the parasitological specimen was taken from the abdominal cavity of the mummy (Slepchenko et al. 2019b). The

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Fig. 2 Siberian native people (Khanty)’s burials at the northern forest-tundra subarctic area (Zeleny Yar) in Priuralsky district of Yamal-Nenets Autonomous Okrug, Russian Federation (Credit: Alexandr Gusev)

preservation status of the 1-year-old infant remain was not good (Gusev 2015). The infant was wrapped in a fur garment, a hide, and a layer of birch bark. The parasitological sample was collected from the place where the infant’s pelvis was believed to have been originally located (Slepchenko et al. 2015). In 1990 to 1998, the South Siberian site of Doge-Bary II, the burial ground of a nomadic Iron Age community, was also investigated by Russian archaeologists (Slavinsky et al. 2018) (Table 1). The burial ground belongs to the Uyuksaglynsk culture that was created by the early nomadic group in Siberia. The estimated date was the fifth to the fourth century BCE. It was presumed that prehistoric contact might have occurred between the South Siberian nomads of the Doge-Bary II and adjacent agricultural areas in China or Central Asia (Slavinsky et al. 2018). In this sense, they were likely influenced by agricultural culture rather than maintaining purely nomadic lifestyle. Among the human remains discovered at the Doge-Bary II burial ground, three mummified individuals were included in archaeoparasitological research (Slavinsky et al. 2018). Finally, the burial grounds Nefteprovod I and II were also found on the bank of the Kan River at Krasnoyarsk region (Slepchenko et al. 2017) (Table 1). Fifteen burials were discovered during the excavation of 2015. The burials were mostly of the Late Bronze Age Karasuk culture (approximately 1500 to 800 BCE), except for

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Fig. 3 Human remains from Siberian native people’s burials (Zeleny Yar burial ground; the twelfth to thirteenth centuries CE) (Credit: Alexandr Gusev)

burial #13 of the Nefteprovod II likely of the middle Bronze Age (2000 to 1000 BCE). The skeleton was found in burial #13. The burial found under an oval-shaped stonework was present about 0.1–0.15 m deep. The skeletonized individual was placed on his (her) right side with the legs bent at the knees. A specimen was collected from the pelvic region as well as sacral foramina. Archaeoparasitological examination was done for the specimen of burial #13 (Slepchenko et al. 2017). During the examinations on the samples of native Siberian nomads, fishermen, and hunter-gatherers, the eggs of Opisthorchis felineus, Diphyllobothrium, and Taenia spp. were found (Table 2). In case of nematode (soil-transmitted – Trichuris trichiura) eggs, however, they were not reported from those specimens except for Doge-Bary II (Slavinsky et al. 2018). The absence of the nematode eggs means that Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis, etc. did not infect the Siberian native nomads, fishermen, or hunter-gatherers until their contacts with Russian immigrants (Slepchenko et al. 2016, 2019c). Slavinsky et al. (2018) also interpreted that the presence of T. trichiura eggs in Doge-Bary II samples might have been due to their consumption of the agricultural foods likely imported from Central Asia oases or even farther from China. This confirms the existence of historical contact between the southern Siberia nomads and adjacent agricultural areas in ancient times (Slavinsky et al. 2018). The archaeoparasitological results mentioned above contrast with the data from modern native peoples who had contact with Russians and were subsequently affected by their sedentary lifestyle. For instance, in 1988–1989, parasitologists

Parasite eggs discovered

Trematoda

Class Nematoda

Parasite eggs discovered Ascaris lumbricoides Trichuris trichiura Hookworm Enterobius vermicularis Strongyloides stercoralis Trichostrongylus spp. Clonorchis sinensis Opisthorchis felineus Paragonimus westermani Metagonimus yokogawai Gymnophalloides seoi ●

●

Different populations Siberian natives: before contact with Russians Doge- Other than Bary Doge-Bary II IIa

Table 2 The ancient parasite eggs found in each Siberian and Asian specimen

●

●

Siberian natives: after contact with Russiansb ●

●

Russian Immigrantdescendantsc ●

●

●

●

●

● ●

●

● ● ●

●

Chinese mummiese ●

●

Korean mummies of Joseon Dynastyd ●

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Taenia Diphyllobothrium sp.

● ●

● ●

●

●

●

●

b

Native Siberians of Niamboito, Kikki-Akki, Vesakoiakha II–IV, Buchta-Nakhodka, Zeleny Yar, Nadym Gorodok, Doge-Bary II, and Nefteprovod II Khanty and Mansi peoples c Russian old-timers or Yeniseisk d Korean mummies (n¼24) of Yongin, Jinju, Sapgyo, Hadong-2, Hadong-1, Sacheon, Gangneung, Dangjin, Mungyeong, Waegwan, PJ SM, Seocheon, Yangju, SN1-2, SN3-7-1, SN2-19-1, SN2-19-2, GJ1-2, Hwasung, Andong, YG2-4, YG2-6, Dalsung, Junggye e Chinese mummies (n¼13) found at Jiangling (Hubei), Changsha (Huibei), Phoenix Hill (Huibei), Jintan (Jiangsu), Hengyang (Hunan), Guangzhou (Guangdong, n¼2), Yangzhou (Jiangsu, n¼2), Fuqing (Fujian), Shaowu (Fujian), Longyan (Fujian), Fuzhou (Fujian)

a

Cestoda

Fasciolopsis buski Schistosoma japonicum

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reported the infection pattern of modern Siberian people at the Khanty-Mansi Autonomous Okrug (Slepchenko et al. 2019c) (Fig. 1). The population is composed of Siberian native groups: Khanty and Mansi. While maintaining traditional lifestyle and dietary habits, Khanty and Mansi peoples contacted Russian immigrantdescendants and lived together at the same place (Slepchenko et al. 2019c). In the copro-ovoscopic survey on modern Khanty and Mansi populations, the parasite eggs of O. felineus, A. lumbricoides, and E. vermicularis were detected (Slepchenko et al. 2019c) (Table 2). The presence of A. lumbricoides and E. vermicularis means that they have different patterns of parasitism compared to Siberian native nomads, fishermen, or hunter-gatherers who did not contact with Russian immigrants. Meanwhile, O. felineus eggs in Khanty and Mansi specimens also show that raw fisheating does not vanish completely even after contacts with Russian immigrants (Fig. 4) because O. felineus infection could be caused by the ingestion of raw or insufficiently cooked fishes (Slepchenko et al. 2019c). In Siberia, parasitologists also examined the specimens of Russian immigrantdescendants (Russian old timers or Yeniseisk in Table 1). The Russian old-timers lived in the settlements around the Khanty-Mansi Autonomous Okrug. They were westernized people but were also adopting the dietary habits of neighboring Siberian natives: raw fish consumption (Slepchenko et al. 2019c). The study of the Yeniseisk Russians was also based on the soil samples obtained from the early seventeenthcentury burials at the Epiphany Cathedral (Yeniseisk) and from a toilet discovered at a nearby eighteenth-century mansion. They were likely the seventeenth- to the eighteenth-century Russian farmers (Slepchenko et al. 2020). In archaeoparasitological study, A. lumbricoides, O. felineus, Taenia, and Diphyllobothrium eggs were found in either the specimens of Russian old-timers (Slepchenko et al. 2019c) or Yeniseisk Russians (Slepchenko et al. 2020) (Table 2). As to the parasitism of Siberian peoples with different sociocultural backgrounds, what attracts our attention the most is a difference in the archaeoparasitological pattern between Siberian natives after contact with Russians and pre-contact native Fig. 4 Raw fish-eating does not completely vanish in Siberia so far (Credit: Dong Hoon Shin)

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peoples. While very few A. lumbricoides or E. vermicularis eggs were detected in the latter specimens, in case of the former, very high egg-positive rates were reported in the species. Especially, finding A. lumbricoides or E. vermicularis eggs in those late twentieth-century Siberian native specimens (Khanty and Mansi) might have been due to the native people’s change in the way of life during the Soviet Union era (Slepchenko et al. 2019c). Most native Siberians began their sedentary life, thus having lived together with the Russian immigrant-descendants in the Soviet period. In brief, higher infection rate of A. lumbricoides and E. vermicularis among the late twentieth-century Siberian native people might have been induced by an increase in person-to-person contacts at overcrowded places of Siberia (Slepchenko et al. 2019c). In this regard, once again, we note Reinhard and Araujo’s assertion (2008, 2012) that serious parasitism could have been caused by the changes in socioenvironmental factors like the emergence of complex societies or evolution in subsistence strategies. All the factors proposed by Reinhard and Araujo (2008, 2012), such as contaminated water sources, concentrated populations, sedentary life, apartment-style living, etc., seem to be also applicable to explain the serious infection of A. lumbricoides and E. vermicularis among the late twentieth-century native or Russian immigrant-descendant peoples in Siberia.

The Joseon-Korean Mummies The archaeoparasitological reports of Korea and China, based on the mummy studies in each country, could also corroborate Reinhard and Araujo’s assertion (2008, 2012) that the difference in parasitism might have been influenced by the societies’ complexity or subsistence strategy. In the case of South Korea, over the past decades, very well-preserved mummies were discovered in the sixteenth- to the eighteenthcentury graves of Joseon Dynasty (1392–1910 CE). Additionally the coprolite specimens from Korean mummies could be an excellent resource for archaeoparasitologists in South Korea (Seo et al. 2014). The details of the archaeoparasitology of Korean mummies (n ¼ 24) are described in another chapter of this book or also in Table 1. Briefly, in the research of Korean mummies, the parasite infection pattern was significantly different from the results of native Siberians mentioned above. Whereas Russian natives who did not contact Russians exhibited relatively a simple set of parasite spp., mainly of Diphyllobothrium or Taenia spp., the sixteenth- to the eighteenth-century Korean mummies showed a larger variety of ancient parasite eggs including A. lumbricoides, T. trichiura, Clonorchis sinensis, Metagonimus yokogawai, Paragonimus westermani, Strongyloides stercoralis, Gymnophalloides seoi, and Trichostrongylus spp. (Seo et al. 2017) (Table 2, Fig. 5). In particular, the Joseon people’s parasitism is also characterized by a lot of soil-transmitted helminths that were not seen among pre-contact native Siberians. The overall positive rates for soil-transmitted helminths in Korean mummies were 58.3% for A. lumbricoides and 83.3% for T. trichiura, respectively (Seo et al. 2017).

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Fig. 5 Ancient Paragonimus westermani eggs found in Korean mummy specimen (Credit: Min Seo)

The extant historical or ethnographic records in Korea are also helpful to understand the parasite infection pattern of Joseon people revealed by mummy study. Actually, in the seventeenth to the eighteenth century, big cities emerged in Joseon society. Therefore, the urban population was unprecedentedly increasing in Korean history. The population density accordingly became higher; and so, the demand for a food in sufficient quantities exploded for the city dwellers’ nourishment. Night soils, human feces collected from latrines, were produced in the cities. These night soils were sold to the farmers working at the suburban area of the cities. The remarkable increase in agricultural productivity, using night soil as a fertilizer constituent, made serious parasite infection among urban people of Joseon dynasty (Kim et al. 2014). High population density in the city also made the Joseon people difficult to find clean water for drinking. According to historical literature, the city dwellers in the Old Seoul City complained about the pollution of the wells that were commonly contaminated by sewage wastewater (Kim et al. 2014). Generally speaking, the sixteenth- to eighteenth-century city of Korea was very vulnerable to serious soiltransmitted parasitism. The archaeoparasitological pattern of the Korean mummies was much similar to those of Russian immigrants or post-contact native Siberians but not to pre-contact Siberian peoples. Considering that the sixteenth- to eighteenthcentury Korea was a highly developed agricultural society, the results mentioned above also support the speculation of Reinhard and Araujo (2008).

The Chinese Mummies In East Asia, in addition to Korean mummies, there has been a research on parasite infection from the mummies discovered on the Chinese mainland. The total number of mummies examined by parasitologists was 13 (Yeh and Mitchell 2016). The results are summarized in Table 1. In Yeh and Mitchell’s review (2016), the

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archaeoparasitological reports on Chinese mummies of different dynasties spanning over 3,000 years were analyzed. The parasitologists found the ancient eggs of A. lumbricoides, T. trichiura, E. vermicularis, C. sinensis, Fasciolopsis buski, Schistosoma japonicum, and Taenia in the specimens (Table 2). Of them, A. lumbricoides, T. trichiura, and C. sinensis were the most commonly detected parasite species (Yeh and Mitchell 2016). In fact, the results from Chinese mummies are very similar to those of Korean mummies and also to those post-contact Siberian peoples in the late twentieth century. Like Korean mummies, the higher level of A. lumbricoides and T. trichiura infection among Chinese mummies was likely due to the contamination of food or drinking water by using human feces as a crop fertilizer (Yeh and Mitchell 2016). Reinhard and Araujo’s argument (2008) that the human parasitism was much serious in agricultural communities than in those of hunter-gatherers could be also proven by archaeoparasitology in China.

Conclusion: Old World Evidence Supporting Reinhard and Araujo’s Hypothesis Using the archaeological specimens, parasitologists have the opportunity to conjecture and confirm the relationship between social complexity and parasitism. In their memorable research about the changing pattern of parasitism in history, Reinhard and Araujo (2008, 2012) compared Colorado Plateau Archaic parasitism to agricultural Puebloan sites, verifying that parasitism was limited in hunter-gatherer archaic societies due to its small band size and high mobility of the group. They noted only one possible factor that might have promoted parasitism in hunter-gatherer society: the consumption of uncooked vertebrate meat or insects (Reinhard and Araujo 2008). Meanwhile, parasitism became serious in the descendant agricultural Puebloan communities of Colorado. The parasitism among Puebloan agricultural society likely depends on worse environmental situations such as concentrated populations, contaminated water sources, sedentary life, unique apartment-style living, the absence of effective sanitation technique, and so on (Reinhard and Araujo 2008, 2012). They also supported their theory about parasitism and social complexity by a comparison of E. vermicularis eggs in the coprolites. Actually, the highest E. vermicularis infection rate was reported from the specimens of walled villages built in rock shelters where a large population lived in the apartment-style communities with poor air circulation, fecal contamination, and crowding (Hugot et al. 1999; Reinhard and Araujo 2008, 2012). It is presumed that parasitism in history might have associated with social complexity, urbanization, and subsequent changes in subsistence strategies (Reinhard and Araujo 2008, 2012). In this chapter, by reviewing parasitological, ethnographic, and historical data from Russian, Korean, and Chinese archaeology, the sociocultural aspects of parasitism in different Eurasian societies of nomads, fishermen, hunters, and agriculturalists can be understood. We confirmed that the parasitism could be limited in the low-complexity societies not contacting agriculture-based western civilization. As to

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the cause of the parasitism rarity in pre-contact Siberia, we also note Reinhard and Araujo (2008)’s argument that parasitism was not serious in case of historical societies with smaller population size, low social complexity, low grade of mobility, etc. In summary, regardless of temporal or spatial perspectives, the argument of Reinhard and Araujo could be also applicable to the archaeoparasitology outside the Americas, proving that serious parasite infection could have occurred more commonly in complex societies than in simple bands or tribes in history.

Cross-References ▶ Bog Bodies and Natural Mummification of Siberia ▶ Joseon Dynasty Mummies of Korea ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies of Song-Ming Dynasty in China ▶ Mummies in Siberia Acknowledgments This research was supported by Basic Science Reseach Program of the National Research Foundation of Korea, the Ministry of Education (2020R1A2C1010708). A part of the present work was also funded by State Task (program XII.186.4, project No. AAAAA17-117050400143-4 of Tyumen Scientific Center SB RAS).

References Gusev AV (2015) Report on the scientific research work “Comprehensive archaeological and anthropological studies of medieval burial grounds near the village of Zelyeny Yar. Priuralsky YANAO district in 2014”. Typography Salekhard, Salekhard Hugot JP, Reinhard KJ, Gardner SL et al (1999) Human enterobiasis in evolution: origin, specificity, and transmission. Parasite 6:201–208 Kim MJ, Ki HC, Kim S et al (2014) Parasitic infection patterns as correlated with urban-rural recycling of night soils in Korea and other East Asian countries: the archaeological and historical evidence. Korean Stud 38:51–74 Kvashnin YN, Tkachev AA (2014) Cult place on the lake Nyamboyto. Anthropol Forum 23:185–194 Poshekhonova OE, Kisagulov AV, Gimranov DO et al (2018) Transformation of Upper Taz Selkup funeral rites according to paleoecological data. J Archaeol Sci Rep 22:132–141 Poshekhonova OE, Razhev DI, Slepchenko SM et al (2019) Dietary strategies of northern selkups in the 18th–19th centuries. Vestnik Archeologii, Antropologii i Etnografii 4(47):121–139 Reinhard KJ, Araújo A (2008) Archaeoparasitology. In: Pearsall DM (ed) Encyclopedia of archaeology. Elsevier, New York Reinhard KJ, Araújo A (2012) Synthesizing archaeology with parasitology in paleopathology. In: Buikstra J, Roberts C (eds) A global history of paleopathology. Oxford University Press, Oxford Seo M, Araujo A, Reinhard K et al (2014) Paleoparasitological studies on mummies of the Joseon Dynasty, Korea. Korean J Parasitol 52(3):235–242 Seo M, Oh CS, Hong JH et al (2017) Estimation of parasite infection prevalence of Joseon people by paleoparasitological data updates from the ancient feces of pre-modern Korean mummies. Anthropol Sci 125(1):9–14

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Slavinsky VS, Chugunov KV, Tsybankov AA et al (2018) Trichuris trichiura in the mummified remains of southern Siberian nomads. Antiquity 92(36):410–420 Slepchenko SM, Ivanov SN (2015) Paleoparasitological analysis of soil samples from the KikkiAkki burial ground of the 17th–19th centuries in West Siberia, Russia. J Archaeol Sci Rep 2:467–472 Slepchenko SM, Reinhard K (2018) Paleoparasitology and pathoecology in Russia: investigations and perspectives. Int J Paleopathol 22:39–44 Slepchenko SM, Gusev AV, Ivanov SN et al (2015) Opisthorchiasis in infant remains from the medieval Zeleniy Yar burial ground of XII-XIII centuries AD. Mem Inst Oswaldo Cruz 110(8):974–980 Slepchenko SM, Ivanov SN, Bagachev AN et al (2016) Traditional living habits of the taz tundra population: A paleoparasitological study. Korean J Parasitol 54(5):617–623 Slepchenko SM, Ivanov SN, Vybornov AV (2017) Taenia sp. in human burial from Kan River, East Siberia. Mem Inst Oswaldo Cruz 112(5):387–390 Slepchenko S, Kardash O, Ivanov S et al (2019a) The Buchta-Nakhodka 2 burial ground: Results of archaeoparasitological and macro-remains investigations of samples from the burial grounds of the 6th–13th century CE on the Yamal Peninsula in Russia. J Archaeol Sci Rep 23:791–799 Slepchenko SM, Ivanov SN, Gusev AV et al (2019b) Archaeoparasitological and palynological analysis of samples from the intestinal contents of a child mummy from the Zeleniy Yar burial ground (12–13th centuries AD). Archaeol Res Asia 17:133–136 Slepchenko SM, Bugmyrin SV, Kozlov AI et al (2019c) Comparison of Helminth Infection among the Native Populations of the Arctic and Subarctic Areas in Western Siberia throughout history: parasitological researches on contemporary and the archaeological resources. Korean J Parasitol 57(6):607–612 Slepchenko SM, Slavinsky VS, Ivanov SN et al (2020) Pathoecology of the town of Yeniseisk in Western Siberia from the 17th and 18th centuries. Quat Int 545:111–118 Yeh HY, Mitchell PD (2016) Ancient human parasites in ethnic Chinese populations. Korean J Parasitol 54(5):565–572

Part V Egyptian Mummies

Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification in Ancient Egypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Significance of Mummification: Becoming an Osiris . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Texts and Other Indirect Insights into Mummification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Mummy of Ameniryirt: Identity, Context, and an Overview of Mummification in Ancient Egypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . New Methods: Toward a More Systematic Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . New Developments in X-Ray Science and CT Scanning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Bioarchaeology of Mummified Human Remains: Toward a More Systematic Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Evolution of Mummification from the Predynastic to Medieval Periods, with New Insights from the British Museum Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Predynastic Period (c. 5500–3100 BC) and Early Dynastic Period (c. 3100–2686 BC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Old Kingdom (c. 2686–2181 BC) and First Intermediate Period (c. 2181–2055 BC) . . . . Middle Kingdom (c. 2055–1650 BC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Second Intermediate Period (c. 1750–1550 BC) and New Kingdom (c. 1550–1069 BC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Third Intermediate Period (c. 1069–656 BC) and Late Period (664–332 BC) . . . . . . . . . . . . . Ptolemaic Period (332–30 BC) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Roman Period (30 BC–AD 395) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Late Antique and Medieval Periods (AD 395–1500) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion: Toward Systematic Methods and Approaches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Grounded in new research from recent analyses of the mummified individuals curated at the British Museum, this chapter provides an overview of the evolution D. Antoine (*) · M. Vandenbeusch Department of Egypt and Sudan, The British Museum, London, UK e-mail: [email protected]; [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_17

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of mummification in the Nile valley. Featuring Ameniryirt, a Twenty-Sixth Dynasty Theban official who lived around 600 BC, it outlines the varying methods employed to preserve human remains in ancient Egypt, as well as the modern approaches used in their analysis. From the natural mummies of the Predynastic Period to those of medieval Nubia, advances in CT-scanning and three-dimensional visualization technology have allowed us to investigate their mummified remains without the need to unwrap them. Many are remarkably preserved and were embalmed with great care and expertise to ensure their survival in the afterlife. A range of approaches – spanning several thousand years – were successfully used to preserve their bodies, with organs treated in numerous ways and great variety in the use of unguents, packing, and wrappings. Bioarchaeological methods are employed to determine their approximate age-atdeath, confirm their sex, and provide new insights into their lives and state of health. Applying new analytical techniques to long-held collections continues to advance our understanding of ancient beliefs and practices, as well as provide a unique understanding of the lives of individuals from the distant past. Keywords

Natural mummy · Mummification · Tattoos · Metastatic carcinoma · Cardiovascular disease · Dual energy CT scanning

Introduction The archaeology and monuments of ancient Egypt are a unique record of the early societies of the Nile valley. However, most of the art and architecture was designed according to strict guidelines and dominated by formal imagery, so it offers limited insights into the daily lives of Egypt’s past inhabitants. Almost all statuary depicts individuals as young and healthy but there is evidence that some people lived into old age. The long-term custom of burying the dead, on the other hand, which has led to the survival of many thousands of human remains, offers direct insights into the people that inhabited these lands, as well as their beliefs. Some were deliberately mummified, others naturally preserved, and owing to the dry and arid conditions prevalent throughout most of Egypt, they have often survived in an excellent state of preservation. Egypt’s dry and hot climate was more conductive to preservation along the Nile valley and Oases, but less so in the Nile Delta, a land of marshes and swamps (Fig. 1). Since the flood plain was usually devoted to agriculture activities, burial lands were often located in the desert. This unique combination of climate and land use allowed for the preservation of mummified individuals. Influenced by temporal, geographical, and socioeconomical factors, the practice evolved over a period of 4000 years (Table 1). The scientific analysis of mummified remains provides information that is rarely accessible in other sources of evidence, including written texts. Methods developed in anthropology and bioarchaeology allow us not only to determine the age at death or biological sex of an individual, they also inform us about important aspects of human biology, diet, the prevalence of diseases, burial practices, past beliefs, and the process of mummification.

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Fig. 1 Map of ancient Egypt. Drawing: Claire Thorne. Image courtesy of the Trustees of the British Museum

Mummification in Ancient Egypt The ancient Egyptians believed that the proper treatment and preservation of the body after death would ensure the continuation of a person’s existence into the afterlife (Taylor 2001). It was thought that the link between body and consciousness could be renewed if the body endured. The intention was to keep the body whole and safeguard it from destruction. Some survived in good condition as a result of natural preservation, while others were intentionally mummified by embalmers. Bodies

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Table 1 Timeline of the main time periods of Egyptian history as divided by Egyptologists. Dates prior to 664 BC are approximate Predynastic Period 5500–3100 BC Badarian period 5500–4000 BC Naqada I 4000–3500 BC Naqada II 3500–3200 BC Naqada III (Dynasty 0) 3200–3000 BC Early dynastic Period 3000–2686 BC First Dynasty 3000–2890 BC Second Dynasty 2890–2686 BC Old Kingdom 2686–2181 BC Third Dynasty 2686–2613 BC Fourth Dynasty 2613–2494 BC Fifth Dynasty 2494–2345 BC Sixth Dynasty 2345–2181 BC First Intermediate Period 2181–2055 BC Seventh and Eighth Dynasties 2181–2125 BC Ninth and Tenth Dynasties (Herakleopolitan) 2160–2025 BC Eleventh Dynasty (Thebes only) 2125–2055 BC Middle Kingdom 2055–1650 BC Eleventh Dynasty (all Egypt) 2055–1985 BC Twelfth Dynasty 1985–1795 BC Thirteenth Dynasty 1795–1650 BC Second Intermediate Period 1750–1550 BC Fourteenth Dynasty 1750–1650 BC Fifteenth Dynasty (Hyksos) 1650–1550 BC Sixteenth to Seventeenth Dynasties 1650–1550 BC New Kingdom 1550–1069 BC Eighteenth Dynasty 1550–1295 BC Nineteenth Dynasty 1295–1186 BC Twentieth Dynasty 1186–1069 BC Third Intermediate Period 1069–656 BC Twenty-First Dynasty 1069–945 BC Twenty-Second Dynasty 945–715 BC Twenty-Third Dynasty 818–715 BC Twenty-Fourth Dynasty 727–715 BC Twenty-Fifth Dynasty (Nubian) 716–656 BC Late Period 664–332 BC Twenty-Sixth Dynasty (Saite) 664–525 BC Twenty-Seventh Dynasty (Persian) 525–404 BC Twenty-Eighth Dynasty 404–399 BC Twenty-Ninth Dynasty 399–380 BC Thirtieth Dynasty 380–343 BC Thirty-First Dynasty (Persian) 343–332 BC Macedonian kings 332–305 BC Ptolemaic Period 305–30 BC Roman Period 30 BC–AD 395

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were interred in individual graves or communal tombs and each settlement had one or more cemetery areas that could remain in use over thousands of years.

The Significance of Mummification: Becoming an Osiris The preservation of the body was an essential aspect of the ancient Egyptian funerary beliefs. Through the process of mummification, the physical remains of the deceased were transformed into a divine image that incorporated the attribute and qualities of the gods (see Ikram and Dodson 1998: 15–21; Taylor 2001: 10–45). Ancient Egyptians believed that this perfect body would serve as the physical base for the spirit aspects of the person, known as the ka and the ba. The ba is usually depicted as a human-headed bird. It was thought to enable the dead to exist in the next world and to travel freely between the world of the living and the realm of the dead. Each individual was also believed to have a ka, the life-force or spirit that also needed the physical form of the body to continue to exist. The ka would be “fed” via offerings of food and drink, ensuring the deceased continued to be sustained after death (Taylor 2001: 92–111). A person’s name was another essential aspect that needed to be kept alive so that the person would continue to exist in the afterlife. This partly explains the repetitiveness of inscriptions stating names, ancestry, and titles on coffins and other funerary belongings. In addition to a physical transformation, the deceased underwent a spiritual journey (Taylor 2001). The notion of rebirth is closely connected with the fate of two gods, Ra and Osiris. The sun god Ra is associated with the idea of rebirth through the daily rise of the sun disk. Each deceased would hope to join the bark carrying the sun god and travel with him. Motifs alluding to the solar journey were often disseminated on funerary objects buried with the deceased as a way of including them into the daily course of the sun. Through the embalming process and the use of funerary masks and other trappings, the deceased also sought an association with Osiris, the God of the Afterlife. According the ancient Egyptian beliefs, Osiris represents the first mummy prepared by the embalming god Anubis. Thus, Osiris was resuscitated by his sister and wife Isis, after being killed and cut into pieces by his brother Seth, the god of chaos. Ancient Egyptians hoped that, by being mummified, they would become an Osiris and resurrect in the afterlife just as the god had. The deceased was also thought to meet Osiris during the transitional path leading the “Field of Reeds,” the Egyptian equivalent to heaven. Judged for his actions on earth in a tribunal presided by Osiris, his heart was weighed against the feather of Maat, the goddess of truth and justice that incarnated the balance between good and bad which formed the core of the ancient Egyptian belief system. The weighting of the heart would happen under the watchful eye of the Great Devourer, usually represented as a crocodilehippopotamus-lion hybrid creature that would – should the deceased fail this test – eat the sinner’s heart and put an end to any ambitions of an eternal life. If successful, the god Osiris would welcome the deceased into the realm of the afterlife. The deceased would be declared maa-kheru, which can be translated as “justified” or more literally “true of voice,” confirming their transition into a divine being and eternal life.

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Texts and Other Indirect Insights into Mummification Egyptian texts that mention mummification are very late in date and focus on the ritual aspects, with few insights into the practical elements (Taylor 2001: 51–76; Ikram and Dodson 1998: 103–108). Images of mummification and actual sets of embalming tools are also very rare (Janot 2000; Taylor and Antoine 2014: 53–59; Fig. 2). Classical authors Herodotus (c. 484–425 BC) and Diodorus Siculus (first

Fig. 2 Scene from the coffin of Djedbastiuefankh (Hildesheim, Roemer-Pelizaeus Museum, Inv. 1954), one of the rare depictions of a mummification being performed. Drawing: Claire Thorne. Image courtesy of the Trustees of the British Museum

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century BC) wrote the clearest descriptions, revealing the existence of several mummification methods. Known as the “Father of History” (already described as such by Cicero; see Luce 1997: 27), the Greek historian Herodotus of Halicarnassus wrote the following account of Egyptian mummification: “The most perfect process is as follows: as much as possible of the brain is extracted through the nostrils with an iron hook, and what the hook cannot reach is rinsed out with drugs; next the flank is laid open with a flint knife and the whole contents of the abdomen removed; the cavity is then thoroughly cleansed and washed out, first with palm wine and again with an infusion of pounded spices. After that it is filled with pure bruised myrrh, cassia and every other aromatic substance with the exception of frankincense, and sewn up again, after which the body is placed in natrum, covered entirely over, for seventy days – never longer. When this period, which must not be exceeded, is over, the body is washed and then wrapped from head to foot in linen cut into strips and smeared on the underside with gum, which is commonly used by the Egyptians instead of glue. In this condition the body is given back to the family” (Herodotus, The Histories II: 86. Translation in de Sélincourt 1972: 160–1). While Herodotus’ account reflects the process as it existed in the fifth century (Peck 1983), Diodorus’ later text, which appears to have been influenced by Herodotus’, adds further insights, particularly the different roles of embalmers (see Taylor 2001: 51–76). Few other written sources explain the actual procedure. These include the Ritual of Embalming that mostly survived as hieratic papyri from the Roman Period and describes the practical and ritual preparation of the body (Töpfer 2015). The Rhind Magical Papyrus, from around 200 BC, also mentions some elements of the embalming procedures (see Ikram and Dodson 1998: 103–106; Taylor 2001: 49–51 and 76–78). The discovery of embalming caches and the analysis of their content (Eaton-Krauss 2007, 2008; Ikram and Lopez-Grande 2011; Hamdy and Fahmy 2018) are also contributing to the study of mummification. However, it is through the examination of the mummies themselves that we can refine our understanding of this unique process and gain detailed insights into how the body was transformed into a divine image that incorporated the attribute and qualities of the gods.

The Mummy of Ameniryirt: Identity, Context, and an Overview of Mummification in Ancient Egypt The Importance of Context: Who Was Ameniryirt? Prior to entering the British Museum collection in 1839, little is known of the history of Ameniryirt. His mummified remains (British Museum EA 6669) and three accompanying coffins (EA 6667, EA 6668 and EA 22811) were purchased from Giovanni Anastasiou (1780–1860), better known as Giovanni Anastasi, a successful Greek merchant based in Alexandria, who acted as the Consul-General in Egypt for Sweden and Norway for almost 30 years (Bierbrier 2012: 19–20). A coffin’s shape and decoration can provide valuable biographical information and often help determine where it was made, with regions of Egypt developing stylistic and

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iconographical trends. The motifs, texts, and scenes decorating Ameniryirt’s three coffins suggest that they were produced in the Theban area. The inscriptions describing Ameniryirt as an official working for the estate of the Divine Adoratrice of Amun Amenirdis confirm its Theban origin. Ameniryirt’s association with Amenirdis also provides a useful chronological reference. Amenirdis was the daughter of king Kashta (about 760–747 BC), who reigned during the TwentyFifth Dynasty (about 716–656 BC) when Egypt was under the rule of the Kushite kings of Nubia (Taylor 2000; Coulon 2018). Ameniryirt was responsible for administrating storage and the incomes of her estate, an important role denoting his high status. Amenirdis and Ameniryirt were not, however, contemporaries. Every time the Divine Adoratrice is referred to on the coffins, it is specified that she is maatkheru or “true of voice.” This expression is generally used to indicate that the person is deceased (see above). Ameniryirt’s coffins also reveal that Amenirdis appears to have already been dead when his father, Mentupaydenu, held the same role as his son. The style of the coffins provides further dating criteria and suggests they were manufactured during the Twenty-Sixth Dynasty (e.g., the green face of the inner coffin), perhaps around 600 BC. The fact that Ameniryirt was mummified and was buried in a set of three elaborately decorated coffins denotes his high status and financial means. Indeed, mummification was the reserve of the elite and he must have belonged to the upper class of the ancient Egyptian society, or at least to the local elite. The inscriptions on the coffins bear a number of titles that provide further insights into his status and work for the estate of the Divine Adoratrice Amenirdis. He held administrative functions and was not a priest, officiating as a “Servant of the income (sedjem-ash en aqw),” “Servant of the entrance of the palace (sedjem-ash en ra en akhet),” “Master of the storerooms (kheri-shenaw),” and “Magazine supervisor (kheri-at)” (Graefe 1981: 26–28). Further contextual information is limited as Ameniryirt’s mummy and coffins were purchased at auction. We have no information on the precise location of his tomb within the Theban region. In addition to coffins, he is likely to have been buried with a range of funerary goods, such as an Osiris figure or shabtis. To date, a wooden funerary stela, now curated at the Museum August Kestner in Hanover, is the only object to have both survived and been identified as belonging to Ameniryirt. It depicts the deceased facing two deities, the solar god Re-Horakhty and the creator god Atum (Graefe 1981: 209–10).

The Mummy of Ameniryirt and an Overview of the Practice of Mummification in Ancient Egypt The arid conditions prevalent in most of the Nile valley provide an ideal environment for the long-term preservation of human remains. Decomposition is usually caused by the actions of enzymes, proteins that are present within the body and function as catalysts that promote specific reactions (see Aufderheide 2003: 41–71). After death, the enzymes start breaking down the soft tissues. Decomposition is also influenced by the bacteria present within the body and from the surrounding environment, as well as insect activity. Halting decay usually involves inhibiting these processes shortly after death by creating an environment in which enzymatic and bacterial

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activity cannot take place. Burial in an extremely cold or arid environment can achieve this naturally, but artificially drying the body can also be a very effective method (see ▶ Chap. 2, “Dried, Tanned, Frozen, Embalmed, Smoked: A Glimpse into Mummification Mechanisms,” this volume). It is unclear when ancient Egyptians first started experimenting with mummification. Rare examples of naturally mummified individuals have been found at Predynastic sites such as Gebelein (around 3500–3300 BC; Antoine and Ambers 2014; Taylor and Antoine 2014: 22–43; Friedman et al. 2018). Buried in shallow pits without coffins and in close contact with the hot sand, their bodies appear to have accidentally desiccated due to the extremely arid desert conditions. Egyptians from this early period may have unearthed natural mummies, when digging new graves or opening old ones, and such chance discoveries may have guided their beliefs toward the need to preserve the body after death (Ikram and Dodson 1998: 108; Taylor 2001; Antoine and Ambers 2014). This may have led to experiments in artificial preservation and early attempts at mummification appear to have taken place from the middle of the fourth millennium BC at Predynastic sites such as Hierakonpolis, around the time the Gebelein bodies were buried (see Jones et al. 2014, 2018). Increasingly elaborate mortuary practices in the centuries just before and after 3000 BC (see Ikram and Dodson 1998: 21–60), with coffins, deeper graves, and longer funerary rituals, would have inhibited natural mummification. This may have prompted the search for alternative methods of preservation. The body of Ameniryirt has been carefully preserved and provides an excellent example of artificial mummification. The techniques used to embalm and wrap Ameniryirt’s body are characteristic of the first half of the first millennium BC. By this time, the process of mummification had become well established throughout Egypt, having begun nearly 3,000 years earlier. The scientific analysis of ancient Egyptian mummies, including that of Ameniryirt, confirms much of Herodotus’ account (see above). The first stage would have been to wash the body soon after death, both a practical necessity and an essential ritual element due to the purifying and life-giving qualities of water (Taylor 2001: 51–63). To stop decomposition, the most perishable internal organs were usually removed. The body was then dried, anointed, filled with packing materials and wrapped in linen. By this process, the body would be refashioned into a divine image. While showing a great deal of variation, approaches also evolved through time, leading up to the pinnacle of Egyptian mummification during the Third Intermediate Period (1069–656 BC). Here, Ameniryirt’s brain was probably removed by using an instrument similar to the hook mentioned in Herodotus’ description. The resulting damage to the delicate bones inside the nose is regularly identified in CT scans, with a route via the left nostril usually favored (Taylor 2001: 53). In Ameniryirt’s case, the embalmer used a very direct route up the left side of Ameniryirt’s nose, carefully breaking the ethmoid and vomer bones along the way and opening a relatively small hole about 1.2 x 1 cm in the base of the frontal bone (approximately 5 cm inside the nasal passage) to access his brain. Despite the very narrow route and limited amount of space, Ameniryirt’s nose and the delicate bones in the lower and upper parts of the nasal cavity (the inferior conchae and the nasal bones, all clearly visible on the CT scan)

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Fig. 3 The mummification of Ameniryirt (British Museum EA 6669). The embalmers used a direct route up the left side of Ameniryirt’s nose to access and remove his brain (top left). His eye sockets were filled with loose textile bundles (top right) and, despite a layer of resin, Ameniryirt’s face can be seen (bottom left). Dense granular packing material and textile were used to fill part of his ribcage and abdomen. An incision of the left flank was used to remove the organs and add packing materials (bottom right). Images courtesy of the Trustees of the British Museum

are intact (Fig. 3). The embalmer’s precision suggests a very good knowledge of human anatomy and reveals the great care and skill with which this procedure was executed. The technique was probably perfected to avoid damaging the face and leave little external visible trace. Alternatively, embalmers would occasionally remove the brain through one of the eye sockets, a hole in the cranium, or via the foramen magnum at the base of the skull (Taylor 2001: 53; Wade et al. 2011; Wade and Nelson 2013a). Although it was not the case for Ameniryirt, plugs were sometimes inserted in the nostrils to reshape the nose after the removal of the brain (see Aufderheide 2003: 236–240). Nonetheless, a significant level of care had been taken to preserve Ameniryirt’s delicate features and, although not a plug, some low-density material appears to have been inserted in his nose as part of the

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embalming process. The scans show that the brain was successfully removed and the skull is completely empty. The embalmer chose not to fill the cranium with textile or resin to aid preservation, as has been observed in other mummies (Wade et al. 2011; Wade and Nelson 2013a), but some of the dura mater, the membrane that surrounds the brain, has been left behind. Some efforts were made to render the body as lifelike as possible. Ameniryirt’s eye sockets were filled with loose textile bundles (e.g., Aufderheide 2003: 241; Taylor and Antoine 2014), although very little packing materials were used to maintain the shape of the face. Unlike examples from earlier periods (see Aufderheide 2003: 236–245), the CT scan shows that the cheeks, mouth, and throat were not filled with textile and resin. Overall, the embalmers were very successful in their attempt to preserve Ameniryirt’s features (Fig. 3). In Egypt’s predominantly hot climate, the internal organs of the chest and abdomen would have been removed soon after death to prevent decomposition, usually via an incision in the left flank (Wade and Nelson 2013b). This was the route taken to remove Ameniryirt’s organs (Fig. 3). To desiccate the body and halt the destructive activity of enzymes and bacteria, the remaining body fluids would need to be rapidly removed. This was occasionally achieved naturally by the hot desert sand, as with the natural mummies from Gebelein (see below), but the artificial process developed by ancient Egyptian embalmers required the use of a salt compound called natron (Ikram and Dodson 1998: 112–113; Taylor 2001: 55–56; Aufderheide 2003: 255–256; Taylor and Antoine 2014: 53–59). Predominantly composed of sodium carbonate decahydrate and sodium bicarbonate, natron occurs naturally as a deposit around dry saline lake beds in certain parts of Egypt. This very efficient drying agent breaks down body fat, and, when exposed to moisture, the carbonate in natron raises alkalinity, creating a hostile environment for bacteria. Dry natron in powder form would have been heaped over the body and, in some individuals, linen bundles full of natron powder may also have been used. Packed inside the empty cavities of the chest and abdomen, the bundles would have helped desiccate the body from within. The corpse would have been left in this state for about 35 days to allow the salt to extract all the fluids and halt decomposition. The 70 days mentioned in Herodotus’ account most probably refers to the duration of the entire process (Peck 1983; Taylor 2001: 56, 76–78), with the second half focusing on “rebuilding” the body. The rebuilding phase would often involve filling empty spaces, such as the abdomen, with a variety of materials to strengthen the body and prevent decay (Aufderheide 2003: 237). Previous studies have shown that a range of materials could be used including sawdust, earth, dried lichen, or bundles of linen (Aufderheide 2003: 252–254; Ikram and Dodson 1998: 160–164; Łucejko et al. 2017). In the case of Ameniryirt, the desiccation treatment successfully dried the body and removed most of the fat, leaving the skin and connective tissues well preserved. Dense granular packing material and textile were used to fill part of the space within the ribcage and abdomen, where the internal organs once were. The packing materials appear to have been inserted via the left flank incision and slope down toward the chest, the uppermost part of which remains empty (Fig. 4). The bulk of the packing is homogeneous but, unfortunately, its exact composition cannot be determined from the CT scan. Its granular structure suggests sand, sawdust, or

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Fig. 4 CT scan visualizations of the mummy of Ameniryirt (British Museum EA 6669). His mummified remains are virtually unwrapped (left to right) and packing materials highlighted in the transparency view (right of center) and longitudinal section (far right). The transparency view also includes the three packages found on top of his lower abdomen, thighs, and knees. Covered by the thick layer of resinous material, they most probably contain the remains of his mummified internal organs. Images courtesy of the Trustees of the British Museum

another comparable material was used. To seal it in, a plug of textile was inserted in the incision (Fig. 3). The CT scan also reveals that the embalmers applied molten resins (often pine, spruce, cedars, Pistacia resin, or bitumen; Taylor 2001; 57–58; Aufderheide 2003: 253; Clark et al. 2016; Łucejko et al. 2017) to the surface of the body. In other mummies, resins were also poured inside cavities such as the skull and chest. These substances would aid preservation and give the body a more pleasant smell, symbolically conferring divine qualities on the deceased (Taylor 2001: 57– 58). The scans show that most of Ameniryirt’s body was covered with such a resinous layer, including his head and face. It does not appear to have been directly poured over his skin. Ameniryirt was first wrapped by what seem to be two layers of textile soaked in resin, leaving large bubbles of air between the two. The substance was probably still in a viscous state when applied to the body, with a large quantity pooling behind Ameniryirt’s back. Of the organs removed by the embalmers, only the liver, lungs, stomach, and intestines appear to have usually been preserved in a manner similar to the body itself (see below). Frequently made into four separate bundles, they were often placed in special containers called canopic jars. However, from the Third Intermediate Period onwards, the dried organs were often returned to the body after

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treatment and placed inside or on top of the mummy. Ameniryirt was mummified after the introduction of this practice, and what appears to be three large flat packages can be seen lying on top of his lower abdomen, thighs, and knees (Fig. 4). They are also covered by the thick layer of resinous material that was applied over his body. Their contents cannot be ascertained using the CT scan images but they are very likely to include the remains of his mummified internal organs. The heart, regarded by ancient Egyptians as the center of intellect and memory, was often – but not always – left in place by embalmers (Taylor 2010: 54; Wade and Nelson 2013b). Indeed, the remains of what appears to be Ameniryirt’s mummified heart are visible on the CT scan images of his chest. The generally accepted idea is that the true function of the brain, which was seemingly discarded during mummification, was not understood (on the various explanations for excerebration, see Aufderheide 2011). Whether or not this organ was preserved in some form is, however, still a matter of debate. In many mummies, the hole used to access the cranium can be relatively small (circa 1–2 cm2 in the case of Ameniryirt) and the delicate bones within the nasal aperture, such as the inferior conchae, show little damage. The removal of the brain appears to have been a very skilled procedure that may have involved some form of liquefaction, as suggested by Herodotus’ text. The remaining liquid or partially devolved brain may have been processed in a manner that makes it hard to detect on CT scans and/or inside canopic jar, particularly once dried. The low density of any remaining brain tissue would make it hard to distinguish from other soft tissues and/or embalming materials, particularly if placed back into the body or combined with other soft tissues within an organ bundle or canopic jar. Throughout the development of the practice of mummification in ancient Egypt, the position of the body shows little variation, most often in a supine position. The most obvious change is the arrangement of the arms and hands. These could either be crossed over the chest, placed along the thighs on either side of the body or over the abdomen so that the hands covered the pubic region. Ameniryirt’s arms were crossed over his chest. Mainly observed on royal mummies during the New Kingdom, this practice became increasingly common in the Late Period onwards. This position is generally interpreted as a gesture associated with Osiris, invoking the resurrection of the deceased in the afterlife. The specific position of the hands – with the left fist clenched and the right hand lying flat on the elbow – is also seemingly linked to Osirian symbolism (see Elias et al. 2014). The final stage in the mummification process was the wrapping of the body (see Ikram and Dodson 1998: 153–192; Taylor 2001: 57–64). From as early as 4000 BC, embalmers were using matting, animal skins, and textiles, but linen wrappings eventually became the preferred material. CT scans show that the bandages around Ameniryirt’s mummy vary in thickness, from 4 cm over the chest area up to almost 12 cm over the neck. Helping to retain the physical integrity of the body, they also carried magical significance. A text known as the Ritual of Embalming describes the ritualistic placement of specific pieces of cloth, fragrant unguents, and oils over the body, as well as the spells invoking the gods which needed to be recited while preparing the mummy (see Töpfer 2015). In some mummified remains, amulets or

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“charms” were also placed in and around the body to magically protect it and provide it with special power (Andrews 1994; Ikram and Dodson 1998: 137–152). Finally, Ameniryirt’s mummy was surrounded by a single piece of textile secured by additional strips of textile. Dyed with tannins and madder, this shroud would originally have been dark pink or red. Now barely visible, the pigments used are, unfortunately, very light sensitive. A net made of faience beads was also placed on top of the shroud. Associating the deceased with Osiris, it is decorated with figures of the four Sons of Horus and a winged sun disk sewn on the chest.

Internal Organs, Canopic Jars, and the Four Sons of Horus Once removed, the liver, lungs, stomach, and intestines were treated separately by the embalmers using the same methods as those employed to preserve the body (see Ikram and Dodson 1998: 276–292; Taylor 2001: 64–76; Aufderheide 2003: 257– 259). The preservation of the organs perhaps reflects the importance of being able to consume food after death, as shown by the placement of food offerings in the tombs and the spells reproduced on many funerary artifacts. Once removed, the organs were separately embalmed with oils and resins, and wrapped in order to be available for all eternity. Often, the four organ bundles were then placed into vessels called “canopic jars.” The first examples date from the Fourth Dynasty (about 2600 BC; Aufderheide 2003: 258). Each jar was magically protected by one of the four Sons of Horus. The head of each of these four gods – mainly known for their association with the internal organs – would generally appear on the stopper of each vessel from the New Kingdom onwards: the human head represents Imsety, the baboon Hapy, the jackal Duamutef, and the falcon Qebehsenuef. As often stated on the jar itself, each one was supposed to guard the specific organ preserved in the bundle so that the deceased could still make use of it in the afterlife. However, the significance of an organ’s preservation and its relation to the gods and to the deceased are still little understood. The number of packages and jars could also vary (see below), which is perhaps an indication that embalmers were not necessarily focusing on the four organ groups that are usually mentioned. Using a range of radiological, histological, medical, and ancient DNA methods, many studies have focused on the analysis of the contents of canopic jars to gain valuable insights into embalming practices and the past prevalence of disease (e.g., Wade and Nelson 2013b; Eppenberger et al. 2018; Senti et al. 2018). A CT scan of some sealed canopic jars (Taylor and Antoine 2014: 60–63) has also revealed that a supplementary layer of protection could sometimes be added around the organs. Those belonging to a woman called Henutmehyt (British Museum EA 51813) had been placed inside miniature coffins. Delicately carved with the image of the goddess Nut spreading her wings, these anthropoid “coffinettes” imitate the cover placed on top of Henutmehyt’s mummy, replicating its function. Protected, the organs were considered to be an incarnation of the mummy in miniature and could also represent the deceased. The rich funerary material found with Henutmehyt suggests that she belonged to a particularly influential Egyptian family and few examples of such coffinettes are otherwise known. They are sometimes associated with royal burials – the coffinettes of Tutankhamun being the most famous.

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Canopic jars were used until the end of the New Kingdom (1550–1069 BC). During the Third Intermediate Period (1069–656 BC), a new fashion for placing the viscera bundles inside the body emerged. In some cases, wax figures depicting the four Sons of Horus were inserted inside the bundles containing the organs (see Taylor 2001: 72–76). By the beginning of the Late Period (664–332 BC), canopic jars were reintroduced, but organ packages were also regularly placed on the body, usually on or between the legs, just as they appear in Ameniryirt’s mummy. Organs were still traditionally placed into four bundles, but this custom was not always respected: Ameniryirt had only three packages, and up to seven can sometimes be found (Taylor 2001: 72). During the Graeco-Roman Period (332 BC–AD 395), organs were sometimes placed back inside the body cavity after desiccation or, in some examples, no efforts were made to remove the organs, particularly during the Roman Period (30 BC–AD 395). In some cases, both the organs – despite being left inside – and body are remarkably preserved (see Antoine and Vandenbeusch 2016).

The Coffin in Ancient Egypt Coffins were produced during the entire dynastic period until Roman times. As today, they had the simple function of protecting the body from external threats such as insects and animals. In addition to being a physical shelter, coffins were also believed to magically protect the deceased, with decorations and shapes used to facilitate their owners’ rebirth (Taylor 2001: 214–243; Ikram and Dodson 1998: 193–275; Taylor and Vandenbeusch 2018). This symbolism is confirmed by the ancient Egyptian name for coffin, “chest of life” or “lord of life,” and by the imagery and texts used to cover its surfaces. Coffins would house the deceased in a new dwelling for eternity, representing the tomb, as well as encompassing the entire cosmos with the sky and the earth (respectively represented by the lid and case), thus recreating the whole universe around the deceased. The notion of rebirth is also complemented by the transformation of the deceased into the god Osiris or by the association with the sun god Ra, both of whom experienced a cosmic regeneration in Egyptian mythology. The mummy was often placed in a set of nested coffins. Their decorative motifs and inscriptions would complement each other, increasing the complex funerary and magical properties. While the external coffin could be rectangular, replicating the shape of the tomb, the inner one usually imitated the anthropoid shape of the deceased’s divinely transfigured body. The shape of coffins, as well as the use of texts and motifs, evolved with time. In the Old (2686–2181 BC) and Middle (2055–1650 BC) Kingdoms, rectangular wooden coffins were the standard, and a pair of eyes through which the deceased could see the rising sun was commonly painted on the side facing east. After the New Kingdom (1550– 1069 BC), the dead were mostly buried in undecorated communal graves, and funerary texts and decorations were often transferred from the tomb walls to the coffins. During the Third Intermediate Period (1069–656 BC), coffin decorations became more complex and, in the Late Period (664–332 BC), innovations in shape and design reflect a change in the role of the anthropoid coffin. During that

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period, coffins started being placed on a pedestal, representing the mummy standing. In spite of the new decorative concept, coffins were still positioned horizontally in the tombs, but may have been held upright during the rituals which took place before the burial. Ameniryirt was buried in three elaborately decorated wooden coffins designed to protect his mummified body for eternity. All three are anthropoid and embody the deceased as an Osiris. Richly decorated, his inner coffin (EA 6668) was gessoed (a white, gypsum-plaster ground layer used to apply pigments) and painted on both internal and external surfaces (see Taylor 2003: 116). With its green skin, Ameniryirt’s face is particularly striking, identifying him with Osiris. The curled beard that ornaments some coffins – usually male – has also a divine significance and suggests the deceased has successfully achieved his divine status. Underneath, the central scene, the mummified deceased can be seen lying on a funerary bier, with canopic jars and other embalming materials underneath the bed. Although we know from the CT scan that Ameniryirt’s organs appears to have been wrapped in three packages that were placed on his mummified body, canopic jars continued to be an essential if somewhat symbolic element of the funerary equipment. Their representation on scenes like this persisted in the Late Period despite organs often being returned to the body, and dummy jars were even sometimes produced as part of the funerary equipment. In comparison with the inner and outer coffins (respectively EA 6668 and EA 6667), the middle coffin (EA 22811) is very simply decorated. On the outer coffin, Ameniryirt newly mummified body is seen placed on a lion-shaped bier being prepared by the embalming god Anubis.

New Methods: Toward a More Systematic Approach In the past, the only way to analyze Ameniryirt’s mummified body would have been to unwrap him, but the emergence of CT scanning in the 1980s has virtually eliminated the need to disturb his coverings. The investigation of mummies now makes use of the latest scanning technology and this noninvasive approach has been employed to study the mummies curated at the British Museum (Taylor and Antoine 2014; Antoine and Vandenbeusch 2016). New findings are used to create a personal profile of each individual, setting them in their historical, geographical, and social contexts, allowing us to explore particular aspects of life or death in ancient Egypt. Insights into their age, beliefs, and the diseases they suffered from help to remind us that all of the mummies were once living people and that, although they died many hundreds of years ago, they still require special ethical considerations (see Antoine 2014). Whether during transport, handling, storage, or display (Antoine and Taylor 2014; Wills et al. 2014), human remains should always be treated with respect, care, and dignity. Although they undoubtedly further our understanding of past human biology and culture, great care and thought must always be given as to the reasons for – and circumstances of – studying and displaying human remains (Antoine 2014; Antoine and Ambers 2014).

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New Developments in X-Ray Science and CT Scanning The Emergence of a New Discipline Before the emergence of archaeology as a scientific discipline in the late nineteenth century, mummies were perceived as curiosities rather than as sources of important knowledge on ancient societies (Ikram and Dodson 1998: 61–102; Aufderheide 2003; Taylor 2010; Taylor and Antoine 2014: 11–17). Even later, up to the mid-twentieth century, when X-rays began to be used, the investigation of mummies would usually involve their unwrapping. In England, this unfortunate trend appears to have been boosted by the explorer Giovanni Belzoni (1778–1823). Both irreversible and highly destructive, most of the mummies unwrapped in this way belonged to private collectors and little information about mummification was gained (Taylor 2010). From the 1880s to the early 1900s, the work and publications of Australian anatomist Grafton Elliot Smith (1871–1937), who unwrapped several royal mummies in Cairo, significantly expanded the understanding of the mummification process (Smith 1912; Ikram and Dodson 1998: 91–102; Aufderheide 2003). Valuable information was also gained from the histological and chemical analysis of mummies – and their embalming materials – by Marc Armand Ruffer (1859–1917) and Alfred Lucas (1867–1945) (see Aufderheide 2003). Developed in the late nineteenth century, radiology only became widely applied to the study of mummies in the 1960s–70s, when several important collections, including the royal mummies in the Cairo Museum and those in the British Museum, were X-rayed for the first time (Dawson and Gray 1968; Ikram and Dodson 1998: 91–102; Taylor 2004; Hawass and Saleem 2016). This noninvasive approach proved very valuable, revealing a wealth of new information without disturbing the bodies or their wrappings. The unwrapping of mummified remains is now rare and multidisciplinary studies focus on minimally invasive sampling. Extracted from damaged areas or incomplete bodies for histological or chemical analyses, these microscopic samples continue to expand our understanding of ancient diseases and embalming practices (Aufderheide 2003). CT Scanning Egyptian Mummies: Data Segmentation and 3D Visualizations Over the last few decades, the use of noninvasive X-ray imaging techniques has transformed the study of mummies. High-resolution three-dimensional (3D) imaging techniques, and in particular Computerized Tomography (CT) scanning, have superseded traditional X-ray machines. A modern CT scanner can analyze an entire body in less than five seconds, generating up to 10,000 two-dimensional X-ray slices (or tomograms) referred to as DICOM (Digital Imaging and Communications in Medicine), with a slice thickness of around 0.3 mm (see Ynnerman et al. 2016). Ideally, the slices should be combined to produce accurate 3D images using the latest generation of specialized volume rendering software developed for the analysis and visualization of CT scan data (e.g., VG Studio Max). With an emphasis on biological accuracy and realism, the 3D models generated are usually referred to as visualizations and take into account the density of what is being observed. Colors can be

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added to aid interpretation. These very detailed and accurate images allow for a better understanding of the mummy’s internal structure and the use of embalming materials, particularly their 3D shape and density. Such software also allows researchers to carefully separate layers – a process called segmentation – so that each tissue or structure can be virtually peeled away in order to observe what lies beneath, or be isolated and studied separately. Although there are some limitations (see Cox 2015; Gerald 2015), the 3D models can also be virtually cut so that crosssections of the body can be analyzed (Fig. 4). Unfortunately, researchers often rely on density differences to virtually distinguish, identify, and separate hard and soft tissues, as well as any embalming materials. Segmentation based exclusively on density can be a source of errors, particularly in areas where tissues and embalming materials are in close contact. This was the case with Ameniryirt, whose skin was covered in resin-soaked textiles (see above). In some areas of his body, the resin had permeated and combined with the skin and textiles, rendering their accurate segmentation difficult, if not impossible. In such areas, manual segmentation, where each element is identified on each of the thousands of DICOM slices, is required. This process often requires hours of processing in order to accurately identify the margins of each tissue or structure on all of the relevant hundreds or thousands of DICOM slices. Only by using this approach can the limits of low density elements, such as the skin, be correctly identified so that, as the bandages are virtually removed, the visualization of the mummified body does not include areas of leftover textile or missing patches of skin. Such errors are, unfortunately, easily missed once color has been added to the 3D rendering. However, this approach is not always available due to time constraints and the high costs of many volume rendering packages. Once delineated, the layers can be virtually removed one-by-one to reveal the embalming materials, bandages, skin, muscles, bones, and internal organs, as well as any objects placed inside the mummies. In the case of Ameniryirt, much of the resin had permeated into the skin, particularly over his face, and could not be fully removed (Figs. 3 and 4). To improve data segmentation, the CT scan visualizations presented in this chapter were generated using VG Studio Max volume rendering software. Most mummies were also scanned using the latest generation of multidetector CT scanners, a Dual Energy CT scanner which uses two X-ray energy sources and two sets of detectors within one scanner (see Taylor and Antoine 2014; Antoine and Vandenbeusch 2016). This allows for a combination of high-energy spectrums (120 or 140 kV) and lower-energy spectrums (80 or 100 kV), with lower settings preferred for lower density mummies (e.g., thinly or unwrapped mummies and non-adults). The combined data offers greater scope for improved segmentation (Ynnerman et al. 2016), with the removal or isolation of specific voxels that contain calcium (Tozer Fink and Fink 2018) and the differentiation of multiple materials (McCollough et al. 2015). Dual Energy scanners also provide additional information about the behavior of tissues or materials at different energies, and thus their composition (Coursey et al. 2010), beyond what is obtainable with single-energy techniques. This can help determine whether the amulets placed within the mummies (or within their wrappings) are made of stone, wax, or metal. Dual spectrums can

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also be achieved by using a single source CT scanner, scanning the mummy twice at different wavelength settings – preferably without moving the mummy – and combining the datasets using volume rendering softwares. The Dual Energy appears to be particularly good at differentiating desiccated soft tissues from low-density bones (Bewes et al. 2016) and has been shown to help reduce manual segmentation times in human mummies by enabling improved semi-automated techniques (Friedman et al. 2012). It can also help with metallic amulets that are sometimes placed in or around the body during embalming and block X-ray radiation from getting to the sensor array, often generating scattering artifacts. The impact of such artifacts on the data gathered can be reduced with state-of-the-art reconstruction algorithms, modified scanning protocols, and multi-energy CT scanning (see Ynnerman et al. 2016). By improving segmentation and increasing the amount and resolution of the data generated, multi-energy scanning allows researchers to create very detailed and clear 3D models, making it possible to capture both the lower and higher density elements without compromising one for the other (e.g., Antoine and Vandenbeusch 2016). Any segmented data can also be 3D printed and shared with audiences and academics (e.g., Taylor and Antoine 2014: 81–90). High-density objects placed inside or around the mummy during embalming, particularly those made of metal, can easily be segmented. Others with densities similar to the skin and surrounding textiles can sometimes be more difficult to separate, particularly if they have been place inside the body and are covered in resin. In such cases, the margins of the objects or amulets need to be carefully segmented so as to avoid the inclusion of any of the surrounding soft tissues, embalming materials or textiles. Without the need to disturb the originals, it is now possible to hold an exact replica in the full knowledge that they are still where the embalmer placed them thousands of years ago.

The Bioarchaeology of Mummified Human Remains: Toward a More Systematic Approach In addition of the detailed analysis of soft tissues, improvements in scanning and visualization techniques now make it possible to unlock valuable biological information found in the skeleton (Taylor and Antoine 2014; Ikram et al. 2015; Antoine and Vandenbeusch 2016; Nystrom 2019). Carefully segmented bones can be analyzed using methods originally developed for the study of skeletal remains by bioarchaeologists, forensic archaeologists, and physical anthropologists. By applying established methods and scoring systems that have been tested on individuals with a known-history (e.g., sex and age), the use of a more systematic and repeatable approach allows for direct comparisons across studies. It also clarifies how the biological profile of a mummified individual was established (e.g., methods used to determine the age-atdeath) and allows for future reinterpretations based on the most up-to-date methods.

Establishing an Age-at-Death The latest generation of CT scanners allows mummified remains to be carefully scrutinized without the need to unwrap them, providing bioarchaeologists with a

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wealth of previously unattainable information. In non-adult mummies, the stage of dental development (e.g., Taylor and Antoine 2014) and the development of the skeleton can be carefully assessed to estimate an age-at-death (e.g., Antoine and Ambers 2014; Antoine and Vandenbeusch 2016). Starting before birth and ending with the completion of the third molar as late as the early 20s, teeth begin forming inside the mandible and maxillae, and subsequently erupt. Not all teeth form at the same time, and the sequence in which they form – apart from the highly variable third molar – is broadly consistent and regular, and can be used to establish a developmental stage (Ubelaker 1989). Radiological studies of living children are used as reference to calibrate these stages and provide an estimate of the age-at-death (Hillson 1996; Hillson 2014). An approximate skeletal age can also be established using both the times of appearance and fusion of bones, as well as the size and shape of growing bones. The various bones of the body have very different growth patterns and timescales of development (see Buikstra and Ubelaker 1994; Scheuer and Black 2000). However, these approaches only provide an approximate age-at-death as both dental and skeletal development can vary between individuals and populations. It is often best to assign individuals to age cohorts such as Preterm (less than 37 weeks), Fullterm (37–42 weeks), Infancy (0–1 years), Early Childhood (2–5 years), Late Childhood (6–10 years), Puberty (11–15 years), and Adolescent (16–19 years). Based on the recommendations set out in Buikstra and Ubelaker (1994) and the groupings discussed in Scheuer and Black (2000: 368–369) and White et al. (2012: 381–385), these subdivisions account for the difficulties in assigning precise ages and allow for comparison between age groups and other published data. In mummified remains, once the skeleton has finished developing and most of the bones are fused, an age-at-death is often best established using age-related changes to the pubic symphysis (Brooks and Suchey 1990; Buikstra and Ubelaker 1994: 21– 37). Located on the front of the pelvis, the surface of this joint has been successfully visualized in CT scans of living patients (Wink 2014), known-age cadavers (Merritt 2018a), and mummified individuals (see below; Taylor and Antoine 2014; Antoine and Vandenbeusch 2016; Hawass and Saleem 2016). This approach offers a more reliable indicator of age than dental wear, which can be influenced by factors other than age (diet and food processing for example). Unfortunately, the pubic symphysis can be difficult to image in some mummified remains, particularly if a metal amulet has been placed immediately over the pubis. Due to the progressive nature of the changes to its morphology and the error range of the scoring method (see Buikstra and Ubelaker 1994: 21–37), it often only allows for an approximate age-at-death and whether a person is most likely to have died as a young adult (20–34 years), middleaged adult (35–49 years), or old adult (50 years or older). Cranial suture closure and changes at sternal end of ribs (see Buikstra and Ubelaker 1994; White et al. 2012: 379–427) have also been used to establish the approximate age-at-death of mummified adults (e.g., Hawass and Saleem 2016). The strengths and limitations of both methods have, however, been the focus of much debate (e.g., Nikita 2013; Muñoz et al. 2018; Ruengdit et al. 2020). Developed for the analysis of skeletal remains, assessing such age-related changes via CT scans (e.g., Moskovitch et al. 2010; Merritt 2018b) may also require modified scoring

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systems and, most likely, broader or differing age interpretations. Their use on mummified remains also has the potential to improve aspects of the methods. The sternal end approach requires the assessment of a specific rib (usually the first or fourth rib), something that is easier to achieve in well-preserved and articulated remains. As noted by Ruengdit et al. (2020), CT scans may similarly improve the disputed reliability of cranial sutures as estimators of age by providing a clearer evaluation of their state of fusion. One such CT study, based on an autopsy sample of 231 individuals with an age range of 19–89 years, was able to distinguish younger individuals (60 years) using the degree of ectocranial suture closure (see Boyd et al. 2015). Further CT scan applications of both methods (e.g., Merritt 2018b) should help clarify their reliability and allow us to develop recording protocols that are more suited to the analysis of mummified remains. Ideally, these should account for the impact of the CT scan resolution on their visibility (e.g., cranial suture closure) and of the embalming processes (e.g., organs removal, packing insertion, desiccation) on the structural integrity of the areas being assessed (e.g., sternal rib end).

Establishing or Confirming the Biological Sex Determining the sex of an individual can be difficult when the soft tissue anatomy is not sufficiently preserved, cannot be clearly imaged on the scan, or is obscured by embalming materials. The pelvis, and to a lesser extent the skull, are good indicators of biological sex (Buikstra and Ubelaker 1994). Estimation is dependent upon observations of multiple skeletal characteristics found on the pelvis, cranium, and mandible. In women, for example, small changes in the angles and shapes of the pelvis – such as a wide sciatic notch or the presence of a sub-pubic concavity – reflect an adaptation to the biological requirements of childbirth (Buikstra and Ubelaker 1994: 16–19; Bruzek 2002; White and Folkens 2005: 385–387). In males, these characteristics differ with, for example, a narrow sciatic notch and sub-pubic angle (e.g., CT scan of Gebelein Man A in Antoine and Ambers 2014). However, due to the often gracile nature of some Nile valley populations, estimation of sex based on cranial traits can be difficult (Godde et al. 2018) and the pelvis should be accorded greater strength. The Life and Death of Ameniryirt: A Bioarchaeological Perspective The CT scans can also be used to estimate the stature of mummified individuals, as well as to investigate their state of health and the embalming techniques used to preserve them. Ameniryirt’s CT scan reveals that, based on modern standard, he was a relatively short man. From the top of his head to the base of his heels, his mummified body measures only 155 cm. To compensate for the desiccating effects of mummification, his height was also estimated using the length of his long bones and based on those measurements (after Raxter et al. 2008), he was most probably taller at around 161–167 cm (stature estimate: 164.3
2.9 cm). The Dual Energy CT scan and 3D visualization software revealed clear changes to the outline and surface of his pubic symphysis suggesting that he was most probably between 35 and 49 years old when he died (see Fig. 5; Suchey-Brooks score 4, Buikstra and

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Fig. 5 Pathological and age-related changes found on Ameniryirt’s skeleton (British Museum EA 6669). The CT scan visualization of his pubic symphysis (bottom center) shows the joint has remodeled (Suchey-Brooks score 4) and he was probably between 35 and 49 years old when he died. The visualization of his pelvis also reveals the presence of multiple irregular osteolytic lesions of differing sizes (upper left), some with well-outlined sclerotic margins (right), characteristic of a metastatic cancer-induced bone response (metastatic carcinomas) from an unknown primary soft tissue cancer that had metastasized to Ameniryirt’s skeleton. Periapical lesions were also found at the end of the roots of one tooth, the upper-left first molar (one lesion shown; bottom left). Images courtesy of the Trustees of the British Museum

Ubelaker 1994: 21–32). Evaluating Ameniryirt’s state of health at the time of death can be difficult as signs of disease are hard to detect in mummified remains (see Aufderheide 2003: 418–499). Many of his internal organs were removed as part of the embalming process (see above) and the surviving organs and mummified tissues have dried, shrunk, and become distorted, often compromising their identification and the detection of any pathological changes. The clear CT scan images of his skeleton provide valuable biological information and, apart from the bones inside his nasal cavity, which were broken by the embalmers to remove his brain, it is relatively intact. Yet, only a limited number of diseases involve bones, and many can take several years before affecting the skeleton (Roberts and Manchester 2005; Waldron 2009). This limits the available evidence, particularly when a disease has yet to spread to the skeleton or only affects organs and other soft tissues (some of which may have been removed or modified by the embalmer). Evidence of physical trauma

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and broken bones can be easier to see but care must be taken to differentiate trauma that take place prior to death (ante-mortem) from any post-mortem changes that may have occurred during embalming, burial, or transport (Rühli and Böni 2000). Hence, detecting and interpreting pathological changes in an embalmed, desiccated, and – when organs have been removed – incomplete body can be difficult, and a cause of death is rarely established. In Ameniryirt’s case, however, the CT scan revealed that he was suffering from a range of pathological conditions at the time of death. His pelvis is riddled with numerous destructive circular and ovoid lesions of varying sizes, several of which are over 2 cm in diameter (Fig. 5). The lesions all appear to be destructive with no detectable signs of repair or extensive new bone formation apart from the presence of a thin sclerotic margin (dense layer of new bone) at the edge of some of the lesions. Several conditions, including many cancers, produce destructive lesions that can affect the pelvis. However, the presence of multiple irregular osteolytic (destructive) and osteoblastic lesions (with well-outlined sclerotic margins of new bone) of differing sizes indicates a metastatic cancer-induced bone response (Disler and Miklic 1999). Metastatic cancers (or metastatic carcinomas) spread from a primary site of origin, where the cancer started, into different areas of the body. With the embalmers having efficiently removed most of Ameniryirt’s internal organs, it is unclear where the cancer may have originated. Despite a lack of direct evidence for the primary site, the presence of numerous pelvic lesions of differing sizes (see Disler and Miklic 1999; Bloem and Reidsma 2012) clearly points to an unknown soft tissue cancer that has metastasized to Ameniryirt’s skeleton. Today, cancers of the breast, lung, prostate, kidney, gastrointestinal tract, and thymus account for most metastases in the axial skeleton (Disler and Miklic 1999), and are a common cause of malignant lesions of the pelvis in individuals over 40 years old (Bloem and Reidsma 2012). With a focus on the pelvis and an absence of detectable lesions in other skeletal elements, the primary tumor may have been in the prostate. Metastatic cancers are rarely described in the archaeological record. The earliest clear example was found at the site of Amara West in Northern Sudan (c. 1200 BC; Binder et al. 2014). Most archaeological examples of cancers to date, including the metastatic carcinoma from Amara West, were observed on skeletal remains and evidence from mummified individuals is rare. Cancer may not have been as prevalent in ancient Egypt as it is today. This probably reflects the impact of modern living, such as smoking and our exposure to modern carcinogens. Cancerous lesions can also be hard to detect in skeletal and mummified remains, and the lower life expectancy of past populations offered less time to develop skeletal lesions (Binder et al. 2014). Ameniryirt’s mummy adds to this scarce body of evidence and, having lived around 600 BC, he provides new insights into the considerable antiquity of cancer. Many of Ameniryirt’s arteries are still present and relatively well preserved. The CT scan revealed that some of them have considerable calcified plaque deposits (atheromas) on their inner surfaces, a disease called atherosclerosis. Ameniryirt’s heart was not removed by the embalmers and what remains of the arteries that feed and immediately surround it, including the coronary arteries, are affected. So are the

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carotid arteries, located in the neck, that supply blood to the brain. No detectable calcified plaque deposits were found in the arteries of his arms and legs. The formation of fatty plaques on the inner walls of arteries is caused by the interaction of multiple genetic and environmental factors. It is usually associated with cholesterol and a diet rich in animal fat, as well as with obesity, but there is also a strong element of genetic susceptibility. The plaques themselves can harden and become calcified, making them detectable on a CT scan by appearing as abnormally dense layers inside the artery. Plaques cause the affected arteries to harden, narrowing the available space, obstructing the flow of blood and damaging organs. Atherosclerosis is a major risk factor in cardiovascular disease. Any detached plaque can cause blood clots, which can in turn block the blood supply to the brain or heart, triggering a stroke or a heart attack. The location of the plaque in Ameniryirt’s arteries suggests he suffered from coronary heart disease, with a partial or totally blockage of the blood flow in the arteries that supply the heart. This may have caused angina (chest discomfort or pain) or may even have resulted in a heart attack. He also appears to have suffered from carotid artery disease, which may have reduced or blocked the supply of oxygen-rich blood to the brain and led to a stroke. As Ameniryirt also suffered from cancer, it is hard to ascertain what he died from, but atherosclerosis – and the resulting cardiovascular diseases – is regarded as the single biggest cause of death in the developed world today, accounting for a third of all deaths (World Health Organisation 2017). Interestingly, as scanning methods improve, arterial plaque is increasingly being detected in ancient Egyptian mummified remains (e.g., Allam et al. 2011). Cardiovascular diseases are not exclusively a feature of the modern world and appear to have been particularly prevalent among some inhabitants of the Nile valley (see Binder et al. 2021). The evidence, however, is biased toward persons of high status who could afford mummification. Most ancient Egyptians were given much simpler burials without any embalming and their soft tissues rarely survived. Owing to their greater wealth, high status individuals may also have had a diet richer in animal fat and a higher prevalence of obesity, exposing them to different risk factors and increasing their chances of developing cardiovascular disease. Consistent with an age of between 35 and 49 years, Ameniryirt’s teeth show signs of advanced dental attrition and most of the crowns are very worn, almost to the roots. Compared to some of the other mummies in the British Museum’s collection, Ameniryirt’s dental health was relatively good in spite of the heavy attrition. Periapical lesions were only found at the end of the roots of one tooth, the upperleft first molar at the back of the mouth (Fig. 5). Located around the tips – or apexes – of the roots, periapical lesions are often the result of a chronic infection that lead to the formation of a dental abscess. The infection develops when bacteria enter the pulp chamber, where the nerves and blood vessels are located, and travels along the root(s) to form a pocket of pus at the apex. These are clearly visible on CT scans as a hollowed area where the bone has remodeled away (i.e., resorbed) to accommodate the growing mass. The lesions at the end of Ameniryirt’s molar roots (the upper molars usually have three roots) may not actually have been abscesses at the time of death (see Hillson 1996: 284–287). Instead, each lesion may have been filled with a less problematic

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granuloma (a mass of tissue produced in response to inflammation or infection) or a cyst (a cavity filled with liquid or semisolid matter). Unfortunately, abscesses, granulomas, and cysts are hard to differentiate from one another on a scan. Severe tooth decay and extreme dental wear are the most common reasons for bacteria entering the pulp chamber, resulting in an infection, the formation of a periapical lesion, and, in most cases, severe throbbing pain. In Ameniryirt’s case, the lesions at the end of his molar are probably the result of a past infection caused by excessive dental wear. A lesion known as Schmorl’s Nodes was also found on one of Ameniryirt’s vertebra (tenth thoracic). Often relatively small, Schmorl’s Nodes are formed when the intervertebral disc (known as the nucleus pulposus) protrudes into the bony tissue of the adjacent vertebra. Clearly visible on the CT scan cross section of the spine, each abnormal protrusion (i.e., herniation) creates a small indentation into the surface of the vertebra. Here, the lesion is also surrounded by a high-density edge called a sclerotic margin, showing that the bone has reacted and some degree of remodeling has taken place during life. Schmorl’s Nodes are very common and usually regarded as minor age-related degenerations.

The Evolution of Mummification from the Predynastic to Medieval Periods, with New Insights from the British Museum Collection Through the analysis of mummified remains and embalming materials, the development of mummification in ancient Egypt is progressively being revealed (Aufderheide 2003; Ikram and Dodson 1998; Peck 1983; Taylor 2001, 2010), with the use of complex and evolving techniques showing both temporal and regional variations and trends (e.g., Wade et al. 2011; Wade and Nelson 2013a, b). Over the past decade, mummies held in the British Museum collection have been the focus of new research using the latest scientific methods (Taylor and Antoine 2014; Antoine and Vandenbeusch 2016). Built up progressively since the Museum’s foundation in 1753, the British Museum curates one of the largest collections of Egyptian mummies outside Egypt. Most were found in the nineteenth century and information about their findspots and context is often lacking (see Taylor 2014). As not all periods are represented in the Museum’s collection, new findings are combined with published data to outline major shifts in the evolution of mummification over time. Influenced by temporal, geographical, and socioeconomic factors, variations are likely to have emerged from practices developed in workshops or by single individuals, most probably through trial and testing.

Predynastic Period (c. 5500–3100 BC) and Early Dynastic Period (c. 3100–2686 BC) Buried in oval or circular shallow graves without a coffin and in close contact with the hot sand, some Predynastic bodies mummified naturally. Often wrapped in textile,

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matting, and/or hides, they were usually laid on their sides in a tightly flexed position. Though few natural mummies from the Predynastic Period have survived to this day, many more are likely to have existed in ancient times and their chance discovery probably guided the beliefs toward the need to preserve the body after death (Ikram and Dodson 1998: 108; Taylor 2001: 79). Seven well-preserved examples from the site of Gebelein, in the southern part of Upper Egypt, are curated at the British Museum. Little is known of their burial context or status (Budge 1920; Dawson and Gray 1968). Their grave goods, burial setting, and the flexed position of the bodies, as well as what remains of their linen, matting, and animal hide coverings, are all characteristic features of the Predynastic burial tradition (Friedman et al. 2018). The radiocarbon results and supporting isotopic analyses on hair and bone sampled from six of the bodies produced dates ranging from 3932 to 3030 cal BC (68.2% probability; Friedman et al. 2018). Dating from the era preceding the country’s unification by the first pharaoh at around 3100 BC, their bodies do not display any apparent signs of the deliberate embalming methods seen in later periods. Of the three that have been CT scanned, none showed evidence of invasive treatments: Gebelein Man A and Gebelein Man B (British Museum EA 32751 and EA 32754 respectively) and Gebelein Woman (EA 32752) (Fig. 6). The remarkable preservation of their skin, hair, and nails is equality matched by the natural mummification of their internal organs, many of which are clearly preserved and visible on the CT scans (see Antoine and Ambers 2014; Taylor and Antoine 2014; Ynnerman et al. 2016; Fig. 6). The excellent survival of their internal organs suggests that their bodies rapidly dried out. The scan from the three Gebelein mummies also revealed the two male mummies were of a young age. Many of Gebelein Man A’s long bones are completing their fusion and he was probably around 18–21 years old when he died. A stab wound through his left shoulder was shown to have fractured his fourth rib, most probably perforating his left lung. With no evidence of defensive wounds or healing, this was the likely cause of death (Antoine and Ambers 2014; Ynnerman et al. 2016). The scans also showed that Gebelein Man B’s pubic symphysis has no or little remodeling and that he was a young adult between the ages of 20–34 years when he died (Taylor and Antoine 2014; Fig. 6). Further studies by Friedman et al. (2018) have also revealed that two of the Gebelein mummies had tattoos. Using infrared imaging, two horned animals were observed on Gebelein Man A’s right arm (Fig. 6). They most probably represent a wild bull and a Barbary sheep, both of which are well-known in Predynastic art. The four small “S” shaped motifs and the linear motif found on Gebelein Woman’s shoulder and right arm (Fig. 6) also have parallels with the imagery of the second half of the Predynastic Period (Naqada IIC-IID; ca. 3500–3300 BC; Friedman et al. 2018). The linear motif closely resembles objects held by figures painted on ceramics of the period and may represent a crooked stave, throw-sticks, or clappers. The S-motif on her shoulder is also a common element of Predynastic pottery decoration, although their significance remains unclear (Friedman et al. 2018). The hair sample from Gebelein Woman produced a radiocarbon age of 3351– 3092 cal BC (2 σ, 95.4% probability) and the sample from Gebelein Man A was

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Fig. 6 CT scan visualization of the Gebelein Man B (British Museum EA 32754) in transparency view to highlight his preserved internal organs (top) and his segmented right pelvis (bottom left), with a clearly visible pubic symphysis. Infrared images of the tattoos found on Gebelein man A (British Museum EA 32751) (center right) and Gebelein Woman (British Museum EA 32752) (center and bottom right). Images courtesy of the Trustees of the British Museum

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dated to 3341–3017 cal BC (2 σ). These results correspond to the latter part of the Predynastic Period, confirming that tattooing was practiced in Predynastic Egypt. Nearly contemporaneous with Ötzi the Iceman, they are some of the oldest preserved tattoos in the world and push back the evidence for tattooing in Africa by a millennium. They also make Gebelein Woman the oldest tattooed female to date and provide new insights into the uses of body art in prehistoric Egypt (Friedman et al. 2018). With the emergence of deeper and more elaborate burials in the second half of the fourth millennium BC, bodies were further removed from the heat of the desert, preventing their natural mummification. Even before the introduction of coffins and tombs, more attention appears to have been given on preserving the body by artificial means. At other Predynastic sites such as Hierakonpolis, the use of linen strips and bundles to wrap and pad the body, as well as traces of resin found on their skin, reveals that early attempts at mummification were taking place from the middle of the fourth millennium BC (Ikram and Dodson 1998: 109; Taylor 2001: 79; Jones 2002). Jones and colleagues have also shown that some form of treatment of the textile or skin may have been used during this period (Jones et al. 2014, 2018). By the Early Dynastic Period, the wrapping of the body appears to have become wellestablished. The disembodied arm from the tomb of King Djer (First Dynasty, around 3000 BC) at Abydos, for example, reveals that limbs were being tightly wrapped in linen and, though skeletonized, it is regarded by many as an early attempt at soft tissue preservation (Aufderheide 2003: 222–223). However, most examples from the First and Second Dynasties show little tissue preservation, with only the bones remaining inside the wrappings. During this period, bodies were often placed in coffins of wood or clay and, as in the Predynastic Period, continued to be flexed, most often on their left side (Aufderheide 2003: 222–223; Ikram and Dodson 1998: 109). Further evidence for the use of resin emerges at Saqqara during the Second Dynasty. Although no soft tissues survived, the bodies had been tightly wrapped in a flexed position and covered in linen impregnated with resin (Spencer 1982: 35; Aufderheide 2003: 222–223). The experimentation seen in Predynastic and Early Dynastic Egypt would continue during the Old Kingdom.

Old Kingdom (c. 2686–2181 BC) and First Intermediate Period (c. 2181–2055 BC) During the Old Kingdom, mortuary practices became more complex (see Ikram and Dodson 1998: 21–60), with longer funerary rituals and the use of coffins, deeper graves, and the building of tombs (pyramids and mastaba) moving the body away from the heat of the desert, all of which inhibited natural mummification. This probably prompted the search for alternative methods of preservation, with an increasing reliance on evisceration and desiccation, and further evidence for the use of natron and liquid resin (Aufderheide 2003: 224–226). Few Old Kingdom mummies have survived to this day and only a limited number of individuals appear to have had access to the skills of an embalmer. Those that have been studied confirm

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that embalming was still very experimental, with varying degrees of success. Though perhaps limited, the use of evisceration was already part of the embalming process. The discovery of viscera in a container found in the tomb of Queen Hetepheres I, mother of King Kheops (fourth Dynasty, c. 2613–2494 BC), suggests that organs were being preserved separately. Their analysis has also revealed the presence of natron (Aufderheide 2003: 226). The body of the queen was, unfortunately, never recovered. During this period, incisions were usually performed vertically on the left flank (Aufderheide 2003: 226). It remains unclear whether the heart was already being left in place. According to Wade and Nelson (2013b), evisceration was a top to bottom practice, which was first seen among the royals and nobles of the Old Kingdom before being practiced on the population who could afford it. It involved the use of packing materials, such as linen and resin. There is, however, little evidence that excerebration was performed in the early part of the period. Removing the brain via the nose seems to have only started around that time, becoming more common practice toward the end of the Old Kingdom (Wade et al. 2011; Ikram and Dodson 1998, 111–112). Initially, the preservation of the skin was variable and most surviving mummies have little soft tissue preservation. Its preservation may, however, not have consistently been sought after and alternative practices, such as that of defleshing or flaming the body, might have also been attempted (see Weser et al. 1998). If anything, the external shape of the mummy was favored using complex packing and modeling made of textile and plaster. Expertly positioned layers of wrappings were used to create a lifelike human shape, with linen padding added under or over the skin (Taylor 2001: 80–81). The body of Ranefer, son of King Snefru from the Fourth Dynasty and found in Tomb 9 at Meidum, was eviscerated and carefully wrapped with linen and modeled using resin. As was often the practice at the time, Ranefer’s mummy was discovered lying on his left side, facing east, with eyes and eyebrows painted on using black, green, and red pigments (Petrie 1892: 17–8; Ikram and Dodson 1998: 110). Unfortunately, his remains were destroyed when a bomb landed on the Royal College of Surgeons in London during World War II. Wrapped in textile, modeled in plaster and painted, the mummy of Waty (or Nefer?), found in a wooden coffin in his Saqqara family tomb, was similarly prepared. The limbs, as well as the fingers and toes of a woman buried in mastaba G 2220 B in Giza (Fourth Dynasty) were wrapped separately. Resin and linen were also inserted to shape her body, face, and breasts, as well as create the appearance of a V-neck dress on the outer layer (D’Auria et al. 1988: 77–78). The dress, following the contemporary fashion, was a common feature of the period used to create a lifelike appearance by the adding clothing (other mummies have also been found wearing tunics and kilts). Her body was also lying on the left side, in an extended position, and had been placed in a wooden coffin. Fashioned in a lifelike manner, embalmers may have endeavored to make them look like statues so that they may live forever (Lacovara et al. 2015). The external appearance of the mummy appears to have been of upmost importance, even if the body itself was not particularly well preserved. The mummy of a man from Gebelein (now in Turin, inv. N. 13,019), dating to Fifth to Sixth Dynasty, was embalmed using multiple layers, with his limbs separately wrapped and his face painted with human

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features. Lying on his side, his body was found fully extended and placed in a stone sarcophagus (Delorenzi and Grilletto 1989, 38 and Table XIX a-b). Toward the end of the Old Kingdom, this naturalistic trend was further developed by using layers of plaster to model the face or the whole body into a human shape. Several mummies with plaster covering their wrappings were, for example, found in tomb G 2037b and appear to date to the Sixth Dynasty (D’Auria et al. 1988: 91–92). While the body was mostly buried in a flexed position in earlier periods, both supine and side-on burials appear during the Old Kingdom, with the body both in semi-flexed or extended positions. Coinciding with the use of full length coffins, the use of an extended position had already been introduced in the Third Dynasty (Taylor 2001: 80 and 218). In coffins, the head would also often rest on a headrest. One such example was found next to a flexed mummy at Gebelein that possibly dates to the Third or Fourth Dynasty (Turin, Museo Egizio N. 13020). With no evidence of treatment other than the wrappings, the bundled body was buried in a wooden coffin along with a headrest (Delorenzi and Grilletto 1989: 39). In a poor condition and heavily conserved, the mummy of a Sixth Dynasty man from Abydos (now in the Michael C. Carlos Museum in Atlanta; Lacovara et al. 2015) has separately wrapped limbs with his hands and feet ties together. He was buried on his right side and in a semi-flexed position (for a similar mummy, found in a wooden coffin with a façade de palais decoration and limbs wrapped separately, see Donadoni Roveri et al. 1995: 29). Due to a lack of preserved mummified remains, particularly those from securely dated contexts, we have relatively little evidence on the practice of mummification in the First Intermediate Period. Of the traditions established in the Old Kingdom, both evisceration and desiccation continued. However, the external appearance of the body switched from being naturalistic and lifelike to being concealed in a cocoon of linen, with a painted cartonnage mask representing an idealized image of the deceased placed over the head (see Taylor 2001: 81). The Ninth Dynasty mummy of Pepyseneb, from Sheikh Farag (tomb 5114, shaft c), now at the Museum of Fine Arts, Boston (inv. 25.1513), provides an example of a style that encapsulates the transition between Old and Middle Kingdoms practices. Eviscerated, but not excerebrated, the body was desiccated in an extended position and wrapped in a large bundle. Lying on his left side, with a cartonnage mask covering his face, he typifies a style that would become increasingly popular and spread across Egypt (Ikram and Dodson 1998, 114; Taylor 2001: 81).

Middle Kingdom (c. 2055–1650 BC) Once again, few relatively well-preserved Middle Kingdom mummies have survived. The mummified head found in the tomb 10A at Deir el-Bersheh, and believed to be from the Eleventh Dynasty, has been extensively researched (e.g., Loreille et al. 2018). Possibly that of Djehutynakht, the owner of the heavily looted tomb, the head has delicately modeled facial features, while the eyebrows are painted, both of which are reminiscent of the Old Kingdom practice. However, unlike most mummies from

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earlier periods, the brain had clearly been removed. Two routes seem to have been used to access the cranium, the nose and the base of the skull. Further adjustments also appear to have been performed, with the removal of zygomatic arches, as well as part of the mandible. The reasoning behind such transformations remains unclear and may have been part of some remodeling or the result of experimentation in the removal of the brain (Chapman and Gupta 2007). Although evisceration had been shown to help with the preservation of the soft tissues in earlier periods, it was not consistently carried out during the Middle Kingdom. This holds true for most mummification methods as, despite the overarching trends of each period, some techniques appear to have been favored by individual embalmers and others not. The Middle Kingdom also sees further evidence for the use of natron and resins. These appear to have been employed to preserve the bodies of six royal women associated to King Mentuhotep II (c. 2055–2004 BC) found at Deir el-Bahari. Tattoos noted on the skin of some of them attest to their preservation despite a lack of evisceration. With no visible incisions on their abdomens, some of their organs are still intact, while others seem to have been dissolved from within. In the latter, an aleo-resin similar to turpentine may have been injected via the anus to help this process, and the same route appears to have been used to remove them once liquefied (Ikram and Dodson 1998: 114). This technique, listed by Herodotus as a cheap embalming method, may have actually been an expensive – and somewhat experimental – treatment reserved for the privilege elite of the royal circle. Not all mummies from the Middle Kingdom show signs of embalming. Sixty bodies found in tomb MMA 507, also at Deir el-Bahari, appear to have been naturally mummified. They are believed to be soldiers who died in battle and many show signs of trauma, fatal wounds, and mutilations. They seem to have initially been buried in the dry hot sand, allowing the rapid desiccation of their soft tissues in a manner similar to the Predynastic mummies from Gebelein (see above), prior to being reburied in their final resting place. This may explain the presence of marks that appear to have been caused by birds of prey, as well as their relatively good preservation without any embalming (Winlock 1945: 7–24). During the Middle Kingdom, mummification was no longer the preserve of the royal family. X-rayed in 1935, the mummy of the Overseer Wah was shown to include scarabs and necklaces. Later unwrapped at the Metropolitan Museum of Art in New York (Winlock 1940), his brain does not appear to have been removed. Although the practice was known during the Middle Kingdom, it was certainly not yet the norm. Organs below his diaphragm – the liver, stomach, and intestines – were removed, while the lungs and heart were left in place. The sheer quantity of linen used to wrap his body is remarkable. In addition to 470 square meters of sheets placed on and in the coffin, a further 375 square meters of textile were recorded during his unwrapping. The wrappings included 75 items of textile of varying quality (Roehrig 2003), as well as a few animals (including a mouse, a lizard, and a cricket!) trapped in the wrappings. Many of these linen sheets and bandages were inscribed, some with the name of the deceased, while others might have belonged to family members or relatives who offered them for his embalming. The use of a gilded mask and the cocoon style wrapping follows on from the style first seen in the

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First Intermediate Period. Unlike Old Kingdom mummies, whose arms tended to be at their sides, Wah’s arms were folded over his chest, with the right over the left, a placement usually seen in the New Kingdom kings and Ptolemaic mummies (Ikram and Dodson 1998: 115). Though methods of organ removal were still variable, the use of resins seems to have become more common. Often combined with linen, they were also applied directly over the skin. The mummy of Senebtisi shows an extreme version of this practice. Dating to Twelfth Dynasty, she was buried on her side, facing east, in an anthropoid wooden coffin in Lisht. Her entire body was covered with resin, which had been poured on top of her after she had been placed in the coffin (Mace and Winlock 1916: 17–18). Her organs had been removed through an incision on the left flank, although the brain had not. The heart seemed to have been wrapped separately and placed back in the body, while her other organs were found in canopic jars. Her limbs were wrapped individually, with wads of linen used to pad the body. In addition to an overall covering, resin had also been used on specific location such as the eyelids, possibly to close her eyes (Smith 1916: 119–120). The use of padding, in the form of rolls of bandages, has also been observed with the earlier mummy of Henu, a high-ranking official who lived during the First Intermediate Period (c. 2181–2055 BC) and whose intact tomb was discovered at Deir el-Bersheh. The paddings had been carefully placed under the outer layers of textile, perhaps as an economical way of improving the general shape of the mummy without surrounding the body with too many layers (preliminary results presented by De Meyer et al. 2019). Although the Middle Kingdom shows some elements of continued development, the limited evidence reveals that different techniques were being used and soft tissue preservation varied, with many bodies reduced to skeletons (Taylor 2001: 82– 83). The modeling of facial features seen in the Old Kingdom continued, while evisceration via the left flank was by now frequently performed, as was the use of natron, and although experimental in some, the removal of the brain became more widespread (Ikram and Dodson 1998: 114–117; Taylor 2001:82–83; Aufderheide 2003: 227–230).

Second Intermediate Period (c. 1750–1550 BC) and New Kingdom (c. 1550–1069 BC) Evidence for embalming practices during most of the Second Intermediate Period is scarce. The presence of canopic jars in tombs dating to Thirteenth Dynasty seems to indicate a continuation of organ removal (Aufderheide 2003: 231). It is not clear whether the new Fifteenth Dynasty rulers, the Hyksos (a group originating from the Middle East), adopted the same funerary practices. Unfortunately, it is not until the Seventeenth Dynasty that further evidence emerges. The remains of King Seqenenre (¼ Taa II) are well known despite their poor preservation. Found in the Deir el-Bahari cache (DB 320), his dramatic death from blows to the head, with clear cranial trauma, is apparent. There is no evidence of brain removal, but the body was seemingly eviscerated and the cavity filled with linen (Smith and Dawson 1924: 83–86).

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In contrast, the body of Unknown Woman B, sometimes identified as Queen Tetisheri, reveals the great care with which the body could be mummified (Ikram and Dodson 1998, 118) and appears to be a continuation of the practices established in the Middle Kingdom. During the New Kingdom, often considered the “classical” period for mummification, standards and techniques reach a new high. The evidence shows that members of the royal family were benefiting from very elaborate embalming techniques but practices differed enormously outside this very elite group. Fortunately, the mummies of many New Kingdom kings were preserved and are identified. Lootings appear to have targeted the royal tombs of the Theban necropolis throughout the New Kingdom, particularly at the end of the period. Several papyri document the robberies and inspections of the Valley of the Kings, as well as ensuing trials. Tombs were restored and mummies were repaired, rewrapped, and reburied in various caches, where they safely remained until the end of the nineteenth century (see Reeves and Wilkinson 1996: 190–207). This work continued into the Third Intermediate Period, when the family vault of the high priest Pinedjem II was annexed for the royal bodies in the Twenty-Second Dynasty. Discovered in 1881, the Deir elBahari cache (DB 320) contained 40 mummies, including 10 kings. In 1898, a further 14 mummies (among them nine kings) were discovered in the tomb of Amenhotep II in the Valley of the Kings (KV 35). Others include the mummy of Tutankhamun, discovered in 1922 in his intact tomb. It is also believed that the body of Ramses I was among a small collection in the Niagara Falls Museum and, from 1999, in the Michael C. Carlos Museum in Atlanta. In 2004, his remains were returned to Egypt (Hawass and Saleem 2016, 1–2). The royal mummies have been the focus of extensive research. Initially studied at the beginning of the nineteenth century (Smith 1912), they were reexamined several times using previously unavailable methods (Aufderheide 2003: 236). Recently, the research has resumed with a comprehensive reanalysis by Hawass and Saleem (2016) using the latest CT scanning techniques and bioarchaeological methods. These studies have shown that, in the New Kingdom kings, brains were being systematically removed from mid-Eighteenth Dynasty onwards. The first three Thutmoses were, however, not excerebrated (Hawass and Saleem 2016: 225). A transnasal excerebration appears to have been favored. In some, resin was poured in the skull, and linen plugs soaked in resin were often used to pack the nose. Eye orbits, as well as the mouth cavity, were usually filled with linen and resin. Apart from the heart, most of the internal organs were also removed, usually through an incision on the left flank. With a diagonal incision starting from the navel, Tutankhamun appears to be an exception. Torsos were then filled with linen, resin, and linen packings. Their arms are normally crossed over their chest, a position that appears intermittently (for Twenty-Fifth Dynasty examples, see Ikram et al. 2018) until the Ptolemaic Period, when it becomes the norm (see Ptolemaic Period). Wrapping techniques for this group of mummies remain elusive as most were not discovered in their original bandages and were rewrapped when their bodies were relocated. The inscriptions left on the shrouds and coffins allowed them to be identified soon after their discovery. In an effort to stop further decomposition and

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prevent damage caused by fluctuating environmental storage conditions, many of these mummies were unwrapped once again in the late nineteenth century. Swiftly unwrapped – the mummy of Ramses II took less than fifteen minutes –, few details were recorded (see, e.g., Maspero 1886a, b). This also limits our understanding of the objects and amulets that would have been inserted in the bandages and on the body, and are no longer in situ. More recent CT scans revealed a few had also been inserted inside the bodies including amulets (wedjat eyes most commonly), beads, and a few pieces of jewelry. An animal bone and some seeds had, incredibly, also been used by embalmers to maintain the shape of Ramses II’s nose (see Hawass and Saleem 2016: 161–162, 232–35). The few non-royal mummies that have been researched reveal a broader range of embalming techniques, although mummies of relatives of the royal family do not dramatically differ. For example, the parents of Queen Tiy, Yuya, and Tuya, show a very similar preparation to that of the kings and queens. Their burials are also consistent with those in the Valley of the Kings and include sumptuous funerary equipment, probably a largess of their stepson King Amenhotep III (Quibell 1908: 68– 73). The bodies found in the tomb of Hatnufer, the wife of Hatshepsut’s architect, tell a different story (Aufderheide 2003: 242). They show no clear evidence of embalming, although one of them was undoubtedly wrapped as a mummy. Their remains document techniques that are similar to those used at Deir el-Medina, the village of workmen who built the royal tombs in the Valley of the Kings. These men prepared small family tombs, vibrantly decorated and densely filled with burial goods, for themselves and their relatives. During the recent study of one such tomb, the mummies of Kha and Merit showed that, under a complex layering of bandages, their bodies had barely been treated (Bianucci et al. 2015). They were not excerebrated or eviscerated. The use of imported pistacia resin was found on both bodies and their soft tissues are relatively well-preserved. However, the CT scan analysis revealed an extensive gap between the soft tissues and the wrappings, suggesting that the body had not entirely dried (if at all) before the application of the bandages and continued to shrink afterwards. Both bodies were adorned with substantial pieces of gold jewelry – some had even drifted in the wrapping due to the space left by the shrinkage. Merit’s long hair (or wig?) was particularly well preserved, and was probably plaited before being carefully placed between two layers of wrappings (see exhibition Archaeologia Invisibile, at the Museo Egizio, Turin, 2019–2020). Recent research at Deir el-Medina has also revealed further examples of tattoos (see above), showing many figurative motifs – including wedjat eyes and animals such as cobras and baboons – spread on the torso of a woman (Austin and Gobeil 2017). The embalmers who prepared the bodies of Kha and Merit, carefully anointing and wrapping them, certainly tried to abide by the coveted preservation of the physical body and its association to the divine and to Osiris in particular. Although the technique was not as complex as that developed for contemporaneous kings, the belief in an eternal life through a preserved body was relatively similar. Despite the simplified procedure, they would still be considered as mummified and perhaps redefin the idea of what mummification may have represented in ancient Egypt. The majority of the population, including the poorest communities, must have shared

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similar funerary beliefs and some of these ideas must have been applied to the treatment of the body when mummification could not be afforded. As revealed by excavations at various Tell el-Amarna cemeteries, simple wrappings using multiple layers of bandages and shrouds in a mummiform shape could also be used, thus broadening the spectrum of embalming methods (Stevens et al. 2020). Individuals were also regularly buried with amulets and other protective elements, including a few with rare examples of decorated coffins. Overall, embalming in the New Kingdom reveals the availability of broader range of materials from abroad, particularly oils and resins (Taylor 2001: 83–86). Soft tissue preservation also improves, with highly skilled embalmers achieving incredible result on the mummies of the kings and queens of the period (see Hawass and Saleem 2016). Now regular features, the extraction of the brain through the nasal passage (and occasionally the back of the head) and evisceration (usually via the left flank) had been perfected (Ikram and Dodson 1998: 118–124). Body cavities were now stuffed with packing materials and resins liberally applied, so much so they would occasionally penetrate into the bones (Taylor 2001: 84–85). The position of the arms also becomes standardized, lying along their sides for most individuals apart from the kings (see above; Ikram and Dodson 1998: 122–124). The embalming methods in the non-royal mummies, however, varied greatly and were often more rudimentary. Organs were often not removed and soft tissues poorly preserved.

Third Intermediate Period (c. 1069–656 BC) and Late Period (664–332 BC) Often regarded as the pinnacle of Egyptian mummification, the Third Intermediate Period builds on the methods perfected in the New Kingdom. With efficient evisceration and drying out techniques now the standard, embalmers began to focus on retaining the natural form of the body by inserting packing under the skin (Taylor 2001: 86–87; Ikram and Dodson 1998: 124–128). The face, neck, chest, back, and legs were filled via the left flank incision, the mouth, and/or additional incisions. The royal mummies of the New Kingdom show that similar fillings had already been attempted on the face (including the cheeks and neck), the torso, and the limbs (Hawass and Saleem 2016: 216–220). Subcutaneous packings were seemingly not a Twenty-First Dynasty invention, as often thought, and the search for the perfect human body shape had already started during the New Kingdom. Although this has never been clearly determined, such fillings appear to have been done after the body had dried. However, this idea may need to be investigated further. It would have made more sense to insert some of the packing before the skin had shrunk and lost its elasticity. Areas such as the neck were, for example, packed with mud or sawdust (Ikram and Dodson 1998: 124). The use of subcutaneous packing peaked during the Twenty-First Dynasty, with Twenty-Second Dynasty mummies revealing a reduction in their use, and a focus on specific areas such as the neck. Genital orifices could also be plugged, most probably to keep abdominal packing inside the body and aid preservation. Molten resin also continued to be poured into

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the cranium after the removal of the brain, as well as into the body cavity (Ikram and Dodson 1998: 124–128). In contrast to earlier periods, the internal organs are often no longer kept in canopic jars and start being placed back in the body cavity. The organs were usually wrapped in linen and wax figures depicting the four Sons of Horus were sometimes placed inside the bundles containing the organs. The number of bundles could vary and up to seven have been noted in some Twenty-First Dynasty mummies (Taylor 2001: 87). In both royal and non-royal mummies, arms were usually left lying alongside the body and hands placed over the genitalia (Ikram and Dodson 1998: 128). The trend for cosmetic treatments continued in the Third Intermediate Period, with the skin of some male and female mummies painted red and yellow, respectively, as well as the introduction of false eyes made of stone or glass (Ikram and Dodson 1998: 127; Taylor 2001: 86–87; Aufderheide 2003: 242–245). Great care was taken, for example, with the mummy of Tjayasetimu, a young temple singer from the Twenty-Second Dynasty (British Museum EA 20744). In her Late Childhood (dental development: 7 years
2 years), her hair is incredibly well preserved and, despite her young age, the CT scan data show no evidence of a sidelock. Instead, thick masses of what appears to be her natural hair had delicately been placed either side of her face, as well as at the back of her head (Taylor and Antoine 2014: 112–133). In contrast to what is commonly attested in the Twenty-Second Dynasty mummies, Tjayasetimu’s brain has not been removed. Her provenance is still debated (between the Fayum and the Theban area) and this differing approach may indicate a northern origin, with most other mummies of this period being Theban. A decline in standards is believed to have occurred over the length of the Third Intermediate Period, particularly after the end of the Twenty-First Dynasty, and by the Late Period, a greater reliance on large quantities of resin emerges (Taylor 2001: 87). The mummy of priestess reveals that mummification standards were still high in the Twenty-Second Dynasty (British Museum EA 25258; Antoine and Vandenbeusch 2016: 112–139). Placed within a cartonnage case, a trend that became widespread during the Third Intermediate Period, her mummified remains are very well preserved. The cartonnage around the feet is unfortunately damaged and the inscription where her name would have been written cannot be read. The remainder of the inscription reveals that she was a priestess (hesyt khenu n Imen or “Singer of the Interior of Amun”) who worked in the temple of Karnak and most probably came from the area around Thebes. The style of the cartonnage and details of its iconography suggest that she lived around 800 BC (Taylor 2014). The CT scan analysis of the skeleton showed that the inferior and superior margins of her pubic symphysis had remodeled with age (Suchey-Brooks score 4), and based on these changes, she was probably between 35 and 49 years old when she died. Post-mummification damage to her feet made it impossible to measure her overall length and based on her long bone measurement, she was around 152 cm tall (151.5
2 cm; after Raxter et al. 2008). Her teeth are also very worn, which appears to be a feature of many of the mummies curated at the British Museum (see Taylor and Antoine 2014). The wear appears to have led to the formation of several periapical lesions, similar to the

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ones found in Ameniryirt’s dentition (see above) and the lower right central incisor is missing, probably lost as the result of an earlier lesion. Her remarkably wellpreserved skin and short hair can clearly be seen on the CT scan visualizations (Fig. 7). Her brain was carefully removed by breaking part of the ethmoid and vomer bones, opening a comparatively large hole (1.9  4 cm) at the base of the frontal bone. No resin was poured into the cranium, which was left empty. As had become the norm in the earlier Twenty-First Dynasty, attempts were made to render her features as lifelike as possible. Spherical textile bundles and false eyes were placed in her eye sockets. Using an incision at the base of the neck, the embalmers also carefully packed her throat and trachea with a large amount of textile and her mouth was then filled with a dense material, possibly resin (Fig. 7). Her organs, removed using an incision on the left side, do not appear to have been placed back in or around the mummy, and may have been preserved elsewhere. To strengthen the empty body cavities, the chest, abdomen, and pelvic area were filled with a disorganized mixture of textile, granular material, resin, and dense packing. This complex packing makes it hard to assess whether her heart was left in place by the embalmers. Various amulets, including a heart amulet and a headrest amulet, a djed pillar, and a scarab were also positioned inside the body (Fig. 7). They do not appear to have been consciously positioned on specific areas and seem randomly placed inside the abdomen, close to the incision made by the embalmers. Small pellets of metal – probably gold – are scattered across the surface of the body, particularly on the face. Resin and layers of bandages were then used to wrap the temple singer. Tamut lived around 900 BC, in the earlier part of the Twenty-Second Dynasty (British Museum EA 22939; Taylor and Antoine 2014: 68–91). As with the priestess, she was probably between 35 and 49 years old when she died. She suffered from cardiovascular disease, with calcified plaque depositions found in her femoral arteries. Also very well preserved, her mummification differs slightly. Her brain was extracted via the right nostril and the empty cranium was then filled with, unusually, textile (see discussion in Wade and Nelson 2013a). To make her appear lifelike, packing materials were used to shape Tamut’s face, with her mouth, nose, and throat filled with dense materials and textile. Artificial eyes made of stone or glass were also added. Her internal organs, divided into four bundles, were repositioned inside her chest, with each one containing at its center one of the Sons of Horus made of what appears to be beeswax (Taylor and Antoine 2014: 84–85). The body cavity was then filled with packing materials and textiles. Two metal plates, each with a wedjat eye or Eye of Horus, were placed over the incision used to remove her organs. Finely incised, the wedjat eye may not have been detected without a dual-energy CT scanner (see above). Each of her finger- and toenails also has metal coverings and, in addition to smaller amulets, four large divine figures made of metal had carefully been placed on her body. Two stola straps, probably made of leather, were also positioned on her chest. The re-analysis of another mummy (British Museum EA 22812b) revealed that they belonged with a set of three coffins made for a woman named Nestawedjat (EA 22812a, EA 22813a, EA 22813b). An exact match between the stains found on the back of the bandages surrounding the left shoulder and those at the bottom of

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Fig. 7 CT scan visualizations and DICOM images from the mummy of a Twenty-Second Dynasty priestess (British Museum EA 25258). Her very-well preserved skin and short hair, as well as false eyes placed in her eye sockets, can be seen on the CT scan visualization (bottom left). Small pellets of metal are also scattered across the surface of her face. Her brain was removed via the nose using a relatively large opening (top left) and an incision at the base of the neck was used to pack her throat and trachea with textile (top right). Her mouth was also filled with what appears to be resin (top left and right). Various amulets were placed inside the body (bottom right). Images courtesy of the Trustees of the British Museum

the innermost coffin confirmed their association (Antoine and Vandenbeusch 2016: 20–53). Originally interpreted as being a male (Dawson and Gray 1968: 9), most probably due to the oblong shape of the internal packing applied to the genital area, a

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CT reassessment established that these were the mummified remains of a female. The style of her coffins indicates that she was from Thebes and lived during the Twenty-Fifth Dynasty, probably around 700 BC, the so-called Kushite Dynasty under the control of rulers from the Kingdom of Kush in modern northern Sudan. Her title, “Lady of the House,” tells us she was married but the inscriptions reveal little else. The CT scans show that Nestawedjat was, by modern standards, a relatively small woman at around 153 cm (152.8
1.9 cm; Raxter et al. 2008) and between 35 and 49 years old when she died. The techniques used to embalm and wrap Nestawedjat’s body are characteristic of the Twenty-Fifth Dynasty. The embalmer used a very direct route to remove her brain, opening a hole about 2.5 x 3.2 cm in the base of the frontal bone (approximately 6 cm inside the nasal passage) and did not fill the cranium. As with other mummies of the period, Nestawedjat’s eye sockets were filled with spherical textile bundles on top of which artificial eyes made of stone or glass were placed (Fig. 8). Packing materials were also used to maintain the shape of the face, and her mouth was filled with textile and resin, accidentally dislodging three teeth. Dense packing material was used to fill much of the space within the ribcage and abdomen, including high-density packing in the upper right chest area (a similar packing can be found in the mummy of the doorkeeper Padiamun, Liverpool M. 14003; Gray and Slow 1968: 21–24). Most of Nestawedjat’s body was covered with a remarkably thick resinous layer, including her head and face. Four large flat packages lying on top of her skin, below the thick layer of resinous material covering her body, can also be seen on the scan. Placed under her hands and on top of her thighs, knees, and lower legs, their contents cannot be ascertained but they are very likely to be the remains of her mummified internal organs. Two unusually shaped amulets were also placed over Nestawedjat’s throat. A single piece of textile surrounds the mummy and is secured by strips, also made of textile, and a net made of beads may have originally been placed on top of the shroud. Also from the Twenty-Fifth Dynasty, around 700 BC, the mummy of Padiamenet (British Museum EA 6683) hints at problems that could sometimes occur during mummification. Although mummified with care and showing good preservation, his head appears to have detached itself during mummification and the CT scans reveal that two wooden poles were used to reattach it. His mummy was also too tall for his cartonnage case, which had to be readjusted in an unconventional manner by covering his protruding feet with extra cartonnage and textile (see Taylor and Antoine 2014: 92–111). From the Twenty-Second Dynasty up to the Late Period, wrappings and cartonnages were sometimes anointed in resin, which could blacken with time (Ikram and Dodson 1998: 129). Although canopic jars were reintroduced during the Late Period, internal organs were often placed on or between the thighs and legs, within the outer wrapping (see Ameniryirt above; Ikram and Dodson 1998: 128– 129). The position of the limbs also begins to change with the arms now found crossed over the chest (Ikram and Dodson 1998: 128–129; see also Ikram et al. 2018 for a Twenty-Fifth Dynasty example). This position was used with Irthorru, a Twenty-Sixth Dynasty priest (circa 600 BC) from the town of Akhmim, around 200 km north of Luxor (British Museum EA 20745; see Antoine and Vandenbeusch

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Fig. 8 CT scan analysis of the Twenty-Fifth Dynasty mummy of Nestawedjat (British Museum EA 22812b). The embalmer removed her brain via the nose and her mouth was filled with textile and resin (top right). Nestawedjat’s eye sockets were filled with spherical textile bundles on top of which artificial eyes made of stone or glass were placed (bottom right). Dense packing material was used to fill the ribcage and abdomen, including high-density packing in the upper right chest area (bottom center). Most of Nestawedjat’s body was covered with a thick resinous layer, including her head and face, with two unusually shaped amulets placed over her throat (top center). Apart from her heart, which is clearly visible in transparency mode (coloured in pink; left), most organs appear to have been removed and the body filled with high-density packing. The separately embalmed organs are probably inside the four large flat packages placed on top of her skin (left). Images courtesy of the Trustees of the British Museum

2016: 82–111). His long bone measurements (after Raxter et al. 2008) suggest that he was around 163 cm tall (162.8
2.8 cm) and was probably between 35 and 49 years old when he died. Consistent with his age, most of his teeth are heavily worn and he had extremely poor dental health. Three of his lower front teeth are missing and the bone around his lower front teeth (left central and lateral incisors) has been lost and, though still present, these extremely worn teeth no longer have actual root-sockets. They are only being held in place by the gums (Fig. 9). Such

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Fig. 9 CT scan visualization and DICOM images of Irthorru, a Twenty-Sixth Dynasty priest from Akhmim (British Museum EA 20745). Most of his teeth are heavily worn, three are missing and the bone around his lower front teeth has been lost, most probably from periapical lesions (top center). His lower left lateral and central incisor no longer have root sockets and are only held in place by soft tissues. Two stone or faience beads were placed over Irthorru’s forehead and a metal amulet can be seen in the back of his throat (bottom right). Irthorru’s brain was carefully removed via the nasal passage and his cranium was filled with resin (bottom center). His nasal cavity was then filled with a homogeneous substance and spherical textile bundles were placed in his eye sockets (top and bottom right). His arms are crossed over his chest and four organ bundles are visible inside his abdomen and chest (left). Images courtesy of the Trustees of the British Museum

extensive bone loss was probably the result of several periapical lesions. The front and right edges of Irthorru’s eighth to tenth thoracic vertebrae also show signs of extensive new bone formation often associated with intervertebral disc disease (Waldron 2009: 42–45). Surprisingly, the embalmers carefully placed in the middle of the bandages a djed pillar amulet – a symbol often associated with the backbone of Osiris – in close proximity to Irthorru’s twelfth thoracic vertebra. However, although placing a djed pillar in this location is rarely known otherwise, it has been found on the mummy of one of Irthorru’s relatives, suggesting an embalming trend rather than a medical treatment. Other amulets were also positioned in and around the body as

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magical charms including an unusual metal amulet 14 mm long and 3 mm in diameter in the back of Irthorru’s throat. Two equally unusual stone or faience beads were also placed over Irthorru’s forehead, an area where amulets are regularly found on mummies from the Akhmim necropolis. Among other more common amulets placed on his upper body, a small wedjat eye can be seen on the back of his right hand. As part of the embalming, Irthorru’s brain was carefully removed via the nasal passage using a small hole about 2.5 x 2.1 cm in the base of the frontal bone. The cranium was then filled with a relatively large quantity of resin (Fig. 9). Perhaps to render his face as lifelike as possible, the nasal cavity was filled with what appears to be a homogeneous substance – perhaps wax or resin – and spherical textile bundles were placed in Irthorru’s eye sockets. His face was then covered with a beautifully gilded mask, which, unusually, does not surround the entire head, as would be expected in ancient Egyptian burials. CT scans reveal that the mask only covers his face and the linen shroud ingeniously hides its absence from the rest of the head. His arms are crossed over his chest, his right hand clenched. Unlike Ameniryirt (also from the Twenty-Sixth Dynasty, see above), whose internal organs were placed on his legs, Irthorru’s four desiccated organ bundles were positioned inside his abdomen and chest. Finally, layers of resin and bandages were used to wrap Irthorru’s body and complete his mummification. Although embalming methods were believed to be in decline by the end of the Late Period, the body of Irthorru confirms the skilled techniques of the Akhmim embalmers and reveals both regional variations and a continuation of relatively high standards in some parts of Egypt. Unsurprisingly, Twenty-Sixth Dynasty mummies from the Theban area show that procedures varied. The mummy of a man, whose name remains unknown, shows the standard removal of the brain but no additional packing in the skull and only a small quantity of linen in the mouth (British Museum EA 22814; Taylor and Antoine 2014: 44–66). A tool – probably made of wood – used by the embalmers to take out the brain seems to have snapped and part of it has been left in the skull cavity. Other mummies attest of very little treatment, especially in comparison to the extensive preparation observed during the Twenty-First Dynasty. The CT scan of another unnamed individual who probably lived during the 26th Dynasty but entered the British Museum in a 22nd Dynasty coffin (British Museum EA 6666), probably from Thebes, reveals that the brain was not always removed, but confirms the extensive use of resin, a feature which will become prevalent during the Ptolemaic Period. As often the case in Twenty-Sixth Dynasty Theban mummies, internal organs were removed and replaced with a granular material that filled most of the chest and abdomen. Unlike the mummy of Irthorru from Akhmim, the organs were not placed back in the chest, but appear to have been preserved in packages above the legs. Many changes – and elements of decline – can be observed between the Third Intermediate and the Late Periods. The careful packing and subcutaneous fillings in vogue during the Twenty-First and Twenty-Second Dynasties tend to disappear. They are replaced with a greater use of resin, applied both directly on the skin and inside the body cavities. The brain is still removed, although not as systematically as before, from the Twenty-Fifth Dynasty onwards (Wade et al. 2011). The organs in

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the thorax and the abdomen are, however, regularly taken out and continue to be preserved separately so as to be placed back on and inside the body (while empty canopic jars continued to be found in tombs).

Ptolemaic Period (332–30 BC) After the Greek (Ptolemies) started ruling Egypt following its conquest by Alexander the Great in 332 BC, mummification not only continued, it appears to have increased. Many mummies of individuals who lived during the Ptolemaic Period – and the subsequent Roman Period – have survived to this day. While Greeks brought their funerary customs, including cremation, these were mainly observed in Alexandria and no noticeable changes seem to have taken place in the rest of the country. Mummification techniques showed some continuation and the considerable use of resin already observed during the Late Period is one of the features of the Ptolemaic Period. Resin was not only being applied on the skin, it was also poured in great quantities inside the body cavities. As seen in the Late Period (see Irthorru above; Fig. 9), the skull was typically filled with liquid resin. However, in spite of the many Ptolemaic mummies still preserved, few have been comprehensively studied. They are often not discussed individually but grouped in the wider Graeco-Roman Period (Aufderheide 2003; Ikram and Dodson 1998). More research is still needed to gain a clearer understanding of mummification practices during this period and clarify the transition from the Late Period to the Roman Period. Questions of dating and provenance greatly restrict such studies. Mummies were often not excavated using modern standards and the location of their discovery is usually not recorded. Their dating is often very broad, limiting any research into changes in embalming practices over time. The mummy of Hornedjitef (British Museum EA 6679), however, is an exception. He is thought to have died in the Theban area around the reign of Ptolemy III (246–221 BC). As a priest of Amun, he was buried with a paraphernalia of objects (including cartonnage trappings, coffins, and papyri), and his body testifies to the high standards of mummification available at the time. In addition to the great number of amulets placed over his body, especially around the neck and on the chest, Hornedjitef was carefully embalmed using an abundance of resin (Filer 1999). The skull, emptied of the brain, was largely filled with resin poured in a liquid form and now solidified. Most of the other organs had also been removed and embalmed separately before being replaced as four packages inside the thorax and abdomen. Hornedjitef’s arms are crossed on his chest, another feature characteristic of Ptolemaic mummies that shows a degree of continuation with the Late Period (see above). Most mummies whose provenance can be determined are from Thebes, as was Hornedjitef, or from Akhmim. The embalming of the Theban mummy of Tarepit (Cairo TR 21.11.16.13) shows close similarities to that of Hornedjitef (Elias and Mekis 2019). Most organs – with the exception of the heart – were removed. The emptied skull was profusely filled with resin, as was the thorax and the abdomen. The latter contained four packages that were probably used to preserve her organs.

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Interestingly, she was also provided with a canopic box (now in Athens). It is possible that it only had a magical role and was kept empty, although it might also have contained other organs that did not find their way inside the body. Many amulets – and several rings – were spread over the body, including a wedjat eye and a double plume near the forehead. Several rolls, possibly of papyri, were placed on the body, including one under the arm. Cordage was used to maintain her arms crossed and her limbs, initially wrapped separately, were covered in more bandages to create the bundle shape. The unwrapping of an anonymous man from Thebes (probably from the early Ptolemaic Period) that took place in Lyon provides further insights on the period’s embalming and wrapping techniques (Goyon and Josset 1988). The homogeneous layer of resin on the inner walls of the cranium suggests that the head was manipulated to evenly distribute the resin when in its liquid state. The bandages, which were listed and studied layers by layers, revealed the use of several shrouds. Multiple knots were also employed to secure the bandages. More surprisingly, the sail of a boat was found – and then reconstructed – among the wrappings, attesting of the reuse of such materials and perhaps suggesting that the deceased or a member of his family owned a boat. It may also have been used to convey the idea of the deceased travelling to the afterlife. Although Ankhhap, a doorkeeper of Amun, was also from the Theban area (British Museum EA 6711), the techniques used to embalm his body show some surprising differences (Fig. 10). Firstly, no attempts were made to remove his brain. Most of the organs from his thorax and abdomen were removed and no packing was used to fill the voids. Unlike some of his contemporaries, resin was not a central feature of his embalming and there is no evidence any had been poured inside his chest. The man’s soft tissues (both skin and muscles) are, nonetheless, remarkably well-preserved, attesting of the skills of his embalmers (although they may have applied small quantities of resins and oils on his skin). The arms of Ankhhap are also crossed over his chest, with the right hand clenched. His limbs were wrapped separately, and then together as a bundle (although very thinly, in some areas no more than two or three layers of bandages are visible). Unlike the other Theban mummies from the period (above), no amulets appear to have been placed on the body or within the wrappings. These differences hint at variations in standards and embalming methods during Ptolemaic Period. They may also indicate differences in status. Ankhhap’s family, who belonged a social class lower than Hornedjitef’s, might not have been able to afford a comprehensive treatment or the supply of expensive amulets, but a simpler and efficient embalming, as well as a beautifully decorated wooden coffin. In 1884, when Gaston Maspero found an intact Ptolemaic necropolis in Akhmim, “he himself saw only five pits and these alone contained about 120 mummies, and there are very many more in the area; he estimates the total number of mummies at 6,000 or more (. . .)” Dawson 1947: 78). Some of these mummies are now in museums around the world. Their analysis reveals that a great quantity of resin was being used, as was observed in the Theban mummies. This is also the case with the mummy of Diptah, dating to the early Ptolemaic Period (Leiden, Rijksmuseum, AES 14; Raven and Taconis 2005: 158–161). Excerebrated using the transnasal method, resin had been poured inside her skull, pooling in several layers. Resin and textiles had also been placed in her orbits, with further resin used to fill the thorax in

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Fig. 10 DICOM images of the mummy of Ankhhap, a doorkeeper of Amun from the Ptolemaic Period (British Museum EA 6711). A layer of resin covers the skin of Ankhhap’s mummy but, unlike many other Ptolemaic mummies, none was poured in his body cavities. Arms and legs were wrapped separately before the body was bandaged in the shape of a bundle. Images courtesy of the Trustees of the British Museum

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which packages – probably contained her organs – had been placed. Arms and legs were wrapped separately and, with the rest of the body, were padded and wrapped in a mummy bundle shape. Her arms are also crossed over her chest. Probably from Akhmim, the body of Hor received a very similar treatment (Leiden, Rijskmuseum, AdS 1; Raven and Taconis 2005: 162–164), although an even greater quantity of resin was used. Despite these similarities, a review of several Akhmim mummies from the Ptolemaic Period reveals a range of approaches were being used to mummify remains (see Elias and Lupton 2008). As noted by Elias and Lupton in their study, the use of differing techniques, especially in the application of resin, may have been driven by social and economic factors. Most of the mummies discussed in this section have their arms crossed over the chest. This is a recurrent feature during the Ptolemaic Period, in Akhmim and elsewhere. However, there are also examples of individuals with their arms placed alongside the body, often with hands on the pubic area. Elias and Lupton (2008) suggest that this position, observed on younger individuals, is age related. Further research into Ptolemaic mummies should help clarify this practice and its significance. Although commonly found on Akhmimic mummies of earlier periods, amulets are more sporadically used during the Ptolemaic Period. The mummy of Nesmin, the priest of Min (New York, Metropolitan Museum of Art 86.1.51; Stünkel and Nankivell 2015) provides, however, a stunning example with 31 amulets identified including a wedjat eye on the forehead (the placement of an amulet on the forehead is a feature already witnessed on mummies of earlier periods; see Irthorru in the Late Period) and strings of amulets on the chest. Ptolemaic mummies from the Northern part of Egypt are less known and studied. Nespamedu’s is remarkable for both the embalming techniques employed and the use of amulets and trappings (Madrid, Museo Arqueologico Nacional 1925/57/1; Pérez-Die et al. 2018). Priest of Imhotep and royal physician, Nespamedu was probably buried in the necropolis of Saqqarah (because of his royal duties, a burial further North, closer to Alexandria, cannot be excluded). The extensive use of resin, poured inside most of his body cavities, is characteristic of the approach developed during the Ptolemaic Period. The canopic packages, the crossing of the arms over the chest and the excerebration follow the practices discussed above. The embalmers, however, also carefully placed several pieces of cartonnage on his body. These amulets in cartonnage, found within layers of textile, may have been produced by the person who had made the external trappings (Pérez-Die and Carrascoso Arranz 2018). As a member of the elite belonging to the royal entourage, the embalming and the trappings elements no doubt represent the best of what could be done at the time. Amulets and objects inserted in the wrappings were not necessarily made of metal, faience, stone or even wax, materials easily visible on CT scan data. As with previous periods, the embalmers would also place less dense organic material that were believed to bring magical protection to the deceased, including garlic and onions found in the orbits or under the sole of the feet. This appears to be the case for the mummy of a woman, now in Poland, dated to the Late or Ptolemaic Period. A bulb of narcissus was identified under the left hand, perhaps because of its analgesic and healing properties (Borysławski et al. 2018). Flowers and other plants, as well as

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pieces of shaped textiles, could also be carefully placed within the wrappings. Often challenging to detect or identify on the CT scan, most examples have been observed in partly damaged and unwrapped mummies. Skilled mummification was not limited to the Nile valley. Well-mummified individuals were recovered from the necropolis of Dabashiya, in the Oasis of Kharga, including a carefully embalmed man with both excerebration and evisceration (see Dunand et al. 2012: 27–28). However, a range of approaches appear to have been used. The mummy of a woman found in the same tomb showed little treatment other than being wrapped (Dunand et al. 2012: 30–31). We do not know whether the two individuals were related, but these extremes confirm the co-occurrence of a range of practices, even within the same archaeological context. Many mummies discussed here date to the early and mid-Ptolemaic Period and, with little change in practices from the first century BC to the Roman Period, it can be hard to distinguish late Ptolemaic and early Roman mummies. A young man (British Museum EA 24800), seemingly from Hawara, lived during this transitional period and his mummy was covered with the fashionable accessory of the time, a cartonnage mask (Fig. 11). Other trappings in cartonnage were also tightly maintained around the body with thin strips of textile. His arms are crossed over his chest but very little resin appears to have been used. His brain was removed through a particularly large nasal opening and the wooden tool used to create it was apparently left inside the skull. Unfortunately, the contents of the chest are mostly disturbed, probably by looters in search for valuable amulets. Although it seems that the chest cavity had never been filled with resin, organs bundles are visible among the misplaced bones. The same is true for the treatment of the skin, which does not have the usual layers of resin.

Roman Period (30 BC–AD 395) After the transition from Greek to Roman rulers, many administrative, economic, and political changes were imposed. As with their predecessors, the new foreign rulers did not try to impose their beliefs. On the contrary, local traditions were encouraged, religious rituals maintained and temples built across the country. By then, the population was multicultural and, in addition to Egyptians, included Greek settlers and Roman newcomers. Traditional funerary practices, anchored for millennia, thrived in this diverse environment and, although some significant changes occur, mummification continued to be practiced. It is difficult to ascertain who within this multicultural society was being mummified. It certainly included Egyptians who could afford it and, most likely, Greek settlers who had been in Egypt for several generations and may have intermarried with the Egyptians. It remains difficult to assess whether some Romans chose to be mummified and there is only one clear example, that of a mask bearing the tria nomina belonging to a man named Titos Flavius Demetrios (Riggs 2005: 21–22). Romans had a tradition of cremating their dead and this is likely to have been the most common practice in Alexandria (Gessler-Löhr 2012: 667). Access to mummification seems, however, to have

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Fig. 11 The mummy of a young man (British Museum EA 24800). The content of his chest have been considerably disturbed, possibly by looters. The large wooden tool found in his cranium was probably used by the embalmers to access the brain. Images courtesy of the Trustees of the British Museum

extended to part of the population that had not previously seemed to be able to afford it. Consequently, a greater number of mummies from the Roman Period have been recovered, possibly because more bodies were embalmed this way. Excavation reports of early archaeologists emphasize that a great many more were originally discovered, only a small percentage of which are preserved today. The search for portraits, transportable goods, and papyri guided excavations for many years, leading to much destruction (Gessler-Löhr 2012: 665–666). Portraits and masks were often detached from the mummy, and the body discarded. Unfortunately, those that did not have such trappings were also often thrown away. The quality of mummification during the Roman Period is often questioned and thought to have been in extreme decline, with a focus on external appearance to the detriment of internal preservation. As already put forward by Gessler-Löhr (2012) and Dunand and Lichtenberg (2019), the issue of decline is not a straightforward one. Practice and techniques diversified greatly, from very little preparation to full mummification. This probably reflected affordability, as well techniques employed

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by specific workshops, and regional and temporal trends. Some mummies from Roman Egypt were sumptuously wrapped, with the bandages skillfully applied, often forming beautiful lozenge patterns. They would sometimes be enhanced with a naturalistic representation of the deceased, painted on linen or wood, or modeled in plaster or cartonnage. However, only a small percentage, perhaps no more than 1 or 2%, were wonderfully ornamented (Gessler-Löhr 2012). This diversity of treatments and practices, both external and internal, makes this period particularly rich, balancing tradition with innovation. Mummies were not only covered with the famous wooden portraits, so intensively naturalistic, but also with painted shrouds, cartonnage, or plaster masks, or a combination of the adornments (Walker and Bierbrier 1997). Portraits and masks appear to reflect a change in beliefs, where the deceased no longer sought an idealized representation of themselves but aimed for a lifelike representation, a Graeco-Roman influence that moves away from traditional ancient Egyptian practices. Interestingly, in the Southern part of the country, particularly in the Theban area, old traditions were maintained for longer, with portraits and naturalistic imagery being more popular further North (with the hundreds of portraits unearthed in the Fayum Oasis, they are sometimes inaccurately referred to as “Fayum portraits”). Theban inhabitants were probably more attached to the old cannons, with greater distance from the center of power in Alexandria giving them more freedom to persevere with old traditions (Riggs 2005, 243–244). Nonetheless, some portraits originate from the Theban area. The mummy of a young man (British Museum EA 6713) is simply wrapped with a plain shroud and adorned with a wooden portrait depicting a teenager (Fig. 12). The young age of the individual was recently confirmed by the CT scan. Based on his skeletal development, he was between 17 and 20 years old when he died (Antoine and Vandenbeusch 2016: 160– 187). His body was remarkably well preserved, with all the organs in the thoracic and abdominal cavities still in place and distinctively visible on the CT data. The embalmers had not made any attempts at removing them – there is no trace of an abdominal incision – but their preservation is remarkable and was perhaps achieved by using large amounts of natron. Small high-density dots visible within the soft tissues may be evidence of the salt’s penetration. Although an attempt to remove the brain was probably made, it was not through the traditional transnasal route but via the foramen magnum. What remains of the brain is covered by a thin layer of resin that was also poured in the cranium via the base of the skull, through the back of the neck. A necklace, made of small circular beads in faience or stone and what could be flowers in wax, adorns the man’s neck and his body was placed on a wooden board. Regularly found within wrappings in Roman Egypt, it is often thought that these boards may have added support in case of decomposition, which can be excluded in the case of the remarkably well-preserved young man. The addition of funerary texts and imagery on the surface of some mummy boards suggests that they also had a religious and protective purpose. It is not known if the wooden board placed within the young man wrappings is inscribed. His portrait depicts him as a young, beardless and rather slim, but the CT scan reveals he was overweight. The cause of his obesity is unknown, although the presence of tooth decay could suggest a fondness for sweet foods. He or his family, however, preferred for him to be remembered as a svelte young man.

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Fig. 12 CT scan visualizations of a young man from the Roman Period (British Museum EA 6713). His soft tissues are remarkably preserved, so much so that most of his internal organs can be identified. A necklace made of small circular beads in faience or stone, and what appear to be flowers made in wax, was placed on his neck. What remains of the brain is covered by a thin layer of resin that was poured via the base of the skull, through the back of the neck (right). Images courtesy of the Trustees of the British Museum

The tomb of the Soter family, discovered in 1820s, possibly in the tomb of Djehutymes (TT 32) in the Theban necropolis (Kákosy 1995), provides further insights. Four generations of the family, named after Soter I, an archons of Thebes, are known (see Herbin 2002). Soter had at least nine children with his wife Cleopatra I Candace, and several of their mummified remains are still preserved today. Among them, Cleopatra II (British Museum EA 6707) lived between AD 100 and 120. Scans of her mummified remains show a skilled embalming with some peculiar features (Fig. 13). An incredible amount of both resin and linen were used to wrap her body, making her much heavier and harder to handle. Her body was probably plunged into a bath of liquid resin, so viscous that it not only penetrated the soft tissues, it can also be seen inside some of her bones. Resins were also applied within the wrappings in three separate layers. This mummification style was also used to preserve other mummies belonging to the family, and largely explains their substantial weight. Cleopatra II’s brother, Petamenophis (now at the Musée du Louvre) was unwrapped in France in 1823, reducing his total weight from 106 kg to a modest 40 kg. The body had been partly gilded, a recurrent trait during the Roman Period (Herbin 2002: 40). Petamenophis, Cleopatra and their relatives were embalmed using very similar techniques, with their brain removed through the nasal passage and organs taken out via an incision on the left side of the abdomen (clearly visible on the body of Petamenophis). There are, however, no traces of organ (or other) packages within the body cavities. Their sister, Sensaos I, now in Leiden (Rijksmuseum von Oudheden, AMM 8-a) received a very similar treatment. The

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Fig. 13 The mummy of Cleopatra (British Museum EA 6707). CT scans reveal she was wrapped in an unusual large number of bandages and that her mummification involved the use of a large quantity of resin. Images courtesy of the Trustees of the British Museum

study of her body also shows that all fingers were bandaged separately (Raven and Taconis 2005: 179–183). Several metal artifacts were found, including two artificial eyes and a snake shaped amulet. Her hair was kept long, embedded in resin. Most mummies belonging to the Soter family were also wrapped in an external shroud, vibrantly painted, showing the deceased face on, surrounded by various traditional Egyptian funerary scenes. Many were then placed in a rectangular wooden coffin. Theban mummies were not always wrapped in a bundle. An unusual group thought to come from the Theban necropolis includes mummies of men, women, and children. Now found in Leiden, Liverpool, Avignon, London, Berlin, and, prior to being lost in a fire, Rio de Janeiro, all nine known so far share a distinctive embalming style. Their arms, legs, and fingers were all individually wrapped and kept separate, with the body being modeled in a life-like shape reminiscent of the style used during the Old Kingdom. Paddings and rolls were also placed on the face and breasts, while the wrappings around their feet were modeled to look like sandals. Most have painted faces, with clearly defined eyes, mouth, and ears. Some are also decorated with the external trappings, kilts, or painted bands. Although their brains and internal organs were consistently removed, the CT scans also reveal variations in their embalming. Their internal packing varies with high-density packages found in the adult male curated at the British Museum (EA 6704), while others remain empty. His packages most probably contain his internal organs, with their high density seemingly due to high resin levels (Taylor and Antoine 2014, 134–149). The use of

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resin in this unusual group of mummies varies but it was mostly applied to the skin and in the abdomen. In none does it reach the levels seen in the Soter group. Regarded as belonging to the same group and most probably prepared in the same embalming workshop, minor variations between the nine mummies suggest embalmers were both adapting and varying their techniques according to individual needs and their own preferences or, perhaps, were guided by specific requests. Many other groups of mummies dating to the Roman Period have been found in the Theban necropolis (see Gessler-Löhr 2012, which also includes mummies that are now lost). The group discovered at Deir el-Bahari by Swiss Egyptologist Édouard Naville, initially mistaken as Christian, was expertly embalmed with evidence of evisceration and gilding on the skin (Riggs 2005: 232–243). One of them was unwrapped in 1956, a few years after having entered the Brooklyn Museum collection, and was recently rewrapped in his original 14 layers of textile (including two tunics), a process that took almost a year (Loos 2010). Hundreds of mummies were also found during the excavation of the Valley of the Queens, including 307 mummies dated to the Roman Period (Macke and MackeRibet 2002: 63–111). Their embalming seems to have been relatively standardized, with 70% of skulls conclusively showing evidence of excerebration (99% through the nasal passage), and almost 80% being eviscerated. The use of balm – a mixture of bitumen, resin and wax of varying proportion – was also noted, but not in the large quantities observed in other part of the necropolis. Three-quarters of the mummies that could be studied had packages in their body cavity that probably contained organs. Although their number varies from one to six, most had a similar cylindrical shape. Soil, linen, and ceramic bowls intentionally placed to protect the pelvis or the packages, or to shape the abdomen, were also found inside the thorax and abdomen. The mouth, eyes, and nose were more rarely packed, while the arms were almost always positioned alongside the body, sometimes held in place with a rope. The use of wooden mummy boards was also observed (seven examples), as well as examples of palm ribs applied to support the body. Multiple layers of fabric of various shapes (e.g., shrouds, bandages, reused pieces of clothing) were also used to wrap the bodies, including some examples of the famous criss-cross lozenge style external bandaging. The lack of amulets within this group is generally consistent with Roman Period mummies. Various small elements – round or rectangular – made of gilded wax were found on some faces, as well as in or on the abdomen, on the navel or on the breast. This large group of mummified individuals reveals a similar (if not totally uniform) approach to embalming and they probably belonged to the same community. Their status and socioeconomical background, however, remain unclear. Extensive Roman Period necropoleis are also located in the Fayum region, where thousands of mummies have been recovered during the 19th century. Of these, few were covered with a portrait or a mask, and therefore deemed worth keeping. In many, only the actual portraits or masks were detached. In spite of this, the large quantity of mummies still preserved presents a great potential for research. Among the 30 mummies from Roman Egypt at the British Museum, ten are likely to be from Hawara, and four were discovered during Petrie’s excavations in 1888–1889. The latter are very well preserved, with beautifully produced portraits and masks that

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suggest they belonged to the wealthy members of the community. The mummy of a man named Artemidoros in the Greek inscription on his outer shroud (British Museum EA 21810) could be one of the mummies that has led to a belief in a complete loss of embalming skills during the Roman Period (Fig. 14). Masterly decorated with an elegant portrait and an ornamented red shroud, X-rays – and more recently CT scans – have shown him to be poorly preserved, with only a disorderly skeleton remaining (Antoine and Vandenbeusch 2016: 168). The mummy of a teenager (British Museum EA 13595), whose beautifully executed portrait was skillfully arranged within bandages in lozenge pattern and gilded studs, provides another example of poor preservation. His body appears to have been in an advanced state of decomposition when it had reached the embalmers and multiple palm ribs were employed to reconstruct him, with several used to attach the head to the rest of the body. Nonetheless, great care had been taken to render him as lifelike as possible. Although the mummification of children was less common, several examples from the Roman Period have been recovered but their treatment may have differed from that of the adults. The societal changes and foreign influences seen in Egypt

Fig. 14 The mummy of Artemidoros (British Museum EA 21810). His stunning external appearance contrasts with the poor preservation of his remains. The body may have been decomposing and multiple palm ribs were employed to reconstruct it. Images courtesy of the Trustees of the British Museum

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during this period may have had an impact on the mummification of children, which appears to have been rare until the Roman Period. Most earlier examples are from high status families (see Tjayasetimu from the Third Intermediate Period above; Taylor and Antoine 2014: 112–133). This perception is perhaps biased by the vagaries of the archaeological record and the discovery of large number of Roman mummies. However, an increase in child burials suggests a change in practices or beliefs, with children usually buried in the same graves as adults. For example, two mummified boys now curated at the British Museum (Fig. 15) were discovered alongside three other mummies (a girl in Manchester Museum, an adult woman in Cairo, and another adult woman now lost). All five may have been from the same family group (see Roberts 1997). The youngest boy was carefully wrapped in many layers of linen creating a cocoon around the body (British Museum EA 22108; Antoine and Vandenbeusch 2016: 140–160). His dental development, clearly visible on the CT scan, indicates he was in his Early Childhood and died when he was approximately two years old (+/ nine months). Most of his internal organs appear to have been removed but the brain, although much reduced, and the skull are both seemingly intact. Perhaps the bones were thought too small and delicate to attempt its removal. Resin was poured on top of the skin, and a small quantity has seeped into the skull via the anterior fontanelle. The head of the boy was tilted forward, a position that repeatedly appears during the Roman Period, particularly in mummified children. It is believed that the raised head conveyed a notion of rebirth and awakening, which is also represented on plaster masks of the same periods. Placed on top of the bandages, it would depict the deceased rising up. In his case, a gilded mask, painted shroud and foot case had been placed over the many layers of bandages and wrappings. Such gilded masks were once believed to have preceded painted portraits. Covered with a very similar shroud on the lower body, the other boy from the same tomb (British Museum EA 21809) had his portrait painted on linen. This suggests that portraits were being used at the same time as gilded masks. The child with the linen portrait, also a young boy in his Early to Late Childhood, was slightly older when he died (around 4 years
12 months). The embalming treatment is very similar to that of the other boy, with his head tilted forward and a thick layering of bandages around the body. The brain had, however, been removed by breaking the ethmoid bone. The cranium is mainly empty with the exception of a small pool of resin at the back. The chest and abdomen are compressed with the dense layers of wrappings, and it is difficult to establish whether all or parts of the organs were removed. Four small objects, perhaps in gilded wax, were placed on the chest and abdomen. Because of their ambiguous shape, these elements are difficult to interpret. Based on their location and shape, they might represent the nipples, the navel, and the genitalia of the deceased. This interpretation fits with the notion of reshaping and preserving the body that was central in Roman Egypt and previously attested in the Theban mummies. Embalming was certainly not exclusive to the Nile Valley and many examples are known in the Oases as well. In addition to the Fayum area, hundreds of mummified individuals have also been found in Kharga and in Dakhla. The excavation of the necropolis of Douch, in the Oasis of Kharga, with more than 700 mummies

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Fig. 15 The mummies of two children from the Roman Period (British Museum EA 22108 on top; British Museum EA 21809 below). Discovered in the same tomb, they were both wrapped using large amounts of bandages so as to form a cocoon around their bodies. Images courtesy of the Trustees of the British Museum

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discovered, has greatly contributed to our understanding of embalming (Dunand 1995). The study of 47 complete mummies showed that, although not always eviscerated, excerebration was more systematic and usually involved the pouring of resin inside the skull and the presence of tampons blocking the nasal passage. The arms were usually placed extended alongside the body and the head variably tilted down towards the chest. Several also had gilding on the skin, although this practice seems to have been reserved for the wealthy. Further variation is revealed in the necropolis of Ismant el-Kharab (Kellis), in the Oasis of Dakhla, where hundreds of mummified remains showed that, unlike excerebration, evisceration was not always practiced (Aufderheide et al. 1999, 2003). A more systematic use of resin seems to have replaced natron as the main preserving agent. The mummies buried there were seldom placed in coffins and none were adorned with painted portraits. Instead, the mummies were covered in red shrouds with gilded masks and foot cases. Some gilding of the skin was also found. The study of sites like Douch and Ismant el-Kharab brings a unique perspective on embalming practices, offering insights from more rural and poorer communities. Surprisingly, their mummification was generally of a good standard, with the use of gilding suggesting an element of social stratification. Data from the Oases help nuance the usual perception that Roman mummification as unskilled and substandard. The notion of a decline since the Third Intermediate Period needs to be revised, with evidence from the Nile Valley and the Oases confirming that embalmers still knew how to perform a high standard of mummification in the Roman Period. However, a range of techniques, probably varying by location and individual, is observed. Different standards were possibly influenced by the social standing and economic means of the family, strongly dictating the level of mummification performed.

Late Antique and Medieval Periods (AD 395–1500) Traditional embalming practices can be observed until relatively late into the Roman Period. Fewer numbers of recorded examples are probably due to their more simplified treatment, a general decrease in embalming, poor preservation or a lack of interest for such mummies by early archaeologists (for a list of archaeological examples, see Dunand 2007: 164–169). The rise of Christianity also greatly impacted beliefs, with the notion of resurrection now in direct conflict with the practice of mummification. Although it mostly stopped after the Arab conquest in AD 642, unsurprisingly some embalming persisted in a country where beliefs in the preservation of the body were long-lasting, especially in pagans and Christians communities. Found both in the Nile Valley and in the Oases, embalming focused on the use of salt/natron that ranged from a thin sprinkling over the body to thick plaques placed on top or within the wrappings (Dunand 2007; Dunand and Lichtenberg 2019). Various unguents and incense were also still in use (Aufderheide 2003: 250). There is, however, no evidence than organ removal was still in practice and though a common feature of earlier periods, resin is no longer being used.

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Wrappings could vary from elements of clothing to shrouds and include, in contrast to Pharaonic Egypt when it was banned, the use of wool. Some individuals are seemingly wrapped in their own clothes (Dunand 1995: 3271–2; Dunand 2007). The mummy of Euphemia (Brussels, Royal Museums of Art and History) is particularly well preserved. Buried in Antinoe, she is known as the embroideress because of the embroidery tools found near her grave. She lived around the sixth century AD (earlier dating has also been suggested, see Van Strydonck et al. 2011) and was wrapped in multiple layers of tunics, linen, and other pieces of fabric, with pillows placed under head and feet. Many examples of natural mummification have also been found during this period, in some cases co-occurring with artificially mummified remains (e.g., at El-Deir, see Dunand and Lichtenberg 2019). Around 50 incredibly preserved naturally mummified bodies were also excavated in the area of the Fourth Nile Cataract in Nubia (modern Sudan) during the rescue campaign preceding the building of the Merowe dam. These mummies, all dating to the Medieval Period (c. AD 600–1500), are part of a larger collection of over a thousand human remains donated by Sudan’s National Corporation for Antiquities and Museums to the Sudan Archaeological Research Society, who in turn donated them to the British Museum where they are now curated. The mummies, as well as the skeletal remains, are currently the focus of an extensive program of research that will provide new insights into the past inhabitants of the Fourth Cataract. The textile clothing and soft tissues of the mummified individuals are well preserved, with no apparent attempt of artificial mummification. All have been passively conserved, allowing for the use of a broad range of analytical techniques (Wills and Antoine 2015). A woman discovered at site 3-J-23, close to the modern village of et-Tereif (British Museum EA 83133), was buried in a pit covered with slabs. Heated by the sun, this enclosed space might have prompted the process of natural mummification. Wrapped in several layers of textiles made of linen and wool, some showing obvious evidence of reuse, radiocarbon dating indicate that she died around AD 655–775 (Antoine and Ambers 2014; Taylor and Antoine 2014: 170–185). The body shows no evidence of embalming; however, her skin and internal organs are remarkably preserved. CT images show that her brain is intact, as well as the lungs, liver, heart, and other organs, which are desiccated but recognizable. She was approximately 158 cm tall and the minimal remodeling to her pubic symphysis shows that she was a Young Adult, most probably between 20 and 34 years old at death. Both of her lower canines appear to have been deliberately removed, probably as part of cultural practices seen elsewhere in Sudanese archaeological sites from the Neolithic onwards. Another remarkable feature is the presence of a tattoo clearly visible on the upper part of her right thigh. The first known example of this practice in medieval Sudan, it takes the form of a monogram, combining and overlapping the Greek letters for Michael, thus connecting her to the Archangel Michael. Saint Michael played an important role in Christian Nubia, featuring extensively in the iconography of the period and often found exquisitely painted on the walls of Sudan medieval churches. It also attests to her Christian faith 1500 years after her death (Antoine and Ambers 2014; Vandenbeusch and Antoine 2015).

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Conclusion: Toward Systematic Methods and Approaches Mummification in ancient Egypt has a complex and varied history but its early development is still relatively poorly understood, in part due to the limited body of published evidence. More research is still needed to explore regional differences over time, while temporal changes would also benefit from further study. The apparent decline in mummification standards after the Third Intermediate Period, for example, is not always noticeable, and the analyses of Late Period, Ptolemaic, and Roman mummies reveal the great care and skills with which individuals could be embalmed. Mummies from the Ptolemaic Period have also been little researched and only recently have elements specific to the period emerged. Despite some of the challenges and limitations of the radiographic methods (see Cox 2015; Gerald 2015), advances in CT-scanning and threedimensional visualization technology are allowing us to analyze mummified remains in greater detail without the need to unwrap them, adding significantly to the available data. Genetic, chemical, and histological analyses are also providing valuable new insights. However, as noted in Nelson and Wade (2015: 942), despite these technological and scientific advancements, a lack of comparable datasets frequently limits research to individual case studies. This approach, often employing differing methods and recording systems, is restricting our broader understanding of local and temporal trends. In order to gain comparable data, researchers need to define how each bioarchaeological profile was established and systematically record mummification practices, thus allowing like-for-like comparisons. So as to gain a more nuanced understanding of whom/why/how individuals were being mummified, synthetic studies that emphasize this comparative approach offer much promise. Broad studies and reviews of the treatment of the brain and organs by Wade and colleagues reveal previously unappreciated variability and complex temporal trends, often varying by status, and possibly by geography (Wade et al. 2011; Wade and Nelson 2013a, b; Nelson and Wade 2015). To better understand the individuals being mummified and the evolution of this funerary tradition, this complex practice needs to be further researched using large-scale comparative approaches that will allow individual findings to be contextualized (Nelson and Wade 2015). Although mummies are now being virtually unwrapped, the wrapping was an essential aspect of transforming the physical remains of the deceased into a divine image (Riggs 2014). Research needs to move beyond focusing on how the body was preserved and explore further the transformational aspects of mummification. With the emergence of more sophisticated imaging techniques, we can gain a better understanding of the how bandages and other embalming materials were elaborately applied to provide the mummy with the attribute and qualities of the gods. By combining disciplines, we can also start building a bioarchaeological profile of each individual, providing insights into aspects of their past lives and go beyond the formal imagery of ancient Egyptian art to reveal the individuals behind the masks.

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Cross-References ▶ Bog Bodies and Natural Mummification of Siberia ▶ Medical Imaging in Mummy Studies ▶ Paleo-oncology and Mummies ▶ Radiology Applications in Mummy Science ▶ The Burden of Arteriosclerotic Cardiovascular Disease in Ancient Populations Acknowledgments This new research was made possible thanks to the help of our British Museum colleagues and the excellent 3D visualization work of Benjamin Moreno (IMA Solutions, Toulouse, France) using the Royal Brompton and Harefield NHS Trust CT scanner on days that did not impact patient use.

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Different Types of Animal Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Brief Overview of the Study of Animal Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Animal Mummies Reveal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . How to Study Animal Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . How Were Animals Mummified . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Socioeconomic Role of Animal Mummies in Egypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . South American Animal Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion: The Future of Animal Mummy Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

While formal animal burials are known from Egypt from the earliest periods of its history, anthropogenically mummified animals are known sporadically from the Old Kingdom (c. 2600–2100 BC) onward, becoming increasingly common in the New Kingdom (c. 1549–1030 BC), and then enjoying an unprecedented popularity from the Late Period (c. 660 BC) until the Christian domination (c. middle of the fourth century AD). Different types of animal mummies are found were produced in Egypt, particularly from the Late Period onward, when they number in the millions. These objects provide an insight into ancient Egyptian religious beliefs, relationship with animals, animal husbandry, the changing environment, trade, economy, medical/veterinary expertise, and scientific knowledge. Animal burials are also known from South America, but are less widespread than those found in Egypt, and tend to be naturally desiccated rather than anthropogenically

S. Ikram (*) American University in Cairo, New Cairo, Egypt © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_18

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prepared. Their roles seem to be slightly different to those of many of the Egyptian animal mummies, but further work on this phenomenon is needed. Keywords

Egypt · Peru · Chile · Offering · Sacred animal · Pet · Food · Natron

Introduction Although the public has known about and been fascinated by human mummies for many years, it is only recently that interest has grown in animal mummies. Animal mummies are known primarily from Egypt, although some examples have been found in South America, notably in Peru and Chile. This chapter will first present Egyptian animal mummies, and then the South American ones. In the nineteenth century Father Ferdinand de Géramb quipped that no one would believe that one had visited Egypt unless they returned “with a crocodile under one arm, and a mummy under the other” – and no doubt a crocodile mummy would trump all souvenirs. Egyptian animal mummies, the artificially preserved bodies of animals, are found in almost all museums of the world, and include a diverse range of creatures, ranging from shrew mice to crocodiles. They, like humans, would be preserved so that their souls, the ba and ka, would live eternally. The act of mummification transformed a creature into a divine being, able to live for eternity. Currently, animal mummies can be divided into six major categories: pet mummies, victual or food mummies, sacred animals, votive animal mummies, false/amalgam/ pseudo mummies, and “other,” which is a diverse group with various interpretations (Ikram 2015a: 1–4; 2019); of course, all these groupings are constantly being refined as more mummies are found in situ and increasing number of texts relating to them and the cults in which they were used are published (Ray 1976, 2011, 2013; Smith et al. 2011).

The Different Types of Animal Mummies The easiest type of animal mummy to understand is that of the pet. Even today people become very attached to their pets and frequently go to great expense in order to bury them, sometimes in a way that the owner and the pet animal can share space in the afterlife (https://www.npr.org/sections/13.7/2017/05/18/528736490/whenwhole-family-cemeteries-include-our-pets; http://blog.sevenponds.com/somethingspecial/new-york-allows-pet-burials-in-human-cemeteries; https://www.telegraph.co. uk/news/health/pets-health/7061716/Owners-pay-to-be-buried-with-their-pets.html; https:// www.theatlantic.com/family/archive/2017/10/whole-family-cemeteries/542493/; all accessed 28 August 2019). The Egyptians were no different. Pet animals such as dogs, cats, monkeys, and gazelles were elaborately mummified and often given their own coffins so that they could be reunited with their owners in the afterlife. In a few

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instances the animals were actually buried with their owners. A man called HapiMin, who lived probably around 300 BC, was found in his coffin at Abydos with his pet dog at his feet (Petrie 1902: 39–40, pls. 8, 9, 79; Fleming et al. 1980: 86). Quite possibly the dog had died of grief shortly after his master’s death, and was thus embalmed and then buried with his master so that they could keep each other company eternally. A high priestess by the name of Maatkare was found with a mummy bundle near her belly in her coffin. Initially the Egyptologists who found her thought that it was a baby and that something scandalous had occurred and that the high priestess had been put to death for having an illicit child. When Maatkare and the mummy bundle were x-rayed in the 1960s, the bundle turned out to be that of a vervet monkey, clearly the high priestess’ pet (Harris and Weeks 1973; Ikram and Iskander 2002: 53, 100). After 100 years Maatkare’s reputation was restored, and we have learned something not only about the ancient Egyptians, but also about the mind-set of the Egyptologists who first examined these mummies. A baboon and a hunting dog were recovered from a tomb in the Valley of the Kings (tomb 50) and presumably were pets of a pharaoh, either Amenhotep II or Horemheb, based on the proximity of these two kings’ tombs to that of the animals (Fig. 1). Queen Isitemkheb D was the proud owner of a pet gazelle that was buried in a gazelle shaped coffin that was placed in her owner’s tomb (Ikram 2000). In other instances, such as that of Senenmut, a horse and a baboon were buried in the courtyard of the tomb (Lansing and Hayes 1935–36), and in Theban Tomb 11–12 a monkey mummy was found in the courtyard as well (personal observation). Probably, if the animal died before or after the tomb owner, it was buried in a nearby spot so that they could enjoy the afterlife together. Victual or food mummies are typically Egyptian and emphasize their idea that you can take everything with you, and that life after death is very similar to life on

Fig. 1 This dog (Egyptian Museum, Cairo CG 29836) was found in a tomb in the Valley of the Kings, tomb 50. It was probably the pet of either King Amenhotep II or King Horemheb. It was eviscerated, desiccated, and wrapped, though thieves have removed most of the wrappings. (Photo: AnnaMarie Kellen/Courtesy Cairo Museum, Animal Mummy Project)

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earth (Fig. 2). These consist of meat offerings, such as ribs, legs of veal, beef shoulders, and even liver that have been prepared to provide sustenance for the deceased in the afterlife. Poultry included ducks, geese, and pigeons – chickens were not common in Egypt until about the second or third century BC. The poultry would be plucked and cleaned, joints of meat skinned and prepared for consumption, mummified, wrapped (presumably without the same prayers that were used for pets and other animal mummies), and then placed in coffinets, mimicking the shape of the joint of meat or poultry, and then sealed so that they could be enjoyed by the deceased eternally. Meat was similarly processed, as if it were ready to be eaten. King Tutankhamun, who died when he was a teenager, had over 40 victual mummies in his tomb – clearly the young ruler was well fed in the afterlife. These mummies tell us what sort of food was thought to be highly desirable and enjoyed forever, as well as providing information about the butchery and jointing of animals. This form of offering was most common in New Kingdom 1549–1069 BC elite burials, although un-mummified food offerings are known from earlier periods (Goodman 1987; Ikram 1995: Appendix II; Ikram 2004, 2013a, 2015b). Sacred animal mummies belong to animals that were worshipped as manifestations of gods on earth. It was believed that certain gods would send their “essence” or part thereof into the body of a carefully chosen animal that was distinguished by being patterned or colored in a particular way, recognizable to the priests of that god (Vos 1998; Ikram 2015a). During its lifetime this animal was worshipped and treated as if it were a god, importuned as an oracle, and after its death mummified and buried in a catacomb with great pomp, while the spirit of the god moved to a new body, much as is believed for the Dalai and other lamas. The most ancient of these animal cults was that of the Apis Bull, sacred to the creator god Ptah of Memphis, and buried at the Serapeum at Saqqara; other bull deities are known from Heliopolis and Armant Fig. 2 A victual or food mummy of a duck wrapped up and placed in a wooden coffinet, originating from the tomb of Yuya and Tijuyu, tomb 46, in the Valley of the Kings. (Photo: Anna-Marie Kellen/Courtesy Egyptian Museum, Animal Mummy Project)

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(Dodson 2015). Rams sacred to Khnum, god of potency, creation and inundation, were buried at Elephantine, while the site of Bubastis housed a cat dedicated to the goddess Bastet (Ikram 2015a). Many such animals lived to an unusually great age due to the care that they received – indeed, many of the rams that were manifestations of the creator god Khnum lived beyond the average age of normal rams, dying when they were over 20 years of age, and had been hand fed with mash as their teeth had worn down entirely (Ikram et al. 2013a) (Fig. 3). Unfortunately, only a few examples of mummies that clearly belong to sacred animals have been found; the majority are of the rams of Khnum, as most other catacombs have been looted. The most common kind of animal mummy is the ex votos or votive offering (Fig. 4). The majority of mummies found in museums throughout the world today fall into this category of mummy. They consist of mummified animals that were dedicated to specific deities. Each god had a specific animal that was their totem or symbol due to some characteristics that it shared with the deity: cats were sacred to the goddess Bastet, goddess of pleasure, love, and beauty, all of which are attributes of a cat; sacred ibises were consecrated to the god Thoth, god of learning, partially because their beaks took the shape of a pen and they were always questing in the mud. These mummified animals were purchased and offered by pilgrims at shrines dedicated to these gods. The mummified animals would present the prayers of the pilgrim to the god throughout eternity, much in the way that votive candles are purchased and burned in churches. Once offered, they would remain in the temple precincts until an annual or bi-annual celebration, possibly attended by thousands of pilgrims, when they would be interred in specially constructed catacombs associated with the temple (Ray 1976, 1978, 2001; Ikram 2015a, b; Nicholson et al. 2015). Fig. 3 A Sacred Mummy of a ram, representing the god Khnum comes from the cemetery of Sacred Rams at Elephantine (Egyptian Museum, Cairo CG 29861). All the bones are fused, and the teeth are all erupted and worn down to a level that it would have been difficult for the animal to survive had soft food not been provided for it. (Photo: Anna-Marie Kellen/ Courtesy Egyptian Museum, Animal Mummy Project)

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Fig. 4 This votive mummy, dedicated to the fertility god Sobek, probably contained the mummified body of a hatchling crocodile (Egyptian Museum, Cairo CG 29712) found in Thebes. Sobek was revered in many places, but primarily the Fayum and Kom Ombo, where several hundred crocodile mummies have been found. (Photo: Anna-Marie Kellen/Courtesy Egyptian Museum, Animal Mummy Project)

Presumably it was thought that a sacrifice of a live flesh and blood creature was more potent and would better command a god’s attention than an offering of stele or image of the creature in stone, wood, or clay. Although animal cults, such as that of the Apis bull, were known from the dawn of pharaonic Egypt (c. 3000 BC; Simpson 1957), their proliferation, together with the institution of votive mummies, started in c. 600 BC and lasted until c. AD 350. This phenomenon might have in part been due to the fact that Egypt was overrun repeatedly by waves of foreign invaders, and thus they turned to an archaistic view of their religious past as a means of unification against the alien rulers, and expressing a cultural and religious identity that was uniquely Egyptian. Additionally, the physical manifestation of a god perhaps gave the Egyptians with a more intense sense of connection with their deities and provided a comforting presence in their midst who could be appeased and would be able to affect their lives. It is notable that many of the animal deities had oracular powers (Kessler 1986; Ray 2001; Ikram 2019). A vast range of creatures were offered: cats, dogs, crocodiles, gazelles, fish of different types, including catfish and Nile Perch, baboons, raptors, ibises, baboons, crocodiles, shrews, and scarab beetles (Ikram 2015a: xvii). By 200 BC all of Egypt had such cemeteries, with millions of creatures being offered. Age ranges of the offered animals, from puppies or kittens or eggs to adult animals, indicate that many creatures were especially bred for this fate. Radiographs of many of these animals have shown that many were deliberately killed, either by strangulation or, in the case of some dogs and cats, by a sharp blow to the head or possibly even drowning (Armitage and Clutton-Brock 1981; Zivie and Lichtenberg 2015; Ikram and Iskander 2002; Ikram et al. 2013b; Nicholson et al. 2020). This seems shocking to a modern sensibility, but perhaps if one were to look at this through the eyes of the ancients it would not be quite so harsh – these animals were sacrificed and would live eternally in the presence of their god in a particularly blessed state. Within the votive offerings, examples of an incomplete mummy have also been found. These are dubbed “false,” “pseudo,” “amalgam,” or “fake” mummies,

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although wrapped to resemble a specific animal, and buried in a catacomb/tomb dedicated to the god to whom they are offered. In this type of mummy, the package, instead of containing an example of the animal that is supposed to be offered, actually enclose remnants of one or several animals, such as a bunch of feathers, or one wing, or a bone from an entirely different species, or mud, or, indeed, nothing at all (Fig. 5). There are various explanations for this phenomenon (Kessler 1986, 1989; Germer et al. 1994; Ikram and Iskander 2002; McKnight et al. 2008; Ikram 2015b, 2019). The cynical one is that the priests were fooling the pilgrims. However, to be more charitable, the Egyptians believed that a part could signify the whole, and that if one said or wrote that something was a particular item, then it magically became so through the statement and prayers. Additionally, as these animals were sanctified as they were mummified and thus transformed offerings, any part that fell off them was sacred, and thus had to be treated with due respect and was carefully wrapped and buried in a catacomb. The final type of animal mummy falls under the general rubric of “other” as these do not fit neatly into any of the above categories. Examples include mixed burials of humans and animals – often dogs (Ikram 2013d). In some cases the animal mummies might act as guardians or amulets, as some scholars have called them (Hartley et al. 2011; Ikram 2019; Kaiser and Ikram 2009). A special rite existed in the Roman period that related to the co-burial of children and canines in the Baharia Oasis (Adam and Colin 2012), which might perhaps explain some of the other canine-human combinations. Of course, it is possible that a burial place was shared sequentially by different species, although the reason for this is debatable and might include a desire to respect the dead, laziness, or have some cultic purpose. In one Theban tomb, animal mummies were carefully placed around a sarcophagus (Rhind 1862). These might have served as “living” images of the Sons of Horus or other funerary deities, placed here in lieu of statues, and might have been regarded as a more powerful means of protecting the deceased than the statues. Fig. 5 False/Pseudo/ Amalgam mummies consisted of a variety of materials, and are generally found among votive mummies. In this case, a wrapped bundle of ibis feathers that was given as a votive offering at Abu Rawash, while other bundles contained complete birds. (Photo: Salima Ikram)

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It is difficult to precisely type animal mummies from museum contexts as often their provenance is unknown or uncertain. The basic means of identifying an animal mummy lie in the find spots. Pets are found with owners, and are few in number. Often they have coffins inscribed with their names. It is easy to identify a Sacred bull as these tend to be found in special burial places, complete with texts, large sarcophagi, and even grave goods (Dodson 2015) (Fig. 6). Votive mummies number in the hundreds, thousands, or millions appear in mass graves or catacombs. Many are in coffins or containers of different qualities (Figs. 7 and 8). However, it is particularly complicated to differentiate between votive mummies, some of the smaller Sacred animals, particularly if they are removed from their fine wrappings and burial equipment, the mummies found under the rubric of “other,” and even pet mummies when decontextualized. It seems that the majority of museum examples are from the most numerous category – votive mummies, although it is always possible that a few that are thus identified fit into another category. More research in current excavations of animal cemeteries should shed light on this matter.

Brief Overview of the Study of Animal Mummies For a long time, despite their vast number and variety, scholars largely ignored animal mummies as they shared the view of the ancient Romans that these objects were quaint oddities and a strange aspect of Egyptian religion. Indeed, thousands of Fig. 6 The Serapeum at Saqqara was the burial place of the Apis bulls. Its underground galleries run for kilometers, consisting of corridors with rooms coming off them, each room containing enormous granite sarcophagi for the bulls. (Photo: Salima Ikram)

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Fig. 7 The Falcon Galleries at Saqqara, with thousands of pottery vessels, each containing a raptor. Pottery vessels were the most common containers, especially for birds, though other animals, such as a group of dogs from Abydos, have also been found in large ceramic jars. The jars were placed in long chambers leading off from an axial gallery and filled with mummies. When the gallery was full it was blocked by stone or mud brick. (Photo: Salima Ikram)

cat mummies, mainly from Bubastis in the Delta, were used as ballast in ships traveling from Alexandria to Europe during the late nineteenth century. Once in Europe the mummies were sold in bulk, broken up, and used as fertilizer (Ikram 2015b; Malek 1993; McKnight and Atherton-Woolham 2015; Daily Graphic, 12 February 1890; Punch, 15 February 1890). Many other examples of animal mummies were collected by nineteenth-century travelers and collectors, and donated to museums when the family became bored of the object, or it started to smell in the damp European and American climates. Eventually, some serious interest was evinced in these artifacts at the end of the nineteenth and the very beginning of the twentieth centuries. However, it was naturalists rather than Egyptologists who initiated the collection, study, and cataloguing of animal mummies. On the whole, Egyptologists regarded animal mummies as a rather strange manifestation of the Egyptian religion, and in the early days of the discipline few scholars wished to engage with these objects and with the phenomenon of animal mummification. The foundation of our information and the basis of the collection of the animal mummies in Cairo’s Egyptian Museum and the Natural History Museum in Lyon, now the Musée des Confluences, two of the world’s largest collections of animal mummies, were due to the initiative and drive of Louis Lortet (naturalist and Director of the Natural History Museum in Lyon), Claude Gaillard (a naturalist), and Georges Daressy (Egyptologist). These naturalists might in part have been inspired by the work of Étienne Geoffroy St. Hilaire (1772–1844), who came to Egypt as a member

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Fig. 8 Many different types of containers were made for animal mummies, including wooden and stone boxes. Cats frequently were buried in containers that mimicked their shape. It is unclear if this coffin (Egyptian Museum, Cairo CG 29773), containing the body of a wild cat, was for a Sacred or votive cat offering as the details of its precise find spot are unknown. (Photo: Anna-Marie Kellen/Courtesy Egyptian Museum, Animal Mummy Project)

of the group of savants who formed a part of Napoleon Bonaparte’s expedition to conquer Egypt between 1798 and 1801. He and his colleagues collected many animal mummies, and studied the contents of the mummy bundles to identify species, speculate about changes in animal morphology/evolution, and the diachronic changes in the Egyptian environment. After Lortet and Gaillard’s work on animal mummies, interest in the subject was minimal and intermittent, and only resurfaced in the 1970s when new excavations yielding animal mummies took place. However, they obtained a better and deeper understanding of ancient Egyptian fauna and environment (Lortet 1901; Lortet and Gaillard 1902, 1903–1909). They also established large collections of these artifact types, as a result of which, the Egyptian Museum was the only museum focusing on an ancient culture that also dealt with its environment (Ikram and Helmi 2002). As time progressed, a few other museums also acquired well provenanced animal mummies through excavations, notably the Louvre, the British Museum, and the Muzeo Egizio in Turin. The main twentieth-century excavations that provided fresh examples of animal mummies and renewed interest in these objects were those of the catacombs at Tuna alGebel and of the Sacred Animal Necropolis (SAN) at Saqqara. The former primarily provided examples of ibis and baboons (Smith 1976; Nicholson and Smith 1996; Nicholson 2015), and the latter of ibis, baboons, raptors, and cattle (Boessneck 1987; Von den Driesch and Kessler 1994; Von den Driesch et al. 2004, 2005; Kessler 1986, 1989, 1998; Kessler and Nur el-Din 2015; http://www.aegyptologie.uni-muenchen.de/ forschung/projekte/tuna/index.html). These excavations, especially the former, focused

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the interest of some scholars on animal cults, but for the most part animal mummies remained in the shadows of traditional Egyptology. It was not until the end of the twentieth century, with the work of Cairo’s Egyptian Museum Animal Mummy Project, directed by the author and N. Iskander (Ikram and Iskander 2002), which coincided with more publications of results from Tuna al-Gebel and the SAN, and the research of Alain Charron (1990, and Ph.D.) on the religious aspects of animal mummies, that animal mummies once more stepped into the spotlight. The Cairo work developed a typology of these mummies, and helped spark media interest, which led to animal mummies gaining higher visibility and gradually becoming a significant part of the world of Egyptology and a focus of public interest. Now excavators, faunal and other specialists, and museums all pay more attention to these artifacts, with many studies being carried out on the material (see below), and many fresh examples coming to light, such as those from Quesna (Rowland et al. 2013), Saqqara (Zivie and Lichtenberg 2015; Ikram et al. 2013b; Nicholson et al. 2015, 2020), Asyut (Kitagawa 2016); Abydos (Ikram 2007, 2013b; Bestock 2013; Abu el-Yazid et al. Forthcoming), Theban Tombs 11, 12 (Ikram and Spitzer in press), and Kharga Oasis (Ikram 2013c; Dunand and Lichtenberg 2005; Dunand et al. 2015).

What Animal Mummies Reveal We study animal mummies now not because they are some type of curiosity, but because they provide a great deal of information about different aspects of ancient Egyptian culture, technology, and history. By identifying and studying the different animals, one can establish what constituted Egypt’s ancient fauna, and gain a better understanding about ancient climate and environment, and diachronic change. Thus, the presence of shrew mummies in Upper Egypt, an area where no shrews are now found, might help track climate change (Woodman et al. 2019) as well as animal acquisition practices. Some animals show signs of disease or trauma, and depending on how these have been treated – or not – we can learn more about veterinary and animal management practices. For examples, some baboons have had broken arms set, while dogs have been hit on the head to control them, or have had limbs splinted so that they can walk, while some creatures suffered from infections and arthritis (Ikram and Iskander 2002; Hartley et al. 2011; Kitagawa 2016). Obviously, the mere fact that vast numbers of animal mummies were produced from the 26th Dynasty until almost the time when Christianity became the state religion tell us a great deal about religious and cultural beliefs of those times. The way in which mummies were produced informs us about the ancient Egyptians knowledge of chemistry and science, and both the embalming materials and bandaging techniques are studied in case they are indicative of diachronic change or contain clues that will help to identify different embalming houses or their geographic locations. The materials used to make millions of mummies provides an insight into ancient Egypt’s trade networks and economy (Ikram 2015c; Charron 2015), and the practice of cults where

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animal mummies were given as votive offerings also informs our understanding of the Egyptian economy in the Late and Graeco-Roman periods.

How to Study Animal Mummies The ways in which one studies animal mummies is very similar to how one studies human mummies (▶ Chaps. 39, “South American Mummies” and ▶ 27, “Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection,” this volume). The main source of evidence is, of course, the mummy itself (Lynnerup 2009; Ikram 2015d, e). Additionally, places in which mummies were manufactured and consecrated, such as the embalming beds at Memphis and the oil and resin smeared space at the mouth of one of the catacombs as Tuna el-Gebel also yield information on mummification (Jones and Jones 1982, 1988; Kessler 1989). As with human mummies (see Antoine, this volume and Ikram and Dodson 1998), there are virtually no ancient Egyptian texts that explain how mummies were made, with the majority of ancient information coming from Greek writers, such as the fifth-century BC historian Herodotus (in Book II of The Histories) and the first-century BC geographer Diodorus Siculus (in Book I of The Library of History). For animals, the only relevant text is the mid-second-century BC Apis Embalming Ritual (Vos 1993; Meyrat 2014). This, however, does not explain how to embalm an animal, but rather, how to wrap it, with specifics provided about the different types of bandages to be used on each limb, prayers to be read, and amulets to be inserted within the linens. The first step in studying an animal mummy is to carry out a macroscopic examination. This includes noting the state of conservation, type of bandaging, quality of bandages, texture of the bandages, color of the mummy and the bandages, other visible presence of any of the embalming materials, the state of the mummy package, and any odors (Ikram 2015e). Then radiography is employed, which includes x-rays as well as CT scans (Fig. 9). The latter provides a more detailed picture of the mummy and its different layers of bandaging, as well as some of the soft tissue that remains. This helps not only in identifying species, but has also shown that animals, like some humans, were provided with food in their afterlife (Wade et al. 2012; Ikram 2017), while the fact that a kestrel might have died from force feeding hints at ways of raising and procuring animals for mummification (Ikram et al. 2015). Increasing amounts of information are being extracted from animal mummies with each improvement in imaging technology. Additionally, the advances in such imaging have enabled us to be able to produce 3-D prints of the skeleton within the wrappings (Du Plessis et al. 2015) (Fig. 10), or amulets that adorn a mummy, which are not only useful to scholars, but add a new dimension to museum displays and public engagement. Chemical test of embalming agents is also productive in identifying the materials used in mummification: natron, resins, beeswax, and oils. Technologies that are being used for this include Fourier transform infrared micro-spectroscopy,

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Fig. 9 A CT-scan’s processed image of the mummy of a Sacred Ibis mummy from the collection of Lisbon’s Museu Nacional de Arquaeologia. (Photo courtesy of the Lisbon Mummy Project and the Museu Nacional de Arquaeologia)

Fig. 10 A 3-D print of a kestrel that was wrapped within a bird mummy bundle (Cape Town Iziko Museum 2575). (Photo: Courtesy R. Slabbert)

gas-chromatography/mass spectrometry, pyrolysis/gas chromatography, enzymelinked immunosorbent assay (ELISA), as well as electron microscopy/energy dispersive x-ray spectroscopy (Ikram 1995, 2013a; Aufderheide 2003; Buckley et al. 2004; Clark et al. 2013; also see ▶ Chaps. 29, “Trends in Use of Organic Balms in Egyptian Mummification Revealed Through Biomolecular Analyses” and ▶ 27, “Mummified Human Remains from Ancient Egypt and Nubia: An Overview and New Insights from the British Museum Collection” in this volume), to name but a few. These too elucidate mummification technology, the embalming agents, their sources, costs, and impact on the economy. DNA is increasingly used to understand the development and distribution of species. Recently the idea that there had been two types of crocodiles in the Nile, as posited by the French naturalist Geoffrey St. Hilaire in 1807, based on the skull morphology of mummified creatures, was explored through DNA investigations

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carried out by Evon Hekkala and her team (Hekkala et al. 2011). They found that indeed there were two types of crocodiles present in Egypt historically – Crocodylus niloticus and C. suchus, the latter being a much less fierce and violent creature than C. niloticus, and therefore easier to maintain and interact with in temple contexts. Now work is being carried out to see when (and if) this species vanished from the Egyptian Nile, as well as trying to understand whether it came to Egypt through trade or was part of the indigenous fauna until its local extinction. The history of the origin and dispersal of domestic cats is also being thus studied (Kurushima et al. 2012; Ottoni et al. 2017), as is the acquisition of animals for mummification (Spigelman et al. 2008; Wasef et al. 2019). Isotopic analysis is also useful for learning where animals came from or were reared, and is a rich source of future investigation, particularly for exotic animals, such as baboons (N. Dominy, W. Van Neer and S. Porcier, personal communications), whose origins might help establish the identity of trading spots such as Punt, as well as the elaborate trade networks that ran throughout Africa. The more precise dating of animal mummies is also a subject that is increasingly researched. Prior to Carbon 14 dating, animal mummies were dated contextually. Often this gave a long range, such as from the Late Period until the advent of Christianity, some 900 years, the time when animal cults were very popular. Some dates could be guessed at from the wrappings: more elaborate, geometric wrappings seem to be a hallmark of the fourth century BC to the third century AD, but this too is an imprecise way of dating, and also encompasses a considerable length of time (Figs. 4 and 11). Now radiocarbon analysis is being applied to mummies in order to establish a better chronology for these (Wasef et al. 2015; Richardin et al. 2017; Bleiberg et al. 2013), though many more samples need to be dated in order to clarify the situation. Experimental mummification is also an extremely useful tool in understanding how mummies were made, what kinds of technologies and materials create specific results, what ways are more effective than others in producing a mummy, and the kind of detritus one might expect in a mummification workshop. Experiments also help archaeologists identify sites associated with the breeding and mummification of animals, as well as to highlight issues that need further study (Ikram 2015f, g; Clifford and Wetherbee 2004; McKnight and Atherton-Woolham 2015).

How Were Animals Mummified Different forms of mummification existed for over 3000 years in Egypt, thus there were many methods applied to the preservation of animals. Furthermore, these varied between species, depending on whether the animal had fur, feathers, or fins, or whether it was a pet, victual, Sacred, votive, or other type of mummy. Desiccation is the fundamental precept of mummification as preserves the form of the body as completely as possible. In an ideal mummification, the Egyptians achieved this by evisceration (but not always), followed out by washing the body cavity with water and palm wine in order to purify it and inhibit bacterial growth. Then the body was

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Fig. 11 A cat mummy wrapped in diamond or lozenge pattern with a modeled and painted head (Egyptian Museum, Cairo CG 29657). (Photo: Anna-Marie Kellen/Courtesy Egyptian Museum, Animal Mummy Project)

packed in powdered natron, a naturally occurring substance that is basically a combination of salt and baking soda (sodium carbonate decahydrate and around 17% sodium bicarbonate together with very small amounts of sodium chloride and sodium sulfate), found in the Wadi Natrun, some 60 km from Cairo and also in the area of ElKab, south of Luxor. It desiccates and de-fats. For humans it is thought that 40 days of desiccation were necessary (Ikram and Dodson 1998; Ikram 2015f). For animals it is possible that there were more variations, as small creatures such as shrews, would not take that long, but larger animals, such as cattle, would take longer, and reptiles and fish might desiccate more quickly than the more meaty mammals, and some animals might not be treated with natron at all. After desiccation the animal would be patted or brushed free of natron and then rubbed with sacred oils, often mixed with aromatic resins. The oils slightly rehydrated the body and gave it some flexibility so that it could be positioned; resins acted as inhibitors to bacterial growth. Both oils and resins were religiously charged, and the latter were obtained frequently from the Levant. The anointing of the body would be accompanied by a series of prayers and then the bandaging of the mummy would take place, also accompanied by a different set of prayers. Linen bandages were the norm – cotton was not known in Egypt until the Roman era. For humans the wrapping was supposed to take 30 days; again, for animals this would have taken different amounts of time depending on their size and the complexity of the mummification. The exterior wrappings of votive mummies are often very complex, using different colored cloth –

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brown and beige primarily, though pink and yellow-beige were also used in the Ptolemaic era (Ikram, Personal Observation) – creating herringbone, lozenge, and coffered patterns (Figs. 3 and 11). The coffering and lozenge patterns are found on human mummies as well, while the herringbone is less common. Quite possibly a skilled subgroup of embalmers were specialized in these sorts of wrappings. Victual mummies are very different to other animal mummies as they were prepared as if they were to be consumed. Thus, for poultry the head, wing-tips, and feet were removed. They were eviscerated, but after mummification the desiccated viscera was returned to the body as has been traditional even in supermarkets so that the consumer can benefit from the liver, heart, and giblets. After desiccation, oils and often imported terebinth resin (Pistacia terebinthus) were used to anoint the processed bird, and then it was wrapped up, often being placed in a coffinet (see Fig. 2) that was also painted with resin (Ikram 1995: Appendix II; Ikram 2004, 2013a, 2015b). Joints of meat were similarly processed so they were ready for consumption – indeed, some might have been boiled or baked before being salted, anointed, and wrapped. Though more mummies need to be studied before one can make generalizations about differences in mummification between species or cemeteries, it is worthwhile mentioning some of the variations that have been noted thus far. While mammals tended to be eviscerated, this was not always the case for smaller animals, such as shrews, and particularly in what have been tentatively identified as mainly Roman votive mummy deposits, such as the Dog Catacombs at Saqqara, evisceration is probably not always the norm. This also holds true for some burials of small bovids as stomach contents survive (personal observation from a Theban example and a few from Saqqara). However, Isitemkheb’s gazelle was as well mummified as an elite or royal human of that era. It was eviscerated, desiccated, stuffed with sand and sawdust, anointed with oils and resins, wrapped in good quality linen, and encoffined (Ikram 2000). Degrees of desiccation also vary, with some animals being well dried and others less so, depending on the amount of time that they have been left in the natron and the amount of natron used. Some mummies contain the discarded pupae of flies, indicating that they were not well covered during mummification and prey to flies as well as dermestid beetles (corroborated by experimental work, such as noted in Ikram 2015g). In the case of some birds, it seems as though some might not have been desiccated, just killed and then dipped into molten oil and resin, before being wrapped (Nicholson 2015). Fish mummies show a limited use of oils and resins on the surface of the body prior to wrapping, in contrast with the birds and some mammals. Crocodile hatchlings are often not eviscerated but generously covered with resinous substances and splinted between pieces of palm rib and then wrapped as the palm ribs give them rigidity and protection. Clearly, more work needs to be carried out on provenanced mummies in order to establish differences in mummification based on mummy type, geographic area, atelier, and the time period of mummification. Economic considerations also have to be taken into account, particularly in the case of votive mummies, as quite possibly the plainer ones were less costly than the more elaborate ones using expensive and imported materials.

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The Socioeconomic Role of Animal Mummies in Egypt Animal mummies not only played a significant role in the belief system of the ancient Egyptians (Kessler 1986, 1989; Ray 2001; Ikram 2015b), as revered gods or offerings to them, but also contributed to their economy and sense of nationalism. The number of animals that had to be bred to fulfill the demand for votive mummies was great. As the Archive of Hor (Ray 1976) documents, many priests were involved in the breeding, feeding, and general upkeep of flocks of ibises that were mummified and offered to the god Thoth; additional birds might have also been raised by villagers living in the environs of the temple, who would have been paid for their birds. This would also have been the case for the cats and dogs, and perhaps also the raptors and crocodiles that were offered to the gods (Charron 1990, 2013, 2015; Ikram et al. 2013b; Ikram 2015c). In addition, the number of embalmers required to produce thousands of mummies would have played a part in the local economy, while the materials they used in mummification, such as natron, oils, and imported resins from the Near East and possibly other parts of Africa, would have boosted national as well as international trade and economy. Indeed, some of the mummified animals, such as the monkeys and baboons would themselves have been the subject of international trade. The temples benefited significantly from the purchase of the completed mummies, and the surrounding villages with their inns and hostelries would have welcomed the vast number of patrons who would have come to attend the festivities surrounding the animal interments (Ikram 2015c; Charron 2015). Thus, it is apparent that animal mummies were pivotal to the emotional and practical lives of the ancient Egyptians, particularly from the Late Period until the Christian era.

South American Animal Mummies The study of South American mummies is a relatively young field as it is only at the end of the twentieth century and the start of the twenty-first century that large numbers of buried animals have been found. As far as one can establish without sufficient chemical tests, these are not true mummies as they have not been anthropogenically created, although they have been wrapped and buried with funerary goods. Peru has yielded a few hundred dog remains, associated with the Chiribaya people (fl. AD 1000–1476), and tentatively dated to AD 1000 and on (S. Guillen, personal communication; Meier 2006), as well as other individual dog burials found earlier in the twentieth century (Aufderheide 2003: 411) (Fig. 12). Many of the Chiribaya dogs were well preserved, some with the fur still remaining, due to the salinity of the soil. These were buried in pits, and many were wrapped in llama blankets, with llama bones and fish skeletons placed near their snouts, so they could eat in the afterlife. Providing food for animal burials is also known for some of the Egyptian mummies, both pet and votive (Malek 1993; Wade et al. 2012; Ikram et al. 2015). It is unclear if the Chiribaya dogs were pets/guardians, or played a role in the religious life of the community, though currently excavators favor the former identification. These animals have yielded important information about the dispersal

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Fig. 12 One of the Peruvian dog mummies where the fur and textile are visible. (Photo: Joanna Motley, courtesy of the Museo del Sitio Pachacamac, Peru)

of dogs in the new world (Leonard et al. 2002), the parasites that they carried (Richardson et al. 2012), as well as an insight into human and canine relations. Other spontaneously mummified dogs have been found at Arica in the Atacama Desert (Allison et al. 1982), dating to 500 BC–AD1500. One animal was buried without grave goods, and another was associated with a human burial. The remaining animals were supplied with food items, cloth, stone tools, and other daily-life objects. These burials were identified as being ceremonial, though the role that the dogs played is unclear. Interestingly, in addition to being pets/guards, dogs were eaten and their skin employed in handicrafts (Aufderheide 2003: 411). In Peru and Chile naturally preserved animals have also been found with human mummies. The types of animals include birds (macaws, parrots, herons, flamingos, and doves)andmammals(dogs,cats,guineapigs,viscachas,llamas,vicunñas,alpacas,anda monkey).Llamaandalpacaburials,commonanimalsintheregion,arealso,likethedogs, associatedwiththeChiribayapeople(Aufderheide2003:411).Whethertheywerepetsor workanimalsisunclear.Whilemostoftheanimalswerelocaltotheregionsinwhichtheywere found, the monkey was not (Aufderheide 2003:411). Here, as in Egypt, the presence of a monkeyisindicativeoftradeinanimals;thismightalsoholdtrueforsomeofthebirdburials. AtleastoneparrotburialfromtheNascaPeriodmightalsohavebeenthatofapet(Aufderheide 2003:417). Some of the South American animal burials were clearly ritual in nature. In Yaral, again a Chiribaya site (c. AD 1150), some 112 naturally desiccated guinea pigs were discovered under the floor of a house. Many were decapitated, and coca leaves were stuffed into the mouths of many of the animals (Rofes 2000); similar instances have been recorded at other sites in Peru (Aufderheide 2003: 416). Coca leaves are frequently found also in the oral cavities of humans from this area, and associated with religious and medicinal use, suggesting that the animals were some form of offering.

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Conclusion: The Future of Animal Mummy Studies Clearly the work on animal mummies and what they tell us has just begun, particularly in South America. In Egypt, using well provenance materials as well as carrying out C14, DNA, and the analyses of embalming materials and isotopes will contribute to the understanding of these objects. Certainly, such broad studies are now being carried out on material from museum collections, such as that of Lyon as well as the Museo Egizio in Turin. These investigations will shed further light on the complex role of these objects in the economic, religious, cultural, and emotional lives of the denizens of Egypt and South America.

Cross-References ▶ Fake and Alien Mummies ▶ History of Ancient DNA Analysis in Mummy Research ▶ Medical Imaging in Mummy Studies ▶ Radiology Applications in Mummy Science ▶ Stable Isotope Analysis in Archaeological Science and Mummy Studies

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Trends in Use of Organic Balms in Egyptian Mummification Revealed Through Biomolecular Analyses

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Placing Chemical Investigations of Mummy Balm in the Cultural Context of Ancient Egyptian Mummification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Materials and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummy Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lipid Extraction and Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Summary of Balm Compositions Analyzed in This Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Variations in Balm Compositions over Time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preparation of Balms and Specific Recipes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Variation on Balm Composition with the Age of the Individual . . . . . . . . . . . . . . . . . . . . . . . . . . . Variations in Balm Composition with the Gender of the Individual . . . . . . . . . . . . . . . . . . . . . . . Variation in Balm Composition with Location on the Body . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Variation of the Chemical Composition with Material Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

The compositions of organic balms from over 70 mummies ranging in date from c. 3500 BC to AD 395, together with ca. 40 undated mummies and a number of canopic jars, have been studied using a combination of gas chromatography (GC), GC-mass spectrometry (GC-MS), GC-combustion-isotope ratio mass spectrometry (GC-C-IRMS), and radiocarbon analysis. Mummies dating to before the Third Intermediate Period (c. 1000 BC) were typically found to be embalmed with only fat or oil, which may have been exogenous to the body in many cases. Investigations of balms from mummies dating from after c. 1000 BC showed R. P. Evershed (*) · K. A. Clark Organic Geochemistry Unit, School of Chemistry, University of Bristol, Cantock’s Close, Bristol BS8 1TS, UK e-mail: [email protected]; [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_9

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them to comprise of fat/oil, beeswax, and/or resin. Pistacia resin, in contrast to coniferous resin, was only identified in a small number of mummies dated to between the Saite and Ptolemaic Periods (c. 700–30 BC). Ruminant adipose fats, nonruminant adipose fats, plant oils, or a combination of these fats and oils were employed in embalming. Steranes and triterpane biomarker analyses for bitumen were performed using selected ion monitoring GC/MS of the saturated hydrocarbon fraction. The earliest example of the use of bitumen identified in this study was in a mummy dated to the Twenty-First to Twenty-Second Dynasties (c. 1064–927 BC), although its use was found to be most prevalent during the Ptolemaic and Graeco-Roman Periods. Quantification of bitumen in mummy balms shows the proportions of bitumen can be high, but balms were never pure bitumen. Combining these findings with those from other studies allowed assessments of variations in the composition of balms according to age, gender, body part, and material type. Most notably, the balms of children and females were found to comprise of fewer ingredients than the adult males. Moreover, it was found that balms collected from the head and limbs were simpler preparations than those applied to the torso. Balms visually identified as “resins” generally contained more ingredients than those applied to bandages and tissues. The major ingredients employed in balms were for the most part probably local to Egypt and cheap (fat/oil and beeswax), although more expensive exotic imported materials (resins and bitumen) were present in a high proportion of balms, especially in those mummies prepared after c. 1000 BC. Keywords

Egyptian mummies · Organic balms · Biomolecular analyses · Gender · Body position · Evolution of embalming · Age · Material type

Introduction The chemistry of mummy balms has intrigued chemists since the mid-eighteenth century, and the work of Lucas (1908, 1911) justifies acknowledgment of him as one of the pioneers of the field. However, it was not until the advent of instrumental methods in that latter decades of the twenty-first century that the molecular dissection of mummy balms became possible. One of the first such studies was undertaken as part of the Manchester Museum Mummy Project. The wrapping of an unprovenanced female mummy (no. 1770) dating to c. AD 380 (Benson et al. 1979) was analyzed using GC/MS to identify beeswax. A little later Storch and Schäfer (1985) analyzed the head, body, body cavity, and wrappings of an unprovenanced and undated mummy from Munich Museum (AS 73B) using a combination of ion chromatography, infrared spectroscopy, X-ray fluorescence spectroscopy, and MS. The head and body were found to be coated with a mixture of oil and beeswax, while the thoracic cavity contained beeswax, oil, bitumen, gum, soda, and fossilized resin. The outer wrappings were coated with beeswax, oil,

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bitumen, gum, soda, and tree resin. The coatings of a number of mummy cartonages of various dates were analyzed by Wright and Wheals (1987) using pyrolysis/MS and found to consist of broadly defined gums, rosins, or waxes. A more precise origin could not be determined because in the absence of chromatographic separation of these complex amorphous materials. The most significant advances in understanding the chemistry of mummy balms began in the 1980s with the increased use of GC/MS. The advantage of this approach is that it is able to provide on-line separation of the complex ingredients and identification via mass spectra. Two groups studied an unprovenanced mummy (Guimet Natural History Museum, Lyon, 90001255) dating from the GraecoRoman Period. Using GC/MS to reveal the presence of steranes and triterpanes, Connan and Dessort (1989) found bitumen in samples taken from the knees, a “balm” from the skull and a sample from the visceral packing. The balms also contained coniferous resin, identified by the presence of dehydroabietic acid, cadalene, retene (indicative of pine pitch) beeswax, fats, or oils and possibly a gum resin. Additional samples from this mummy were re-examined by Mejanelle et al. (1997). The methanolic extract was analyzed by GC/MS and was found to contain derivatives of aromatic acids, including vanillic and gallic acids and inositols. The source of these compounds was suggested to be a vegetable tannin. This is the first hint of tannins in a mummy, supporting the association between the processes used in leather making in the Roman Period and embalming. Examinations of Late Period mummies, mummy boards, and coffins by Rullkötter and Nissenbaum (1988) using GC/MS revealed the presence of bitumen biomarkers. The source of bitumen present in the mummy and mummy boards was identified as the Dead Sea (Harrell and Lewan, 2002) whereas the coffin (British Museum, 24906), which dated to c. 900 BC, had a different distribution of steranes and triterpanes, indicating that the source of this bitumen was Gebel Zeit on the Gulf of Suez. Given its proximity to Egypt and the early date for the coffin, this is perhaps unsurprising. In a further study carried out on a number of mummies ranging in date from the Nineteenth Dynasty (c. 1298–1187 BC) to the Graeco-Roman Period by Connan and Dessort (1991), showed the balm from these mummies contained a mixture of coniferous resin and bitumen. The bitumen was identified as originating from either the Dead Sea or Hit in Iraq based on the distribution of steranes and triterpanes. Quantifications based co-injected deuteriated standards suggested bitumen comprised 3–80% of the balm. The presence of bitumen in a mummy dating to c. 1298–1187 BC included in this latter study is the earliest example of bitumen in a mummy balm. The feet of a mummified child dating (World Heritage Museum, University of Illinois) from the Graeco-Roman Period were studied by a team in Illinois (Proefke et al. 1992). Using GC/MS, diterpenoids characteristic of a degraded coniferous resin: dehydroabietic acid, 7-oxodehydroabietic acid, and 15-hydroxy-7oxodehydroabietic acid, were identified. n-Alkanes ranging between C19 and C33 (max C23) with no odd-over-even predominance, were identified as originating from bitumen. No information about the steranes and triterpanes was reported, however, and it is possible that these n-alkanes were the result of contamination with paraffin

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wax, which was used in early post-excavation treatments (Petrie 1920). The skeletal muscle tissue from a Ptolemaic mummy (Staatliches Museum Ägyptischer Kunst, Munich) was analyzed using GC/MS and found to contain pistacia resin as the major component of the balm (Kaup et al. 1994). The high concentration of noroleanone indicated that the resin had been heated intensely. An oil of turpentine from the Aleppo pine (Pinus halepensis) was also identified from the presence of monoterpenoids including α-pinene, cymene, and limonene. This mummy was re-examined by Koller et al. (2005) using solvent extraction, followed by GC/MS where the presence of cedar wood tar oil was confirmed by the identification of compounds derived from phenol, guaicol, and naphthalene using GC/MS. Balms from a human mummy and two animal mummies were analyzed by Mejanelle et al. (1996) using GC/MS. The balms of these mummies were found to contain fatty acids indicating the presence of a fat/oil and hydrocarbons, characteristic of beeswax, and terpenoids, indicating the use of a resin. The earliest evidence for artificial embalming was found by a German team who examined an Old Kingdom (c. 2663–2195 BC) male adult, Idu (Roemer-Pelizaeus Museum, Hildesheim, 2639; Koller et al. 1998; Weser et al. 1998). This mummy, however, was soaked in paraffin when excavated, evident in the high concentration of hydrocarbons present, which the author acknowledged would hinder subsequent chemical analyses. Samples of bone contained cyclic alcohols and coniferous diterpenoids, identified using GC/MS, which were thought to derive from wood tar. The methyl esters of these diterpenoids are usually minor components in fresh resin and their high abundance in the samples analyzed indicated that the resin had been subjected to a heating process, although other markers of high heating such as retene (Evershed et al. 1985; Robinson et al. 1987; Connan and Nissenbaum 2003) were not detected. Samples taken from the thoracic cavity of a Third Intermediate Period mummy (c. 1064–656 BC; British Museum, EA74303) were examined using GC/MS (Serpico and White 1998). These were found to contain dehydroabietic and 7-oxodehydroabietic acid and retene, indicating a pine pitch. The absence of any degradation products from cis-abienol excluded the possibility of the resin originating from fir. Other samples from the body cavity and the skull were found to contain pistacia as revealed by the presence of oleanonic and moronic acids and their derivatives, and beeswax in addition to the pitch already identified. A sample of resin from a canopic jar was found to contain dehydroabietic acid and 7-oxodehydroabietic acid, indicating the presence of coniferous resin. Thirteen mummies dating from 1100 BC and AD 400 were analyzed by Connan (1999, 2002) using GC/MS. All the balms analyzed were found to contain coniferous resin and the majority were found to contain fat. Beeswax was identified in six balms and bitumen originating from the Dead Sea and Iraq in eight balms. The “resin” from another Ptolemaic Period mummy (Archaeological Museum, Kraków) was analyzed by Klys et al. (1999) using Fourier transform infrared (FTIR) spectroscopy. This “resin” was identified as copal based on the frequency of absorption and comparison with reference standards. Beads taken from the eyes of two animal mummies (Manchester Museum, 22948 and 22949), dating to c. 1800 BC, were analyzed using Fourier transform Raman spectroscopy (Edwards et al. 1999) and identified as

29

Trends in Use of Organic Balms in Egyptian. . .

657

being largely keratotic in composition (possibly being made from horn). GC/MS of the solvent extract of the bead identified a balsam coating. The use of these spectroscopic methods, particularly in identification of “resins,” must be viewed with caution as the results from previous analyses indicated that balms are generally composed of a mixture of materials which have undergone extensive chemical alteration over time. These changes and the presence of a mixture of materials are likely to lead to inaccurate conclusions. A Saite Period (c. 664–525 BC) mummy, Merneith (Institute of Egyptology, University of Pisa) was analyzed by an Italian team (Colombini et al. 2000). Triterpenoids including moronic and oleanonic acids were identified in the balm taken from the left side of the thorax, indicating the presence of pistacia resin. The identification of oxidized and dehydrogenated products including norolean-17-en-3one indicated that the resin had been strongly heated. Vegetable oil, identified by the presence of fatty acids, beeswax, identified by n-alkanes and wax esters, and bitumen identified by steranes and triterpanes, were also present in low concentrations in the balm. Four mummies found in the Dakhleh Oasis and dating to the Graeco-Roman Period were examined and found to be embalmed using coniferous resins, beeswax, and bitumen. The distributions of the steranes and triterpanes of bitumen from two balms were characteristic of Dead Sea bitumen, while the distributions from the other balms indicate another, unidentified, source (Maurer et al. 2002). Systematic analyses of provenanced and dated human mummies dating from the Old Kingdom to the Graeco-Roman Period (c. 2686 BC–AD 395) was carried out by Evershed and coworkers (Buckley 2002; Buckley and Evershed 2001; Buckley et al. 1999, 2004). Analyses were performed using a combination of chromatographic techniques: GC/MS, thermal desorption-GC/MS and pyrolysis-GC/MS. Identified in these samples were combinations of fats and oils, balsam, coniferous and pistacia resin, and beeswax. Unfortunately, the earliest and rarest mummy examined, that of the Old Kingdom male adult (Bristol Museum, H640), was contaminated by paraffin wax, probably as a result of a post-excavation treatment. The analysis of several animal mummies dating from the Twenty-Third to Thirtieth Dynasties (c. 897–342 BC; Buckley and Evershed 2001; Buckley et al. 2004) identified similar materials in the balm compared to the balms of human mummies, indicating that the animals included in the study were treated to the same standard as contemporary human mummies, finding supported by the recent work of Lucejko et al. (2017). The first use of thermal desorption and pyrolysis GC/MS techniques to study mummy balms (Buckley et al. 1999) demonstrated the advantages of these techniques over the standard solvent extraction methods are that very small amounts of sample can be investigated, which is of considerable value where sample sizes are limited and minimal sample preparation is required. However, the amount of information gained using this technique is often less than that obtained from the conventional solvent extraction techniques. Six mummies were included in a study by Tchapla et al. (2004). One of these was previously analyzed by Connan and Dessort (1989) and Mejanelle et al. (1997). The balms taken from this mummy (Ptolemaic male adult; Guimet Museum of Natural History, Lyon, 90001255) were analyzed using GC/MS, and found to contain fat, beeswax, coniferous resins, and vegetable tannins. Castor oil, identified based on the

658

R. P. Evershed and K. A. Clark

presence of ricinoleic acid, was found in approximately half the samples analyzed. Balms from a Middle Kingdom male child (c. 2066–1650 BC; Guimet Natural History Museum, Lyon, 90001626) and a Twenty-Fifth to Twenty-Sixth Dynasty male adult (c. 752–525 BC; San Lazaro Monastery, Venice) were found to contain only vegetable oil, whereas the balm from a Twenty-First Dynasty male adult (c. 1064–948 BC; Museum of Natural History, Perpignan) was found to contain fat and beeswax. The balm from an Eighteenth Dynasty female adult mummy (c. 1549– 1328 BC; Georges Labit Museum, Toulouse) was found to contain a gum resin, identified by the presence of mannose, galactose, and glucose. The embalmed viscera in a canopic jar (Guimet Museum of Natural History, Lyon, 90002013), reportedly belonging to Ramses II, were found to contain pistacia resin. Although this object was inscribed with the cartouche of Ramses II it is possible that the vessel was reused in antiquity, as it contains a heart and it is known that the heart of Ramses II was not removed, thereby placing doubt on the origin and importance of this identification. The embalming material from the Eighteenth-Dynasty tomb of Saankh-kare at Deir el-Bahari (Metropolitan Museum, New York, cemetery field no. 26225) was analyzed using GC/MS by Koller et al. (1998, 2005). This material was found to contain no evidence for di- and triterpenoid components. Sesquiterpenoids, napthalenes, azulenes, and phenolic compounds including cresols, xylenols, and guicols were identified, indicating a cedar wood tar oil origin. However, these compounds are not limited to cedar oil and could be from material derived from other coniferous species (Mills and White 1994). GC/MS was used to identify frankincense in a “resinous” sample from the tomb in Dahshour of Sat-mer-Hout, (Victor Loret Egyptologic Institute, Lyon, L41) based on the presence of α-boswellic acid, β-boswellic acid, and their acetate derivatives (Mathe et al. 2004). This is the first identification of frankincense in a Pharonic period funerary setting, although it was earlier identified in material from Qasr Ibrim, Nubia (c. AD 400–500; Evershed et al. 1997; van Bergen et al. 1997) using the same criteria. Among the most recent reports of the chemistry of mummy balms is that of Lucejko et al. (2017) who analyzed balms from ten mummies and storage jars from the Old Kingdom to Copto-Byzantine period. Their principle conclusion was that the most abundant constituents of the balms were fats, oils, waxes, conifer resin, pitch, mastic resin, castor oil, and bitumen. A purportedly new approach was presented the quantification of bitumen in mummy balms which serves to demonstrate the difficulties associated with using sterane and triterpene biomarkers as basis for quantification, primarily due to their variable concentrations in the source bitumens (Clark 2007). A recent study overcame this problem, finally resolving the question of the quantitative importance of bitumen in mummification in the largest reported investigation of Egyptian mummies. This was achieved by combining sterane and triterpene biomarker analyses with radiocarbon determinations (Clark et al. 2016). Analyses of 91 materials comprising balms, tissues, and textiles from 39 mummies dating from ca 3200 BC to AD 395. GC-MS SIM (m/z 217 and 191) showed no detectable bitumen use before the New Kingdom (ca 1550–1070 BC). However, bitumen was used in 50% of New Kingdom to Late Period mummies, rising to 87%

29

Trends in Use of Organic Balms in Egyptian. . .

659

of Ptolemaic/Roman Period mummies. Despite the widespread incidence of bitumen use in latter periods, quantitative determinations using a novel 14C analyses reveal that even at peak use balms were never more than 45% w/w bitumen, the major proportion being fats, oils, resins, beeswax, etc.

Placing Chemical Investigations of Mummy Balm in the Cultural Context of Ancient Egyptian Mummification The details of mummification obtained from the methodical study of the physical remains in conjunction with surviving images and texts are invaluable to the understanding mummification practices in ancient Egypt. While there was clearly an evolution in the practice of mummification over a period of four millennia, which can be observed by the physical changes to the process, it is unclear exactly what the major sources of influence that led to the refinement of embalming techniques might be. It is likely that the idea of preserving bodies to ensure passage into the Afterlife came from the observation of natural mummification of bodies in desert sands. However, it can be hypothesized that subsequent developments were stimulated by observations of the preservative properties of many natural products and techniques involved in food preservation and leather production. Beyond this, many other factors would have influenced the particular style of mummification of individual burials: that is, status and economy (including practicalities of scale), gender, religious influences and fashion, availability of ingredients (strongly influenced by trade routes and political stability), idiosyncratic variants of individual embalmers (including experimentation). The investigations of the embalming ingredients have so far been presented in a piecemeal fashion with single mummies or small numbers of mummies being considered in publications. Despite the many studies this approach has failed to lead to the natural ingredients used in mummification being systematically assessed terms of some of the critical cultural drivers, that is, the changes that occurred through ancient Egyptian history and variation by the age, sex, and status of the individual, the location on the body, or type of material the balm was applied to. In this chapter we attempt to bridge this gap in understanding. We present here extensive new chemical compositions on previously unstudied mummy balms. Combining them with previously published reports of mummy balm allows us to compare the themes and trends in the chemistry of balms from over 100 mummies to be compared. The categories chosen for discussing the mummy balms reflect outstanding questions in the field of mummification that cannot be answered other than by chemical means. The external physical treatment of a mummy, the treatment of the viscera, the way the body was wrapped, and the amulets included among the wrappings are all known to have undergone significant changes over ancient Egyptian history (Ikram and Dodson 1998). The quality of preservation of the body also appears to change, in some periods mummies are generally better preserved than at other periods. During the New Kingdom to the end of the Twenty-First Dynasty

660

R. P. Evershed and K. A. Clark

(c. 1549–948 BC) the treatment of the body in terms of the bandaging applied and the quality of the remains is thought to be at its best and this period is known as the height of mummification or the classic phase of mummification (Quirke 1992; Ikram and Dodson 1998). During this period the embalmers made the body look as lifelike as possible, whereas previously the body was prepared to look like an idealized person. Adult and child mummies are clearly distinguishable because of the difference in their size. In ancient Egypt, children are depicted as miniature adults with the attributes of childhood such as nakedness and a single sidelock; they became involved in the adult world, however, at a relatively young age. Child mortality in ancient Egypt was relatively high; approximately one-third of children did not reach their first birthday and almost half died before their fifth birthday (Janssen and Janssen 1990). There is evidence to suggest that if child mortality was relatively high in a society, strong attachments would not be formed and therefore children would not be considered as complete members of society (Aries 1962). This would indicate that in death, children, and adults were not treated in the same manner. However, there is only limited evidence of physical differences in the external treatment (bandaging and treatment of the viscera) between adults and children. Recent excavations at the Deir el-Medina cemetery, for the skilled workers and artisans connected with building and decorating the tombs in the Valley of the Kings during the Eighteenth to Twentieth Dynasties (c. 1549–1064 BC), there is evidence that the death of a child did not go unnoticed and that children were provided for in the Afterlife with food and pottery vessels (Meskell 1999b). The children from all socioeconomic levels were treated minimally, compared with contemporary adults, in terms of the embalming treatment and wrappings until the later Graeco-Roman Period when child mummies are as intricately wrapped and elaborately treated as the adults (Meskell 1999a). In ancient Egyptian society, women had the same legal and economic rights as men, of the same social class (Capel and Markoe 1996). Again, there is limited physical evidence that the bodies of males and females were treated differently in death, although the arms were often crossed in different ways for males and females (Gray 1972). Evidence from Deir el-Medina indicates that, in death, the genders were not considered to be equal. The majority of grave goods in a family tomb belonged to the deceased male and this distinction between men and women increased with higher status and wealth. This disparity suggests that a woman’s destiny in the afterlife was dependent on that of her spouse (Meskell 1999b). Variations in the treatment of males and females can be seen in the mummies of wealthy individuals and their wives at Deir el-Medina; the male mummies of Kha and Sennefer dated to the Eighteenth Dynasty (c. 1549–1328 BC) were well wrapped; however, their accompanying females Merit and Nefertiry were poorly wrapped and therefore found in a worse condition. Differentiation between genders in the lower social classes was less obvious (Meskell 1998, 1999a). Different parts of the body had different significance and importance to the ancient Egyptians. For example the Rhind Bilingual Papyri (British Museum,

29

Trends in Use of Organic Balms in Egyptian. . .

661

10188; Birch 1863) indicates that the number seven was important; sacred oils were to be applied to the seven openings of the head (eyes, ears, nostrils, and mouth). This idea is continued to the 17 members of the god (i.e., the corpse): “seven openings of the head, four sons of Horus (internal organs), two legs, two arms, one front torso and one back giving a total of seventeen.” The Cairo and Louvre papyri entitled “The Ritual of Embalming” (Sauneron 1952) specify that the head was to be anointed with frankincense and packed with aromatic spices and the rest of the body anointed with an unguent. Then, the head was wrapped in linen and sealed with “thick oil” or resin. Further instructions are given for the hands and legs. If these parts of the body had a special significance for the ritual of embalming, then maybe the balms applied to these body parts were themselves different, to reflect the ritual aspects of embalming. It is thought that wax was used in sealing the eyes, nose, and evisceration wound (Adams 1988). Finally, the types of sample, tissues, bandages, and “resins” removed from mummies may also have undergone different treatments. These differences may be related to the variations in treatment of the different parts of the body. In order to explore the above phenomena in greater depth we determined the chemical compositions of balms from a large number of Egyptian mummies encompassing the entire period of the development and practice of mummification. Over one hundred samples of balms were studied from mummies from museum collections mainly in the United Kingdom and Europe. The full list of samples is detailed in Table 1.

Materials and Methods Mummy Samples All archaeological samples were generously provided by: Prof. Rosalie David and Dr Trish Lambert-Zazulak of Manchester University Museum; Sue Giles of Bristol Museum; Julia Gresson of Auckland War Memorial Museum, New Zealand; Dr John Taylor of The British Museum, London; Dr Maarten Raven of the Rijksmuseum van Oudheden, Leiden, The Netherlands; Faye Kefalonis, Norwich Castle Museum; Dr William van Haarlem, Allard Pierson Museum, Amsterdam; Prof Emma Rabino-Massa, Museum of Archaeology and Ethnography, Turin, Italy; Vicky Turner of Durham Oriental Museum; Drs Salima Ikram, Muhammed Saleh and Nasry Iskander from the Cairo Museum. Descriptions of the samples are given in Table 1, which details the type of sample taken from each mummy, categorized by location on the body and listed in chronological order. Where possible samples were taken from a number of locations on the body and included different material types (i.e., bandages, tissues, and “resins”). Mummies were chosen so that the full history of the practice of mummification was represented. Male, female, and child mummies were sampled in order to investigate age and gender differences.

Female adult

Adolescent

230 BM 32752

232 BM 32753

173

170 171 172 TUR Drawer 520 Female adult

169 TUR Drawer 528 Adult

Mummy Male adult

Museum and No. number 231 BM 57353

Predynastic Period c. 3200 BC

Predynastic Period c. 3200 BC

Date Predynastic Period c. 5000– 3000 BC Predynastic Period c. 4000– 3000 BC Predynastic Period c. 4000– 3000 BC

S

S

S

S

Gebelein

Gebelein

Gebelein

Gebelein

Davide (1972)

Davide (1972)

Dawson and Gray (1968)

Dawson and Gray (1968)

Location Dating method Provenance Reference Head S Gebelein Dawson and Gray (1968) Tissue from lower back

Torso

Table 1 Collection of balms assembled for this study together with details of locations for the samples analyzed herein

Tissue from sole of right foot

Tissue/ bandage from heal of right foot Tissue, knee end, tibia (black)

Limbs Tissue/ bandage from thigh

Tissue light Light bone Bandage from piece with fur

Other

662 R. P. Evershed and K. A. Clark

115

114

113

112 TUR

180

Predynastic Period c. 3200 BC Predynastic Period c. 3200 BC

Predynastic Period c. 3200 BC

Female adolescent Old with dress Kingdom 2410–2195 BC

179 TUR Drawer 535 Adult

178 TUR Drawer 517 Female adult

175

174 TUR Drawer 522 Adult

R

S

S

S

n.d.

Gebelein

Gebelein

Gebelein

Davide (1972)

Davide (1972)

Davide (1972)

Davide (1972)

Tissue from right temporal area

Tissue from left frontal -parietal area

Tissue from skull

Tissue from inner side right leg

Tissue from right leg

Bandaging from top of right hand Tissue from palm

Bandaging from lower leg Tissue from lower leg

(continued)

29 Trends in Use of Organic Balms in Egyptian. . . 663

Male adult skull, Meryrehashetef

Male adult, Heny

233 BM 23425

Fifth Dynasty c. 2355– 2195 BC Middle Kingdom c. 2066– 1650 BC

Mummy Date Female adolescent Old with dress Kingdom 2410–2195 BC

229 BM 55725

120

119

118

117

Museum and No. number 116 TUR (cont)

Table 1 (continued)

S

S

Asyut

Fayum

Dawson and Gray (1968)

Dawson and Gray (1968)

Tissue from orbit of left eye, near nose

Location Dating method Provenance Reference Head R n.d. Davide (1972)

Bandages on torso

Torso

Tissue from right forearm

Limbs Tissue from inner side right forearm

Tissue

Dust & fiber fragments from left leg Dust from upper part of torso & below coffin

Other

664 R. P. Evershed and K. A. Clark

MAN 21471

Male adult, Khnumnakht

78

77

76

75

NMS 1909.527

Female adult

143 47 NMS 1909.527.2 Alabaster jar

73 74

S

Thirteenth to R Seventeenth Dynasties 1650 BC Thirteenth to R Seventeenth Dynasties 1650 BC

Twelfth Dynasty c. 1994– 1781 BC

Qurna

Qurna

Rifeh

Petrie (1909)

Petrie (1909)

Murray (1910), David (1979)

(continued)

“Resinous” material from bottom left of coffin “Resin” Impregnated tissue from debris “Polymerized” fat on front and middle Fragment from debris in newspaper

Muscle tissue “Resin”/body tissue? Bandage/tissue Red/orange “resin” contents

29 Trends in Use of Organic Balms in Egyptian. . . 665

85

84

152 153

79

154

150 151

LIV 1976.159.267

NMS 1909.527

Museum and No. number 149 NMS 1909.527

Table 1 (continued)

Head

Child

Mummy Female adult

New Kingdom c. 1549– 1064 BC

S

Thirteenth to R Seventeenth Dynasties 1650 BC

Thebes

Qurna

Gray and Slow (1968)

Petrie (1909)

Bandaging from head Skin/“resin” back/top of left side of head

Location Dating Date method Provenance Reference Head Thirteenth to R Qurna Petrie Seventeenth (1909) Dynasties 1650 BC Torso

Limbs

Other Textile/fatty material Textile/tissue Stained bandaging Stained bandage from cloth doubled under body “Resin”? On inside of coffin bottom Bone/cartilage Stained bandaging

666 R. P. Evershed and K. A. Clark

CAI CG5109

64

63

BRI H5074

188 RMO 33

225 BM 51812 226

224 BM 48001

65

216 RMO 54

S

S

C

S

Twentieth to S Twenty-First Dynasties c. 1200– 1000 BC Male adult, Twentieth to S Djedkhonsiufankh Twenty-Fifth Dynasties c.1186–656 BC

Head of Khonsuhotep

New Kingdom c. 1549– 1064 BC Beef ribs meat Eighteenth mummy from Dynasty tomb of Yuya and c. 1386– Tjuiu 1349 BC Female adult, Nineteenth Henutmehyt Dynasty c. 1250 BC Meat mummy Nineteenth Dynasty c. 1250 BC

Hand

n.d.

Thebes

Thebes

Thebes

Thebes

n.d.

Dawson et al. (2002)

Raven and Toconis (2005)

Taylor (1989, 1999) Taylor (1999)

Raven and Toconis (2005) Quibell (1908)

Tissue/ “resin”/ bandage

Black tissue from left hand side of chest Black bandage from feet

Blackened bandaging from palm

(continued)

Black “resin” from rear of inner coffin Skin from duck Tissue from goat? Leg

Black/brown stained bandaging

29 Trends in Use of Organic Balms in Egyptian. . . 667

MTB G6

37a MTB G44

36

148

147

146

145

Museum and No. number 144 BRI Ha7386

Table 1 (continued)

Male adult, (Glasgow)

Mummy Male adult, Horemkenesi

Third Intermediate Period c. 1064–656 BC S

n.d.

Dating Date method Provenance Twenty-First S Deir el Dynasty Bahri c. 1064–948 BC

n.a.

Location Reference Head Dawson et al. (2002)

Black bandage package-

Torso “Resinous material” from left hand side of spine “Resinous material” from left hip/spine Head of right femur muscle tissue Bandage/ tissue from right calf Bandage from left ankle Black bandage back left hand

Limbs

Other

668 R. P. Evershed and K. A. Clark

Calf victual mummy

Head of a female adult

Male adult

192 RMO 38

234 BM 6660

MTB G32

39

185 CAI CG29852

MTB G20

38

37b MTB G44

Twenty-First S Dynasty c. 1064–948 BC Third S Intermediate Period c. 1064–656 BC Twenty-First S Dynasty c. 1064–948 BC n.d.

n.d.

n.d.

Dawson and Gray (1966)

Guilard and Daressy (1905) Raven and Toconis (2005) Black tissue from left hand side of jaw bone

blackened “resin” Black bandage packagebandage Black material front abdomen Black bandage & tissue right upper arm

(continued)

Blackened “resin” from stomach area

Bandages

29 Trends in Use of Organic Balms in Egyptian. . . 669

53

52

62

61

50

49

48

BRI Ha7563

BRI H6140

NZ

Museum and No. number 33 MTB 5681

Table 1 (continued)

Child (BRI)

Male Child

Female adult

Mummy Cornell mummy (Penpi)

Late or Ptolemaic Periods c. 727–30 BC S

Twenty-Fifth S Dynasty c. 743–656 BC

n.d.

n.d.

Dawson et al. (2002)

Dawson et al. (2002)

Location Dating Date method Provenance Reference Head Twentieth to S Thebes n.a. TwentyThird Dynasty c. 897–715 BC Third R n.d. n.a. Embalming Intermediate resin from -Saite head Periods c. 850–575 BC

Tissue from right shoulder

Torso

Bandage from left knee Tissue from right ankle Bandaging from left hip

Limbs

Coating on base interior coffin Flake from base exterior coffin

Other “Resin”

670 R. P. Evershed and K. A. Clark

MTB 528/1

Male adult, Pediamun Ipuwer

Adult, Asttayefnakht

LIV 1953.72

MTB 400

13

Female adult

Male adult, Besenmut

86

110 111

109 NOR

27

26

25

23 24

22

TwentySixth Dynasty c. 664–525 BC TwentySixth to TwentySeventh Dynasties c. 664–404 BC TwentySixth Dynasty 650 BC S

S

S

Twenty-Fifth S Dynasty 700 BC

n.d.

Thebes

Saqqara

Akhmim

n.a.

Gray and Slow (1968)

Dawson (1929)

n.a.

“Resin” from inside of cartonage at back of head

Blackened bone/“resin”

Tissue from right foot

Tissue/ bandaging from left scapula region

Trends in Use of Organic Balms in Egyptian. . . (continued)

Skin with nineteenthcentury varnish

Darkened bandages 1 Bandages 2 Bandages 3

External debris bandage, tissue Red/orange “resin”

Bandaging

29 671

AP 10.842

15

14

209

208

Date TwentySixth Dynasty 650 BC

Ptolemaic Period c. 332–30 BC

Late Period c. 525–332 BC Head and feet of a Late Period female adult c. 525–332 BC

Female head

MTB 4158/3347 Female mummy (Greek)

207 RMO 48

93

32

30 31

29

Museum and No. number Mummy 28 MTB Female adult, 528/SLA50.1928 Panesittawy

Table 1 (continued)

S

S

S

n.d.

Thebes

n.d.

n.a.

Raven and Toconis (2005)

Koens (1998)

Black tissue/ bandage Black “resin” Black “resin” Bandaging from foot

Location Dating method Provenance Reference Head S n.d. n.a.

3rd core past left shoulder 1st core mid post thorax

Torso 2nd core above mid post thorax Package right thorax

Tissue near hip bone

Limbs

Tissue & bandage

Bandage

Other

672 R. P. Evershed and K. A. Clark

BRI H7212

BRI H5543

NMS 1956.352

58

59

80

81

BRI Ha7385

54

Ptolemaic Period c. 332–30 BC

Young male adult Ptolemaic Period c. 332–30 BC Female adult right Ptolemaic foot Period c. 332–30 BC Right foot PtolemaicGraecoRoman Periods c. 332 BC– AD 395 Female adult Ptolemaic Period c. 332–30 BC

MAN 7700/5275 Head

42

S

S

S

S

S

Thebes

Thebes

Thebes

n.d.

n.d.

Sheridan (2000)

Dawson et al. (2002)

Dawson et al. (2002)

Dawson et al. (2002)

David (1979)

“Resinous” material from amulet on neck Stained bandaging from right hand side of neck

Bandage/ tissue under left hand side of jaw bone Black “resin” coated outer bandages

Black bandaging from ankle

Black tissue from ankle

(continued)

29 Trends in Use of Organic Balms in Egyptian. . . 673

Male adult, Djehor

Adult

228 BM 29782

Female adult

Mummy Fur from a votive mummy (dog)

227 BM 29776

187

186 RMO 13

Museum and No. number 184 CAI CG29760

Table 1 (continued)

Ptolemaic Period c. 332–30 BC

Ptolemaic Period c. 332–30 BC

Date Ptolemaic Period c. 332–30 BC Ptolemaic Period c. 332–30 BC

S

Akhmim

Dawson and Gray (1968)

Location Dating method Provenance Reference Head S Saqqara Guilard and Daressy (1905) S Thebes Raven and Toconis (2005) Tissue from left hand side of top of skull S Akhmim Dawson and Gray (1968) Black “resin” coated bandages from left shoulder Black “resin” coated bandages from left hand side of neck

Bandaging from right hand side of upper torso

Torso

Limbs

Other Fur

674 R. P. Evershed and K. A. Clark

Male adult with folded arms

121 TUR Pravv 540

83

182 82 NMS 1911.210.3 Female child

181

123

122

Male adult with prosthetic hand

124 DUR 1999.32.1 S

GraecoRoman Period c. 30 BC– AD 395 S

PtolemaicR GraecoRoman Periods 100 BC–AD 395

PtolemaicGraecoRoman Periods c. 332 BC– AD 395

n.d.

Asyut

Luxor

Sheridan (2000)

Davide (1972)

Birch (1880), Gray (1966)

Darkened bandaging under right breast Darkened bandaging under right shoulder

Blackened “resin” on stomach

Black “resin” coated outer bandages from right hand side of upper arm Bandages from tip left foot Stained bandages from leg Stained bandages from sole left foot

(continued)

Pale bandaging

29 Trends in Use of Organic Balms in Egyptian. . . 675

Mummy Male child

Child

Adult

Head of a female child

Head of a female adult

Museum and No. number 125 DUR 1985.61

133 DUR 1999.52

158 UP 4

189 RMO 34

190 RMO 35

Table 1 (continued)

Date GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395 S

S

S

S

Saqqara

n.d.

n.d.

n.d.

Raven and Toconis (2005)

Raven and Toconis (2005)

n.a.

Birch (1880)

Bone from left hand side of jaw bone

Black tissue inside neck and hair

Blackened bandaging inside neck

Location Dating method Provenance Reference Head S Luxor Birch (1880)

Interior of mummy

Torso Stained bandages from left hand side

Limbs

Other

676 R. P. Evershed and K. A. Clark

Head of a female adult

195 RMO 41

18

206

205

GraecoRoman Period c. 30 BC– AD 395

Mediaeval

MTB 5599/S212 Nubian natural

204 RMO 47

GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395

GraecoRoman Period c. 30 BC– AD 395 GraecoRoman Period c. 30 BC– AD 395

Head of a male adult

Head of a female adult

200 RMO 44

201

Head of a male adult

199 RMO 43

197

Head of a male adult

193 RMO 39

C

S

S

S

S

S

Nubia

n.d.

n.d.

n.d.

Thebes

n.d.

n.a.

Raven and Toconis (2005)

Raven and Toconis (2005)

Raven and Toconis (2005)

Raven and Toconis (2005)

Raven and Toconis (2005)

Black tissue/ “resin” Black tissue from neck Blackened tissue Bandaging base of neck Modern wax mount

Black tissue/ “resin” Black “resin” on hair Black tissue/ “resin” and bandaging

Black tissue/ “resin”

Skin from upper back (continued)

29 Trends in Use of Organic Balms in Egyptian. . . 677

16

5b

5a

4

2

142

141

139 140

No. 19 20 137 138

n.d.

n.d.

n.d.

–

n.d.

–

n.d.

–

n.d.

Amsety canopic jar Hapi canopic jar

n.d.

–

n.d.

Nubian natural

n.d.

–

n.d.

Nubian natural

n.d. n.d.

– –

n.d. n.d.

Nubian natural Nubian natural

Provenance Nubia Nubia n.d. n.d.

Dating method C C – –

Date Mediaeval Mediaeval n.d. n.d.

Mummy Nubian natural Nubian natural Nubian natural Nubian natural

MTB 7700/9430 Canopic jar

Museum and number MTB 5599/S217 MTB 5599/S81 UWO NAT637-5 UWO 24I3-B165 UWO NAT657-5 UWO 24I3-B175 UWO 24I3-B135 UWO 24I3-B405 MAN 7700/ 11103 MAN 7700/4963

Table 1 (continued)

n.a.

n.a.

n.a.

n.a.

n.a.

n.a. n.a.

Location Reference Head n.a. n.a. n.a. n.a. Torso

Limbs

Black resin from sides Black “resin” from base of lid Linen and lump from jar-“resin” Linen and lump from jar-bandage Blackened textile with tissue/“resin”

Skin

Skin

Skin Skin

Other Skin Skin Skin Skin

678 R. P. Evershed and K. A. Clark

Left Foot

Right hand

Female left hand

MAN 7700/ALI

BRI H537

BRI Ha5546

46

55

56

Hand & arm

n.d.

n.d.

n.d. Thebes

Memphis

–

–

n.d.

44b 45 MAN 7700/ 1977.1161 n.d.

n.d.

–

n.d.

44a MAN 7700/7740 Head

–

n.d.

–

n.d.

n.d.

n.d.

–

n.d.

–

n.d.

–

n.d.

n.d.

40a MAN 7700 / Head 2145 (11729) 40b 41 MAN 7700/ Head 22940 43 MAN 7700/SAL Head (Salford)

Eton canopic jar

n.d.

MTB 1363/ ECM1564a

–

35

Dawson et al. (2002)

David (1979) Dawson et al. (2002)

David (1979)

David (1979)

David (1979) David (1979)

David (1979)

n.a.

Black “Resin” Bandage “Resinous” lumps Black tissue from left hand side base chin & inside skull Clear “resin” Bandage Tissue from right hand Tissue from heal Black tissue/ Bandage from finger Black bandage from finger (continued)

Tissue, bandaging/ “resin”

29 Trends in Use of Organic Balms in Egyptian. . . 679

Miscellaneous bandaging

AP

90

Child head

AP 13.009

94

Male head

Male head

AP 13.010

AP 13.011

96

97

95

Head

AP 10.841

92

91

Guilt left foot

BRI Ha5459

Mummy Hand

60

Museum and No. number 57 BRI Ha5545m

Table 1 (continued)

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

Date n.d.

n.d.

n.d.

–

n.d.

–

–

n.d.

n.a

–

–

n.d.

–

Koens (1998)

Koens (1998)

Koens (1998)

Koens (1998)

Dawson et al. (2002) Koens (1998)

Black tissue/ bandage Black tissue from outside head Black tissue from under jaw Black bandage behind ear Black tissue from back/ side head

Location Dating method Provenance Reference Head – n.d. Dawson et al. (2002) Torso

Limbs Black tissue underside wrist Brown bandaging from sole

Dark bandaging Light bandaging

Other

680 R. P. Evershed and K. A. Clark

Left foot

Left foot

Canopic jar

Adult Adult

AP 8.418b

99

100 AP 8.418a

155 UP 1

157 UP 3 159 TUR 14406 (033.064)

164 TUR 2

Adult

Adult

162 TUR 1

163

Adult

161 TUR 14.389

160

Hand

AP 8.418b

98

n.d.

n.d.

n.d.

n.d. n.d.

n.d.

n.d.

n.d.

n.d.

n.d. n.d.

– –

–

n.d.

n.d.

n.d.

–

–

n.d.

–

n.d.

n.d.

–

–

n.d.

–

Davide (1972)

Davide (1972) Davide (1972)

n.a. Davide (1972)

n.a.

Koens (1998)

Koens (1998)

Koens (1998)

Bandage behind knee Tissue from right knee

Bandage on left thigh Tissue from left upper arm

Black tissue top side of wrist Black tissue from ankle Black tissue underside heal

Trends in Use of Organic Balms in Egyptian. . . (continued)

Blackened bandaging

Stained outer bandaging

“Resinous” Contents Bandage

29 681

191 RMO 37

183 CAI 15+4/24+1

Cat Shaped sarcophagus Head of a female adult

Adult

176 TUR 3 (drawer)

177

Adult

Mummy Adult

167 TUR Pravv 545/14428 168

166

Museum and No. number 165 TUR Pravv 569

Table 1 (continued)

n.d.

n.d.

n.d.

n.d.

Date n.d.

n.d. n.d.

–

n.d.

–

–

n.d.

–

Raven and Toconis (2005)

n.a.

Davide (1972)

Davide (1972)

Blackened bandaging top of head

Tissue, top of head, under bandaging

Location Dating method Provenance Reference Head – n.d. Davide (1972) Torso

Bandaging from near big toe Tissue from near big toe

Limbs

“Resinous” lump

Other Bandaging, pile of bandages on top in box Bandaging underneath attached to mummy Bandaging thorax

682 R. P. Evershed and K. A. Clark

Head of a female adult

Head of a female adult

Head of a male adult

Left hand of an adult

Left hand of a female adult

Hand of an adult

198 RMO 42

202 RMO 45

203 RMO 46

210 RMO 49

211 RMO 50

212 RMO 51

213

Head of a male adult

194 RMO 40

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

n.d.

–

–

–

–

–

–

–

Raven and Toconis (2005) Raven and Toconis (2005) Raven and Toconis (2005) Raven and Toconis (2005) Raven and Toconis (2005) Raven and Toconis (2005) Raven and Toconis (2005) Hair and tissue/ “resin”/ bandaging Blackened tissue from neck

“Resin” coated bandaging from neck Black “resin”/ bandage

Black bandaging from thumb Scrapping of black “resin”

Black tissue from wrist

Tissue from wrist

(continued)

29 Trends in Use of Organic Balms in Egyptian. . . 683

Hand of a child

Head

Adult

215 RMO 53

217 RMO F2004/ 12.2

218 RMO Grey 7

n.d.

n.d.

n.d.

Date n.d.

Dating method Provenance Reference – n.d. Raven and Toconis (2005) – n.d. Raven and Toconis (2005) – n.d. Raven and Toconis (2005) – n.d. n.a.

Key: n.d. ¼ not determined; n.a. ¼ not available; C ¼ context; R ¼ radiocarbon; S ¼ style

219

Mummy Hand of an adult

Museum and No. number 214 RMO 52

Table 1 (continued)

Tissue from neck, bandaging fragments

Location Head

Bandaging from upper torso

Torso

Bandaging from sole of right foot

Tissue from wrist

Limbs Tissue from wrist

Other

684 R. P. Evershed and K. A. Clark

29

Trends in Use of Organic Balms in Egyptian. . .

685

Lipid Extraction and Analysis The analytical methods used have been extensively described in previous publications (Evershed et al. 2002; Buckley and Evershed 2001; Buckley et al. 1999, 2004; Clark et al. 2013), with the overall protocol employed summarized in Fig. 1. The approach achieved identification of the major organic solvent-soluble components of the balms, including fats, oils, resins, and beeswax, by means of gas chromatography (GC), GC/mass spectrometry (GC/MS), and GC-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Bitumen in mummification was determined by means of GC/MS with selected ion monitoring (SIM) and radiocarbon analysis as described by Clark et al. (2016).

Results Summary of Balm Compositions Analyzed in This Study A summary of the composition of the new balms analyzed is given in Table 2. A full description of the analyses leading to these compositions is given in Clark (2007). Table 2 gives details of the proportion of lipids from each ingredient present in the balm (red indicates portion of fat/oil in balm; blue, beeswax; green, coniferous resin; orange, pistacia resin; black, bitumen; gray, no bitumen). The bitumen present is not included in the total composition of the balm because of the difficulties in determining this accurately, as described in Clark et al. (2016); however, an indication of the quantity of bitumen in the balm is given in proportion to the biomarker concentrations. Analysis of the compositions of balms reveals a number of interesting features. Firstly, in balms that contained two or more ingredients, the percentage composition of those ingredients in the balm varied between each of the different balms analyzed; in most cases the major ingredient of the balm is fat/oil, although there are also a number of examples of beeswax or resin being the major ingredient. For example, the blackened bandaging from a female hand (BRI Ha5546; Fig. 2), contained 95% coniferous resin and the bandaging from the torso of the female adult (c. 332–30 BC; RMO 13; Fig. 3), consisted of 66% beeswax. However, in the majority of balms, the major ingredient was fat/oil, which is unsurprising given that it was widely available and relatively cheap. Beeswax was identified in 37% of all mummy balms, of these, 1–69% of the balm was composed of beeswax, although higher concentrations were observed in coffin coatings; the majority of these balms contained between 5% and 50% beeswax. Resins were identified as a component of 25% of the mummy balms, in the majority of cases accounting for less than 25% of the balm, the other 75% comprising of fat/ oil or a mixture of fat/oil and beeswax (except the resin portion was higher in bandaging from a female hand (BRI Ha5546) described above, 95%, bandaging from the beef ribs meat mummy (CAI CG5109), 46%, and a bandaging from a foot, (BRI H5543), 31%). The lower incidence of resin in balms suggests that resin was a

Fig. 1 A schematic of the adopted analytical protocol

686 R. P. Evershed and K. A. Clark

29

Trends in Use of Organic Balms in Egyptian. . .

687

Table 2 Summary of percentage compositions of major balm ingredients of mummy balms analyzed herein Mummy

Museum number

Date

Location

Male adult Female adult Adolescent Adult

BM 57353 BM 32752 BM 32753 TUR Drawer 528

c. 5000-3000 BC c. 4000-3000 BC c. 4000-3000 BC c. 3200 BC

Female adult

TUR Drawer 520

c. 3200 BC

Adult

TUR Drawer 522

c. 3200 BC

Female adult Adult

TUR Drawer 517 TUR Drawer 535

c. 3200 BC c. 3200 BC

Female adolescent with dress

TUR

2410-2195 BC

Male adult skull, Meryrehashetef Male adult, Heny Male adult, Khnumnakht

BM 55725

c. 2200 BC

Tissue/bandage from thigh Tissue from lower back Tissue/ bandage from heal of right foot Tissue, knee end, tibia (black) Tissue light Light bone Bandage from piece with fur Tissue from sole of right foot Bandaging from lower leg Tissue from lower leg Tissue from skull Bandaging from top of right hand Tissue from palm Tissue from left frontal -parietal area Tissue from right leg Tissue from right temporal area Tissue from inner side right leg Tissue from inner side right forearm Bandages on torso Tissue from right forearm Dust & fibre fragments from left leg Dust from upper part of torso & below coffin Tissue from orbit of left eye, near nose

BM 23425 MAN 21471

c. 2066-1650 BC c. 1994-1781 BC

NMS 1909.527.2 NMS 1909.527

1650 BC 1650 BC

Child

NMS 1909.527

1650 BC

Head

LIV 1976.159.267 RMO 54 CAI CG5109

c. 1549-1064 BC c. 1386-1349 BC

Tissue Muscle tissue ‘Resin’/body tissue? Bandage/tissue Red/orange ‘resin’ contents ‘Resinous’ material from bottom left of coffin ‘Resin’ Impregnated tissue from debris ‘Polymerised’ fat on front and middle Fragment from debris in newspaper Textile/fatty material Textile/tissue Stained bandaging Stained bandage from cloth doubled under body ‘Resin’? On inside of coffin bottom of one end Bone/ cartilage Stained bandaging Bandaging from head Skin/‘resin’ back/top of head to left Blackened bandaging from palm Black/brown stained bandaging

BM 48001

c. 1250 BC

Black ‘resin’ from rear of inner coffin

BM 51812

c. 1250 BC

RMO 33 BRI H5074

c. 1200-1000 BC c. 1186-656 BC

BRI Ha7386

c. 1064-948 BC

MTB G6 MTB G44 MTB G44 MTB G20 MTB G32 CAI CG29852 RMO 38 BM 6660 MTB 5681

c. 1064-656 BC

Skin from duck Tissue from goat? Leg Tissue/ ‘resin’/ bandage Black tissue from left hand side of chest Black bandage from feet ‘Resinous material’ from left hand side of spine ‘Resinous material’ from left hip/spine Head of right femur muscle tissue Bandage/tissue from right calf Bandage from left ankle Black bandage back left hand Black bandage package- blackened ‘resin’ Black bandage package- bandage Black material front abdomen Black bandage & tissue right upper arm Bandages Black tissue from left hand side of jaw bone Blackened ‘resin ‘from stomach area ‘Resin’

NZ

850-575 BC

Alabaster jar Female adult

1650 BC

Hand Beef ribs meat mummy Female adult, Henutmehyt Meat mummy Head of Khonsuhotep Male adult, Djedkhonsuiafnkh Male adult, Horemkenesi

Male adult, (Glasgow)

Calf victual mummy Head of a female adult Male adult Cornell mummy (Penpi) Female adult

Male Child

BRI H6140

c. 1549-1064BC

c. 1064-948 BC c. 1064-656 BC c. 1064-948 BC c. 897-715 BC

c. 743-656 BC

Child (BRI)

BRI Ha7563

c. 727-30 BC

Male adult, Besenmut

MTB 528/1

c. 700 BC

Female adult

NOR

c. 664-525 BC

Male adult, Pediamun Ipuwer

LIV 1953.72

c. 664-404 BC

Embalming resin from head Coating on base interior coffin Flake from base exterior coffin Bandage from left knee Tissue from right ankle Bandaging from left hip Tissue from right shoulder Tissue/ bandaging from left scapula region Bandaging Tissue from right foot External debris bandage, tissue Red/orange ‘resin’ Blackened/Burnt? Vertebrae Hot ‘resin’? Darkened bandages 1 Bandages 2 Bandages 3 ‘Resin’ from inside of cartonage at back of head

Bitumen present

% composition# 100 100 No extractable lipid 100 No extractable lipid 100 100 100 No extractable lipid 100 No extractable lipid No extractable lipid 100 100 100 100 100 100 100 100 100 100 Paraffin wax No extractable lipid 100 100 100 100 100 95 5 100 100 100 100 100 No extractable lipid No extractable lipid No extractable lipid No extractable lipid No extractable lipid No extractable lipid 100 6 46

48

100

21

No extractable lipid 100 100 96 4 No extractable lipid 100 100 100 100 100 23 2 29 2 16 15 30 5 27 5 No extractable lipid 100 98 2 100

75 69 69 65 69

96 7

n.q. (0.6) 8

4 93

22

78

n.q.

83

17 100

81

19

n.q.

100 100 100 100 100 89

n.q. 3

100 100 100 99 58

8

33 22

1 42

n.q.

688

R. P. Evershed and K. A. Clark

Table 2 (continued) Mummy

Museum number

Date

Location

% composition#

Adult, Asttayefnakht Female adult, Panesittawy

MTB 400 MTB 528/ SLA50.1928

c. 650 BC c. 650 BC

100 100 100

Female head Head and feet of a female adult

AP 10.842 RMO 48

c. 525-332 BC c. 525-332 BC

MTB 4158/3347

c. 332-30 BC

MAN 7700/5275 BRI Ha7385 BRI H7212 BRI H5543

c. 332-30 BC c. 332-30 BC c. 332-30 BC c. 332 BC395 AD c. 332-30 BC

Skin with 19th C varnish 2nd core above mid post thorax Package right thorax Bandage Black tissue/ bandage Black ‘resin’ Black ‘resin’ Bandaging from foot Tissue & bandage Tissue near hip bone Bandage/tissue under left hand side of jaw bone Black ‘resin’ coated outer bandages Black tissue from ankle Black bandaging from ankle

Female mummy (Greek) Head Young male adult Female adult right foot Right foot Female adult

NMS 1956.352

Male adult with Prosthetic hand Fur from a votive mummy (dog) Female adult

DUR 1999.32.1 CAI CG29760

c. 332 BC395 AD c. 332-30 BC

RMO 13

c. 332-30 BC

Male adult, Djehor Adult

BM 29776 BM 29782

c. 332-30 BC c. 332-30 BC

Male adult with folded arms

TUR Pravv 540

100 BC-395 AD

Female child

NMS 1911.210.3

c. 30 BC-395 BC

Male child

DUR 1985.61

Child

DUR 1999.52

Adult

UP 4

Head of a female child

RMO 34

Head of a female adult

RMO 35

Head of a male adult

RMO 39

Head of a female adult

RMO 41

c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD c. 30 BC395 AD n.d. n.d.

Head of a male adult

RMO 43

Head of a female adult

RMO 44

Head of a male adult

RMO 47

Amsety canopic jar Hapi canopic jar

MAN 7700/11103 MAN 7700/4963

Canopic jar Eton canopic jar

Head Head (Salford)

MTB 7700/9430 MTB 1363/ ECM1564a MAN 7700/2145 (11729) MAN 7700/22940 MAN 7700/SAL

Head

MAN 7700/7740

n.d.

Hand & arm

MAN 7700/ 1977.1161 MAN 7700/ALI BRI H537 BRI Ha5546 BRI Ha5545m BRI Ha5459 AP

n.d. n.d. n.d. n.d. n.d. n.d. n.d.

AP 10.841 AP 13.009

n.d. n.d.

AP 13.010 AP 13.011 AP 8.418b AP 8.418b

n.d. n.d. n.d. n.d.

Head

Left Foot Right hand Female left hand Hand Guilt left foot Miscellaneous bandaging Head Child head Male head Male head Hand Left foot

n.d. n.d. n.d. n.d. n.d.

n.q.

78

22

100 98 100 100 94 96

4

2

6

88

7

52

5 1

47 89 69

‘Resinous’ material from amulet on neck Stained bandaging from right hand side of neck Black ‘resin’ coated outer bandages right hand side of upper arm Fur Bandaging from right hand side of upper torso Tissue from left hand side of top of skull top Black ‘resin’ coated bandages from left shoulder Black ‘resin’ coated bandages from left hand side of shoulder/ neck Bandages from tip left foot Stained bandages from leg Stained bandages from sole left foot Blackened ‘resin’ on stomach Pale bandaging Darkened bandaging under right breast Darkened bandaging under right shoulder Stained bandages from left hand side half way up body Blackened bandaging inside neck

Bitumen present

11 31

23

59 100

18

42

62

0.3 (27) n.q. n.q. n.q.

58 No extractable lipid

34

66 100

38

62 16

59

16 3

25

No extractable lipid 51 22 8 26 43 14 69 26 No extractable lipid No extractable lipid 100

27 66 42 5

88

Interior of mummy

12

100

Black tissue inside neck and hair

100

Bone from left hand side of jaw bone

99

1

2

Black tissue/ ‘resin’

94

6

2

Black tissue/ ‘resin’ Black ‘resin’ on hair Black tissue/’resin’ and bandaging

91 100 100

Black tissue/ ‘resin’ Black tissue from neck Blackened tissue Bandaging base of neck- modern contamination Black ‘resin’ from sides Black ‘resin’ from base of lid Linen and lump from jar-‘resin’ Linen and lump from jar-bandage Blackened textile with tissue/‘resin’ Tissue, bandaging/ ‘resin’

9

96 100

4

59

41

25

n.q. 0.06 0.3

75 72

20

8

100 78

20 13

30

44 77 79

Black ‘Resin’ Bandage ‘Resinous’ lumps Black tissue from left hand side base chin & inside skull Clear ‘resin’ Bandage Tissue from right hand Tissue from heal Black tissue/ Bandage from finger Black bandage from finger Black tissue underside wrist Brown bandaging from sole Dark bandaging Light bandaging Black tissue/ bandage Black tissue from outside head Black tissue from under jaw Black bandage behind ear Black tissue from backside head Black tissue top side of wrist Black tissue from ankle

10

84

9 25 23 19 16

100 98 100 90 92

2

17 3 26

10 8 Paraffin wax

78 76

5

22 24 95 100 No extractable lipid 88 12 90 10 62 38 87 13 90 10 71 14 15 100 100 94 6

n.q. 56

n.q.

29

Trends in Use of Organic Balms in Egyptian. . .

689

Table 2 (continued) Mummy

Museum number

Date

Location

% composition#

Left foot Canopic jar Adult Adult

n.d. n.d. n.d. n.d.

100

Adult Adult

AP 8.418a UP 1 UP 3 TUR 14406 (033.064) TUR 14.389 TUR 1

Adult Adult

TUR 2 TUR Pravv 569

n.d. n.d.

Adult

TUR Pravv 545/14428 TUR 3 (drawer)

n.d.

Cat Shaped sarcophagus Head of a female adult Head of a male adult Head of a female adult Head of a female adult Head of a male adult Left hand of an adult Left hand of a female adult Hand of an adult

CAI 15+4/24+1

n.d.

Black tissue underside heal ‘Resinous’ Contents Bandage Bandage on left thigh Tissue from left upper arm Stained outer bandaging Bandage behind knee Tissue from right knee Blackened bandaging Bandaging, pile of bandages on top in box Bandaging underneath attached to mummy Bandaging thorax Tissue, top of head, under bandaging Bandaging from near big toe Tissue from near big toe Resinous’ lump

RMO 37 RMO 40 RMO 42 RMO 45 RMO 46 RMO 49 RMO 50

n.d. n.d. n.d. n.d. n.d. n.d. n.d.

Blackened bandaging top of head ‘Resin’ coated bandaging from neck Black ‘resin’/bandage Hair and tissue/ ‘resin’/bandaging Blackened tissue from neck Tissue from wrist Black tissue from wrist

RMO 51

n.d.

Hand of an adult Hand of a child Head Adult

RMO 52 RMO 53 RMO F2004/12.2 RMO Grey 7

n.d. n.d. n.d. n.d.

Black bandaging from thumb Scrapping of black ‘resin’ Tissue from wrist Tissue from wrist Tissue from neck, bandaging fragments Bandaging from sole of right foot Bandaging from upper torso

Adult

n.d. n.d.

n.d.

72 33

5

28 61 6 No extractable lipid 100 No extractable lipid No extractable lipid No extractable lipid No extractable lipid No extractable lipid 85 15 No extractable lipid No extractable lipid 100 No extractable lipid 69 26

70 56 77 67

Bitumen present

No extractable lipid 30 2 42 23 100 33 No extractable lipid

n.q. n.q.

No extractable lipid No extractable lipid 100 No extractable lipid 99 1 100 100

Key: n.d. = not determined; #, % composition of balms calculated from mass of lipids as a portion of the solvent soluble extract; §, % composition of bitumen not indicated as part of the whole balm, due to the difficulties in determining an accurate quantification, described in Clark et al. (2016); number within shaded area is the % composition of that ingredient, number in brackets (bitumen) was determined from radiocarbon analysis; red indicates portion of fat/oil in balm; blue, beeswax; green, coniferous resin; orange, pistacia resin; black, bitumen; grey, no bitumen.

more expensive ingredient than either fat, oil, or beeswax, because these ingredients would have been available from local sources, whereas resin would have been imported, as the resin-producing trees did not grow in Egypt (Serpico and White 2000). Similar variations in balm composition were observed by Buckley and Evershed (2001) (Table 2). Bitumen was identified in 39% of the mummy balms analyzed for bitumen biomarkers. Semi-quantitative estimates of the biomarkers indicate that balms could be composed of 0.1–62% bitumen, which is a similar range to beeswax. A smaller number of more precise quantitative estimates of bitumen concentrations based on 14C analysis suggested a somewhat similar range in bitumen concentration, that is, 0–45% bitumen (Clark et al. 2016). Like resins, the majority of bitumen was imported into Egypt, making it more expensive than fats/oil, hence, its presence in such high concentrations in balms is noteworthy. The overall composition of balms varied widely in the individuals and materials investigated; however, similarities were seen between the balms applied to different parts of an individual mummy. The compositions of the TLEs of the balms removed from a variety of locations on the body of the male adult mummy (Glasgow) are shown in the chromatograms in Figures 4 and 5. It can be seen that the compounds present and their relative concentrations are almost identical between the different locations, where the mean for the percentage of fat/oil is 69.4% (σ ¼ 3.6%),

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Fig. 2 Partial gas chromatogram of the trimethylsilylated TLE of a sample of blackened bandaging from a female hand (BRI Ha5546), indicating the high abundances of oxidized dehydroabietic acid derivatives compared with free fatty acids. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation. IS are internal standards

beeswax, 23% (σ ¼ 6.5%), and resin, 5.8% (σ ¼ 5.3%). The balms that show the largest difference are those from the package from inside the body, which suggests that additional beeswax and/or resin was added when the package was prepared. The distribution of n-alkanes of the beeswax also differs between the balms taken from the package compared with those from the other locations on the body. This is further evidence of a difference between the balms, possibly arising from the further addition of beeswax applied to the packaging, a different preparation method or protection from loss through environmental factors or microbial action. However, it would appear that the rest of the mummy was prepared using one “pot” of balm rather than different balms applied to specific areas, given the similarities of the TLE and the percentage composition of the ingredients. The balms sampled from the legs and torso of the Graeco-Roman adult mummy with the folded arms (100 BC–AD 395; TUR Pravv 540) also contained the same mixture of ingredients, namely, fat/oil, beeswax, and resin. The relative abundances of the compounds present in the TLE (Figs. 6 and 7) indicate that the balm applied to this mummy was also from a single mixture, to which further ingredients were added during the embalming process, creating differences in the proportion of the ingredients measured at each location; the sample of pale bandaging contained a much higher beeswax content than the other balms and the stained bandaging from the foot

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Fig. 3 Partial gas chromatogram of the trimethylsilylated TLE of a sample of the bandaging from the bandaging from the torso of the Ptolemaic female adult (c. 332–30 BC; RMO 13), indicating the high abundances of wax esters compared with free fatty acids. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation; WEx are wax esters of chain length x; and HWx are hydroxy wax esters of chain length x. IS are internal standards

containing less. The distribution of wax esters of beeswax from this pale bandaging also differs from that identified in the other bandaging, which is further evidence for the use of additional beeswax or a different preparation method. The mean of the percentage composition between the different sampling locations of fat/oil is 35% (σ ¼ 26%), beeswax 43% (σ ¼ 26%), and resin 22% (σ ¼ 5.6%). An adult mummy dating to the Fifth to Sixth Dynasties (2410–2195 BC; TUR) was also sampled in a number of locations (Figs. 8 and 9). In this mummy, only fatty acids and their degradation products were identified, indicating that the balm contained fat or oil, although the fatty acids are possibly derived from the body tissues. An important difference in these samples lies in the relative abundances of the fatty acids. For example, the concentrations in the bandaging were much lower than in the tissues. The tissue from the inner side of the right forearm contains a high concentration of C16:0, relative to the other tissues (Fig. 10). These differences indicate that this mummy was possibly not treated with the same balm all over the entire body, that is, different fats and oils were used on different locations. However, it is possible that these differences arise because of the differing human fatty acid compositions of tissue around the body (Chapter 3 and Bereuter et al. 1996; Makristathis et al. 2002), or that these areas were afforded more protection from the surrounding environment. Similar compositions of balms from different locations on the body is not the norm. The samples of tissue, bandaging, and resin from the Twenty-Third to Twenty-

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Fig. 4 Comparison of partial gas chromatograms of the trimethylsilylated TLE of balms from different locations of the Third Intermediate Period male adult (c. 1064–656 BC; MTB G6, 20, 32, 44) showing the similarities between the extracts. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation; ALx are n-alkanes of carbon chain length x; DHA is dehydroabietic acid; and Wx are wax esters of C16:0 fatty acid (palmitic acid) with carbon chain length x. IS indicates internal standards

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Fig. 5 Comparison of the composition of the balms taken from a number of locations on the Third Intermediate Period male adult (c. 1064–948 BC; MTB G6, 20, 32, 44), showing the similarity of the balms from the various locations. The colors indicate the presence of a commodity, and except for bitumen, the % composition in the balm (red ¼ fat/oil, blue ¼ beeswax, green ¼ coniferous resin, black ¼ bitumen)

Fifth Dynasty male mummy Besenmut (c. 700 BC; MTB 528/1) varied greatly in composition (Figs. 11 and 12). The sample of resin was found to contain a mixture of fat/oil, beeswax, pistacia resin, and bitumen, whereas the tissue and bandaging contained only fat/oil and bitumen. Additionally, the δ13C values of the C16:0 and C18:0 fatty acids from “resin” indicate that they originated from a ruminant adipose source, whereas those of the bandaging indicate a mixture of ruminant adipose and non-ruminant adipose or plant oil, or plant oil on its own. The differences between these balms indicate that different mixtures of balms, perhaps indicating a specific ritual significance or higher class of mummification, was used to treat the parts of this body.

Variations in Balm Compositions over Time The mummies investigated cover the entire period over which the ancient Egyptians were known to prepare them (c. 3500 BC–AD 395), thereby enabling a timeline of the variations of the composition of balms to be constructed. This is demonstrated by displaying each mummy as a box spanning the date range for that mummy. Each of

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Fig. 6 Comparison of partial gas chromatograms of the trimethylsilylated TLE of balms from different locations of the Graeco-Roman adult mummy with folded arms (100 BC–AD 395; TUR Pravv 540) showing the similarity of balms applied to different body locations. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation; ALx are n-alkanes of carbon chain length x; DHA is dehydroabietic acid and Wx are wax esters of C16:0 fatty acid (palmitic acid) with carbon chain length x; HW are hydroxy wax esters of carbon chain length x. IS indicates internal standards

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Fig. 7 Comparison of the composition of the balms taken from a number of locations on the Graeco-Roman male adult (100 BC–AD 395; TUR Pravv 540), showing the similarity of the balms from the various locations. The colors indicate the presence of a commodity, and except for bitumen, the % composition in the balm (red ¼ fat/oil, blue ¼ beeswax, green ¼ coniferous resin, gray ¼ no bitumen, white ¼ no extractable lipid)

the commodities investigated in Chapters 3–6 in Clark (2007) is included in order to allow changes in use of the commodity to be recognized (Fig. 13). In addition to the mummies analyzed in this study those examined in other studies are included to obtain as complete a view of the changes in mummification as possible. By comparing the changes in compositions of the balms over time a number of trends are apparent. The first is the simplicity of balms applied before the Third Intermediate Period (c. 1000 BC). Mummies from the Predynastic Period to the New Kingdom appear to be very simply embalmed, with only fat/oil or not embalmed at all, in which case the fatty acids and their derivatives detected are the result of fats derived from the body itself. Balms applied to three mummies from this period contain coniferous resin, either on its own or possibly mixed with fat from a ruminant source (Koller et al. 1998; Buckley and Evershed 2001), whereas one mummy contains a mixture of coniferous resin and bitumen (Connan and Dessort 1991; Connan 2002). The only mummy to be more elaborately embalmed, the beef ribs meat mummy (c. 1386–1349 BC; CAI CG5109) from the Eighteenth Dynasty high status burial of Tjuiu and Yuya, contains a mixture of ruminant fat, beeswax and pistacia resin (Clark et al. 2013). This mummy cannot be directly compared with other human mummies due to the high status, but could possibly be seen as avantgarde, as it contains the mixture of ingredients routinely identified in lower status human mummy balms over two centuries later.

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Fig. 8 Comparison of partial gas chromatograms of the trimethylsilylated TLE of balms from different locations of the Fifth to Sixth Dynasty female adult with dress (2410–2195 BC; TUR) showing the difference between the balm extracts. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation. IS indicates internal standards

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Fig. 9 Comparison of the composition of the balms taken from a number of locations on the Fifth to Sixth Dynasty female adult (2410–2195 BC; TUR), showing the similarity of the balms from the various locations. The colors indicate the presence of a commodity, and except for bitumen, the % composition in the balm (red ¼ fat/oil, gray ¼ no bitumen)

The major modification to ingredients applied in balms occurred toward the end of the New Kingdom and the start of the Third Intermediate Period. This is the period that is often described by Egyptologists as the “height of mummification,” where the physical treatment carried out to the body, such as evisceration and bandaging and the preservation achieved, is considered to have reached their peak (Quirke 1992). The introduction of beeswax and resins into the balms toward the end of this period suggests that there were modifications to the chemical treatment of mummies, in addition to the physical treatment, which might explain the improved preservation of many mummies from this period. This is the first time that a connection between the changes that occurred to the balm, the improved preservation of the mummy and the height of mummification has been identified. The coincidence of both the chemical and physical “height of mummification” suggests that during this period mummification had undergone significant changes, possibly through experimentation, culminating in a “standard” balm. Interestingly, beeswax and resin were not always present together in the same balms, although the majority of balms from mummies dated to the Third

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Fig. 10 Variation of the fatty acid concentrations between the different locations on body of the Fifth to Sixth Dynasty female adult with dress (2410–2195 BC; TUR), an indicator of limited application of balms or different degrees of degradation between the locations on the body

Intermediate Period onwards (after 1000 BC) contain fat/oil, beeswax, or resin (see discussion below). In addition, bitumen was included more frequently in balms dated to after the Third Intermediate Period (c. 750 BC), most likely due to the increased availability of bitumen through trade routes with the Near East (Clark et al. 2016). As mentioned above both resins and bitumen were likely to have been imported into Egypt as they were either not available in Egypt or the imported material was a preferred product. The major trade routes into ancient Egypt are shown in Fig. 14 (Shaw 2000), which encompasses the areas where the majority of resins and bitumen can be sourced. These trade routes were in operation from during the Old and Middle Kingdoms but were extensively exploited from the New Kingdom (c. 1549–1064 BC) onwards. Ancient Egypt’s increasing trade with its neighbors would provide an explanation for the availability and employment of resins and bitumen in embalming after this period. The balms do not, however, appear to have undergone the same decline as the physical treatments after the height of mummification; the mixture of fat/oil, beeswax, and resins, with the later introduction of bitumen, continued to be included in balms to the end of the Graeco-Roman Period when embalming ceased to be carried out. This suggests that the use of these materials had proved to preserve the mummies sufficiently and therefore further modification was not required. The additional religious and symbolic associations that may have surrounded the materials applied to the mummy, such as the protection afforded by the use of bee products, may also have contributed to their continued utilization, even though the standards of mummification had declined.

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Fig. 11 Comparison of the composition of the balms taken from a number of locations on the Twenty-Third to Twenty-Fifth Dynasty male adult, Besenmut (c. 700 BC; MTB 528/1), showing the similarity of the balms from the various locations. The colors indicate the presence of a commodity, and except for bitumen, the % composition in the balm (red ¼ fat/oil, blue ¼ beeswax, orange ¼ pistacia resin, black ¼ bitumen, gray ¼ no bitumen)

Preparation of Balms and Specific Recipes Although it is interesting to examine the changes in use of the individual ingredients of balms, it is also important to assess the changes in the composition of the balm (Fig. 15). When the mummy balms are compared according to the mixture of ingredients, other patterns become apparent: the very earliest mummy balms are very simple, containing only one detectable ingredient, i.e. a fat or oil. During the Second Intermediate Period and New Kingdom (c. 1750–1064 BC) the balms become more complicated, with the introduction of resins. The end of the New Kingdom and Third Intermediate Period sees the introduction of a third ingredient, beeswax. The addition of the fourth ingredient, bitumen, appears at the end of the New Kingdom, and it becomes a regular ingredient during the Late Period. The balm-like deposits seen on Predynastic mummies have only been found to contain a

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Fig. 12 Comparison of partial gas chromatograms of the trimethylsilylated TLE of balms from different locations of the Twenty-Third to Twenty-Fifth Dynasty male mummy Besenmut (c. 700 BC; MTB528/1), indicating the difference between the balms. FAx:y are fatty acids where x is the carbon chain length and y is the degree of unsaturation; ALx are alkanes of carbon chain length x; Wx are wax esters of C16:0 fatty acid (palmitic acid) with x carbon chain length. IS indicates internal standards

Trends in Use of Organic Balms in Egyptian. . .

Fig. 13 Timeline showing the changes of the materials used in mummy balms from the Predynastic to Graeco-Roman Periods. Key: (a) mummies examined in this study; (b) Buckley and Evershed (2001); (c) Rullkötter and Nissenbaum (1988); (d) Connan and Dessort (1989, 1991); (e) Proefke et al. (1992); (f) Kaup et al. (1994); (g) Mejanelle et al. (1997); (h) Koller et al. (1998); (i) Serpico and White (1998); (j) Connan (1999, 2002); (k) Colombini et al. (2000); (l) Maurer et al. (2002); (m) Tchapla et al. (2004)

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Fig. 14 Map indicating the major import trade routes into ancient Egypt. (Adapted from Shaw 2000)

Fig. 15 Comparison of the frequencies of compositions of balms

fat or oil, thus they may not be balm and most likely originate from the body itself and not from a deliberately applied substance. The statistical significance of the number of components in the balm can be tested using the χ2 test. This compares the observed frequency with the expected frequency

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for a set of variables (such as the components in a balm). The calculated χ2 value determines whether some variables occur more often than others, and if their distribution is statistically significant. Therefore, the lower the value of χ2, the more random the distribution, whereas higher χ2 values indicate a deliberate discrimination among the variables. If the χ2 exceeds the value of the “test statistic,” there is a low probability that the observed distribution is a random distribution. The numbers of balms displaying a specific composition are displayed in Fig. 15. The highest proportion of the mummy balms contained fat/oil, although this is likely to be affected by the number of tissues analyzed and the detection of human fat, rather than exogenous fat/oil. From the mixtures of other embalming agents, those occurring most often are a mixture of fat/oil, beeswax and resin > fat/oil, beeswax, resin, and bitumen > fat/oil and resin. Statistical analysis of the frequency at which the different mixtures occur (χ2 tests excluding mummies with no applied balm and fat/oil because they do not necessarily indicate the deliberate application of a balm) gives χ2 ¼ 115 (d.o.f. ¼ 13). This value exceeds the critical value of 29.8 (P ¼ 0.005) and therefore indicates that the probability of the observed distribution being the result of there being no difference in the way that the mixtures were used is less than 0.5%. We must conclude, therefore, that there was a deliberate variation between balm mixtures employed. The proportions of ingredients was also expected to have varied over time. By comparing the percentages of the various ingredients used in balms over time (Fig. 16), it can be seen that, while the amount of fat/oil used in the balm decreased, the amount of beeswax increased at almost the same rate. The amount of resin in the balm also increases with time but not at the same rate as that of beeswax. In all but one case, the proportion of resin never accounts for more than 40% of the balm, whereas the proportion of fat/oil and beeswax in the balm varied more widely. Bitumen was not included in these calculations because of the difficulties in accurately determining the bitumen concentration in balms.

Variation on Balm Composition with the Age of the Individual The balms from a number of child mummies were investigated in this research and studies carried out by other authors (Proefke et al. 1992; Buckley and Evershed 2001; Tchapla et al. 2004). A graph comparing the frequency of occurrence of mixtures of ingredients in balms from children and adults is displayed in Fig. 17. This plot shows that balms from children are generally simpler than those from adults. No balms from children contained a mixture of four ingredients and only a few contained three ingredients (fat/oil, beeswax, resin and bitumen). A large proportion of the balms from children comprise fat/oil and resin, which occur more frequently in the balms from children than those from adults. A χ2 test of the combinations of ingredients used in children again shows that the frequency of use of fat/oil and resin is significant (χ2 ¼ 34, d.o.f. ¼ 13), whereas the frequency of use of fat/oil, beeswax, and resin in adult balms is also significant (χ2 ¼ 125, d.o. f. ¼ 13). The probability that these mixtures were used in preference to others is greater than 99.5%.

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Fig. 16 Variations in percentage composition of (a) fat/oil, (b) beeswax, and (c) resin in balms over time

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Fig. 17 Comparison of the frequencies of compositions of balms from adult and child mummies

Variations in Balm Composition with the Gender of the Individual Consideration of the balms from the different genders (Fig. 18) shows that male mummies appear to be treated more elaborately than their female counterparts. The highest proportion of balms from female mummies contains only fat or oil, with fewer female mummy balms containing two or more balm ingredients than balms from male mummies. A significant proportion of male mummy balms contain fat/oil, beeswax, and resin or four ingredients (fat/oil, beeswax, resin, and bitumen) whereas the proportion of female mummies with this combination is much lower. A χ2 test of the ingredient combinations used in male mummy balms shows that the occurrence of three ingredients (containing fat/oil, beeswax and resin) is also significant (χ2 ¼ 169, d.o.f. ¼ 13) at 95.5%. Another difference between the genders is the incidence of pistacia resin in human mummy balms. Pistacia resins has been detected in balms from four female mummies (Serpico and White 1998; Colombini et al. 2000; Buckley and Evershed 2001), one male mummy Besenmut (MTB528/1; this study), and one where the gender was not reported (Kaup et al. 1994). Given that pistacia resin has only been identified in a small number of mummies, this gender differentiation is interesting and suggests a possible (unknown) significance for the use of pistacia in balms used on female adults. However, the limited incidences of pistacia use means that firm conclusions surrounding its use cannot be drawn.

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Fig. 18 Comparison of the frequencies of compositions of balms from male and female mummies

Variation in Balm Composition with Location on the Body The balms applied to different parts of the body may have varied because of different rituals and associations given to each part of the body. The samples studied were divided into those from the head, torso, and limbs. When all the balms from different locations are compared (Fig. 19) there appears to be some variation according to the location even though individuals showed no variation. Balms containing fat/oil account for the highest proportion of all balms from all the body locations. However, mixtures of fat/oil, beeswax, and resin dominate balms from the torso and limbs, whereas very few balms from the head contain this mixture. Balms from the head are almost equally likely to contain a mixture of fat/oil and beeswax or resin, a mixture of fat/oil, resin, and bitumen or a mixture of fat/oil, beeswax, resin, and bitumen. χ2 tests of the frequency of occurrence of balm mixtures from the torso and limbs indicate that the number of balms containing fat/oil, beeswax, and resin is significant at the 95.5% level (limbs: χ2 ¼ 38, d.o.f ¼ 13; torso: χ2 ¼ 52, d.o.f ¼ 13). Similarly for the head, the number of balms containing fat/oil and beeswax or resin, a mixture of fat/oil, resin and bitumen or a mixture of fat/oil, beeswax, resin, and bitumen is also significant (χ2 ¼ 41, d.o.f ¼ 13).

Variation of the Chemical Composition with Material Type The type of sample, tissue, bandage, or “resin” may also display variations in the composition of applied balms. Fig. 20 shows that the tissues are dominated by fat/ oil; however, this is often likely to derive from the body itself rather than the deliberate application of fat/oil to the body. A high proportion of the bandages

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Fig. 19 Comparison of the frequencies of compositions of balms from different locations on the bodies of human mummies

Fig. 20 Comparison of the frequencies of compositions of balms from material types

contains only fat/oil, which is more likely the result of the deliberate application, although as some bandages analyzed were directly in contact with the body, so the fat/oil identified may also originate from the body. Other than fat/oil, a high proportion of tissues, and bandages were also found to contain a mixture of fat/oil,

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Fig. 21 Variations of compositions of “resinous” outer coatings

beeswax, and resin. The samples that are visually described as “resin” are dominated by three mixtures, either fat/oil, fat/oil, and resin or fat/oil, beeswax, resin, and bitumen. The number of different combinations of embalming agents used in “resins” and tissues is greater than that seen in bandages. A number of mummies studied dating from the Third Intermediate Period to the Graeco-Roman Period were covered by a thick dark layer of ‘resin’ on the outer bandages. Despite all the balms being similar in appearance, that is, hard black resinous coatings, their composition varied considerably (Fig. 21). The sample from the earliest mummy considered dating to the Twenty-First Dynasty (c. 1064– 948 BC; BM 6660) mostly comprised fat/oil. In the later mummies the composition varied from a mixture of fat/oil and beeswax; fat/oil, beeswax, and resin; fat/oil, beeswax, and bitumen and fat/oil, beeswax, resin, and bitumen. The proportions of the commodities comprising the balm also varied between the mummies. This difference in the “resinous” coatings of outer bandages serves to highlight the need for caution when assigning the ingredients used in the balm based on their visual appearance alone.

Discussion Comparison of the compositions of balms from mummies studied here combined with those previously reported has revealed a number of important trends in the materials used in the balms and the embalming process, despite the significant individual variation present in the balms. Many of these trends indicate the development of the embalmer’s technique and the increasing complexity of embalming ritual, which is also highlighted by the morphological differences of embalming, that is, the physical treatment of the body, including the removal of the viscera. The increasing complexity of ingredients present in the balms around 1000 BC was a major development in the embalming practice. The coincidence of the addition of more ingredients to the balm with the reported height of mummification, together

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with the changing proportion of the ingredients used, is further evidence of the development of the embalming practice over ancient Egyptian history. Egyptologists have hypothesized such a development because of the increasing elaboration of the bandages and physical treatment of the body (Quirke 1992; Ikram and Dodson 1998), but this is the first time we have chemical evidence from the balms to support this idea. Analysis of the cemetery at Deir el-Medina also indicates that there was a major shift in the burial practices and beliefs between the Eighteenth (c. 1549–1328 BC) and Nineteenth Dynasties (c. 1298–1187 BC), from a representational focus on the living world to an emphasis on the next world (Meskell 1999a). During the Eighteenth Dynasty the objects left with the dead reflected the daily life activities of the individual: food, work tools and musical instruments. However, during the Nineteenth Dynasty there was a gradual shift to objects that had a magical or ritual element: magical texts, amulets, and shabtis. In addition to the differing objects present in the tombs, there is also evidence to suggest that there was a major change in the methods used to preserve the bodies, which also reflects the changes in the mortuary sphere. During the Eighteenth Dynasty, simple body treatments with little or no embalming were evident; the bodies were simply wrapped rather than dried with natron and eviscerated. In the following Nineteenth Dynasty there is a major change in the way the bodies were treated, viscera were removed and evidence for the use of natron and “resin” has been found. These methods were however, not new innovations as a small number of the Eighteenth Dynasty mummies were treated in this way. The elaborate preparations carried out on the body where therefore deemed necessary to preserve its integrity in the Afterlife. The reasons that are suggested by Meskell (1999a) for this sudden and dramatic shift in burial practices are that, following the major upheaval of the Amarna period at the end of the Eighteenth Dynasty, there were new ideas about life and death, particularly that to cope with and control destiny, a good death and Afterlife would have become important. These rituals were required so that mortals would become gods, literally becoming Osiris. The decline in the mummification techniques during the Late and subsequent periods is not reflected in the composition of the balms. A decline is suggested because mummies dating after the height of mummification, that is, after the TwentySecond Dynasty (c. 900 BC), are not so skillfully or carefully treated as before, in terms of the bandaging and other treatment of the body. However, the burials at Deir el-Medina indicate that the elaboration of the body that occurred during the Nineteenth Dynasty continued through to the Graeco-Roman Period, with each culture having its own ideas about the body in death (Meskell 1999a). This would suggest that as the body was still required for the Afterlife, and therefore the chemical treatment continued after the “height of mummification.” Between these periods there was perhaps a shift in focus for mummification, particularly from the physical treatment of the body, such as evisceration, and efforts to make the body look lifelike and a focus on the appearance of the final mummy, for example wrapping of the mummy was intricately performed. The increasing democratization of mummification during the Ptolemaic and Graeco-Roman Periods inevitably decreased the amount of time the embalmers spent on each body, thereby reducing the quality of

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physical treatment, although the chemical treatment was not altered possibly, due to the effectiveness of results achieved. There also appear to be differences in the balms used on different classes of individual (age and gender). In general, the balms from children contained fewer ingredients than adults; however, if children are compared with contemporaneous adults, there are some examples of the same treatments applied to adults and children. Females were also treated more simply than males, with the majority of balms from female mummies containing fat/oil and fewer balms containing more complex mixtures of ingredients. A high proportion of male mummy balms investigated contained a mixture of fat/oil, beeswax, and resins. Even though females were generally treated more simply than males, one of the more exotic ingredients identified in balms, pistacia resin, and it is identified more frequently in balms from female mummies than male mummies, possibly indicating a preferential treatment of these females for a hitherto unknown reason. The narrow period of time where pistacia was used in balms (c. 700 years) adds to the enigmatic nature of this commodity. Similar differences between the treatment of males, females and children were seen at Deir el-Medina (Meskell 1999a, b). It has not been possible to distinguish between the different classes of mummification described by Herodotus (Herodotus trans. De Sélincourt 1996) as information about the location of the viscera is the major difference between the different classes, which was not detailed for a number of the mummies examined. Additionally, the archaeological evidence suggests that the treatment of the viscera does not always indicate class; for example the mummy of Kha (Eighteenth Dynasty, c. 1549–1328 BC) still contains his viscera, although the goods which accompany him indicate that in life he was wealthy while the contemporary burial of a male was stuffed with rags, which suggests that the organs were removed, although his burial was not as prestigious (Meskell 1999a). One difference that may indicate the different classes of mummification is the variation in balms. Of the mummies where it was possible to take multiple samples, the extracts of balms analyzed from two male adults dating to the Third Intermediate and Graeco-Roman Periods were identical between the sampled locations on the body, indicating the balm came from one pot. However, the balms sampled from different locations on another male mummy, Besenmut (c. 700 BC; MTB 528/1), were different. This was also the only male mummy to contain pistacia resin and possibly indicates this was a high status individual and the different treatments applied to the different body parts (and material types) may indicate a preferential treatment. It is not possible to determine conclusively whether these mummies were treated differently because they received a different class of embalming or whether different embalmers each used their own methods, based only on the results of three mummies. More general trends regarding the differences in the treatment of different parts of the body become evident when all the mummy balms analyzed are compared. This showed that balms from the torso are more likely to contain fat/oil, beeswax, and resin than those applied to the head and limbs, which are most likely to contain only fat/oil. Other mixtures frequently identified on the head are fat/oil and resin or beeswax, fat/oil, resin, and bitumen and fat/oil, beeswax, resin, and bitumen. On

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limbs, fat/oil, beeswax, and resins also occurred frequently. These differences in the treatments of body parts indicates that there were biases in the specific treatments applied to the body, perhaps involving the application of the sacred oils to the head and the limbs to re-animate the body, as described in papyri describing the rituals surrounding embalming (Birch 1863; Sauneron 1952). Variations in the balms can also be seen on the different ingredients associated with embalming, tissues, bandages, and “resins.” Fat/oil was identified most often on tissues, although this is most likely corresponds to fat from the body and not the intentional application of a balm. Fat/oil was also identified on many bandages; however, this may also be from the body if insufficient desiccation had occurred when the body was wrapped. Fat/oil, beeswax, and bitumen were also identified in significant proportions in tissues and bandages. “Resins” most frequently consist of fat/oil and resin or fat/oil beeswax, resin, and bitumen. The difference in the composition of these balms again indicates that there were different stages to the embalming process and the embalmers treated bandages and tissues differently.

Conclusions In this contribution we present the largest synthesis of the organic chemistry of Egyptian mummy balms, tissues, and bandages yet presented. In addition to presenting substantial new data on the chemical compositions of mummy balms we also integrate data from previously published reports. By doing this we have opened up the opportunity for assessing for the first time a number of critical trends and themes in the evolution of embalming practices in ancient Egypt. The key findings of this are: (i) Balms become increasingly complex over time. Initially, they were very simple, with more ingredients added during the Third Intermediate Period (after c. 1000 BC). This coincides with the period known as the “height of mummification.” (ii) Balms from children are simpler than those from adults, generally only containing one or two of the major ingredients rather than three or four, as frequently found in adult mummies. (iii) Balms from female mummies are simpler than those from male adults, containing fewer ingredients than the males. However, pistacia resin, an ingredient only found in a small number of balms, has been identified more frequently in female mummies than male mummies. (iv) Tissues were found to contain mainly fat/oil, whereas bandages and “resins” contained more ingredients and were more variable in composition. The “resins” applied to the outer bandages of different mummies, although appearing to be visually identical, can be very different in composition. (v) The composition of the balms changed markedly over time; the percentage of fat/oil decreased as the proportion of beeswax increased, while the proportion of resin remained relatively constant after its introduction, never consisting of more than 40% of the balm.

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(vi) The increasing complexity of the balms over time, particularly the introduction of new ingredients i.e. beeswax, resin and bitumen, at a time that coincides with the height of embalming demonstrates an important new parallel development in the overall evolution of the embalming practice. Acknowledgments We thank John Taylor (British Museum); Marteen Raven (Rijksmuseum van Oudheden, Leiden, The Netherlands); Rosalie David and Trish Lambert (Manchester museum and tissue bank); Sue Giles (Bristol Museum); Vicky Taylor (Durham Oriental Museum); Julie Greeson (Auckland War Memorial Museum, New Zealand); Willem van Haarlem (Allard Pierson Museum, Amsterdam); Emma Rabino Massa and Raffaella Bianucci (Università degli Studi di Torino); Faye Kalloniatis (Norwich Castle Museum); Joanna Hayward (Liverpool museum); Kathy Eremin (National Museum of Scotland); and Muhammed Saleh and Nasry Iskander (Cairo Museum) for provision of samples. We thank NERC for studentship to K.A.C. and funding MS facilities, Ian Bull and Rob Berstan fortechnical assistance, and Tom Higham of the Oxford Radiocarbon Accelerator.

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Part VI Mummies in Europe

Life and Diseases of the Neolithic Glacier Mummy “Ötzi”

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Andreas G. Nerlich, Angelika Fleckinger, and Oliver Peschel

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Detection of the Body . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dating of the Mummy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Individual Age . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Anthropological Examination, Body Size, and Stature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Type of Mummification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Provenance and Genetic Origin of the Iceman . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Nutrition of the Iceman . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Evidence for Physical Activities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Infectious Diseases and Diseases of Internal Organs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Intestines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pulmonary Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Vascular Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Non-traumatic Skeletal Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Diseases of the Teeth and the Oral Cavity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Abnormalities of Hair . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Systemic Infectious Disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Potential Therapeutic Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Sequelae of Traumatic Injuries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Old Healed Trauma . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Trauma Acquired Several Days Before His Death . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Prof. Peschel is additionally the leader of the conservation team for the mummy of the Iceman, Bolzano, Italy. A. G. Nerlich (*) Institute of Pathology, Academic Hospital Munich-Bogenhausen, Munich, Germany e-mail: [email protected] A. Fleckinger South Tyrol Museum of Archaeology, Bolzano, South Tyrol, Italy O. Peschel Institute of Legal Medicine, Ludwig-Maximilians-University Munich, Munich, Germany © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_19

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The Mortal Arrow Shot . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Further Possible Perimortal Lesions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Possible Scenarios of the Iceman’s Life and Death . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Iceman’s Life . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ötzi’s Final Days – The Scenario of His Death . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Final Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

The unusual find of a very well-preserved human adult male mummy in an Alpine glacier in 1991 was the starting point for a unique series of scientific analyses that resulted in a concise reconstruction of life, diseases, and death of the Neolithic individual (dating to c. 3350–3100 BCE). Not only his basic anthropological information, genetic origin, nutritional supply, and physical activity pattern were investigated, but also numerous diseases, possible therapeutic treatment, and a sequence of traumatic injuries have been identified. The c. 45-year-old man was highly mobile, well-nourished with a balanced animal-faunal diet, suffered from intestinal parasitic infections (Trichuris trichiura) and gastric bacteria (Helicobacter pylori), gallbladder stones, pulmonary anthracosis, mild to moderate arteriosclerosis (of the media-type), mild focal osteoarthrosis, and cervical/ lumbar spondylosis. Besides old-healed rib fractures, he had acquired a deep hand wound from stabbing several days before his death; lastly, a lethal arrow strike to the back of his chest wall led, most probably, to laceration of the subclavian artery with extensive bleeding. Whether he also suffered from severe brain trauma by falling on his head, and systemic infectious disease (Lyme’s disease; Borrelia burgdorferi) is not yet clear. The extensive analysis of his belongings, found nearby, add significant further information to the Iceman’s life. It remains to be seen what further information can be obtained on this unique mummy. Keywords

Neolithic mummy · Intestinal infections · Arteriosclerosis · Osteoarthrosis · Trauma sequels

Introduction The mummified corpse of the Neolithic glacier mummy, popularly termed “The Iceman” or even more simply “Ötzi,” is one of the most intensely investigated human mummies ever seen. A Pubmed and Google Scholar search for both terms reveals several hundred publications in scientific journals on this unique historic individual, and there are many more in books, newsletters, etc. This enormous interest comes from both the unusual age of the mummy and the exceptional preservation; its astonishing state of mummification has allowed scholars to perform, for nearly 30 years, numerous investigations on it using different techniques and a variety of approaches.

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Fig. 1 The Iceman (© Südtiroler Archäologiemuseum/Eurac/ Samadelli/Staschitz, with permission)

Besides his body (Fig. 1), the Iceman had numerous belongings with him; clothes, everyday items, weapons, etc. Research on these findings offered significant insights into the daily life of a Middle European Neolithic dweller. The combination of examination of the mummy and study of the clothes, tools, and artifacts has had a synergistic effect on the scientific out-put. In this state-of-the-art-review, we mainly focus on the investigations of the human body and touch, only briefly, on the significant findings associated with his belongings. To some extent, “The Iceman” has developed to be a test bed for the use and technical optimization of novel analytical techniques – an issue that will continue in the future.

Detection of the Body An unexpectedly intense summer warming determined an unusual extensive recession of most alpine glaciers, particularly the glacier of the Schnalstal/Val Senales (South Tyrol, northern Italy); this led to the exposure and discovery, on September 19, 1991, of a mummified body by two German mountain hikers (Henn 1992; Zissernig 1992). A few days after the recovery and transport of the corpse to Innsbruck University, it became clear that the male mummy was that of a Neolithic individual

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Fig. 2 Detail view of the Iceman’s face (© Südtiroler Archäologiemuseum/Eurac/ Samadelli/Staschitz with permission)

that had lived several thousand years ago (Fig. 2). The body was found very close to the route from Val Senales/Schnalstal in the south to the Ötztal in the north, at an altitude of approximately 3200 m.a.s.l. The place where “Ötzi” was found (which was eventually shown to lie approximately 90 m south of the Austro-Italian border on the Italian side) suggests that the Iceman was on his way north. The mummy was located within a small rim of the surrounding rock. This geographical peculiarity seems to have protected the body from being incorporated into the major moving part of the glacier, whose movement would otherwise have caused serious to complete destruction of the remains. Because of the position in which it was found, it is currently assumed that the cadaver remained in the place where he fell several thousand years ago. In addition, close to the Iceman, a great deal of his equipment was found, including most of his clothes and several pieces of his “technical equipment.”

Dating of the Mummy Following the transfer to Innsbruck University and initial macro-morphological investigations, several samples from both the mummy and his equipment (organic compounds) were radiocarbon dated. Repeated use of calibrated radiocarbon dating in various specialized laboratories showed that the Iceman lived 5,100–5,350 years before present (i.e., c. 3100–3350 BCE) (Bonani et al. 1992). This indicates that the

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Iceman lived in a time period at the early dawn of civilization, as seen in several high cultures such as Mesopotamia or ancient Egypt.

Individual Age The Iceman’s age at death was determined histologically on the basis of the remodeling rate of cortical bone (Gaber and Künzel 1998). This analysis indicated he was between 35 and 50 years old with the most likely age as 46 years old. This is in accord with the degree of tooth abrasion (Seidler et al. 1992), although the radiological investigation of the skull sutures suggested a younger adult age (Seidler et al. 1992). The currently “accepted” age of the Iceman is around 45 years old (+/ 5 years).

Anthropological Examination, Body Size, and Stature When the body was found, the mummy had a body length of 158 cm and a weight of 13.3 kg (Seidler et al. 1992; Gaber and Künzel 1998) (On a re-examination in 1992, Bernhard determined the precise body height as 1590 mm (Bernhard 1992). Later extensive anthropological evaluation of the body and of the CT-scan dataset (Ruff et al. 2006) concluded that Iceman’s stature was around 158 cm, thus suggesting that no major post-mortem shrinkage had occurred. This data contrasts to later nonproven statements of a body length of 154 cm on first examination and 160 cm on recalculation by bone length determination (Zink 2016), which may suggest mild postmortal shrinkage. The estimated living body weight was calculated at around 61 kg (Gaber and Künzel 1998). The body has been regarded as short, but relatively wide or stocky, suggesting high mobility over rough terrain. The skull provides a hyper-orthognate facial form; cranial measurements and the cranial capacity were well within known Neolithic Age ranges (Seidler et al. 1992). The wear of several of the Iceman’s tools strongly suggest that he was right-handed (Wierer et al. 2018).

Type of Mummification Despite the major reduction in body weight – most obviously due to postmortem dehydration – the Iceman is not a “dry mummy,” but still rather a “wet mummy,” i.e., the body still contains significant amounts of fluid. Therefore, this “ice mummy” represents a particular type of natural mummification. This peculiarity is also reflected by the fatty acid composition and its alterations in the Iceman. Accordingly, Mayer et al. (1997) and Makristathis et al. (2002) identified the subcutaneous fat tissue of the mummy to reveal changes that represent a partial transition into adipocere with incomplete transformation. This is well in line with the excellent preservation status of the body, including the superb conservation of various internal organs.

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The exact mechanism of mummification of the Iceman still remains unclear, in particular whether the corpse was covered immediately after death with snow and/or ice or if it was first exposed to open-air drying with subsequent freezing. Similarly, it is still not known whether the Iceman was stored within the ice for the whole period of approx. 5,200 years or if there were some intermediate periods of free exposure. However, recent investigations on the highest South Tyrolean glacier (Ortler/Alto dell’Ortles) (Gabrielli et al. 2016) suggests that all Alpine glaciers were continuously covered by ice for the last approximately 7,000 years. It is clear, however, that the body must have “dried out” superficially. Well in accord, a histological analysis of a skin sample (Nerlich et al. 2003) shows complete loss of the superficial epidermis and some adipocere. Furthermore, a strand of hair and several fingernails were found besides the mummy but no longer attached to it (Seidler et al. 1992). Finally, an electron microscopic investigation of the Iceman’s connective tissue by atomic force microscopy (Janko et al. 2010) showed very wellpreserved collagen fibers with slightly reduced mechanical fiber resilience, suggesting some type of “freeze-drying” of the tissue. This is well in line with previous immunohistochemical studies that identified specific epitopes of various collagen types to be preserved, although with reduced staining intensities (Wick et al. 2001).

Provenance and Genetic Origin of the Iceman The analysis of the genetic material of the Iceman significantly profited from recent progress in ancient DNA (aDNA) analysis. Initial studies in the 1990s (Handt et al. 1994) were restricted to fragments of the mitochondrial aDNA allowing the Iceman to be placed with contemporaneous Central to Northern Europeans. A further study by Rollo et al. (2006) extended this work and connected the Iceman to the haplogroup K1, which is focused in modern Southern Alpine populations, suggesting a local maternal origin of the ancient individual. This was confirmed (and extended to haplosubtype identification to K1f) in subsequent studies (Ermini et al. 2008; Endicott et al. 2009). Meanwhile, technical progress allowed the analysis also of paternal Y-chromosomal aDNA including common autosomal SNP variations (Sikora et al. 2014). Thereby, a genetic relationship was established between the Iceman and recent and ancient Sardinian populations, suggesting a paternal ancestry in a common European population that may have been conserved in isolated communities such as in Sardinia or Corsica. Further whole-genome investigations (Keller et al. 2012) confirmed the common relationship between the Iceman and present-day inhabitants of the Tyrrhenian Sea region. Additional biological information of these investigations indicates individual characteristics such as brown eye color, blood group 0, and lactose intolerance, such as enhanced genetic risk for arteriosclerotic vascular disease (see below) (Keller et al. 2012). These molecular genetic studies parallel the investigation of various stable isotope compositions that provide additional information about the Iceman’s origin.

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Strontium and lead isotopes suggest a local origin on the basis of local isotope mapping (Müller et al. 2003). Accordingly, the Iceman spent his infancy and childhood (up to the end of his first decade) in the lower Pustertal/Val Pusteria or the upper Eisacktal/Valle Isarco, which are regions relatively close to the final place of retrieval. In his further life, the individual might have migrated during adulthood locally, but not further. These findings are further corroborated by the content of the Iceman’s intestines where faunal residues, in particular pollen, also indicate South Tyrolean origin. Accordingly, the Iceman spent his last 48 h or so on the Southern slope of the Alps. The investigation of the Iceman’s belongings also point to local South-Alpine origin of various goods and items (Wierer et al. 2018). Likewise, the Iceman’s toolkit displays typological characteristics of the Northern Italian tradition. The copper material of his axe and flint stones further suggest a close relationship to the socalled Remedello-culture (Lippert et al. 1995), a copper age population that has been termed according to its main location close to Brescia/Northern Italy. In consequence, the availability of those items also strongly supports a Southern Alpine origin for the Iceman. The material for his flint stones and the stone dagger obviously came from the Lessinian mountains and the Lombard basin, which are north/ northwest of present-day Verona (Wierer et al. 2018).

Nutrition of the Iceman Extensive studies were conducted on the Iceman’s nutrition based on several factors. Firstly, the abrasion of the teeth was interpreted not only to be a consequence of age but also of a nourishment containing hard substances, such as seen in early civilization individuals that used flour prepared in stone mills. The addition of small debris from those mills (but also other sources, such as in desert regions by sand, e.g., Egypt) (Nerlich et al. 2000) may lead to major abrasion of the teeth. Secondly, information of the Iceman’s nutrition came from a stable isotope analysis of a hair strand that contain signatures of the ingested diet. In this regard, especially stable nitrogen and carbon isotopes are of value. With respect to the Iceman, stable isotope investigation on hair in an initial analysis was interpreted as indicating a primarily vegetarian diet (Macko et al. 1999). This, however, was questioned in a reevaluation (Dickson et al. 2000) coming to the conclusion that the diet was omnivorous. This was confirmed by the investigation of the content of the lower intestinal tract (Dickson et al. 2000). Thirdly, there was the analysis of the content of the lower (Dickson et al. 2000) and upper (stomach, Maixner et al. 2018) gastrointestinal tract. The large bowel content revealed numerous herbs of different crops, especially ancient forms of wheat and barley (mainly “einkorn”), mixed up with various cereals, fibers, hairs, cereal glume fragments, pollen grains, etc. Besides the morphologic identification of the intestinal content, ancient DNA analysis also showed the presence of various animal DNAs, which could be identified as deer, ibex, and goat (Rollo et al. 2002). This confirms that “Ötzi” ate a mixed diet; he was not a vegetarian. Lastly, a very

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recent thorough analysis of the stomach contents (Maixner et al. 2018) detected not only typical animal muscle fibers but also fat-containing connective tissue fragments and plant particles from bran and glumes of the Triticum/Secale type. Molecular investigation further identified the animal material as Capra ibex and Cervus elaphus (red deer). The fat residue content was surprisingly high at 46 +/ 19% of the stomach content. The further identification of charcoal particles suggests further food processing such as by smoking. Beyond the direct and indirect investigation of nutrition and intestinal contents, there exists indirect evidence for direct or indirect disturbances of nutritional supply (or its metabolism). Of particular interest was the detection of several fingernails found close to the mummy. These showed three slightly lighter transverse lines (Mariani Costantini et al. 1994). Similar tissue features are known as so-called Beaufort lines. They indicate significant physical and/or nutritional stress. The lines can be attributed to distinct time periods and the Iceman’s nail revealed nutritional deficiencies at approximately 8, 12, and 16 weeks prior to his death. Thus, we have evidence that the Iceman suffered from at least three periods of absolute or relative malnutrition during the last period of his life. Beyond these features during his final weeks, there is some evidence for possible impairment (also of potentially nutritional supply) during the Iceman’s childhood and adolescence. The radiological examination revealed a few faint Harris lines in the proximal and distal metaphysis of both tibiae (Murphy et al. 2003). Unfortunately, no further analyses of these lines were undertaken nor reevaluated on subsequent CT scans, so nor the timing of their formation with respect to the Iceman’s age has been determined.

Evidence for Physical Activities The thorough analysis of the body also revealed evidence for physical activity. This mainly manifests in the skeleton, either by signs for articular wear (osteoarthrosis) or enhanced bone formation at the sites of muscular attachment to bones (enthesiopathies). As early as in 1992, Zur Nedden and Wicke described radiological evidence for mild to moderate degenerative changes in the right hip and the right ankle. Additionally, the vertebral bodies of the cervical and lumbar spine revealed degenerative changes. Subsequent examinations were summarized in 2003 by Murphy et al. (2003) who described, in their thorough radiological analysis, a reduction of the articular space of the right hip (the left hip was difficult to evaluate due to major post mortem changes that happened during recovery of the body from the ice). The coincident subarticular osseous sclerosis in the anterior joint, and slight osteophyte formation of acetabular and femoral sides of the joint, support the diagnosis of minor osteoarthrosis. Furthermore, the initial radiological investigations (zur Nedden and Wicke 1992) had also noted some calcification in the periarticular soft tissues of both knees, which were also attributed to degeneration.

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The further more precise evaluation also detected minor degenerative changes in the apophysis of the facet joints of the vertebral bodies of the cervical (C4-C5) and lumbar spine (L5) where a tiny osteophyte was seen at the superior endplate of this lumbar vertebral body, and minor apophyseal joint osteoarthrosis was detected. All other vertebral bodies – including the complete thoracic spine – were regarded as “remarkably normal with excellent mineral content” (Murphy et al. 2003) ruling out any major spondyloarthritic degeneration. The transitional lumbosacral segment additionally revealed a Schmorl’s node which represents a developmental, but not degenerative change. Recently, Kean et al. (2013) reinvestigated the mummy’s musculoskeletal findings. They assumed that the Iceman had suffered from various ligament injuries, which might have been treated medically by tattooing (see below). There was some speculation that rheumatoid diseases could be present but this has not been subsequently confirmed. Accordingly, the Iceman had undergone some minor osteodegenerative changes (at the age of c. 45 years), which were, however, restricted to the lower extremities and parts of the spine. Therefore, it is fair to assume that he was very mobile without any significant medical disorders to affect his life style or occupation. This is supported by the findings of normal cortical bone thickness and obviously unaltered mineralization (Murphy et al. 2003). These latter studies further indicate that: “the cortices of the femora, tibiae and fibulae were thicker than what is typically encountered in modern populations. The lineae asperae were well developed and prominent . . . the antero-posterior diameters of the tibiae and fibulae were much greater than the medio-lateral diameters . . . and suggest presence of powerful lower-extremity musculature.” (Murphy et al. 2003). These morphological features support the notion that the Iceman was a very active hiker, but not otherwise subjected to specific occupational activities or stress.

Infectious Diseases and Diseases of Internal Organs Since he was discovered, major interest has been on the identification of the Iceman’s health conditions. Currently the Tyrolean iceman is the most investigated ancient human corpse in the world. The studies indeed revealed several pathological conditions that may have impaired the Iceman’s life. In this respect it seems important to distinguish between the patho(physio)logical change and its potential implication on daily life in the Neolithic period. The latter can only be speculation about the consequence of the actual diseases the Iceman had. The pathological changes can be divided into several affected (“organ”) systems: the intestines, the lungs, the vasculature, skeleton and teeth, and finally the hair, plus systemic infectious diseases.

The Intestines The availability of a completely preserved intestinal tract, including stomach, upper and lower bowel, is unique. The investigation of the contents provides

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circumstantial information, part of which has already been described under the chapter of “Nutrition.” Beyond the mere composition, there is evidence for specific infections that may have affected the Neolithic man. Unfortunately, mummification and long-term storage render the Iceman’s tissues difficult to investigate microscopically. The only histological study that had access to various internal tissues (Hess et al. 1998) describes major adipocere formation, which is in contrast to subsequent studies on isolated tissues, such as skin (Nerlich et al. 2003, 2009), which was excellently preserved. Furthermore, the initial histological analysis described the usual paraffin embedding technique as “poor and of uncertain stainability” so that they had to use epoxy resin embedding which does not permit subsequent immunohistochemical studies. Similarly, Maixner et al. (2016) was unable to investigate stomach mucosa histologically and to identify specific structures, which may be important for the interpretation of molecular observations. In this respect, future studies, with adequately adopted technical procedures, may provide more information. Already in the very first studies on the intestinal content, numerous eggs typical of the whipworm Trichuris trichiura have been identified (Aspöck et al. 1996; Dickson et al. 2000; Fig. 3). This parasite results from poor sanitation and reduced hygienic conditions. The worms are ingested, then multiplicate within the intestines, and may lead to diarrhea. Interestingly, the Iceman had some “remedies” against intestinal parasites with him (see below).

Fig. 3 Egg of the intestinal parasite Trichuris trichiura such as that found in a sample from the Iceman’s intestines (colon) (reproduced with the kind permission of Prof. Aspöck; see Aspöck et al. 1996)

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Very recently, a further intestinal infection was identified by the presence of bacterial aDNA of Helicobacter pylori in the stomach contents (Maixner et al. 2016). Using extensive molecular analyses, the authors identified this bacterial species and subtyping indicated presently as an “Asian type” known strain. Furthermore, the analysis of the stomach contents identified pro-inflammatory proteins, including fragments of calprotectin, a protein typically associated with intestinal inflammation. Accordingly, there is evidence that the Iceman was not only a pathological bacterial carrier (approximately 90% of all infected individuals are asymptomatic), but indeed may have suffered from clinical symptoms, typically, an active gastritis. Furthermore, the identified strain is currently considered one of the more aggressive bacterial strains. In addition to the mere identification of this pathogen, the attribution of this infection to a particular (and currently termed) “Asian strain” proposes that a revision of the spread of Helicobacter during human history is warranted (Megraud et al. 2016). Finally, the Iceman suffered from three gallbladder stones (Gostner et al. 2011). Similar stones are frequently asymptomatic and therefore it remains uncertain whether the Iceman suffered from any symptomatic disease, which may has manifested as recurrent colic and/or biliary inflammation or jaundice.

Pulmonary Pathology In an initial histological study of the shrunken lungs Hess et al. (1998) reported only a transformation of the tissue into adipocere; therefore, it was non-informative. A more specific investigation (Pabst and Hofer 1998) on minute lung tissue samples, however, revealed, beyond the diagenetic changes, significant deposits of dark carbon-containing particles (anthracosis), like those typically seen as inhaled fine coal particles originating from open fires. These particles were, at least in part, associated with tiny birefringent crystals, such as in inhaled dust particles. In turn, however, the lack of tissue calcification reasonably rules out that the Iceman suffered from one of the most widespread infectious diseases of numerous ancient High cultures, e.g., in Egypt, tuberculosis. However, Murphy et al. (2003) reported on “pleural adhesions in the right hemithorax that prevented total collapse of the right lung.” Accordingly, the Iceman must have suffered from a period of pleuritic inflammation a considerable time before his death. The cause of the inflammation remains unknown.

Vascular Pathology The radiological studies provided clear evidence that the Iceman suffered from arterial calcification affecting several vascular locations (Murphy et al. 2003). These were both carotid arteries at the sella turcica (intracerebral location), the left common carotid artery in the neck, the distal aorta, and the right iliac artery. Interestingly, there was no clear evidence for calcification of the coronary arteries

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or in the peripheral arterial vasculature system, such as in the upper or lower extremities. Additionally, none of the several radiological/CT-studies describe calcification of small arterial vessels (microangiopathy). It is also of great surprise that the large vessels, in particular the thoracic and upper intestinal aorta, do not reveal major calcification. This points to a particular type of arteriosclerosis (of the media-type) frequently associated with chronic renal insufficiency. Whether this was already present in Neolithic times or not remains unknown. In addition to the physical evidence for focal calcifying arteriosclerosis, the metagenomic investigation of the Iceman (Keller et al. 2012) provided data suggesting a genetic predisposition to arteriosclerosis.

Non-traumatic Skeletal Pathology In addition to multiple signs of trauma (see below), the mummy reveals subtle changes indicating metabolic disturbance associated with injury. The histological investigation of cancellous bone shows impaired bone remodeling with disturbed ossification of small bone trabeculae suggestive of mild osteomalacia (Nerlich et al. 2005a). This disease results from either a lack or a metabolic disturbance of vitamin-D metabolism, and/or its precursors. In advanced stages there may be a loss of bone mass, which is, however, excluded in the Iceman by the radiological analysis (Murphy et al. 2003). However, the observation of mild osteomalacia on a microscopic level (Fig. 4) fits with the above described parasitosis. This in turn implies that the parasitosis must have been active, and thereby possibly also clinically relevant.

Diseases of the Teeth and the Oral Cavity Initial inspections of the Iceman’s oral cavity only revealed the aforementioned dental abrasion (obviously due to the nutrition), a diasthema (congenital gap Fig. 4 Histological image of a cancellous bone biopsy (hip); the sample shows an undecalcified preparation with hypomineralized lamellar areas of bone remodeling highly suggestive of vitaminD-deficiency (MGG staining, in blue: densely mineralization; in violet: hypomineralized bone,  400; Credits: A. G. Nerlich)

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between the upper incisors), and congenital lack of the third molars (Murphy et al. 2003). On careful reevaluation, several more pathologies have emerged including dental caries, periodontal bone loss (periodontitis), and dental trauma (Seiler et al. 2013). These observations indicate considerable consumption of carbohydrates (in accord with the nutritional studies, see above) and poor oral hygiene. The Iceman possibly suffered from painful dentition. However, despite these diseases, and the significant abrasion, there is no evidence for (premature) tooth loss, which is highly prevalent in contemporaneous high culture populations such as Egypt where this is found at much younger ages (Nerlich et al. 2000). Also, there is a complete absence of so-called dentogenic abscesses, resulting from advanced abrasion and/or carious lesions, which also prevail in other ancient populations.

Abnormalities of Hair During the extensive archaeological examination of the Iceman’s resting place several items, including personal belongings, were found near the mummy; among these were several bundles of hair. Most of them could be identified as deer and goat; however, some were indisputably attributable to human scalp hair (Wittig and Wortmann 1992). These were excellently preserved and found to be dark-brown in color. In addition, the authors detected Trichorrhexis nodosa at a considerably high number. This disorder is characterized by thickening or weak points (“nodes”) that cause the hair to break off easily. It may have a genetic basis, but appears to be precipitated by environmental factors, or occasionally, hormone disorders, such as hypothyroidism. At present, there exists no understanding whether the observed hair anomalies had any pathological association/effect or not. In addition, Brothwell and Grime (2002) analyzed the Iceman’s hair for the presence and distribution of heavy metals. These authors report enhanced levels of copper and arsenic in the Iceman’s hair; with arsenic being a common impurity to copper ore. They concluded that the Neolithic individual was involved in copper working. However, one cannot assume that these heavy metal constituents indicate toxic contamination and therefore any health impairment.

Systemic Infectious Disease Finally, the metagenomic analysis of the Iceman showed genetic material suggestive for a systemic infection by Borrelia burgdorferi (Keller et al. 2012). This data interpretation, however, has recently been questioned. On reevaluation of the published data, Ames et al. (2013) identified a nonpathogenic bacterial strain of Borrelia. It is therefore questionable whether the Iceman may have suffered from borreliosis (Lyme disease), a chronic disease with various clinical manifestations, such as recurrent painful articular episodes, fatigue, paralysis, heart disease, and many more.

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Potential Therapeutic Treatment Besides the paleopathological identification of various diseases, the thorough investigation of the mummy provides evidence for “therapeutic” interventions that might have been used by the Iceman to obtain relief. It was a surprise that the archaeobotanic examination of the Iceman’s belongings revealed two small specimens of a particular fungus, attached by two small leather ropes to each other (Capasso 1998) (Fig. 5). This was identified as Fomitopsis betulina (formerly: Piptoporus betulinus, Birch polypore), which has known anti-parasitic (anti-helmintic) properties (Pleszczynska et al. 2017). In other words, this fungus represents a recently identified remedy that destroys intestinal parasites when an extract of the fungus is consumed. Accordingly, we can assume that the Iceman had an effective remedy that may have allowed him an occasional removal of his parasitic load. Nevertheless, the aforementioned mild osteomalacic osseous reaction suggests that the treatment was not fully protective, or even completely preventive, for the parasitosis. In addition, the aforementioned extensive investigation of the Iceman’s gut identified the presence of the bracken fern Pteridium aquilinum (Oeggl et al. 2007; Maixner et al. 2018) that contains carcinogenic, but also anti-helmintic components which may have been used by the Iceman following detoxification of the fern, e.g., by extensive washings or other pretreatments. So, possibly the Iceman used also this substance as a further remedy against his parasitic infection. Beyond these potential remedies for treatment of his assumed intestinal problems, the Iceman contains numerous tattoos at various parts of his skin, which have repeatedly been associated to “medical treatment” of joint problems (Capasso 1993; Dorfer et al. 1999). Recently, the application of multispectral light to the mummy’s surface increased the number of proven tattoos to 61, which can be divided into 19 regional groups (Samadelli et al. 2015). Most tattoos are present as short parallel lines with those at the right knee and left ankle forming a perpendicular cross. The location of several of

Fig. 5 The two pieces of the birch polypore (Fomitopsis betulina) that were found in the Iceman’s belongings (© Südtiroler Archäologiemuseum/Harald Wisthaler, with permission)

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these tattoos are associated with osteodegenerative lesions, such as the lumbar spine and the right hip and knee, and several of the so-called acupuncture meridians, suggesting therapeutic application resembling techniques of modern-day acupuncture. These included meridians associated also with the treatment of intestinal problems, including gallbladder, liver and bowel. Although these theories are very intriguing, their interpretations have to be taken with great care: besides the lack of any evidence that such a type of well-developed “therapy” by tattooing/acupuncture really was perceived at that time period, it is noteworthy that the osteoarthritic lesions of the lumbar spine are minor; at the right knee only some soft tissue calcification has been detected, but no degeneration of the joint is present. The left hip has been so heavily destroyed during the recovery of the mummy that no statement can be made on any joint changes or on the overlying skin tattoos. Furthermore, the fact that the Iceman obviously was highly mobile (see below) argues against significant restrictions by joint/vertebral disease.

Sequelae of Traumatic Injuries Finally, the Iceman’s mummy reveals numerous sequelae of traumatic injuries, part of which can be attributed to the robust maneuvers needed during the recovery process from the ice (detailed listed by Murphy et al. 2003). Likewise, the left humerus was broken because the corpse did not fit into the transport coffin. These accidents have to be separated from several injuries that may have happened after his death when the remains became encased in the glacier (also enlisted by Murphy et al. 2003). Besides these clearly, or at least very likely, post-mortal injuries, the body shows trauma of various ages of recovery.

Old Healed Trauma The CT scans showed completely healed rib fractures of the left 7th and 8th ribs. These were located along the posterior axillary line and were arranged linearly so that the Iceman may have suffered from one or several episodes of thoracic trauma during his lifetime, although the pattern of the fractures favors a single event (Murphy et al. 2003). All these lesions were well-healed indicating they happened long before his death. Similarly, an isolated defect in the left little toe that presented with a faint marginal sclerosis, points to an old-healed lesion. Type and morphology suggest the sequela of healed frostbite (Murphy et al. 2003).

Trauma Acquired Several Days Before His Death The detailed examination of a small hand wound (Fig. 6) led to the identification of a deep soft tissue injury with signs of significant bleeding into the connective tissue along with a superficial defect of the adjacent bone of the second right metacarpal

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Fig. 6 Wound at the right hand of the Iceman (© Südtiroler Archäologiemuseum with permission)

Fig. 7 Histological image of the hand wound showing bleeding residues (yellow) with interspersed hemosiderin-positive macrophages (blue) (Prussian blue staining,  600; A.G. Nerlich)

bone (Nerlich et al. 2003). Since we had identified histologically transformation into hemosiderin-containing macrophages (Fig. 7), an event that requires active transformation of cells – this lesion must have happened several days before the Iceman’s death. Recent studies show minor hemosiderin formation to usually occur after more than 3 days (Nerlich et al. 2005b) indicating the age of this lesion.

The Mortal Arrow Shot In 1998, with the transfer of the mummy to South Tyrol, a reexamination of the body including a CT-scan revealed the presence of a foreign body in the left chest above the second rib (Gostner and Egarter Vigl 2002). Because of its shape (21  17 mm) and radiodensity, it was clear that the foreign body represented an arrowhead, most probably made of flint (Fig. 8). A small skin wound was also detected on the Iceman’s back, on the dorsal side of the left shoulder, which was identified as the

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Fig. 8 CT-scan 3Dreconstruction showing the foreign body in the left axilla with the typical radiomorphological characteristics of an arrowhead (radio-dense) (© Südtiroler Archäologiemuseum/ Regionales Krankenhaus Bozen, with permission)

entry point of the arrow (Fig. 9). A small puncture biopsy of this entry point revealed signs of fresh bleeding with excellently preserved red blood cells, but without evidence of remodeling or hemosiderin formation (Nerlich et al. 2009). Therefore, this entry point proved to be a perimortal wound. Furthermore, the close proximity of the arrow to large blood vessels of the arm, strongly suggests that any of those structures may have been affected, leading to extensive bleeding. This is further corroborated by the identification of a post-laceration pseudo-aneurysm of the left subclavian artery (Pernter et al. 2007). In consequence, this finding strongly suggested that Ötzi had died from severe bleeding following a lethal assault with an arrow, although the arrow shaft is missing.

Further Possible Perimortal Lesions On the CT scans, the otherwise very well-preserved brain revealed a heterogenous attenuation of signals that did not follow anatomical structures. Recently, two brain biopsies were taken under video-assistance (Maixner et al. 2013). Using a broad panel of techniques, these authors claim to have identified red blood cells and coagulation-associated proteins suggesting perimortem brain trauma. However, a critical evaluation raises serious concerns about these conclusions: blood is in brain tissue and is also abundantly present within blood vessels. The enhanced proteomic expression of various proteins usually requires reaction times beyond the perimortal period. Accordingly, it remains an open discussion whether the Iceman really suffered from massive skull trauma, possibly by falling on his face after the arrow hit, or if the changes are only postmortal taphonomic ones.

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Fig. 9 Entry point of the arrow at the left scapula (© Südtiroler Archäologiemuseum with permission)

In addition, there are some further spot-like areas of dark coloration of the skin on the Iceman’s back in the paravertebral region, which have been investigated histologically and immunohistochemically (Nerlich et al. 2009). Out of the several spots, only one provided minor suffusion that may have occurred perimortem. The other spots were nonreactive.

Possible Scenarios of the Iceman’s Life and Death The extensive investigation of the mummified body offers considerable information about the life, diseases, and death of the individual. Although several items are still unsolved, plausible scenarios can be established:

The Iceman’s Life Genetic analyses and isotopic investigations provide circumstantial evidence that Ötzi was born and grew up on the Southern Alpine slopes, possibly in the Passeier or Eissack Valley, which are close to the final resting place of the mummy. He was part of the local population. The Iceman was highly mobile, which is clearly manifested in his lower extremities’ musculature and their well-developed attachment sites to the bones. His belongings comprise several valuable items, including a copper axe attributable to the South Alpine Remedello culture. The flint stones and the arrow heads in his possessions came from the Lessinian mountains north of Verona. Although we have no specific information on the Iceman’s earlier years, several, although faint, Harris lines suggest several periods of ill health when he was younger. At the time of his death, the Iceman was well nourished, obviously eating typical local products. The contents of the stomach and the upper and lower bowel further

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indicate high mobility, since it contains three meals that can be attributed to various levels of altitude. The nutritional composition was balanced with faunal and animal products; dental caries indicates considerable carbohydrate consumption. The presence of gallbladder stones is well in line with mild to moderate arteriosclerosis, which may result both from genetic predisposition as well as cholesterol-rich nutrition. The Iceman also carried intestinal parasites and had Helicobacter pylori in his stomach that may have caused health problems. The parasitosis seems to have been present for some time since the Iceman showed mild osteomalacia. He had potential therapeutic herbal remedies with the birch polypore and bracken fern, in his belongings. All other scenarios, particularly as to his profession and the possible reasons for his frequent travels, are highly speculative and shall not be pursued here.

Ötzi’s Final Days – The Scenario of His Death The analyses on the Iceman’s intestinal content allow a reconstruction of the final 48 h or so of Ötzi’s journey. The composition of the intestinal content suggests that this took place during spring or early summer (Oeggl et al. 2007). Accordingly, he moved from a higher altitude to a lower one, and then up again to the place where his life ended. He took his last meal only shortly before his death. Before these ascents and descents, the Iceman must have suffered from a severe wound to his hand, which he had acquired around 3–5 days before his death. The type of lesion is compatible with a defensive movement against a stabbing weapon. Having arrived at the glacier, at approximately 3,200 m altitude, the Iceman was attacked by an arrow that hit him from behind. The arrow hit his chest lacerating the left subclavian artery. This led to extensive subacute bleeding, which finally may have caused his death. Possibly, the Iceman fell onto his face receiving severe brain trauma with terminal bleeding. Since the arrow shaft is lacking it has been hypothesized that the attacker tried to withdraw the arrow but only managed incompletely. Anyway, the Iceman died at the glacier where he was naturally mummified after being covered by ice and snow until the unusual recession of the glacier uncovered his body.

Final Remarks Although there have been numerous and extensive scientific examinations performed on the Iceman, further investigations will hopefully extend our knowledge on life and disease in the European Neolithic era. In this regard, this mummy’s completeness and preservation, and all his belongings, offer a unique and extraordinary insight into the life and times of a prehistorical man from over 5,000 years ago. Acknowledgments The authors are indebted to Prof. Simon Donell for language correction of this manuscript.

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . What Is a Mummy? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Spontaneously Mummified Corpses: Some Biological Considerations . . . . . . . . . . . . . . . . . . . . . . . . Anthropogenic Mummification of Human Corpses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Some Historic Aspects of Mummification in Post-antique History: The Origin of Church and Crypt Mummies in Mediaeval to Modern Periods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies in Crypts and Catacombs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Italy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Germany . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Austria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Czech Republic . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Hungary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lithuania . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Other Countries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions and Outlook . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

There exist numerous crypts and catacombs in churches and cemeteries that harbor mummified human remains. Due to burial customs these are most frequently detected in European countries, mostly as the result of spontaneous (natural) A. G. Nerlich (*) Institute of Pathology, Academic Hospital Munich-Bogenhausen, Munich, Germany e-mail: [email protected] R. Bianucci New Mexico Health Enhancement and Marathon Clinics Research Foundation, Albuquerque, NM, USA Warwick Medical School, Biomedical Sciences, University of Warwick, Coventry, UK Legal Medicine Section, Department of Public Health and Paediatric Sciences, University of Turin, Torino, Italy © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_20

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mummification. Beyond those mummies, occasionally artificial mummification has led to permanently preserved bodies. These were performed as either dry or wet type of embalming. All mummified human bodies may represent important information about life, living conditions, diseases, and causes of death in historic populations; they represent a “bio-archive.” Although many mummies seem to be present in numerous locations, only a small number of them have been as yet investigated scientifically. This scientific analysis has been performed by various techniques: anthropological examination, CT scans and/or X-rays, histology and isotope analysis, molecular studies for human and microbial DNA, paleobotany, and many more. In this chapter, the available observations from numerous locations in different countries have been compiled. In most instances, only data from series of mummified bodies have been collected; single mummies have only occasionally been considered. The series come from Italian and German churches, several findings from Austria, the Czech Republic, Hungary and Lithuania, and isolated cases from almost every country. Keywords

Crypt · Catacomb · Interdisciplinary study · Wet embalming · Dry embalming

Introduction Most people may associate the term “mummy” only with preserved human bodies from arid desert zones of the earth, such as the Sahara, the Gobi, the Atacama Deserts. Less well perceived is the fact that numerous mummies are also present in crypts and catacombs of European, and occasionally American and Asian churches and/or cemeteries. Finally, several bodies of “important individuals,” even from recent times, have been preserved as mummies, mostly for political reasons. This holds particularly true for the human remains of important politicians such as Lenin in Russia, Ho Chi Minh in Vietnam, or Maria Eva (Evita) Duarte de Perón in Argentina (Reiblich 2015). Until now the crypt and catacomb mummies have only been described occasionally, and rarely in a systematic way. One of the few exceptions is the Joseon Dynasty mummies (South Korea) that have been very thoroughly investigated; they are described in detail by Dong Hoon Shin and coworkers in a separate chapter. Beyond these examples, there exists, unfortunately, up to now no registry of important findings. However, few researchers have tried to compile this information and so it remains incomplete. Likewise, the doyen of paleopathology, Arthur C. Aufderheide, reported in his book “The Scientific Study of Mummies,” a collection of mummies from all over the world (Aufderheide 2003). This now requires substantial updating. This review does not fill the gap completely but may stimulate further work in this important field of research. Although most of these mummified corpses are more recent by far than those from ancient Egypt or from the Peruvian and Chilean deserts, they still represent a highly interesting bio-archive of their respective time periods and areas. Before we can go in detail in a more systematic manner, we have to discuss the origin of mummified corpses plus a brief overview of the history of crypt and

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catacomb mummies. This is necessary, since there are two different ways crypt mummies can be formed; spontaneously, i.e., due primarily to particular microclimatic conditions without human agency or through a formal embalming process.

What Is a Mummy? This question is not so easy to answer, since there exists a sequence of tissue preservation in dead bodies. At one extreme is preservation of residues without soft tissue that includes the skeleton (bone including teeth, but otherwise no soft tissue), to the other extreme where the body is intact and complete, but mostly shrunken with not only preserved skin, tendons, ligaments, joints, bones and teeth, but also various internal organs, musculature, and connective tissues. Between these two extremes, there are many potential intermediates, i.e., only the preservation of the soft tissue of a particular body region together with otherwise skeletonized remains, or the preservation of only selective soft tissue components, such as only skin or tendons and ligaments but no other soft tissues or internal organ remnants. In actual practice, crypt and catacomb mummies are always more than a mere skeleton (although within a crypt there may exist numerous skeletons between more or less complete mummies) and complete mummies are mostly found in either very particular places with unique environmental microconditions (in spontaneous natural mummies), or exceedingly fine preservation in artificial mummies due to the excellence of the embalmer. The scientific value of mummies as bioarchives, is of course very dependent on the amount and type of tissue present. In order to overcome any bias with the comparison of mummies of very different completeness and preservation, Aufderheide and colleagues (Wittmers et al. 2011) provided a scoring system to evaluate an individual observation. Furthermore, recently, a similar soft tissue score has also been presented for CT-analyses by Panzer et al. (2015). In the future and in order to make observations and results more transparent and comparable, all researchers in the field are urged to use such scoring reference systems when they describe their material.

Spontaneously Mummified Corpses: Some Biological Considerations The usual decay of human cadavers is the result of an endogenous dissolution of cells and tissues by the liberation of degrading enzymes and the destructive action of microbes, particularly bacteria, via the activation of degrading enzymes, but also the destruction of human tissues by parasites, etc. Under normal circumstances, every human (and animal) individual degrades at the molecular level as soon as the regular function of the circulation/metabolism of living cells stops. Every process that blocks, or even only retards, the destructive action of endogenous enzymes or bacteria, parasites, etc., may prevent tissue decay and thereby may

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promote spontaneous mummification. Since the endogenous and exogenous destruction systems are dependent on humidity, the deprivation of fluid therefore may lead to the conservation of cadavers. Another important factor for major decomposition processes is oxidation, and thereby the presence or absence of oxygen. This removal of fluid may come from a dry, arid climate, but may also be the result of a continuous air stream that steadily reduces the humidity and, possibly even more importantly, removes humidity from the corpse’s surface. In consequence, a significant constant airflow in a crypt, with constant air temperature that avoids water condensation, may dry a human body more rapidly than it is destroyed. This usually requires (at least partially) open (or ventilated) niches or coffins. Cold room temperature may additionally retard destruction since most enzymes and bacteria are more active under warm than cold climatic conditions. Further factors that may promote spontaneous mummification are the material that surrounds the dead body: humic acids, which come from either wooden chip fillings of the coffin or the wooden coffin itself, may also interfere with enzymes and bacterial growth. Accordingly, even airtight coffins with such a filling can produce a mummy by blocking enzymatic destruction and adsorbing fluid from the body. Furthermore, oxidative processes may stop when oxygen is completely consumed in such an airtight sarcophagus. Finally, individual factors, such as size or amount of human body fat may accelerate or retard the process of human decay and may be relevant for body conservation, e.g., by the formation of adipocere: a special transformation of fat tissue into a stable and degradation-resistant form. Taking all these factors into account, even in the hot and usually more humid zones of the earth, spontaneous mummification of humans may occur (Figs. 1 and 2).

Anthropogenic Mummification of Human Corpses Besides the artificial mummification that has been practiced in various cultures, such as ancient Egypt, the Canary Island (Guanches), and some Southern American cultures (including the Chinchorros) (Aufderheide 2003), the artificial conservation of cadavers has a long tradition in Western European cultures. However, embalming was restricted to the wealthy part of the population, mostly to the aristocracy, some clerics, and (particularly in later periods of Modern Times – beginning with the eighteenth century) to tradesmen and (wealthy) citizens. As will be discussed subsequently, the technique of artificial mummification developed over the centuries. Generally, it has to be divided into dry and wet embalming techniques, depending on differences in the methods and resources used. The aim of anthropogenic mummification was the preservation of the dead body, either for logistic, political, or religious reasons. The principle of the ancient Egyptian embalming technique was, firstly, the removal of the major internal organs (including the brain) knowing that those organs promote endogenous (enzymatic) and exogenous (microbial) destruction, followed by an enhancement of desiccation (by the application of the chemical natron), and finally, the sealing of the body

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Fig. 1 The miraculous history of St. Magnus (Basilica St. Mang, Füssen, Germany; seventeenth century). The undestroyed body of the Saint was found several decades after his death, possibly a spontaneous natural mummification within the church crypt? (Photo: A. G. Nerlich)

surface using chemicals such as bitumen, oils, beeswax, and linen bindings (Ikram and Dodson 1998). This type of embalming, termed dry embalming, was principally started in the High Mediaeval period (Magnus 1839). However, there were some differences from the ancient Egyptian practice. Likewise, the embalmers did not use natron to enhance desiccation. This may be because natron was naturally found in Lower Egypt but was not readily available in Europe; as was the same reason applied to for bitumen, or other tar products. This was compensated for by the use of surface “balms” containing alcohol, oils, and extracts of various herbs; these may have also been used by the ancient Egyptians. Finally, the emptied body cavities, which were occasionally filled by the ancient Egyptians with textiles and/or natron packages, were covered or filled with numerous herbs, roots, and several more natural products that have antiseptic (antimicrobial) function in European practice (Charlier et al. 2013; Bianucci et al. 2016). Over time, and especially with the discovery of blood circulation by William Harvey, a significantly different method of embalming came into practice. Termed “wet embalming,” it used the vascular system as the route for the introduction of

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Fig. 2 1781 Copper plate engraving by the Francesco Danieli illustrating the mummified corpse of Emperor Frederick II Hohenstaufen (1194–1250) at the first exhumation (Danieli 1874)

fixative fluids. These fluids thereby disperse throughout the body, from tip to toe. Mostly heavy metal solutions, such as arsenic, mercury or lead, were used as fixatives. These metal compounds have both a strong antimicrobial and tissue stabilizing effect (Magnus 1839; Johnson et al. 1990). All attempts to preserve mummies are accordingly dependent on the type of embalming, i.e., in dry mummification the ongoing exclusion of fluid preserves the tissue. In wet embalming the opposite occurs, the embalmed body must be continuously treated with the fixative, which may be diluted, washed out, or destroyed by chemical reaction, so that it may lose its fixative potential.

Some Historic Aspects of Mummification in Post-antique History: The Origin of Church and Crypt Mummies in Mediaeval to Modern Periods While the conservation of human bodies was a constant practice in several ancient cultures, probably most widely spread in ancient Egypt, the active conservation of human bodies stopped in the European–Mediterranean regions with the rise of the Roman Empire, and later on, with the advent of the Christianity; although ancient Egyptian Christians, for example, still continued to perform embalming during the Roman Period. The main reason for the change within the Roman Empire was the

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Roman cult of cremation of the dead. While Christianity did not support the destruction of the dead human body, its belief in the transcendence of the human soul to heaven, and the loss of meaning of the human body in the afterlife, stopped all efforts to conserve the cadaver. In consequence, there was no general custom to practice human body conservation in Mediaeval Europe (Johnson et al. 1990). Following the disappearance of ancient Egyptian embalming practices in Greece and the Roman Empire, only the Early Arabic world had retained the knowledge of how to conserve cadavers. There is now scanty evidence that embalming was practiced during the Merovingian (Bianucci et al. 2016) and Carolingian (Schleifring et al. 2019) periods in Mediaeval Europe. However, with the rise of travelling, at least during Crusades and the beginning trade expeditions beyond Europe, the necessity to transport dead human bodies, or at least some parts of them, over long distances required artificial preparation (Brenner 2014). Particularly, during the crusades of the twelfth till fourteenth century, high ranked individuals that had died in the Holy Land (or on their way to the Near East) were more and more transported back to their home countries, either to verify the death of an important person, or to provide a physical memory on an elite individual. Initially performed as the Mos Teutonicus (Charlier et al. 2020a, b), i.e., the boiling of the dead body in order to separate the skeleton from soft tissues and transport it over long distances and during poor climatic conditions. Inner organs were removed prior to boiling and were treated with aromatic plants as preservatives; this raw type of “conservation” was banned in 1299 by Pope Bonifatius VIII. Although this ban was not very strictly kept, the meaning of the Mos Teutonicus rapidly declined (Magnus 1839; Johnson et al. 1990). In consequence, the need for alternative ways to conserve a cadaver increased. The contact with the Near East opened the way to transfer ancient medical knowledge to European universities including the method of cadaveric conservation such as used by ancient Egyptians. Likewise, already by the thirteenth to fourteenth century, it was generally accepted that the removal of fluid/humidity from the dead body was the most essential factor during embalming. The Persian physician Rhazes (865–925 CE) had developed techniques for the preservation of dead bodies (Magnus 1839; Schmitz-Esser 2014). Since several ingredients were difficult to be procured, he modified the ancient Egyptian method by use of an alcoholic balm for the surface treatment instead of the bitumen application. Also, the body cavity treatment was modified by the application of various herbs and root instead of filling with natron. The transfer of the Arabic-Persian knowledge to Central Europe lead to the conservation of high ranked peers, such as it has been exemplarily documented for Gertrud von Hohenberg (around 1225–1281 CE), wife of the Roman-German King Rudolf von Habsburg and founding mother of the Habsburg dynasty in the Holy Roman. Gertrud died in 1281 in Vienna where she was embalmed. Her subsequent transport to Basle took approximately 4 weeks. Since it was planned to expose the body before the official inhumation in Basle, the embalming procedure was essential. Unfortunately, we have no report on the success of the conservation (Nerlich et al. 2019).

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A second and also exemplary case is that of Emperor Friedrich III (1415– 1493 CE) who had died in Linz/Upper Austria 2 months after a leg amputation due to severe arteriosclerosis. Since the Emperor had to take leave from his country, the embalmed body traveled for several months through parts of his country until he was buried in Vienna, together with his amputated leg (Nerlich et al. 2019). The Renaissance, with the rediscovery of antique knowledge, also facilitated the handling of dead bodies and the embalming techniques (all of the type of dry embalming; see above) developed. Likewise, the famous French surgeon Ambroise Paré (1510–1590) embalmed several members of the French royal family, including Kings Henri II, III, and Charles IX (Magnus 1839; Nerlich et al. 2019). He copied the ancient Egyptian method by removing all internal organs, including the brain (via a circular opening of the complete skull), but added to this technique by also incising deeply into the musculature and soft tissue of the extremities. He then filled the body with numerous herbs, roots, and faunal ingredients. The skin was treated with turpentine and etheric oils then tightly wrapped in linen bindings. His method was applied all over the European continent (Penicher 1699; Hawlikvan de Water 1989). Likewise, King Henri IV of Navarre, the first Bourbon king of France, was embalmed using Pare’s method (Fig. 3). During the French Revolution when all royal tombs at Paris were plundered, his body was kept in such a perfect condition that the mummy was exposed for some time to the public. Most

Fig. 3 King Henri IV on his deathbed in 1610. (Engraving by R. Dunkarton, Wikimedia. commons.org, GNUFDL)

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interestingly, in 2010, Philippe Charlier and colleagues succeeded in positively identifying a very well-preserved mummy head to be that of Henri IV (Charlier et al. 2010). As already stated before, with the detection of the blood circulation by William Harvey (1578–1657 CE), the way was open for an alternative technique for embalming: the “wet embalming.” Frederik Ruysch (1638–1731 CE), a Dutch anatomist, was one of the first to develop a very successful method for conservation that used the injection of heavy metal-containing fluids through the vasculature in order to block endogenous as well as exogenous decay (Sucquet 1872). Although Ruysch never published the exact composition of his fluid, several examples of the excellence of his technique can be admired today, such as in the Kunstkamera of St. Petersburg/Russia, where some examples are on show (Fig. 4). While wet embalming provided excellent results in the small bodies of children and infants, the long-term results of adult embalming seem to have been far poorer. Additionally, the handling of the highly toxic fluids was not simple for the embalmer. Nevertheless, the technique was cheaper than dry embalming. In consequence, both dry and wet embalming were used for more than 200 years (Brenner 2014). During the eighteenth century numerous researchers tried to modify the wet and dry embalming techniques with differing results and unpredictable outcome. Nevertheless, the different embalming procedures were expensive so their application was mainly restricted to noblemen and members of the higher clergy. Furthermore, Fig. 4 Example of embalming by Frederik Ruysch: malformed newborn conjoined twins in the collection of the Kunstkamera/St. Petersburg. (Photo: A. G. Nerlich)

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trained embalmers, mostly surgeons together with pharmacists, were rare, and mostly only available in larger cities or royal or noble residences. Since both wet and dry embalming required numerous ingredients, only larger pharmacies were able to provide a ready stock of these substances, which was essential since the embalming procedure had to start as soon as possible after death in order to provide adequate results (Hawlik-van de Water 1989; Schmitz-Esser 2014). By the beginning of modern times (i.e., since the eighteenth century), the need for embalmers even increased, since then not only wealthy tradesmen and citizens increasingly died away from their hometowns, but also wanted to be buried close to their relatives. While the “academic” embalmers refined their techniques more and more, heavy metal conservations were abolished for forensic reasons; the use of heavy metal embalming solutions could have obscured poisoning (Gannal 1841). Finally, the detection of formaldehyde, a natural acid with strong fixative function, provided a technically adequate solution (Brenner 2014). In the meantime, for low cost burials, or for just the stabilization of a dead body for a short period, e.g., for a laying-out period or short-term transport, alternative techniques came up. In the eighteenth century England, carpenters and undertakers detected a novel commercial field, the so-called “tar-and-saw-dust-embalming,” i.e., filling a dead body with an adsorbent, such as saw dust, and/or the treatment with a chemical, such as tar, in order to stabilize the body for a limited period of time (Greenhill 1705; Litten 1991). This was a new commercial field. There was of course a serious debate by conventional embalmers, i.e., surgeons and pharmacists, as to the poor outcome of those measures. As will be shown later in this review, a simple dead body conservation was indeed undertaken in the eighteenth and nineteenth century burials, at least occasionally. The limitation of this low-cost embalming to the group of undertakers furthermore led to an almost complete ignorance of this practice in the scientific literature. Eighteenth and nineteenth century undertakers conveyed their knowledge only by oral instruction, not by written documents. Only the aforementioned complaint of academic embalmers witnesses this practice (Greenhill 1705) – which additionally may have been executed very differently. At least in Central Europe the low-cost embalming was applied since that period (e.g., in Italy: Selmi 1872). Finally, the development of wet embalming in the late nineteenth and beginning twentieth century by the infusion of formaldehyde solution into the vasculature was revolutionary (Brenner 2014). The method was technically very easy and the costs were moderate. The formaldehyde solution could be stored for a long time and thereby was readily available. Only in those cases where the vasculature was severely injured, such as in major arteriosclerosis, did the technique fail. Furthermore, it was realized that the formaldehyde fixation changed the bodies into a brown color, an effect that was counteracted by the addition of red colors (e.g., eosin) into the fixation solution. In those cases, where, for ethical reasons, a very high-level result in terms of a naturally looking “sleeping” cadaver was desired, modification of the formalin-technique was developed and applied. The most prominent modification was that of the Sicilian self-taught taxidermist and embalmer Alfredo Salafia (1869–1933 CE). He is renowned for having embalmed several corpses of prominent individuals (PiombinoMascali 2009; Piombino-Mascali et al. 2011). Salafia’s handwritten memoires were discovered in September 2007 (Piombino-Mascali 2009; third edition: 2012). Fourteen

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years after the discovery, no full-length independent review of the handwritten manuscript has been published, although extracts taken from Salafia’s original manuscript are presented in their original Italian (Piombino-Mascali, 2009; third edition 2012, pp. 52, 53, 54, 55, 57, 58) and English (Piombino-Mascali et al. 2009, pp. 356, 357). Piombino-Mascali (2009) reported that the memoires contained various chemical formulae including one that is believed to be the so-called “Perfection Fluid”: a mixture of zinc sulfate with chloride, glycerin, formaldehyde, and alcohol with salicylic acid (Piombino-Mascali 2009, third edition: 2012, p. 55). The exact procedure could not be reproduced on a modern corpse due to the unavailability of zinc salts (https://www. youtube.com/watch?v=phbUj4j6dJI&feature=youtu.be). To this end, the original Salafia’s embalming fluid was not used to treat the cadaver. No exact experimental reproduction of Salafia’s embalming procedure, in terms of use of the exact same formula, has been performed to date.

Mummies in Crypts and Catacombs Most crypt and catacomb mummies can be found in Europe and only occasional examples have been recorded in North and South America, in the Middle East, and in Asia. No reports exist on similar mummies in Africa and Australia/Oceania. However, a comparison of the reports between the compilation by Aufderheide (2003) and recent reports, strongly suggests that there may be many more cases that remain unknown. As detailed below and summarized in Table 1, within the European continent, the most extensive investigations have been performed in Italy, Germany, Austria, the Czech Republic, Hungary, and Lithuania with only relatively few examples in other countries, such as England, France, Scandinavia, Spain, and Portugal, and several other Eastern European countries. This may change in the future when crypts and burial places are more thoroughly investigated in other countries. Catacombs usually are large subterranean constructions of varying size containing labyrinthine corridors with or without cave-like enlargements. They were built to harbor usual burials, but were occasionally restricted to family members or specific social groups such as monks. In contrast, crypt burials are much more limited in space and mostly restricted to a specific family or a particular group. The crypts and catacombs contain mostly a variety of skeletonized and/or mummified remains. Both naturally mummified and artificially prepared mummies may be found side-by-side. Burials of high noblemen or members of the high clergy may be surrounded by less important or even unidentified members of their families, members of a social community, or foreigners. Of course, the results of recent investigations may provide more extensive and more precise results and observations than previous examinations. Furthermore, it is obvious that the investigation of mummified human remains requires much more interdisciplinary collaboration than the analysis of skeletons. In addition, it is evident that the complexity of dried human remains requires significantly more medical, and not only anatomical, knowledge than the examination of bones and teeth. We will now present major examples of catacomb and crypt burials. A tentative list of findings currently available is given in Table 1. This list contains, with very few exceptions, only series of individuals; isolated case reports are typically not

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Table 1 Summary of crypt and catacomb mummies series and their scientific work-up Time period Number 1599–1940 2081

Anthropol x

“Rosalia Lombardo”

1920

nr

Sicily

Local Church

1773–1858 26

x

Savoca

Sicily

Capuchin Church

1795–1876 17

x

Italy

Novara d. S.

Sicily

Local Church

1849–1873 6

x

Italy

Sicily

Local Church

1610–1900 48

x

Italy

Santa Lucia del Mela Comiso

Sicily

Local Church

18–19th

x

Italy Italy

Randazzo Gangi

Sicily Sicily

Santa Maria Chiesa di San Nicolò-Cripta dei Preti

1524–1863 135 18–19th 64

Italy

Altavilla Irpina Campania

Local Church

17–19th

>100

x

Italy

Naples

Campania

15–19th

31

x

Italy

Navelli

Abruzzo

San Domenico Maggiore Local Church

17–19th

>100

x

Italy

Rocca di Papa

Lazio

Local Church

16–18th

>100

x

Italy

Ferentillo

Umbria

18–19th

16

x

Italy

Florence

Tuscany

16–18th

39

x

Italy

Tuscany

11–19th

26

x

Italy

Badia Pozzeveri Arezzo

Tuscany

S. Stefano/ Museo delle mummie Mausoleo Medici San Pietro di Pozzeveri S. Francesco

14–18th

9

x

Italy

Urbania

Marche

Chiesa dei Morti 17–19th

18

x

Italy

Roccapelago

Chiesa S. Paolo

16–18th

13

nr

Italy

Venzone

Emilia Romagna Friuli

Cappella San Michele

14–18th

40

x

Country Italy

Location Palermo

Region Sicily

Designation Catacombe dei Cappuccini

Italy

dto

Sicily

Italy

Pirano

Italy

1

50

x x

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Mummies in Crypts and Catacombs

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Paleomolec Paleomolec X-ray CT Autopsy Histology Isotopes human microb Paleobotany References x x nr nr x nr x nr PiombinoMascali et al. (2011) x x nr nr nr nr nr nr PiombinoMascali et al. (2011) x nr nr nr x nr nr x PiombinoMascali et al. (2011) x x nr x x nr nr nr PiombinoMascali et al. (2011, 2015a) nr nr nr nr x nr nr nr PiombinoMascali et al. (2011) nr nr nr nr nr nr nr nr Bessa et al. (2018) x nr x x nr nr nr nr Gaeta et al. (2007) x x x nr nr nr nr x Lo Gerfo (2011) x nr nr nr nr nr nr nr Micciché et al. (2014); Beckett et al. (2020); Bianucci et al. (2021) nr nr nr nr nr nr nr nr Fornaciari (1998) x nr x x x nr nr nr Fornaciari (1998) nr nr nr nr nr nr nr nr Fornaciari (1998) nr nr nr nr nr nr nr nr Fornaciari (1998) nr nr nr nr nr nr nr nr Fornaciari (1998) x

nr

x

x

x

nr

x

nr

nr

nr

nr

nr

nr

x

nr

nr

nr

nr

x

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

x

x

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

Fornaciari (1998) Fornaciari et al. (2016) Fornaciari (1998) Fornaciari (1998) Petrella et al. (2016) Fornaciari (1998)

(continued)

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Table 1 (continued) Region Mecklenburg Mecklenburg Mecklenburg Bremen

Designation Schelfkirche Local Church Gruft Mirow Bleikeller

Time period 1665–1813 1560–1631 1773–1933 17–18th

Germany Berlin

Berlin

Parochialkirche

1701–1867 571

x

Germany Mankin

Brandenburg

Local Church

1637–1900 15

x

Germany Illmersdorf Germany Lüneburg

Brandenburg Niedersachsen

Local Church 1696–1740 11 Äbtissinnengruft 1634–1838 11

x x

Germany Bodenstein Germany Sulzbürg

Thüringen Bayern

Familiengruft Local Church

1824–1927 8 1617–1740 8

x x

Germany Wackerstein Germany Sommersdorf

Bayern Bayern

Familiengruft Castle Church

1816–1859 5 17–19th 10

x x

Germany Attel

Bayern

Local Church

1705–1760 19

x

Austria

Vienna

Wien

1560–1784 4000

x

Austria

Vienna

Wien

1363–1784 Thousands

nr

Austria

Vienna

Wien

1618–1990 Dozens

nr

Austria Austria

Michaeler Kirche St. Stephan’s Catacombs Capuchin Church Local Church Local Church

Waldhausen Oberösterreich St. Thomas am Oberösterreich Blasenstein Altlichtenwarth Niederösterreich Local Church

1450–1782 3 1756 1

x x

16–18th

66

x

114

x

Country Germany Germany Germany Germany

Austria

Location Schwerin Wolgast Mirow Bremen

Number 17 10 21 8

Anthropol x x x x

Cech Klatovy Republic Cech Mnichov Republic

Bohemia

Local Church

17th

Bohemia

Local Church

1710–1820 30

nr

Cech Brno Republic Hungary Vacs

Moravia

17–18th

nr

Comitat Pest

Lithuania Vilnius

Vilnius

Capuchin Church Dominican Church Dominican Church

Lithuania Vilnius

Vilnius

Basilian Church

41

1729–1790 265

x

18–19th

x

500

1717–1788 74

x

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Mummies in Crypts and Catacombs

755

X-ray x x x nr

CT x x x nr

Autopsy nr nr nr nr

Histology nr nr nr nr

Isotopes nr nr nr nr

Paleomolec human nr nr nr nr

Paleomolec microb nr nr nr nr

Paleobotany nr nr nr nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr nr

nr nr

nr nr

nr nr

nr nr

nr nr

nr nr

nr nr

x x

x x

x x

x x

x x

x nr

x x

x nr

x x

x x

x nr

x nr

x x

x x

x nr

x nr

x

x

nr

x

x

nr

x

nr

nr

nr

nr

nr

nr

nr

nr

x

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

Bouchal and Schaub (2013) Wunn (2015)

x x

x x

x x

x x

x x

nr nr

nr x

nr x

Nerlich (2020) Nerlich (2019)

nr

nr

x

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

nr

Großschmidt et al. (2014) Subert et al. (1993) Parsche (Personal communication) Bihl (2019)

x

x

x

x

nr

x

x

nr

x

x

x

x

nr

x

x

x

nr

nr

nr

nr

nr

nr

nr

x

References Ströbl (2011b) Ströbl (2011b) Ströbl (2011b) Uhlenhuth and Weidanz (1914) Jungklaus and Vick (2011) Jungklaus and Vick (2011) Scheven (2011) Jungklaus and Vick (2011) Nerlich (2020) Lösch et al. (2008) Nerlich (2016) Alterauge et al. (2017) Nerlich et al. (2015) Rainer (2011)

Szikossy et al. (2015) PiombinoMascali et al. (2015b) PiombinoMascali et al. (2017b)

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included here, although there are a few exceptions. A major focus will be upon the different levels of scientific analysis, indicating the applied techniques, and may provide a perspective for the future investigation of crypt and catacomb burials.

Italy One of the oldest and most extensive subterranean burial places is the “Catacombe dei Cappuccini” of Palermo (Sicily, Italy) (Piombino-Mascali et al. 2011; Ströbl 2011a). This Capuchin monastic burial place in Palermo was used after the foundation of the monastery in 1534 CE. The first use of the crypt is dated to 1599; documentary sources suggest that the first mummified friar was placed in the Catacombs was Fra Silvestro da Gubbio who had died on October 6th of that year. The monks progressively enlarged the existing subterranean burial place into a large catacomb with several corridors. With time, there was an increasing need for further space for additional burials. This led to the construction of additional corridors. During the seventeenth century, only very few individuals beyond the monastic community were incorporated into the catacomb. However, in the eighteenth centuries, it became a more widely used privilege to be buried in the Capuchin catacombs. Accordingly, by 1732 CE, an additional complex was built that was required to contain the numerous burials. It is worth remarking that most of the corpses were left clothed; when dried out some of the bodies were exposed in a standing and some in a lying position. Accordingly, the corridors were filled with more and more dead bodies standing in line as if waiting in a row (Fig. 5). A survey in 2011 indicated that the crypt housed 1252 bodies and 600 wooden coffins, some of which were empty (Piombino-Mascali et al. 2011). The extent of conservation varies widely from well-preserved corpses in their Sunday best to skeletonized residues. A review listed a total of 1182 entombments in a standing or lying position with an additional 899 skulls and 60 coffins (out of the 1252 bodies) (Piombino-Mascali et al. 2011). Out of these only 3.4% were found to be excellently preserved and further 8.8% to be adequately well preserved. The remaining 87.8% of burials were in poor condition (Ströbl 2011a). In the eighteenth and nineteenth centuries, the Palermo catacombs were increasingly used as a burial place for nonclergymen. In consequence, the expansion of the catacombs included more and more women and children of various ages. They continued in use until the middle of the twentieth century. Natural mummification was obtained thanks to the so-called “colatoi,” funerary rooms whose function was to accelerate the process of mummification through desiccation (Fornaciari et al. 2010). The “colatoi” could be either vertical or horizontal. A sitting “colatoio” was a funerary chamber with a series of niches placed along the walls and provided with masonry seats; each seat had a hole in the center. The corpse was placed into a sitting position and the fluids produced by putrefaction process were passed through the hole which, in turn, was connected to a drainage canal. The horizontal “colatoi” were small rooms (approximately, 4 m  5 m-sized cells) provided with horizontal wooden or ceramic grilles. The corpses were placed

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Fig. 5 Historic view of one of the corridors of the Capuchin catacombs with numerous mummies – and a large number of coffins (historic postcard, c. 1920). Currently, the corridors are lined by grilles. (Credit: A. G. Nerlich)

on the grilles and left there until the dehydration process was completed. Ventilation was guaranteed by simple airflow systems and drainage systems were present for the putrefaction liquids. The rooms were sealed for about 9 months. Subsequently, the dried body was washed in vinegar, dressed, and then exposed to air. Although not recorded, there is some evidence that some of the bodies were dipped in lime or arsenic, if death occurred during an epidemic (Fornaciari et al. 2010). Among the later burials there are several with clear evidence of embalming, with one example of a very elaborate conservation; the mummy of the little Rosalia Lombardo. This unique mummy will be discussed later. In previous times, this extensive burial place had scarcely been subjected to scientific investigation. Accordingly, there is little information available about the physical and health status of the individuals when they were alive, and any diseases that they may have suffered. Only very recently, the “Sicily Mummy Project” led by Dario PiombinoMascali began analyzing the corpses in a more orderly and scientific way. However, the extensive size of the catacombs and the poor state of conservation of most of the bodies may prevent a thorough investigation of this enormous “bioarchive” of Sicilian history. Started in September 2007, the Sicily Mummy project has provided, up to now, few isolated reports on CT scan analyses (Panzer et al. 2010, 2013), paleonutrition, paleobotany, entomology, and bacteriology (Piombino-Mascali et al. 2011; Pinar et al. 2014). The most excellently preserved mummy is the body of Rosalia Lombardo who died at the age of 2 years of bronchopneumonia in 1920. According to PiombinoMascali (2009; third edition: 2012) Rosalia’s father asked the taxidermist and embalmer Alfredo Salafia (see above) to embalm her body. The corpse was laid in an airtight coffin lined by lead. The result of the embalming was excellent and the infant appears as if having fallen asleep (Fig. 6).

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Fig. 6 The mummy of Rosalia Lombardo that was embalmed by Alfredo Salafia in 1920. (Wikimedia, Wikimedia.commons.org, GNUFDL)

A recent extensive CT-scan investigation (Panzer et al. 2013), technically difficult since the lead lining of the coffin caused interference with the scan, showed excellently preserved internal organs that were slightly shrunken, but otherwise in anatomically correct position. In the lungs, condensations were seen confirming the necroscopic report following which broncopneumonia was the ultimate cause of her death. The CT scan did not show evidence of a single point injection in the femoral artery using a gravity injector, the technique usually recommended by Salafia. Neither was there evidence of other intra- or inter-muscular injection sites (Panzer et al. 2013). No biochemical investigations have been so far published to allow scholars to clarify whether the embalming substances used to treat Rosalia’s body corresponded to Salafia’s formula (“Perfection Fluid”) nor if he had applied a paraffin treatment to her face to soften and smooth the skin. Following Panzer et al. (2013), it seems very likely that the paraffin treatment was used since, for a long period, Rosalia had a florid face. The Rosalia Lombardo’s case has given rise to many discussions. While some scholars maintain that the mummy remained untouched for almost 90 years (Piombino-Mascali 2009), other authors indicate that her body was restored over the decades (Lanza 2010). Family members indicated that her dresses and ribbons were changed several times (Lanza 2010; La Stampa 2011). Furthermore, Lanza (2010) suggests that her coffin was changed at least twice. What appears clear is that, after February 2008, Rosalia’s face started to show evidence of deterioration. Her face is considerably darker and dehydrated, and her hair and textiles have a lighter

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color, implying progressive oxidation (Panzer et al. 2013). To stop the process of deterioration, the coffin containing her mummy was transferred to a novel highlytechnological coffin with a 100% nitrogen atmosphere that should reduce the lowlevel oxidative destruction of the body (Samadelli et al. 2019). There are further Italian crypt burials that have been investigated scientifically, both on the island of Sicily and on the mainland. Likewise, crypt mummies have been detected in Piraino, a small city close to Messina (Sicily) where 26 naturally mummified bodies, including a skeletonized individual, have been identified (Piombino-Mascali et al. 2017a). Another collection of mummified remains is found in the church of Savoca, a small town near the Sicilian north-eastern coast. In this crypt, 17 mummies and another 17 wooden coffins have been found (Fornaciari and Giuffra 2006; Piombino-Mascali et al. 2015a). However, several coffins were reported to be empty. Except in one, all the mummies were naturally formed. The crypt burials date to between 1795 and 1876. They have been investigated undergoing X-rays imaging; gout and degenerative spine disorders were detected (Piombino-Mascali et al. 2015a). Further church crypts in Sicily also contain mummified remains of various quality and quantity. Likewise, the local church of Comiso (Province of Ragusa) contains 50 mummified burials, which, so far have only been partly investigated (Gaeta et al. 2007). These showed natural mummies with several pathologies including arteriosclerosis, goiter, splenomegaly, and parasitic infestation. Two natural crypt mummies (MSA 1 and MSA 2) were identified in the seventeenth century church of Sant’Anna in Modica di Sicilia (Ragusa Province). Computed tomography (CT) showed that the late eighteenth- to early nineteenth-century natural mummies belonged to two males (one middle-aged and the other elderly); both had primary pulmonary tuberculosis. MSA 2 further showed a 36 mm  15 mm incision in the fourth right intercostal space consistent with either a traumatic or iatrogenic pneumothorax (Ventura et al. 2016). Pulmonary tuberculosis was also identified in a twentieth-century natural mummy found in the nearby town of Scicli (Ventura et al. 2012). A further extensive study has been performed on crypt mummies in the church Santa Maria di Randazzo (Province of Catania) (Lo Gerfo 2011). Anthropological investigation with further particular analysis of isolated specimens by CT scanning and endoscopy were performed on 135 individuals (92 males, 24 females, 12 subadults, and 12 undetermined). While some corpses were naturally mummified, in some cases traces of substances pointing to an embalming procedure were found (straw, string, metallic particles, fragments of textiles, cotton, laurel leaves (Laurus nobilis), butcher’s broom (Ruscus aculeatus), chestnut peel (Castanea sativa Mill), oak (Quercus spp.), rose leaves, and rose petals (Lo Gerfo 2011). The church of Chiesa Madre di S. Maria Annunziata in the small village of Novara di Sicilia houses six spontaneously mummified bodies dating to between 1849 and 1873 (Piombino-Mascali et al. 2011). Five mummies (1849–1873) belonging to clergymen have been placed in an upward position, whereas one mummy (1868), wearing a dark cloak, lies in a coffin placed on the ground. The mummies show an extremely poor state of conservation and need urgent restoration. Some very

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preliminary results have come from the investigations of the Capuchin church (built in 1610) and annexed crypt of Santa Lucia del Mela (Messina, Sicily) (Bessa et al. 2018). Another crypt (Cripta dei Preti) associated with the church San Nicolò of Gangi houses 64 mummified individuals (clergymen and laymen) dated to the eighteenth to nineteenth centuries. Macroscopic morphological investigations were initially carried out (Micciché et al. 2014). Recently paleoimaging and bioarchaeological analysis were performed on 36 of 64 bodies (Beckett et al. 2020; Bianucci et al. 2021). Other Sicilian sites contain mummified human remains, e.g., Burgio (Province of Agrigento). Forty-nine mummies dated between the eighteenth century and the first years of the twentieth century were uncovered in 1994. These mummies are part of the Museum of the Mummies of Burgio (Di Cristina et al. 2007); while the crypt and the robes of the skeletonized bodies underwent restoration, no anthropo-paleopathological investigation has been published so far. In addition to the Sicilian mummies, continental Italy also has numerous crypts that have been partly investigated, such as the extensive analyses by the group of Fornaciari of the Aragonese mummies in the crypt of San Domenico Maggiore in Naples and the mummies/skeletons of the Medici family at the Cappelle Medicee in Florence. In a compilation of crypt burials, Fornaciari (1998) refers to a large number of church crypts that house only one mummy. Likewise, in 1998, when his overview was undertaken, 65 places with single mummies and 11 places with multiple mummies have been reported in Italy. Since then, it can be assumed that many more cases and/or new investigations with, possibly, additional techniques may have been identified and performed. In this regard, this survey can only provide representative data. In Naples, the Aragonese royals and other aristocrats were conserved for “eternity” (Fornaciari 2006; Marinozzi et al. 2011). In a detailed study, these mummies were thoroughly investigated using a broad panel of techniques. In total, 31 Italian Renaissance mummies from the Basilica of San Domenico Maggiore revealed a wide spectrum of paleopathological diagnoses including; infectious disease (smallpox, hepatitis, condyloma, syphilis, and pneumonia), metabolic disorders (obesity, atherosclerosis, gallstones), articular (DISH) and neoplastic (colon adenocarcinoma and skin carcinoma) pathologies. The four mummies from Napoleonic times were less informative (Marinozzi et al. 2011). Besides a few unidentified individuals, most of the investigated bodies could be attributed to particular individuals with ten of the royal family and several other higher court noblemen. The well-preserved mummy of Maria d’Aragona, Marquise of Vasto (1503–1568), showed, on the left arm, an oval cutaneous ulcer with the morphological characteristics of treponematosis, a venereal infection. Furthermore, the mummy showed a large pedunculated arborescent neoplasm of the right inguinal region that was identified as condyloma acuminatum, a papillomavirus-induced squamous lesion; a venereal wart (Fornaciari et al. 2003). Serological investigations showed that Isabella di Aragona, Duchess of Milan (1470–1524) suffered from zoonotic visceral leishmaniasis (ZVL), which would account for her well-documented recurrent fevers and subsequent death (Lanzirotti et al. 2014). There has been speculation that the most likely source of the mercury in Isabella

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was an oral anti-syphilitic treatment. However, there is no ancient medical report to support the notion that Isabella suffered from venereal treponematosis. The mummy of Ferrante I d’Aragon, King of Naples (1431–1494 CE), revealed an adenocarcinoma extensively infiltrating the muscles of the inner pelvis with a typical mutation of the K-ras gene (Ottini et al. 2011). The alimentary milieu of the Neapolitan court of the fifteenth century, with its abundance of natural alimentary products, may well explain this mutation. These and other diseases, such as a case of cirrhosis, some cases of anthracosis, and traumatic conditions including a mortal stab-wound, can illustrate the disease spectrum of the wealthy of the Italian Renaissance. In 2004, the Medici Project, a multidisciplinary research of the study of the 49 burials of the Medici in San Lorenzo, Florence, was officially launched. Currently 20 tombs, including the burials of 9 children, have been investigated thoroughly (Fornaciari et al. 2008, 2009). The bodies of the Medici were treated before burial, as was customary for the political and economical prominence of these personages (Pieraccini 1986). However, the status of conservation of most of these burials is poor. This comes from repeated extensive earlier investigations. The traces of soft tissues have disappeared; in fact, the bodies are currently skeletonized, despite them being originally almost all artificial mummies (Lippi 2011). Nevertheless, 11 out of 20 individuals showed signs of autopsy and/or embalming (Fornaciari et al. 2008; Giuffra et al. 2011). Further crypt mummy series (Fornaciari 1998) have so far been investigated in Altavilla Irpina (more than 100 corpses, mostly skeletonized), Navelli and Rocca di Papa (also each more than 100 burials), Ferentillo (16 mummies), Urbania (18 bodies), Roccapelago (13 mummies; Petrella et al. 2016), Badia Pozzeveri (26 skeletonized mummies; Fornaciari et al. (2016), and Venzone (40 corpses). The latter are of particular note since it is assumed that in this location the action of a particular fungus, Hypha bombycina Pers. caused accelerated dehydration and thereby preserved the bodies (Aufderheide 2003). The mummies of Venzone have already been scientifically investigated from the middle of the eighteenth century (i.e., in 1829; Marcolini 1831) (Figs. 7 and 8). A modern scientific study adds to these older data (Baggieri and Di Giacomo 2002). The mummies from Urbania have been further investigated by Beckett et al. (Unpublished manuscript). The gallery of 18 mummies at the Church of the Dead included 11 males and 7 females. The age range was approximately 20 to greater than 70 years old. The bodies were generally light brown with the exception of mummy #8, said to be Vincenzo Piccini, whose surface appearance was white. The display position was consistent with the burial position with regards to the placement of the arms along the sides and with the females having their arms over the pelvic region. The arms were found to be crossed over the chest in three mummies. Internal preservation was generally good. Radiographic and endoscopic data revealed a variety of paleopathological conditions. Among the pathologies demonstrated were congenital dislocation of the hip, scoliosis, severe thoracic kyphosis, a small bladder stone, a large bladder stone, arthritic changes, and pulmonary pathology. Evidence of cardiovascular disease was seen with calcification of the aorta and other

762 Fig. 7 One of the Venzone mummies; earliest representation of the spontaneous mummy of a hunchback (in Italian: il gobbo) (Marcolini 1831)

Fig. 8 Right: some of the Venzone mummies are currently on display in the local museum. (Credit: A. G. Nerlich)

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vessels, as well as calcification of the vas deferens associated with diabetes. Possible trauma was noted in the case of Mummy #12 with a penetrating chest wound. Trichocephalus trichiura ova, an intestinal parasite that is commonly acquired through the ingestion of eggs found in dry goods such as grains and beans, were associated with Mummy #18. Beckett et al. (Unpublished manuscript). Chemical analysis of various skin samples from mummies 1, 2, 3, 4, 8, 13, 15, and 16 showed high levels of calcium, the main component of limestone, and may have played a key role in the natural mummification of these individuals. The overall state of preservation was good to fair in that the integument and, in most cases, internal organs and structures were still identifiable. The heads of the majority of the mummies were less well preserved than the remainder of the body. The three mummies who were said to have been artificially mummified, Mummies #1, 2, and 8, were the least well preserved. Beckett et al. (Unpublished manuscript) When Arthur C. Aufderheide (2003) collected data about mummies from all over the world, he noted that the number of crypt and catacomb mummies in Italy was significantly high. Accordingly, he posed the question as to the possible reasons and therefore suggested that a combination of the enormous number of large churches and cathedrals and climatic conditions might be the answer. It might be interesting to look at other European countries with different conditions.

Germany In comparison to Italy, Germany has a very different climate, mostly moderately warm and much wetter, and, at least in the Protestant North of the country, fewer churches, or at least fewer large churches exist. Therefore, it is of interest to see how many crypt mummies exist there. A recent compilation of data shows only a few locations with crypt mummies and a low number of burials in those places. However, it must be stated that the systematic scientific investigation of crypt mummies in Germany has begun much later than in Italy. In the North of Germany, several crypts have been methodically investigated by archaeologists and anthropologists, however, without application of further analytical investigations such as systematic CT-scans, molecular, or chemical analyses. The studies revealed mostly very well-preserved bodies that were naturally mummified. The eight cadavers of the “Bremer Bleikeller” (in coffins made of lead) are mentioned as excellent examples of natural mummies. Although the exact reason for this mummification is still a matter of debate, the high concentration of lead in the environment of the location is associated with the rapid desiccation and conservation of the corpses (Uhlenhuth and Weidanz 1914). As an isolated finding, the spontaneously mummified corpse of Christian Friedrich von Kahlbutz (1652–1702) is noteworthy. His very well-preserved mummy lies in a small North German local church; there exist numerous theories about the formation of the body and its history (Aufderheide 2003).

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Similarly, the burials of several church crypts in Mecklenburg-Vorpommern, in particular in Schwerin, Wolgast and Mirow, are naturally mummified and still clothed. The exact causes of death, or any further information about life habits or diseases, are unknown (Ströbl 2011b). Several extensive anthropological investigations of burials in Berlin and Brandenburg confirmed the generally good conditions for conservation (Jungklaus and Vick 2011; Scheven 2011). However, from the largest series in the crypt of the Parochialkirche in Berlin, only parts of the mummies are still present; typically, due to the losses during World War II when the majority of the burials were destroyed. Again, further information about life and diseases is scarce and does not seem to be consistent. In contrast, several mummy projects in the Southern parts of Germany, although mainly covering relatively small numbers of individuals, have been performed in much more depth, particularly with respect to the application of interdisciplinary techniques. A very recent, and still on-going study on a small series of a noble crypt in Thüringen (Bodenstein) (Author’s personal work) of four complete mummies and a further four skeletonized mummies, revealed, not only the molecular identification of historically known individuals, but also the unique observation of a sclerosing bone disease in four of the eight individuals, suggesting an hereditary bone disease that is still being investigated. Other crypt burials of complete mummies (Wolfstein, Wackerstein, Sommersdorf, and Attel) cover a broad range of aristocratic and higher clergy individuals in conditions of very good preservation in parts (Fig. 9). Molecular analysis on the Wolfstein mummies (Lösch et al. 2008) showed that some of them were infected by tuberculosis. Otherwise, stable isotope analysis revealed that they were of a good nutritional status. The investigation of an aristocratic family crypt in Sommersdorf also contributes interesting findings which range from presumed individual identification to physical properties and paleopathological findings (Alterauge et al. 2017). Similar has been found in a series of mostly skeletonized mummies from the small monastery of Attel in Bavaria. Although the monastery was not part of the very wealthy Episcopal seed in the region, the nutritional status of the monks was very Fig. 9 The mummy of Friedrich Wilhelm von Wolfstein (1716–1728) in the crypt of the local church of Sulzbürg/Bavaria. (Photo: A. G. Nerlich)

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good, as confirmed by the finding of typical gout in two individuals (Nerlich et al. 2015). Finally, a still ongoing project on the small family crypt at Wackerstein (Bavaria, near the river Danube), built by the Bavarian General Wilhelm von Jordan in 1836 for his family and a close friend, provides an ample insight into life, disease and the causes of death in all five individuals that were found mummified and in excellent preservation (see Figs. 10, 11, and 12). The thorough analysis uncovered typical features of long-standing tuberculosis in two individuals, one with a large

Fig. 10 The mummy of the Royal Bavarian General Wilhelm von Jordan. (Photo: A. G. Nerlich)

Fig. 11 The mummy of the wife of Wilhelm von Jordan, Violante nee Countess Sandizell. (Photo: A. G. Nerlich)

Fig. 12 The mummy of Carolina von Jordan. The evisceration is evident by a longitudinal cut in the bowel. Otherwise the body is excellently preserved. (Photo: A. G. Nerlich)

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cold tuberculous abscess in the retro-peritoneum that he had obviously had for several decades (Nerlich 2016). A second mummy, which also revealed long-term post-tuberculous pulmonary emphysema, died of metastatic prostate carcinoma, as confirmed by CT analysis, bone histology, and immunohistochemistry. Besides the natural mummies, artificially embalmed corpses can occasionally be found. One of the most interesting findings was the mummy of eldest daughter of the Jordan family, Carolina von Jordan. She was born in 1815, but died 1 year later in Naples. She was embalmed according to the Aragonese protocol and brought to Bavaria as a mummy. Her state of conservation is excellent. The thorough interdisciplinary investigation uncovered a detailed “story” of her history (Nerlich 2019). The analysis covered, besides X-rays, CT-scans, histology, and isotope studies for nutritional conditions and both human and pathogen aDNA analysis for family relations and infectious disease identification. In addition to the interdisciplinary scientific analysis, this study provides clear evidence that the inclusion of an intense and thorough investigation of historical information leads to a precise reconstruction of historic events. While the crypts in Germany are mainly not completely investigated, a further problem is the lack of comparability of mummy studies between different study groups. In this regard, the establishment of distinct criteria for such analyses seems pivotal.

Austria The study of catacomb and crypt mummies in Austria was almost exclusively focused on the capital of the country, Vienna. There exist two large catacombs – and a smaller one – that contain the majority of the burials during historic times. These contain the burials in the city until Emperor Joseph II abolished inner-city ones and forced the population to use cemeteries that were outside of the city. Accordingly, the catacombs of the St. Stephan’s Cathedral had several levels that extend deep below the basement (Fig. 13). Part of these catacombs is still open, and part has been filled up with debris, particularly following the World War II. It has been estimated that several tens of thousands of cadavers have been buried there, of which at present only a small part still exists (Bouchal and Schaub 2013). St. Stephan’s catacombs also contain burials of various ducal families. Furthermore, the intestines of Royal family members have been deposited in a designated section of the catacombs (Bouchal and Schaub 2013). Despite this, there exists no evidence of any scientific examination of the burials and we have no information about any of the individuals buried there. The second largest catacomb of Vienna city is that of the church St. Michael (also called the “Michaeler crypt”) very close to the Hofburg (the old imperial residence in central Vienna) (Rainer 2011). Due to its close location to the imperial residence, as early as 1508, the Emperor Maximilian I abolished the use of the church’s cemetery resulting in the monks digging a system of catacombs below the church to enable

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Fig. 13 St. Stephan’s Cathedral catacomb: a visit to the burial place in 1872 (from Die Gartenlaube 1872). Note that several coffins are opened containing mummified human remains

further burials to take place. This ended up with a labyrinthine system of halls and vaults. Parts of these burial places were reserved for high aristocratic families. There exists a crypt for the church’s clergy, and other parts were used by lower aristocrats. All depended on the costs of maintenance of the crypts and vaults. The Michaeler crypt (Rainer 2011) was in use until 1784 when, as occurred everywhere in Austria, the Emperor Joseph II abolished burials within towns and cities forcing people to use out-of-town cemeteries. Due to the climatic conditions, with a well-established ventilation system, part of the burials in the Michaeler crypt spontaneously mummified, although the major part of the burials decomposed into skeletal remains. Likewise, when Ekkehard Kleiss viewed the crypt in the later 1970s, he found a diversity of tissue preservation from complete mummies with near perfect soft tissue cover to disintegrated skeletons. Only in some occasional cases is there evidence for artificial embalming techniques, such as the removal of internal organs (Aufderheide 2003). In recent years, changes in the ventilation system, due to modification of the environment, has led to a dramatic increase in moisture levels within the crypt leading to a progressive destruction of the biomaterial. In addition, an infiltration of the crypt by a particular beetle (Genus and species) that destroys the wooden

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coffins has contributed to the destruction. Meanwhile, attempts to control air moisture and other conservation methods have rescued part of the burials, including some very well-preserved mummies (Rainer 2011). These have been investigated by anthropological techniques but no further examination seems to have been performed. Besides these large catacomb burial places within the city of Vienna, there exists a very particular crypt in another Viennese church, the Capuchin church, where in the imperial crypt 12 Austrian Emperors, 19 Empresses, and numerous members of the imperial family have been buried. This particular practice started in 1618, and was in use until the late twentieth century (Wunn 2015). Outside Vienna, only a few places are known to contain mummies (Baumgart and Stark 2003). Out of these few the three mummies of the monastery of Waldhausen (Oberösterreich), and the near-by buried mummy of the air-dried chaplain at St. Thomas am Blasenstein, are of particular interest, since all four have recently been investigated in more detail (Nerlich et al. 2019). The three mummified corpses of the Waldhausen monastery are only partly preserved. The mummy in St. Thomas am Blasenstein is almost complete. The interdisciplinary investigations provided the surprising evidence that one of the Waldhausen corpses and the mummy from St. Thomas had been embalmed by a unique technique; the dead bodies had been stuffed with various materials; textiles, wooden chips, and branches and leaves. Furthermore, the addition of zinc sulfate may have promoted desiccation and thereby tissue stabilization. While no evisceration cut was detectable on the body surface, there was clear evidence that the mummies had been prepared via a transanal approach. In the Waldhausen mummies, several had been affected by severe and longstanding tuberculosis of the lungs. The mummy from St. Thomas am Blasenstein was also infected, and obviously died of an acute hemorrhage of a tuberculous lung cavity, which was still visible during the examination (Nerlich et al. 2019) (Fig. 14). Beyond these few examples, occasional investigations have also been done on crypt mummies in other places in the country (Großschmidt et al. 2014). Also, ongoing research is gathering more and more information on mummy findings in local populations.

Fig. 14 The air-dried chaplain from St. Thomas am Blasenstein (Oberösterreich). The very wellpreserved mummy has been prepared by stuffing the body with textiles, wooden chips, and branches and leaves, via a transrectal approach. (Photo: A. G. Nerlich)

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The Czech Republic Close to Austria, and until World War I connected in the single state of the AustroHungarian Empire, crypt mummies are also present in the Czech Republic. The most famous crypt is that in the small Bohemian town of Klattovy where a total of about 114 mummies have been found in the local church. These have all been of spontaneous origin and are very well preserved; obviously due to excellent ventilation conditions (Subert et al. 1993). They cover the burials of the local clergy, but also local aristocratic family members and other citizens of the town. The crypt was renovated in the early 2000s and the mummies have been restored. Some of the bodies have been investigated by anthropological examination, but no data are available about other specialized investigations. Beyond the Klattovy mummies, there exist several other places where mummies are stored in crypts. However, data on these places, as well as the bodies, are very scarce and have not been examined in detail. Likewise, mummified corpses have been described in the crypt of the Capuchin monastery at Brno, where 41 mummies out of approximately 205 burials have survived (Bihl 2019). These date to the seventeenth to eighteenth century and are assumed to cover local aristocracy and citizens of Brno (Fig. 15). Similarly, several other churches contain mummified corpses in their crypts (e.g., at Broumov or Mnichov). These seem to be natural mummies without evidence of artificial conservation (F. Parsche, Personal communication). It can be assumed that many more crypt bodies with mummification can be expected. However, since the number of churches and crypts are not even known, no scientific investigation has been carried out.

Hungary Similar observations on crypts and crypt mummies have been made in another part of the former Austro-Hungarian Empire, Hungary, where there seem to be similar Fig. 15 One of the spontaneously mummified bodies of the small church of Mnichov, Bohemia, showing an excellently preserved corpse. (Photo: A. G. Nerlich)

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types of burial rites in use as in Austria and the Czech Republic. There is, however, one large exception from the largely unexplored church crypts, the crypt of the small city of Vac in the northern part of Hungary (comitat Pest) (Szikossy et al. 2015). The crypt was rediscovered by chance in 1994 and contains approximately 265 burials of which 166 individuals can be identified named according to the detailed data that has been retrieved from the local death registry. Besides these historical investigations the Vac mummies have been subjected to extensive analyses using various techniques, such as CT scans and, particularly, molecular investigations. More specifically, the analysis for tuberculosis infection has been applied to numerous samples from various individuals. The most surprising result is not only a proven high TB infection prevalence at 69.8% (Fletcher et al. 2003), but, even more importantly, the evidence for infection with multiple TB strains. The pattern suggests genetic factors influence the risk for both infection and the occurrence of symptoms (Kay et al. 2015). In this regard, the molecular studies on the Vac mummy population are extremely helpful for modern TB and infection disease research. Further investigations on this population uncovered detailed demographic data with the life expectancy of males at 44.6 years and females at 38.8 years, precise dental pathology and evidence for the sequelae of trauma. Furthermore, isolated cases shed interesting light on autopsy techniques. The unique case of the selective removal of the heart post mortem in the mummy of a nun, a case of sectio in mortua, a postmortem Caesarean section, in a 26 years old female, and the identification of familial relationships (Szikossy et al. 2015). The studies on the Vac mummies have not yet finished and it has to be expected that more important data will be recovered from this unique bioarchive.

Lithuania Similar to other Central and Northern European countries, there exist several crypts in East European countries with human mummies. While little is known about crypt mummies in strictly Catholic eastern countries such as Poland, recent investigations reveal interesting findings in Lithuania. Up to now, two reports describe on investigation of corpses in two large church crypts in the Dominican church of the Holy Spirit in the country’s capital Vilnius. Out of a total of approximately 500 burials, 23 mummies have been examined in detail (Piombino-Mascali et al. 2015b); 15 adults and 8 subadults with signs of natural mummification and evidence for multiple pathological conditions more precisely diagnosed in some using CT-scans, which included dental pathology, arteriosclerosis, and obesity. The second church crypt that has been investigated in Vilnius is that of the Basilian church of the Holy Trinity where 74 burials, including 56 skeletons, 13 poorly mummified bodies, and 5 mummies, have been analyzed (Piombino-Mascali et al. 2017b). All mummies are of natural origin except for one individual that shows filling of the chest and abdomen with small wood chips. Due to limited preservation, there are as yet only limited results and observations. The majority of individuals were males, suggesting the burials are of members of the order of St. Basilius. There is one major

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exception, a sample from the mummy of a child that underwent molecular identification of the oldest genome of the smallpox virus (Variola virus) (Duggan et al. 2016). Ongoing research will probably provide significantly more data.

Other Countries In a few other European countries, isolated cases of mummified corpses have been described associated with crypts and catacombs. In Switzerland, in Basel, the mummy of a Franciscan friar had been placed into the Orders’ museum suggesting artificial mummification by application of mercury (cinnabar) on its surface (Aufderheide 2003; Kaufmann 1996). In France, isolated crypt mummies have been noted in the church of St. Thomas in Strasburg. The mummy there was identified as the Count de Nassau, a spontaneous mummy of approximately 600 years of age (Aufderheide 2003). In Spain in the Basque region, mummified remains of six local Saints have been described (Aufderheide 2003). During the 2011 restoration works in the central nave of the church of the Assumption of Our Lady, known as “The Piquete,” in the village of Quinto, which is about 50 km southwest of Zaragoza, Spain, the remains of 70 individuals were uncovered (Bianucci et al. 2018). Of these there were 32 mummified bodies, four of which were investigated with CT scans. The presence of pressure erosion and distortion of the upper thoracic spine, the cause of which may well represent a neurenteric duplication cyst, was identified in a child aged between 7 and 8 years (Loynes et al. 2018). In Ireland in the crypt of St. Michan’s church, Dublin, the spontaneously mummified body of a late mediaeval crusader is interred along with several other mummies (Aufderheide 2003). The report by Pulz et al. (2011) describes a series of mummified clerics in a crypt in Croatia. Finally, in non-European countries, mummies have also been identified in crypts and catacombs. For example, the mummies from Guanajuato, Mexico, are now housed in a special museum. The spontaneously mummified bodies had been discovered by chance during construction work at the local cemetery where they were found well-preserved in their coffins. Approximately 100 mummified bodies have been exhumed (Aufderheide 2003). Similarly, a recent report describes some findings on a series of 40 mummies from the nineteenth century in the monastery of Santo Domingo at Mexico-City (Leboreiro Reyna and Mansilla Lory 2008) which, however, provides only some anthropological and historical information; no other investigations seems to have been done on them.

Conclusions and Outlook This report presents a current overview of crypt and catacomb mummies that have mainly been found in European churches and cemeteries. Both natural, as well as embalmed, mummies have been identified. The latter group can further be

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subdivided into those with the classical types of dry or wet embalming; the dry embalming includes evisceration by opening the abdomen and/or the chest and removal of the inner organs. However, recent studies also provide evidence that also simple forms of conservation have been used, possibly only to preserve the bodies for a limited period of time, e.g., for transport or lying-in-state of the cadaver. Simple embalming did not use elaborate techniques to accelerate desiccation or chemical interference of microbial tissue destruction. It just introduced textiles, wood chips or twigs, and leaves into the dead body to stabilize the tissue and desiccate the body from within. This “tar-and-sawdust” embalming was established at the beginning of eighteenth century when undertakers looked for new business opportunities. Additionally, this type of conservation was much cheaper than the traditional embalming methods, and it could be performed without the help of surgeons and pharmacists. It is surprising to see how many churches and cemeteries, with crypt burials, contain human remains in a mummified state. In this respect, this compilation of locations of mummies must be incomplete. It is to be hoped that the ongoing research efforts will fill in the present gaps in knowledge. Besides just increasing the places under investigation, and thereby the number of mummies examined, the rising range of analytical techniques is of major importance. Today, it is no major problem to transport a mummy to a hospital unit equipped with a modern CT scanner. Furthermore, the application of modern techniques in histology, isotope chemistry, and molecular biology (looking at sequencing human and microbial ancient DNA), broadens our knowledge significantly. These important bioarchives hold information that persists and adds to the knowledge gleaned from written sources, and may allow us to understand about the life, diseases, and deaths of important historical personages. Acknowledgments We are indebted to Professor Simon T. Donell who generously proofread and edited this chapter.

References Alterauge A, Kellinghaus M, Jackowski C et al (2017) The Sommersdorf mummies – an interdisciplinary investigation on human remains from a 17th to 19th century aristocratic crypt of southern Germany. PLoS One 12:e0183588 Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Baggieri G, Di Giacomo M (2002) Le mummie di Venzone: morfologia, radiologia e Tac. Bollettino dell'Associazione Mummie di Venzone, Anno XXXII Baumgart G, Stark H (2003) Der ewig Leib. Mumien in österreichischen Sammlungen und Grüften. Verlagshaus Ärzte, Wien Beckett R, Conlogue G, Bravo A et al (Unpublished manuscript) Paleoimaging survey of eighteen mummies from the Church of the Dead, Urbania, Italy: a paleopathological analysis Beckett R, Conlogue G, Viner M, Saleem SN, Said A-H, Piombino-Mascali D (2020) A Paleoimaging Study of Human Mummies held in the Mother Church of Gangi, Sicily: Implications for Mass Casualty Methodology. Forensic Imaging https://doi.org/10.1016/j. fri.2020.200416

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Samadelli M, Zink AR, Roselli G et al (2019) Development of passive controlled atmosphere display cases for the conservation of cultural assets. J Cult Herit 35:145–153 Scheven A (2011) “Du wirst uns unvergeßlich sein . . .” Eine besondere Grablege im Land Brandenburg: die Mumiengruft Illmersdorf. In: Sörries R (ed) Geschichte und Tradition der Mumifizierung in Europa. AGFD Kassel, pp 79–102 Schleifring JH, Galassi FM, Habicht ME, Rühli FJ (2019) Autopsing history: the mummy of Charlemagne (c. 747–814 AD), father of Europe. Econ Hum Biol 32:11–17 Schmitz-Esser R (2014) Der Leichnam im Mittelalter. Einbalsamierung, Verbrennung und die kulturelle Konstruktion des toten Körpers. Jan Thorbecke Verlag, Ostfildern Selmi F (1872) Enciclopedia di chimica scientifica e industriale, ossia dizionario generale di chimica, Vol. Quinto. Torino, Unione Tipografico-Editrice Torinese, p 76 Ströbl A (2011a) Die Catacombe dei Cappuccini in Palermo. Geschichte und Hintergründe einer einzigartigen Gruft. In: Sörries R (ed) Geschichte und Tradition der Mumifizierung in Europa. AGFD Kassel, pp 11–24 Ströbl R (2011b) Schwerin – Mirow – Wolgast. Die drei großen Herzogsgrüfte in MecklenburgVorpommern. In: Sörries R (ed) Geschichte und Tradition der Mumifizierung in Europa. AGFD Kassel, pp 49–64 Subert F, Dvorak V, Spalek E et al (1993) Problems of contemporary mummy conservation. Charles University, Prague Sucquet JP (1872) De l’embaumement chez les anciens et chez les modernes. Delahaye, Paris Szikossy I, Kustar A, Guba Z et al (2015) Natürlich mumifizierte Leichname aus der Dominikanerkirche in Vac, Ungarn. In: Wieczorek A, Rosendahl W. Mumien – Der Traum vom ewigen Leben. Reclam Verlag, Darmstadt, pp 355–367 Uhlenhuth P, Weidanz O (1914) Die biologischen Methoden im Dienste der anthropologischen Forschung. Zschr Morphol 17:671–698 Ventura L, Pensiero V, Caruso C, Voi G, Fornaciari G (2012) The natural mummies from the church of Santa Maria della Consolazione in Scicli (South-Eastern Sicily). In: 19th European Meeting of the Paleopathology Association, Lille, 27–29 Aug 2012 Ventura L, Pensiero V, Romeo F, Causarano A (2016) Paleopathology of the natural mummies from the church of Sant’Anna in Modica (South-Eastern Sicily). Pathologica 8:293–294 Wittmers L, Aufderheide AC, Buikstra J (2011) Soft tissue preservations system: applications. Int J Paleopathol 1:150–154 Wunn I (2015) Mumien in Klöstern und Kirchen – Mönche, Päpste und Fürsten. In: Wieczorek A, Rosendahl W. Mumien – Der Traum vom ewigen Leben. Darmstadt, pp 165–174

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Contents Introduction – How Our Story Began . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . History of Vác . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Dominican Church of Vác . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Under the Dominican Church . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Crypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Loretum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ossuary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Crypt Entrance I . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Crypt Entrance II . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Vent Holes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Discovery of the Crypt . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Mummification Mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Church and Civic Archives – Life Stories of Individuals and Families of Special Interest . . . Antónia Tauber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Annamaria Beer – The 95-Year-Old with Past Tuberculosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tuberculosis in the Hausmann Family and Beyond . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Evidence of Colon or Bowel Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A Case of Renal (Kidney) Stone . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A Case of Gout . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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H. D. Donoghue (*) Centre for Clinical Microbiology, Division of Infection and Immunity, University College London, London, UK e-mail: [email protected] I. Pap · I. Szikossy Department of Anthropology, Hungarian Natural History Museum, Budapest, Hungary M. Spigelman The Kuvin Center for the Study of Infectious and Tropical Diseases and Ancient DNA, Hadassah Medical School, The Hebrew University, Jerusalem, Israel Division of Infection and Immunity, Centre of Clinical Microbiology Royal Free hospital UCL London, London, UK Department of History and Archaeology, Macquarie University, NSW, Australia © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_21

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Two Young Men with Developmental Abnormalities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Death of a Mother and Child (Postmortem Caesarean Section) . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Director and Teacher at the Institute for the Deaf . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Molecular Analysis for Tuberculosis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Extent of Tuberculosis Infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Aims: This chapter aims to investigate the 265 eighteenth-century human remains interned in a sealed crypt within the Dominican Church in Vác, Hungary, between 1731 and 1838, including their lifestyle, occupation, and modes of death. In particular, this chapter aims to study the natural history of tuberculosis in the preantibiotic era related to age at death, sex, and body site and to characterize the infecting organisms. Church and civic records provided details of the individuals buried in the crypt. Methods: Bodies were examined by naked eye or radiographic examination, initially by X-ray, then via CT analysis. Several selected individuals were examined for noninfectious conditions, including a nun with a severe spinal deformity, two young men with developmental abnormalities, and an elderly woman with gout. Skeletal and naturally mummified tissues were examined for the presence of Mycobacterium tuberculosis DNA using specific molecular markers and techniques. Results: The crypt individuals included wealthy citizens and clergy. Overall, specimens from 176/265 (66.4%) individuals were positive for tuberculosis, with 36/67 (53.7%) positive aged 80 years. The examination of samples has continued after the publications in 2003 until the present day, including reexamination of the early samples, plus examination of additional samples taken in subsequent years. The latest data show that with the exception of infants and children under the age of 5 years, 75% of females and 81% of males buried in the crypt show molecular evidence of tuberculosis. However, there is limited evidence of tuberculosis being the direct cause of death.

The Va´c Mummy Project: Investigation of 265 Eighteenth-Century Mummified. . .

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Fig. 17 Number of individuals by age at death and presence or absence of M. tuberculosis (Credit: Helen Donoghue)

Analysis of the effect of sampling site was performed. Pulmonary disease was defined as in chest contents, including lungs, pleura, and ribs. Extrapulmonary sites were predominantly abdominal or pelvic contents – including upper and lower abdominal wall, the sacral region, spine, and a few samples of skin, hair, and other soft tissue. There were no significant differences in the proportion of positive samples obtained from pulmonary or abdominal samples, nor from chest tissue (51.0%; 73/143) compared with ribs (48.5%; 49/101). There was, however, a higher proportion of M. tuberculosis-positive samples in calcified pleura (74.1%; 20/27) compared with ribs or chest tissue, p < 0.05. In 91 individuals, samples were examined from a single site – ribs or tissue. In this case, 46/91 (50.5%) were M. tuberculosis-positive. However, when multiple samples were examined from the same individual, the proportion that had at least one M. tuberculosis-positive sample was significantly greater, rising from 73.1% (49/67) where two samples were taken ( p < 0.01), to 87.2% (41/47) where three or more samples were taken, p < 0.001. Samples were taken from both pulmonary and extrapulmonary sites in 101 individuals. Pulmonary tissue was positive in 64 cases (63.4%); 38 (37.6%) at that site only, and in 26 (25.7%) positive in both chest and abdomen. A further 15 (14.9%) were positive only in extrapulmonary tissue. In one such individual, pathological changes were noted on a cervical vertebra (Donoghue et al. 2011, Fig. 2). In 18 individuals, both pulmonary and extrapulmonary tissues were strongly M. tuberculosis-positive, and of these, 12 showed gross pathological signs of tuberculosis; these included cachectic appearance in mummified remains (Video 2),

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being extremely small for age, calcified lesions visible by radiograph or visual examination (Video 2), and deformities of the spine (Donoghue et al. 2011, Fig. 3). The TbD1 deletion was demonstrated in the DNA extracts from 27 individuals, thus demonstrating that they were infected specifically with M. tuberculosis rather than Mycobacterium bovis or any other species in the M. tuberculosis complex. Samples from 29 individuals gave partial or complete spoligotypes; 24 of these were sufficiently complete to confirm that M. tuberculosis was the infecting organism (Donoghue et al. 2011, Fig. 4). Compared to modern clinical isolates, the spoligotypes showed few, if any, spacer deletions. However, care was needed to distinguish deletions from poor DNA preservation. Using both methods, DNA extracts from 33 individuals were shown to be of M. tuberculosis.

Discussion Due to the precautions that were taken and the specificity of the PCR reaction, we assume that individuals positive for M. tuberculosis were indeed infected. This is supported by the correlation between pathological sites and the detection of M. tuberculosis in several cases. This collection of human remains has been studied since the early 1990s and confirmed by microbiology, radiology, coffin inscriptions, and church and civic records. The first reports of the detection of M. tuberculosis DNA in archaeological remains (Spigelman and Lemma 1993; Salo et al. 1994) have been followed by many publications, including specimens dating from the Neolithic predairying era (Hershkovitz et al. 2008), preanimal domestication in ancient Syria (Baker et al. 2015), ancient Egypt (Zink et al. 2007), prehistoric Britain (Mays and Taylor 2003), and pre-Colombian America (Arrieza et al. 1995; Braun et al. 1998; Donoghue 2016). Most of the Vác burials are infected with M. tuberculosis, indicating the high exposure of the population to tuberculosis. Despite this, there were many infected individuals who lived to middle or late age, a clear indication of the importance of host immunity in controlling the disease. The time, during which the crypt was in use for burials, coincided with a fairly prosperous and peaceful period. The individuals were from a prosperous neighborhood, and according to their clothing, artifacts, and contemporary archives, many were professionals or skilled artisans. These factors would have contributed to their resistance to the disease. Newborn infants and young children are more vulnerable due to their naïve immune system (Lewis 2017), and it was noteworthy that there were cases of congenital and primary tuberculosis in this population. Disseminated tuberculosis was detected in all adult groups, and the spread from pulmonary to extrapulmonary sites was similar to that noted in a retrospective analysis of a more recent group of historical tuberculosis cases from the preantibiotic era (Slavin et al. 1980). Other authors have noted the significantly higher proportion of tuberculosis infection in males (Hudelson 1996; Holmes et al. 1998; Hamid Salim et al. 2004). The distinction between biological and social explanations for this observation has proved difficult. The present study indicates a genuine higher infection in males in

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eighteenth-century Hungary, but this may be related to different social habits between women and men, leading to greater exposure of the latter. Having demonstrated that it was possible to identify cases of active tuberculosis, a preliminary whole genome study was undertaken to examine host susceptibility and resistance genes that determine the NRAMP protein and killer-cell immunoglobulin-like receptors (KIRS), in the presence and absence of tuberculosis. (Spigelman et al. 2015) The aim was to seek any correlation between KIR isotypes that inhibit natural killer (NK) cell activity and specific HLA class 1 allotypes. A type 2 variation of the promoter region was found in every patient infected with tuberculosis. The comparable recovery of M. tuberculosis DNA from bone and mummified tissue is in contrast with findings from studies of mammalian DNA (Lassen et al. 1994). We have suggested previously that this may be due, at least in part, to the greater stability of the GC-rich mycobacterial DNA, and the persistence of the lipid-rich bacterial cell wall. The survival of M. tuberculosis in formalin-fixed tissue (Gerston et al. 2004) is an indication of the protective effect of the cell wall. As the majority of archaeological remains are skeletalized, it is useful to be able to confirm that ribs are excellent specimens for M. tuberculosis DNA examination. However, the data show that evidence of M. tuberculosis infection is more likely to be detected if multiple samples, including tissues, are examined. Therefore, examination of totally skeletalized remains may lead to an underestimation of the extent of infection. The higher proportion of pleural samples positive for M. tuberculosis DNA confirms earlier findings (Fletcher et al. 2003a; Donoghue et al. 2011) and reflects the pathological changes associated with a chronic tuberculous infection. Lesions were rarely noted; only one or two of the ribs sampled showed any sign of a lesion. Ribs positive for M. tuberculosis indicate contiguous spread from an infected lung and pleura, because the inner surface of the bone was sampled. It is known that M. tuberculosis DNA can be found in ribs without lesions (Baker et al. 2015), and one group concluded there was no relationship between visceral surface rib lesions and tuberculosis (Mays et al. 2007). The present study confirms that the use of M. tuberculosis-specific PCR provides a more accurate estimate of the extent of infection with M. tuberculosis, than can be obtained by gross pathology alone. In the better-preserved samples, further characterization of the mycobacterial DNA was possible. Where a result was obtained, only M. tuberculosis was identified, neither Mycobacterium bovis nor any other member of the M. tuberculosis complex. This agrees with other archaeological findings (Pap et al. 1999; Arrieza et al. 1995; Braun et al. 1998) and supports the latest hypothesis on the evolution of the M. tuberculosis complex. M. tuberculosis is a distinct lineage, and both ancient and modern lineages of M. tuberculosis can be identified from ancient Egypt (Zink et al. 2007; Donoghue 2016). There is very little evidence of M. bovis in historic human populations, but it has been identified in human remains from the Iron Age in South Siberia, where pastoralists over-wintered with their animals (Taylor et al. 2007). The high numbers of the Vác eighteenth-century population shown to have tuberculosis have encouraged others to undertake further investigations. Lemma et al. (2008) investigated whether any of the human remains still contained viable M. tuberculosis. It is known that the mycobacterial cell wall is lipid-rich and highly

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resistant to desiccation. M. tuberculosis is also known to withstand exposure to embalming fluid and formalin. Guinea pig inoculation and resuscitation factor were applied. There were a few positive results using PCR but no signs of growth. Hendy et al. (2016) performed a proteomic analysis of seven mummified lung tissue samples from Vác, including five that were reported positive by DNA analysis (Fletcher et al. 2003a; Donoghue et al. 2011). Four peptides were identified that had unique matches to the M. tuberculosis complex, but none were unique to M. tuberculosis sensu stricto. The impact of tuberculosis is significantly enhanced by comorbidity. An example is that of a male infant, aged 1.5–2.5 years, who not only had tuberculosis but also had a rare form of leukemia, Langerhans cell histiocytosis (LCH) (Spigelman et al. 2006). There were rib lesions with no evidence of healing, on ribs petrous temporal bone and pelvic bone. LCH is believed to be a neoplasm, and the peak incidence is in children aged 1–3 years. Tuberculosis causes greater mortality today than at any other time in history, approximately 1.5 million people per year. It is estimated that one-third of the world population is infected, and the highest burden is in adult men (WHO 2019). It is crucial, therefore, to increase our understanding of the interactions between M. tuberculosis and its host. In Europe between the seventeenth and the nineteenth centuries, tuberculosis caused one in four deaths. Human remains from Vác enable the study of this host/pathogen interaction with historical strains of M. tuberculosis, unaffected by the selection pressure caused by the introduction of antibiotics. Acknowledgments A grant from the Wellcome Trust (UK), the Wenkart Foundation (Australia), and the Ancient Ills Modern Cures Fund, Sydney, Australia, funded preliminary work, led by Mr Leon Fink. During the years of the research, several Hungarian funds supported the work: NKA (Hungarian Cultural Fund), OTKA (Hungarian National Science Foundation), T023671 (Interdisciplinary mummy project I), OTKA 61155 (Interdisciplinary mummy project II), and NKFI 125561 (Tuberculosis and evolution). Dr Thomas Wenkart of the Wenkart Foundation and Professor Daniel Spira, from the Hebrew University Medical School, Jerusalem, participated in the endoscopic examinations at the Natural History Museum, Budapest, Hungary. The endoscope was kindly loaned by the Karl Storz Company and its CEO, Dr H. C. Mult. Sybill Storz. The earlier contribution to the project by Dr Helen Fletcher is gratefully acknowledged. Dr G. Michael Taylor (Imperial College London), Dr Adri van der Zanden (Apeldoorn, The Netherlands), and Dr Hamidou Traoré (London School of Hygiene and Tropical Medicine) kindly carried out the spoligotyping. Excellent technical assistance was provided by the following students: Sheima Abdulla Ahmed, Farah Aladin, Lynn Biderman, Rebecca Butt, Anneli Cooper, Hussein Farah, Poonam Gokal, Lauren Goldschmidt, Jaffer Sufi, and Ria Cickers. Tibor Bolcskei and Marian Ciuca kindly translated the eighteenth-century Hungarian and Latin texts.

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Bog Bodies

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Bog Bodies - Context . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Geographic Distribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Historical and Cultural Contexts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Potential Intents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientific Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Defining a Peat Bog . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Characteristics of Peat Bogs Impacting Body Preservation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Common Characteristics of Bog Bodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Analytical Challenges in Bog Body Research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Case of the Weerdinge Couple . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preservation Challenges . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

This chapter reviews the unique group of mummies known as the bog bodies. The reader is provided with a review of the geographical distribution of bog bodies as well as the associated historical and cultural contexts. While some bog bodies were likely victims of accidental drownings, the chapter explores the possible alternate reasons why an individual may have been placed in a bog. The science of peat bogs is discussed with a focus on those characteristics of the bog environment that impacts bog body preservation. The varied characteristics of bog bodies are described with regards to the decalcification of bony structures and R. G. Beckett (*) Biomedical Sciences, Bioanthropology Research Institute, Quinnipiac University, Hamden, CT, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_22

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soft tissue preservation. Given the changes experienced by the bog bodies, analytical challenges are explored. The bog bodies known as the Weerdinge Couple are presented as a case study demonstrating the impact of mummification in the bog, the analytical challenges, the possible cultural implications, and the variability of preservation. The chapter concludes with a discussion of the challenges associated with the preservation and conservation of bog bodies. Keywords

Bog Bodies · Peat bogs · Paleoimaging · Spontaneous mummification · Human sacrifice

Introduction Background As we consider the varied concepts and constructs of human and animal mummification, the bog body is truly a unique phenomenon. We first need to place the bog bodies into their appropriate classification in order to better understand not only the processes involved but also the cultural considerations surrounding these mummies. Dr. Arthur Aufderheide (2003) suggests a taxonomy of mummification which includes four distinct classifications. The first of these is anthropogenic (artificial) mummification in which the human remains are intentionally subjected to a process with the intent of preserving the remains and maintaining them in their human shape and form. Egyptian mummification using evisceration and natron application is the typical example used to describe mummies in this classification. There are many examples of intentional mummification around the world. The smoked bodies of Papua New Guinea and the Ibaloy of the Philippines are just a few such examples. The next major classification is that of spontaneous (natural) mummification. In this classification, there is no external treatment of the remains and mummification is a result of the natural forces of nature. The contextual environment in which spontaneous mummies are found plays a critical role not only in the preservation of the body but also on the mechanism of that preservation. For example, a natural mummy produced in a cool dry environment such as a crypt below a religious building would be mummified through a process of desiccation where the body fluids are wicked into the burial garment and eventually evaporate into the surrounding air. Without an aqueous environment, the natural processes of decomposition are impeded or halted resulting in mummification. Another example of an environment where natural mummification may occur is in areas where the ambient temperature is such that bacterial action on a corpse is halted, such as the high-altitude Inca mummies from the Andes or Ötzi from the Alps. In contrast to these examples, natural mummification can also occur in wet environments where the conditions are such that the forces of decomposition are stopped resulting in a preserved body. Two examples of wet environments are earthy environments or graves where the soil is

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alkaline resulting in saponification while the other is a wet environment that has an acidic pH as is seen in peat bogs. The latter results in a completely different type of spontaneous mummification when compared to the processes involved with mummification in a dry arid location, a cold high-altitude context, or a wet burial in alkaline soil. Thus, the bog bodies represent a unique group of mummies within the spontaneous taxonomic classification. It is important to note that Aufderheide (2003) offers a sub-classification within the spontaneous mummification category. The classification of spontaneous enhanced is distinguished from spontaneous mummification in that these mummies are produced when a culture realizes that conditions in certain locations, like a cave or mountain top, produce natural mummies. The culture then decides to inter their deceased in these or similar locations to affect the preservation of their dead. Even though there is intent, there is no actual treatment of the body itself. This distinction, spontaneous versus spontaneous enhanced, is an important one to make as we consider the possible reasons behind the formation of bog bodies. Given that the bog bodies are discovered many years after their deposition into a bog environment, there seems to be no attempt at spontaneous enhanced mummification in that whoever placed the body in the bog had no intention to preserve the body. In this chapter, we will explore bog body mummification first from a contextual framework regarding where bog bodies have been found as well as a discussion of the possible motives for relegating a body to a burial in a peat bog. We will then describe the natural processes involved in the production of a bog body through a review of what a peat bog is and its associated chemistry. We will then examine the preservation stages of bog body mummification as well as the limiting factors involved. Finally, we will describe specific bog body case studies as well as the preservation challenges associated with these bodies once removed from the peat environment.

Bog Bodies - Context Geographic Distribution It would seem that bog bodies would be found anywhere that there are peat bogs. This is not the case however. The geographic distribution of discovered bog bodies has been in northwestern Europe. Countries in which bog bodies have been found include Denmark, Germany, Sweden, Norway, Poland, the Netherlands, the United Kingdom and Ireland (van der Sanden 1996). Although there are peat bogs in North America, no bog bodies have been discovered there (Gill-Frerking 2015) (Fig. 1). One exception is at the Windover Archaeological site. The Windover Archaeological site is a National Historic Landmark in Brevard County near Titusville, Florida, USA. This is an early archaic site (8000–5000 BCE) characterized as a muck pond of primarily peat. One hundred sixty-eight skeletal remains have been excavated at the site with preserved brain tissue found in several of the skulls. While these are not the typical findings that researchers would refer to as bog bodies, the fact that they were discovered in the peat layer at the bottom of a muck pond and that

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Fig. 1 Global distribution of peat lands. (Retrieved from- https://www.grida.no/resources/12546, Map by Levi Westerveld – used with permission)

some of the brain tissue was preserved would qualify these individuals as bog discoveries (Gill-Frerking 2015). Bogs themselves are found in even a wider distribution pattern. The world’s largest peat bogs are found in the Western Siberian lowlands of Russia. These wetlands cover over a million square kilometers (386,102 square miles). In North America the largest peat bogs are found in the Hudson Bay lowland area and the Mackenzie River basin. In the Southern hemisphere, the Magellanic moorland in southern most Chile and Argentina covers around 44,000 square kilometers (17,000 square miles). The Republic of Congo also boasts a large peat bog. No bog bodies have been discovered in these locations, however, with the exception of Western Siberia (Maslyuzhenko and Shilov 2006). This case is discussed in a separate ▶ Chap. 34, “Bog Bodies and Natural Mummification of Siberia” in this volume. Although we do not know the actual number of bog bodies discovered, there are estimates, with some likely quite accurate, available from geographic locations that are worth noting here. In Ireland about 130 individuals have been recorded (O’Floinn 2006). Turner (1995) reports 140 from England, Scotland, and Wales. In the northern Netherlands, in 2011, the reported number was 31. There are reported to be around 120 finds in Germany (Eisenbeib 2003; Gebühr 2002). While other geographic areas have reported finds, it is thought that only about 45–53 individual bog bodies remain in a preserved state. While there are hundreds of bog bodies that have been discovered across varied geographic locations, it is well beyond the scope of this chapter to offer detail on each individual case. With that said we will discuss some specific cases of bog mummification later in this chapter.

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Historical and Cultural Contexts While bog bodies demonstrate a wide geographical distribution, these bodies are also widespread chronologically. Successful carbon dating indicates that the earliest known bog body was around 8000 BCE with the most recent being from the Second World War era, about 1945 CE. Given the broad range of chronologic distribution it is helpful to place the bodies classified in to temporal ages. One such classification approach places bodies into three broad time frames including (1) the Mesolithic (~10,000 to 8,000 BCE) – Bronze age (~3,200 to 600 BCE in Europe), (2) the iron age (beginning around 600 BCE in Northern Europe), and (3) the Medieval (~500 to 1500 CE) to Modern period (Fischer 1998; Aufderheide 2003). The fact that there have been bog bodies found across this wide range of prehistory and history has served as an impediment to researchers and prevents creating categories or groups from which to draw broader conclusions as they relate to a specific cultural context. The chronological distribution necessitates considering each body as an individual case. With that said, many of the individual cases do hold cultural significance in terms of historical known traditions of a given region. Further, the relationship between an individual and a culture of a region or period may be implied from the burial style, clothing, and associated grave goods. A further exploration of the cultural implications of the varied bog body finds will be presented in the following section regarding potential intents. The first reported discovery of a bog body was in Shalkholz Fen in Germany 1640 CE (Nield 1986; Brothwell 1996). In 1747 and 1781, two more bog bodies were found (Gmelch and Gmelch 1980). The Second World War bog body, that of a Russian pilot, was discovered in 1998 (Fischer 1998). Old Croghan man was discovered in Ireland in 2003 as well as the Clonycavan man. In 2011, a bog body dating to around 2,000 BCE was discovered on Bord na Móna land in Co Laois, Ireland. The actual number of bog bodies that have been discovered will likely never be known. Alfred Dieck began to catalogue bog bodies first in Germany, then worldwide. Dieck’s first count was 465 in 1958, followed by 1354 in 1972, and finally more than 1850 in 1986, the year of his death (Dieck 1986; Turner and Briggs 1986). Although ambitious, Dieck’s efforts have been largely discredited due to his broad criteria for inclusion, while some bodies included in his registry were only body parts or never even existed (Fischer 1998). The first studies regarding bog bodies were conducted in the 1870s by Johanna Mestorf, a German archaeologist (van Vilsteren 2004). Mestorf developed a catalogue of the known mummies and interpreted the bodies as being murder victims. There are certainly other reasons why it is not probable that we will know the number of bog bodies that have been discovered. It is possible that bog bodies that were discovered were never reported. Bog bodies have been typically found during peat cutting operations and if a body was discovered, it may have been reburied or destroyed due to superstitious traditions (Gill-Frerking 2015). Heather Gill Frerking (2015), a scholar on the study of bog bodies, also notes that once peat harvesting

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became mechanized, bodies may have simply been ‘processed’ along with the peat. Additionally, there are likely still bog bodies to be found. Another factor that complicates the issue of cataloging an accurate number of bog bodies is that many exist on paper only. These have been called “paper bog bodies” in that the bodies may exist in church, municipal, or museum records, but the physical body itself is lost to time, deterioration, or research attrition. The bodies may have also been “misplaced” and are yet to be rediscovered (van der Sanden 1996, 2013; Gill-Frerking 2015).

Potential Intents There is much speculation regarding the reason individuals ended up in a peat bog and eventually became mummified. It would seem that inhumation into a bog would be an act of finality as the bodies were never intended to be seen again. Given that bog bodies represent a very long-time span, have been discovered in a variety of geographic contexts, and cross-varied cultural periods, it is not possible to group all bodies into a unified theory of purpose or intent for interment in a peat bog. Given the high degree of variability in temporal and geographic contexts, researchers have theorized the bog body phenomena based on specific time periods, geographic locations, and what is known about that time in place through historical records. Keep in mind that no actual written records exist for those bodies from Antiquity. The major theories regarding why individuals were found in a bog range from the funerary practices of the prevailing culture to torture of an enemy or criminal. Specific motives include cultural burials, accidents, murder victims, suicide, punishment, ritual sacrifices, and individuals who were social misfits. It is highly likely that some of the bog bodies were accidental drownings occurring as someone attempted to navigate the bog environment. Peat bogs are notoriously hazardous and it would be quite probable that a person simply lost their balance or tripped, fell in the bog waters and drown. There they remained until they were discovered years or centuries later. In the case of accidental drowning in a bog and, if they were in fact preserved, one would expect that while the body may have been clothed there would be no associated grave goods. Bodies may have been interred in bogs as a part of the typical burial pattern seen in a given region. The usual deaths of a village or township may have been placed in an area that was unusable for farming or pasturing such as in a bog. In these routine burials associated with standard mortuary practices one may expect to find evidence of not only clothing but other grave goods. Criminals may have used the bogs to hide or otherwise dispose of a body that had been murdered. One modern case of murder highlights the possibility that the behavior may have occurred in past times as well. In 1984 a human head was recovered from a bog near Lindow close to Manchester, United Kingdom. A local man confessed to murdering and dismembering his wife’s body 20 years earlier. He stated that he hid the body parts in the bog. He was tried and convicted of murder. The recovered bog head was radiocarbon dated to be around 1740 years old. Even

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though this was not his wife’s head the conviction held. The story does suggest that the bog was a place used to put things, or in this case people, where they would never be found (Aufderheide 2003). Bog bodies may also represent individuals who committed suicide. While this is highly speculative, the possibility should be considered. The distraught person would wander into the bog, end his or her life through a self-inflicted wound or by ingesting poison and be swallowed up by the bog. Another theory regarding the reason for bodies being buried in a bog is that of some superstitious belief. Several bog bodies have been discovered who had been tied to the bog with wooden stakes or with large rocks. This may have been an attempt to keep the dead person’s ‘spirit’ from moving around. In some cases, a stake was found to be piercing the chest symbolically destroying or slaying the spirit. It cannot be known for certain if superstition was the motivating factor in these cases. Yet there is a recurring theme that suggests that there was a fear that for certain individuals, be they suicide, victims of violence, murder or accident peace would not be found in a typical grave. The spirit of these deceased individuals needed to be rendered harmless through desecration of their bodies, removing any clothing and cutting their hair and finally disposed of in the bog. However, these bodies may have been a type of punishment burial for deviations from societal norms or for crimes committed (Glob 1965: Fischer 2007). There is ample evidence that some of the discovered bog bodies show signs of inflicted violence (Glob 1965; Aufderheide 2003; Fischer 2007). Ropes still around the necks of some bog bodies have been well preserved by the bog. The position of the ropes suggests hanging or a garroting of the individual. Stab wounds are also found in some of these cases. It is important to note that it is not possible to determine if these penetrating wounds were inflicted before or after the death of the individual. The Roman historian Tacitus writes in 98 CE that the Gauls would drown individuals in bogs for crimes committed against the state or civility. This lends credence to the postulation that, at least in some cases, the interment of a body in a bog was the result of criminal punishment. If this were a common punishment it would have served as a deterrent to criminal activity (Tacitus 98 CE/ 1981). The most common theory regarding the use of the bog as a place of inhumation is that of ritualistic sacrifice to a deity. The wounds inflicted in those bodies theorized to have been the result of criminal punishment are the same types of wounds that supports the ritualistic sacrifice theory. Stabbing, blunt force trauma to the skull, chest or extremities, and garroting are cited as evidence of votive offerings to the gods. The Celtics would often use watery sites for votive offerings of material goods. It could be then that, in some cases, the bog was considered a special place, perhaps even sacred (Bradley 1990). Aufderheide (2003) reports that the stomach and intestinal contents of some Danish bog bodies show pollens and seeds suggesting a death period of late winter or early spring. Iron age Danish groups would hold fertility festivals at this time of year which involved human sacrifice hoping to appease the gods resulting in a strong agricultural season. The associated bog bodies were found to have been individuals with delicate features, no callouses, and smooth

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fingernails suggesting that they were not likely criminals but rather persons selected from within their normal societal group. Given the various theories offered above, and considering the broad temporal context represented by the bog bodies, it is clear that no one general theory explains the intent behind all of the bog bodies discovered. Each bog body must be considered as an individual case with careful consideration given to whatever historical context is available. The archaeological record as well as past and present environmental conditions must be considered in the attempt to reconstruct the life and death of these unique mummies.

Scientific Perspectives Defining a Peat Bog A peat bog is a unique ecologic region covering only about 1% of the earth’s surface. Peat bogs are not evenly distributed across the globe with the highest concentrations found in Ireland, Northern Europe, and Western Siberia (Fig. 2). Peat is produced from partially decomposed plants in water. Peat exists in several forms based on its constituent plant material. Common descriptors include bog peat made of mosses, meadow peat made up of grasses, forest or wood peat made primarily of decomposing trees, and sea peat made up of seaweed (Neilson et al. 1939). Not all peat is created equal when considering the preservation qualities. Depending on the

Fig. 2 A typical peat bog. (Retrieved from: https://siberiantimes.com/ecology/opinion/features/ f0099-new-warning-about-climate-change-linked-to-peat-bogs/)

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Fig. 3 Sphagnum moss is the critical biomass that creates the chemistry require for bog body preservation. (Retrieved from: https://www. gardeningknowhow.com/ garden-how-to/soil-fertilizers/ sphagnum-moss-vssphagnum-peat-moss.htm)

location and the associated topography a peat bog can be considered a lowmoor bog which is flushed with groundwater flowing from higher ground or a highmoor bog with is watered by rainfall. The major difference in these two bogs is that the lowmoor peat bog contains minerals and salts that tend to buffer the bog yielding a pH of around 5.5 to 6.5. Although slightly acidic, this pH range does not correlate with preservation of human or animal tissues. The highmoor peat bog, because of the rain water, has a lower pH range of 3.2 to 4.5. This pH range as well as having the lower concentrations of nitrogen and phosphorous when compared to the lowmoor bogs, creates an environment suited for the growth and spread of Sphagnum moss (Fig. 3). Due to its growth characteristics, sphagnum moss gives the bog a raised surface thus such bogs are referred to as raised bogs. The Sphagnum layer compresses the peat below to the degree that molecular oxygen only reaches to about 30-50cms. Below that level the peat is anoxic to the point where aerobic bacteria have a hard time surviving. It is important to note these Sphagnum bogs are those with the potential to preserve the soft tissues of humans and other animals (Aufderheide 2003). Peat has been harvested as a fuel for centuries (Fig. 4). Even today peat is cut to be used as fuel. Cut peat is often stored stacked like bricks (Fig. 5). It is fairly efficient as a fuel falling between coal and wood. It is during peat cutting procedures that most of the bog mummies have been discovered. As stated earlier, with the mechanization of peat harvesting one has to wonder how many bog bodies are never discovered but rather processed along with the peat, stacked, and used as fuel.

Characteristics of Peat Bogs Impacting Body Preservation The characteristics of a peat bog that may be related to the preservation of human and animal remains are varied. First, the pH of the peat bogs where mummies have been found is around 5.5–6.5 (Aufderheide 2003). This mild acidic characteristic would impede the activity of some microorganisms. While the peat bog becomes anoxic, aerobic bacteria are deprived of their necessary supply of oxygen. It is important to

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Fig. 4 Harvesting peat. (Photo courtesy of author)

Fig. 5 Peat used for fuel are stacked like bricks. (Photo courtesy of author)

recall that anaerobic bacteria can also decompose human tissue. So, the anaerobic conditions and the pH, while having some impact on decomposition, cannot fully explain the mummification forces seen in a peat bog. This brings us back to the characteristics of Sphagnum. Upon death, Sphagnum releases a polysaccharide component called sphagnan, which possesses chelating properties that bind metal ions. As a result of this process, some metals are no longer available to bacteria, depriving them of an important source of nourishment. This lack of nourishment slows down microbial activity. Additionally, calcium chelation results in this element being leached out of the bones, which become pliable or even dissolved, although this is not a necessarily uniform phenomenon. Another chemical process which takes place within the bogs is the “Maillard reaction” which involves sphagnan, ammonia, amino-acids, and polypeptide amino-groups. The humic acids

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resulting from this biodegradation, which are among the major organic substances of the bog pit, are extremely resistant to microbial attack due to the lack of nitrogen which becomes unavailable to microorganisms (Painter 1991). Importantly, the abovementioned reaction also creates cross-linking of the collagen fibers, which expels water from the interstices and confers resistance to bacterial attack. Sphagnan and its holocellulose also inhibit the activity of the enzymes secreted by bacteria, further contributing to the human remains preservation (Aufderheide 2011). Tannins are sometimes present in bogs, and it is plausible to believe that they also play a role in the mummification process (Painter 1991). Similar to the tanning effect, the crosslinking which results in the expulsion of water from the tissues makes them more resistant to decomposition. These and the substances involved in the Maillard reaction confer a brown color to the bodies. Taphonomic observations reveal that preservation of the connective tissue is the most common structure seen in bog bodies. Other characteristics of preserved bog bodies include decalcified bones, the body surface deprived of the epidermis, and little or differential muscle preservation. The skin collagen and keratin of hair and nails are very well preserved. Hair color, however, is modified as melanin oxidizes to a lighter tone. As far as the internal organs are concerned, preservation involves those with a mainly collagenous content, such as the lungs and the intestinal walls (Brothwell and Gill-Robinson 2002). Not all bodies that end up in a peat bog are preserved. Several environmental factors impact the process described in the above paragraphs. If the peat watery environment is diluted from heavy rains or floods with non-peat waters, the preserving constituents of the bog are of a lower concentration. The chelation potential would be impeded by foreign metal ions from these waters. The pH too may be altered in these conditions. The result would be a skeletonized body rather than soft tissue preservation with decalcified skeletal features. In these cases, the body may in fact be completely dissolved. Indeed, skeletons are discovered in bogs and are often considered bog bodies along with their mummified counterparts.

Common Characteristics of Bog Bodies While it is true that each bog body presents with some degree of variation in preservation there are some commonalities worth noting. The most common feature in bog bodies is that they present with decalcified bones and teeth. Recall that, due to the Sphagnum, the peat environment at the level where the bodies are found is under considerable pressure. Thus, once the bones have been sufficiently decalcified the remaining soft tissues are compressed by this weight giving the bog body the appearance of having been crushed. The thorax is usually collapsed as is the cranium. This can be problematic when examining a bog body in that collapsed anatomical features distort the spatial relationship of organs and structures within the body. A collapsed skull may have the appearance of being crushed prior to the death of the individual. This may be misinterpreted as the cause of death when in fact it is a pseudo-trauma brought on by the compression forces of the bog associated with the loss of structural integrity from the decalcification process.

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There is often excellent preservation of finger and toenails, keratin in the hair and the dermis. Any perimortem lacerations or wounds are generally well preserved and have been used to interpret the death scenario of individual mummies. It is important to recall that it is not possible to determine if a wound was inflicted premortem or postmortem, thus most such wounds are classified as perimortem in nature (Asingh and Lynnerup 2007; Fischer 2007; Lynnerup 2007). Internal organs are preserved with considerable variability. This makes inter-bog body comparisons difficult to say the least. Generally, those structures having a greater degree of collagen are more likely to be preserved. These structures include the lung, pericardial sac, intestinal wall, and ligaments. In contrast, structures with less collagen and more epithelial cell components tend to not be as well preserved. These would include the kidneys and the liver which are often shrunk in size, spatially misplaced, and compressed (Aufderheide 2003). An interesting series of experiments were conducted in an attempt to better understand the preservation forces associated with bog bodies. Heather Gill-Robinson (2002) used pig analogues which were placed in bogs in the United Kingdom. The pigs were exhumed from the bogs at predetermined intervals which began at 6 months and continued to 3 years from the time of initial placement. The pigs that came out of the bog at the six-month mark were found to have decalcified ribs to a great degree and the skin had begun to darken. Those pigs that came out at the 3-year mark showed a greater degree of bone decalcification and were darker than the pigs exhumed at the 6-month mark. The researcher reported an interesting observation in that the degree of preservation appeared to be related to the level of water with those in more watery bogs showing the greatest degree of soft tissue preservation.

Analytical Challenges in Bog Body Research Given the unique nature of the preservation forces associated with bog bodies, several challenges arise. The lack of homogeneity among individual bog bodies creates a barrier to inter-mummy comparisons. While it is true that several preservation features such as the decalcification of bone and teeth are shared among the majority of these mummies, many other variables impact both the presentation status and, thus, the interpretability of the information derived. The bodies represent varied time periods and varied climate patterns or cycles. Even today, the impact of climate change, in particular the acidification of rain water, will potentially impact the preservation status of bog bodies yet to be discovered. Coupled with modern land use variations that often include the draining of bogs for alternate use or development, agricultural run-off has a potential of altering the chemical nature of these bogs. Thus, the peat environment would be rendered less hospitable to the preservation of any bog bodies in their midst. Land use change, global weather cycles and climate change will all influence the taphonomy of bodies interred in the affected bogs. Changes in their environments may influence or reactivate the decomposition processes. In most cases, a bog body is from an era or is associated with a culture that did not keep written records of events. This is a major limiting factor when attempting to

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develop the contextual framework which would allow for a more complete bioarchaeological analysis. Artifacts are rare as are complete knowledge of cultural aspects such as religious rituals. Lacking the necessary archaeological data analysis of the bioarchaeology of care is also rendered inapplicable. When faced with a bog body that has been deformed by the decalcification of skeletal features and the lack of useful paleopathological information, it is nearly impossible to reconstruct the lifeways of these individuals. It is quite difficult to determine if any orthopedic impairments existed or if any disease of organ systems were present. A few exceptions exist. One is the Yde Girl who was determined to have scoliosis. Another case is that of Zweeloo Woman who had short forearms (Bianucci et al. 2012). Aschbroeken Man had inhibited growth as a child and a poorly healed right humerus (van der Sanden 2013). The degree to which a lifeway scenario can be developed is dependent entirely on the remaining anatomic material. In one case, a mostly skeletonized bog body with only a small amount of skin remaining was discovered in Uchter Moor near Nienburg Germany. However, enough of the skeletal elements allowed researchers to develop a profile of the individual. It was determined that the skeleton was that of a 16–21-year-old female and had poor dental hygiene. The biostress markers on the bony articulations suggest that she had a hard life. Growth arrest plates, or Harris lines, indicate that she suffered from bouts of malnutrition and chronic inflammation. Her spine showed an abnormal curvature and a benign tumor at the base of her skull was observed. There were also two healed skull fractures likely caused by blunt force trauma (Bauerochse et al. 2008). The body is known as Moora and was carbon dated to the seventh century BCE. The cause of death could not be determined and it is generally assumed that she lost her way in the bog where she died. One of the primary tools in bioarchaeological research is that of paleoimaging (Beckett and Conlogue 2010; Beckett and Conlogue 2020). Applying multimodal imaging methods to mummified remains can yield a great amount of data allowing researchers to develop a “biography” of sorts of the individual being studied. These data are then analyzed by various experts to establish a diagnosis or interpretation by consensus. The decalcified bones associated with bog bodies challenge researchers to extend the boundaries of normal imaging parameters in order to derive images that are useable for interpretations. Standard radiography needs to use a low kVp in order to pick up any semblance of remaining bone. One such case related to field imaging challenges will be discussed later in this chapter. Even if standard radiographs can be coaxed into producing an image, the superimposition of shadows from more dense soft tissue structures reduces the interpretability of the images. Computed tomography (CT) scans are challenged in that the compressive aspect of the decalcified bog body disrupts the spatial relationship of internal organs as well as a distortion of expected morphology. These variations can lead to misinterpretation of data as well as misidentification of structures. The post processing of CT data can, however, help in presenting the images in a perspective more closely resembling what the structure would appear like without the distortion brought on by pressure over time buried in a bog. Thus, post-processed CT data and 3D rendering can piece the puzzle back together aiding in the assessment of supposed injury patterns or other

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anthropological concerns such as age at death or sex determinations (Lynnerup 2015). Paleoimagers unfamiliar with the unique characteristics of bog bodies or lacking skills to properly adjust sophisticated imaging technology can limit the information derived. In the study of mummified remains analysis of the dentition can yield important information about how the individual interacted with their environment. A variety of dental conditions inform researchers about diet, presence of periodontal disease, age at death, periods of growth arrest, cavities and abscesses associated with the individual. This, in turn, may suggest societal features such as status. In the case of bog bodies, the enamel is often partially or entirely removed rendering study of the dentition less than helpful. Considering the injury patterns seen on some of the bog bodies, it is safe to assume that some of these wounds may have resulted as the death event of the individual. It is not possible to say with any degree of certainty if wounds such as a penetrating stab or slashing were inflicted pre mortem or post mortem. The best that can be said is that the wounds occurred around the time of death (perimortem). If one adopts the ritual sacrifice theory or the punishment theory of bog body interment, those cases in which garroting or hanging were involved may have been the death event. When more is known about the cultural norms associated with the location of the find and the socio-spiritual practices as well as the temporal context, determinations may be less generalized. An interesting case is that of the Grauballe Man. Grauballe Man was found to have had a “blow to the head” injury which was for many years interpreted as an attempt to keep him from escaping a ritualistic sacrifice. Later post-process CT scans with 3D renderings put the images of the skull bones back together, and it was revealed that there had been no blow to the skull but rather a deformed skull appearance due to the softened decalcified bones subjected to the compressive forces of the peat (Lynnerup 2015). Of critical importance to mummy science is the study of ancient disease patterns present in the mummified remains of past peoples. Paleopathological analysis is a critical component in the understanding of lifeways of mummified individuals or larger population studies of related groupings of mummies. Given the aforementioned limitations to skeletal analysis due to partial or complete decalcification, skeletal anomalies are often lost to the forces of the bog. As mentioned, some bog bodies are skeletonized in which case skeletal analysis of paleopathological conditions such as bony lesions, tumors, past traumas, and biological stress factors can be determined. Although soft tissues are preserved in many bog bodies, these soft tissues and/or organs are usually distorted, smaller, and often misplaced within the body. The paleopathologist then must consider interpretations with great caution. Another important aspect of mummy research involves the analysis of any remaining stomach or intestinal material that may suggest what was the most recent foods consumed by the individual. In addition, coprolite analysis for paleoparasitological analysis can yield additional information regarding a given case. Several interesting studies have been conducted on bog bodies related to the contents of the alimentary system with a few described here. First, the Tollund Man (circa 220 BCE)

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had ingested a porridge of gruel of barley, flax, false flax, and knotgrass approximately 12 h prior to his death (Fischer 1984). Parasitological analysis showed that the Tollund Man had whipworm and mawworn infestations. Another bog body, The Lindow Man (circa first century CE) suffered from parasitic worms and his last meal was likely unleavened bread made from wheat and barley and cooked on an open fire. Perhaps one of the more interesting cases related to intestinal contents was that of Grauballe Man (circa 310 BCE). Grauballe Man’s final meal consisted of a vegetable porridge with some meat (Fischer 1984) with traces of ergot, a toxin that can bring on hallucinations. The presence of ergot in Grauballe Man’s stomach contents led researchers to speculate that the ergot represented evidence of ritualistic sacrifice. This theory persisted until the contents were reexamined and it was determined that the level of toxin was within safety limits and would not have been enough to affect his metal status (Dicks 2003). An important note is that the contents of each of these examples suggest that they were interred in the winter months meaning that the temperature of the bog was such that mummification was more likely. Another significant case was that of the parasitological analysis of the liver sections and intestinal wash of the Zweeloo Woman (Searcey et al. 2013). The Zweeloo Woman was discovered by peat cutters in Zweeloo, Netherlands. Upon anthropological examination her age at the time of her death was 35–50 years (Bianucci et al. 2012). Radio carbon dating places the Zweeloo Woman in the Roman Perion (Searcey et al. 2013). One important finding from this study was that the liver and kidneys were preserved to the degree that paraffin sectioning for histological and parasitological analysis could be conducted. These organs are often so poorly preserved that they hold less analytical value (Aufderheide 2003). In this study, researchers were able to identify the eggs of D. dendriticum in the liver parenchyma sections. The significance here is that this represents the first true record of human dicrocoeliasis, a remarkable find (Searcey et al. 2013). While the analytical challenges surrounding the study of bog bodies are many, the attempts continue to be made supported with the advancement in both technology and technique. The greatest contribution to the study and understanding of bog bodies will continue to be the placement of individual bog bodies into their appropriate temporal and cultural contexts. While some writings exist such as the Roman historian Tacitus (56 CE–120 CE) and Ptolemy (c100 CE – c170 CE) (Aufderheide 2003), it is possible that their explanations of the observed death rituals of punishment and sacrifice may have been biased in an attempt to ensure expansion of the Roman Empire. These writings influence the interpretive thinking of modern scientists as they continue to unravel and attempt to understand the meanings behind the occurrences of bog bodies.

The Case of the Weerdinge Couple Much has been published about the various bog bodies that have been found. Bog bodies such as the Tollund Man, Grauballe Man, Lindow Man, Meenybraden Woman, and the Yde Girl and others have been presented in detail in a variety of

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other sources (Aufderheide 2003; Briggs and Turner 1986; O’Floinn 1995, Turner 1995; van der Plicht et al. 2004;). Here we will focus our discussion on the case of the Weerdinge Couple in order to illustrate the challenges associated with bog body research. The Weerdinge Couple was discovered in 1904 by a peat-cutter named Hilbrand Gringhuis in the Bourtanger Moor near Weerdingerveen, Drenthe, the Netherlands (van der Plicht et al. 2004) (Fig. 6). Once discovered, Gringhuis notified the local constable who helped bring the bodies out of the bog. The bodies were moist and flat so one was placed on top of the other, rolled up like a carpet, and transported to the police station and later to the Drents Museum (inv. No. N 1904/VII2) where the bodies remain today (Mummy Road Show 2002). As with all bog bodies, once removed from the watery bog environment, they dry and become brittle making handling very difficult and thus difficult to research. Given the variance in their stature, the mummies were initially thought to be the remains of a man and a woman. They were given the names of Darby for the taller of the two assumed to be male, and Joan for the presumed shorter female. The taller of the two had recognizable genitalia while the shorter mummy did not. In addition, both of their crania had been completely dissolved by the bog forces leaving shriveled bags of skin where their heads were with the taller mummy having more tissue remaining (Figs. 7 and 8). The Weerdinge Couple have been radiocarbon dated with the results indicating the bodies were from about 160 BCE and 220 CE (van der Plicht et al. 2004). The taller mummy exhibited a severe chest wound to the left mid-anterior thoracic cage starting at approximately the fourth rib and tracking downward to about the phrenic margin. Macroscopic examination showed that the edges of the wound were sharp indicating that the wound was inflicted at or around the time of death. The intestines are protruding out of the wound site and visible on the outside of the chest (Fig. 9). It has been speculated that this perimortem injury was possibly a ritualistic sacrifice. Strabo, a Roman historian, reported that Iron Age Europeans Fig. 6 The Weerdinge Couple at the Drents Museum, Assen, the Netherlands. Note the fragile appearance of the bodies as well as the flat presentation brought on by the weight of the bog layer. (Photo courtesy of author)

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Fig. 7 The shorter mummy showing dissolved cranial bones and a collapsed appearance with very little soft tissue remaining. (Photo courtesy Larry Engel, used with permission)

Fig. 8 The taller mummy showing dissolved cranial bones and a collapsed appearance with a greater degree of soft tissue remaining. (Photo courtesy Larry Engel, used with permission)

would eviscerate in this manner in order to read the entrails in an attempt to divine the future (Strabo 10 CE/ 1977). In 2002, paleoimaging and DNA analysis was conducted on the Weerdinge Couple (Mummy Road Show 2002). Standard radiography was conducted on site at the Drents Museum using 8 X 10 inch Polaroid Instant Film as the image receptor (Fig. 10). Given that the skeletal components of the bodies were decalcified by the bog forces, as is the case in most bog bodies, standard radiography was challenged even at the lowest X-ray output settings. In order to decrease the output further than the instrument was designed for, researchers placed aluminum over the X-ray tube where the X-rays were emitted (Fig. 11). This allowed the researchers to visualize several of the decalcified bones and soft tissue where the integument had folded over onto itself (Fig. 12). Radiographs were able to demonstrate the ‘shell’ of the remaining bones in the forearm that crossed over the chest of the taller mummy (Figs. 13 and 14) as well as a few ‘ghosts’ of bones on the foot of the smaller

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Fig. 9 Intestines protruding from the wound of the taller mummy. (Photo courtesy of author)

Fig. 10 Smaller mummy being positioned for an X-ray using Polaroid Instant film as the image receptor. (Photo courtesy of author)

mummy. It was interesting in that these radiographs demonstrated that the decalcification was variable even in the same body. The smaller mummy had no detectable bones in the thorax or spine (Fig. 15). Both mummies were transported to a local hospital for a computed tomography (CT) scan. Neither of the mummies showed signs of internal organs or any remaining calcium in the bones.

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Fig. 11 Aluminum sheet (arrows) affixed to the X-ray tube to reduce the amount of X-rays allowing for image capture. (Photo courtesy of author)

Fig. 12 Shorter mummy: Polaroid X-ray image demonstrating the lack of visible bony structures. The skin folds are faint but present due to low density. (Photo courtesy of author)

The taller mummy had an opening in the right upper mid-clavicular thoracic region. This allowed researchers to introduce an endoscope into the thoracic cavity. The endoscope revealed the outline of ribs that were so decalcified that it did not show up on X-ray. The endoscope images also revealed what appeared to be the heart (Fig. 16). It is important to note that on both standard radiograph and CT scan no rib outlines or organs were visualized. This suggests that multimodal imaging

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Fig. 13 Taller mummy: Left forearm resting across the chest (arrows). (Photo courtesy of author)

Fig. 14 Taller mummy: Polaroid X-ray image showing some remaining calcium in the radius and ulnar. The X-ray demonstrates the variable degree of decalcification in the same body. (Photo courtesy of author)

approaches to bog mummy research must be considered in our attempts to extract useful information from these challenging cases. Samples were taken from each of the two bog bodies, a toe from the smaller mummy and a part of the olecranon from the taller body. The samples were sent to the Paleo-DNA lab in Thunder Bay Ontario, Canada, where researchers were able to extract usable data DNA. The results confirmed that both of the individuals were male and that they were not maternally related. This case demonstrates the challenges associated with bog body research. The decalcification of the bones along with the misshapen and displaced soft tissue features make even the usually most straight forward anthropological questions difficult to answer. Characteristics such as sex, stature, dental assessment, paleopathologies, biomechanical stress indicators etc. all become elusive. As this case

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Fig. 15 Smaller mummy: Polaroid X-ray of upper thorax and what remains of the head (arrow). There are no bony structures present. Skin folds can be seen. (Photo courtesy of author)

Fig. 16 View screen of endoscope showing and organized structure that is what appears to be the heart (arrows). (Photo courtesy of author)

demonstrates, some information can be derived and researchers studying bog bodies work tirelessly to discover what information they can in an attempt to tell the story of these mummified time travelers. Whether they were social outcasts, outlaws or sacrifices, the bog bodies remain as mysterious as the forces that have mummified them.

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Preservation Challenges As previously discussed, the peat bog is a watery environment and bodies subjected to that environment undergo a variety of changes ultimately reaching a state of stasis with their aqueous tomb. The chemistry of the bog and the undisturbed nature of bog body findspots, both in their isolation and in the depth at which bog bodies are found, contribute to the maintenance of the shape and size of the body. Once disturbed or exhumed, the preservation forces are disrupted. At exhumation, the bodies are moist and pliable but once the water begins to evaporate they become brittle, stiff and shrink in size (Lynnerup 2015). At exhumation, the bodies that are still moist are subject to alterations in their appearance by a variety of factors. Since most of the bodies are discovered during a peat harvesting operation, the machinery may damage the body in a manner that extremities, if present, can become disarticulated, displaced, or even lost. Once the body is handled in its immediate post peat stage, even the fingers, hands, or tools of the those bringing the body out of the peat can leave surface impressions altering the appearance lending interpretations to be questionable (Krebs and Ratjen 1956). These post-exhumation changes to the bodies create unique challenges for those who hope to research them further or who hope to maintain them for educational purposes in museum displays. Bodies discovered prior to World War II often desiccated to the degree where meaningful research could not be continued. At that time, the drying out of the bodies was the only way to preserve them. The body becomes fixed in the position that they dried out in. This desiccation also alters the tissues and the identifying features of tissue type, even within a single body part. For example, some bones maybe more decalcified than others as illustrated in the Weerdinge Couple case described above. In the post-World War II era, successful methods of preserving bog bodies have found their way into curatorial efforts. The Borremose Woman discovered in 1948 in Denmark was immersed in a formaldehyde solution and kept in a wet environment (Lynnerup 2015). The head of the Tollund Man discovered in 1950 was preserved in two manners. The body was placed in various fluids including formalin, acetic acid, and alcohol and then allowed to dry. Tollund Man’s head was treated quite differently. The head was first placed in alcohol to drive the bog water out of the cells. The alcohol was then replaced with the solvent toluol. The toluol was replaced with liquid paraffin which could be mixed with wax. Bee’s wax has a low viscosity and would replace the paraffin within the cells. The attempt was successful yet the head was about 12% of its original size (Fischer 2007; Asingh and Lynnerup 2007). The head is still preserved but the displayed body and head at the Silkeborg Museum is a recreation of the original to demonstrate what the body looked like at its discovery. Grauballe Man discovered in 1952 went through a different process of preservation. Conservator G. Lange-Korbak used a tanning method to preserve this bog body. The body was kept submerged in a solution of one third fresh oak, two thirds oak bark, and 0.2% of a disinfectant, Toxinol (Moesgaard Museum 2020a). After 18 months, the body was removed from the solution and rinsed in a bath of 10% Turkish-red oil in distilled water to dissuade loss of size (Asingh and Lynnerup

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2007). In the early 2000s, the Moesgaard Museum conservators created a nitrogen filled glass display case for Grauballe Man. This case is a climate-controlled case designed to maintain a constant environment in which oxygen, humidity, and temperature are precisely regulated so any breakdown of soft tissues could be minimized. The glass is also coated with a UV absorbent film (Moesgaard Museum 2020b). The current method of preserving bog bodies is freeze drying. Freeze drying allows the water to sublimate reducing drying stresses (Wills et al. 2014). The Lindow Man and the Meenybraddan Woman, as well as many others are conserved in this manner (Aufderheide 2003; Mulhall 2010). The examples provided here are a testament to the efforts made to preserve these bog bodies for academic study and educational enrichment of the public at large. Each bog body is a rare find and the unique natural forces of peat bogs have made them involuntary eye witnesses to their life and times.

Conclusion The bog bodies are a unique group of mummies that represent various time periods and cultural practices. They provide valuable insight into the lives of past peoples as bioarchaeologists attempt to reconstruct the lifeways of the individuals and the world they interacted with. They hold important information that helps to create a more accurate understanding of humankind’s journey through time. The nature of the process of mummification associated with the bog bodies present researchers with challenges not present with other naturally mummified bodies. These challenges are both contextual as well as methodological making information difficult to extract. New methods specific to bog body research need continued development. Once the bodies are removed from the watery bog, their preservation becomes an urgent concern. The need for rapid conservation has been mitigated to some degree through the practice of freeze drying the bodies, allowing researchers access for appropriate scientific study. Given that the bog bodies are usually discovered during peat harvesting, it is difficult to predict when and where another body or group of bodies will be found. Many bog bodies have been lost to time either through deterioration, mishandling, or misplacement. In addition, as urban development encroaches on bog land and other land use changes necessitate draining wetlands less bog bodies are likely to be discovered. Any bodies that are exposed would begin to deteriorate once out of their watery tombs. In conclusion, researchers must continue to develop new ideas and methods regarding the study of bog bodies. Further, researchers must continue to explore new preservation ideas and manage conservation efforts while still serving to educate the public. The study of bog bodies will no doubt continue with these efforts in mind as we attempt to learn more from and more about these truly unique time travelers.

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Cross-References ▶ Bog Bodies and Natural Mummification of Siberia

References Asingh P, Lynnerup N (eds) (2007) Grauballe man. An iron age bog body revisited. Archaeological Society Press, Jutland Aufderheide AC (ed) (2003) The scientific study of mummies. Cambridge University Press, Cambridge, MA Aufderheide AC (2011) Soft tissue taphonomy: a paleopathology perspective. IJJP 1:75–80 Bauerochse A, Habmann H, Püschel K (eds) (2008) Moora – the girl from the Uchter moor: an iron age bog body from Lower Saxony. Leidorf, Rahden Beckett RG, Conlogue GJ (2010) Paleoimaging: field applications for cultural remains and artifacts. CRC Press, Boca Raton Beckett RG, Conlogue GJ (eds) (2020) Advances in paleoimaging: applications for paleoanthropology, bioarchaeology, forensics, and cultural artifacts. CRC Press, Boca Raton Bianucci R, Brothwell D, van der Sanden W, Papageorgopoulou C, Gostner P, Pernter P, EgarterVigl E, Maixner F, Janko M, Piombino-Mascali M, Mattutino G, Rühli F, Zink A (2012) A possible case of dyschondrosteosis in a bog body from the Netherlands. J Archaeol Low Countries 4:34–67 Bradley R (ed) (1990) The passage of arms. Cambridge University Press, Cambridge, MA Briggs CS, Turner RC (1986) A gazetteer of bog burials from Britain and Ireland. In: Stead IM, Bourke JB, Brothwell D (eds) Lindow man: the body in the bog. Cornell University Press, Ithaca, pp 181–195 Brothwell D (1996) European bog bodies: current state of research and preservation. In: Spindler K, Wilfing H, Rastbichler-Zissering E, zur Nedden D, Nothuerfter H (eds) Human mummies: a global survey of their status and techniques of conservation. Springer, New York, pp 161–172 Brothwell D, Gill-Robinson H (2002) Taphonomic and forensic aspects of bog bodies. In: Haglund WD, Sorg MH (eds) Advances in forensic taphonomy: method, theory, and archaeological perspectives. CRC Press LLC, New York, pp 119–132 Dicks L (2003) Mystery in the mire. New Scientist 177(2382):38–41 Dieck A (1986) The status of European bog body research in 1986 as well as presentation of material about anthropological and medical evidence. Telma 16:31–168 Eisenbeib S (2003) Bog bodies in lower Saxony: rumors and facts. An analysis of Alfred Diecks sources of information. In: Bauerochse A, HaBmann H (eds) Peatlands, archaeological sites-archives of nature-nature conservation-wise use, Proceedings of the Peatland conference 2002 in Hanover, Germany, Rahden, Leidorf, pp 143–50 Fischer C (1984) Bog bodies of Denmark. In: Cockburn A, Cockburn E (eds) Mummies, disease, and ancient cultures. Cambridge University Press, Cambridge, MA, pp 177–193 Fischer C (1998) Bog bodies of Denmark and northwestern Europe. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, diseases, and ancient cultures, 2nd edn. Cambridge University Press, Cambridge, MA, pp 237–262 Fischer C (2007) Tollund man; the gift to the gods. Hovedland, Silkeborg Gebühr M (2002) Bog bodies in Schleswig-Holstein. Association for the Promotion of the Archaeological State Museum, Schleswig Gill-Frerking H (2015) Bog bodies. In: Cardin M (ed) Mummies around the world. ABC-CLIO, LLC, Santa Barbara, pp 31–36 Gill-Robinson H (2002) This little piggy went to Cambria, this little piggy went to Wales: the tales of twelve piglets in peat. In: Mathieu J (ed) Experimental archaeology: replication past objects,

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behaviors, and processes, British archaeological reports, international series 1035 (BAR S1035). Archaeopress, Oxford, pp 111–126 Glob PV (1965) The bog people; Iron age man preserved. New York Review Book, New York Gmelch SB, Gmelch G (1980) Ireland’s bountiful bogs. Nat Hist 89(11):48–57 Krebs C, Ratjen E (1956) The radiological examination of the peat-bog body from Grauballe. Kuml:138–150 Lynnerup N (2007) Mummies. Yrb Am Assooc Phys Anthropol 50:162–190 Lynnerup N (2015) Bog bodies. Anat Rec 298(6):1007–1012 Maslyuzhenko DN, Shilov SN (2006) On the discovery of a mummified human head of the era of the early Russian settlement of the forest-steppe of Tobol. Materials of the XIV international scientific seminar, Krasnoyarsk-Omsk, 2006. S 279–282 Moesgaard Museum (2020a) A new life for an old body. https://www.moesgaardmuseum.dk/en/ exhibitions/permanent-exhibitions/grauballe-man/a-new-life-for-an-old-body/a-pickled-bogbody-grauballe-man-s-conservation/. Accessed 17 Aug 2020 Moesgaard Museum (2020b) Displaying a bog body. https://www.moesgaardmuseum.dk/en/ exhibitions/permanent-exhibitions/grauballe-man/grauballe-man-today/displaying-a-bog-body/. Accessed 17 Aug 2020 Mulhall L (2010) The peat men from Clonycavan and Oldcroghan. Br Archaeol 110:34–41 Mummy Road Show (2002) Death in a bog. National geographic channel. Engel Brothers Media, Engel Entertainment, New York Neilson WA, Knott TA, Carharet PW (eds) (1939) Webster’s new international dictionary of the English Language, 2nd edn. Unabridged. G & C Merriam Co. Springfield, Massachusetts Nield T (1986) An iron age murder mystery. Sciences 4:4–6 O’Floinn R (1995) Ireland. In: Turner RC, Scaife RG (eds) Bog bodies. British Museum Press, London, pp 221–234 O’Floinn R (2006) Appendix 1: supplementary list of irish bog bodies noted since 1995. In: Bermingham N, Delaney M (eds) The bog body from Tumbeagh. Bray, Wordwell, pp 217–227 Painter TJ (1991) Lindow man, Tollund man and other pet-bog bodies: the preservation and antimicrobial action of Sphagnan, a reactive glycuronnoglycan with tanning and sequestering properties. Carbohydr Polym 15:123–142 Searcey N, Reinhard KJ, Egarter-Vigl E, Maixner F, Piombino-Mascali D, Zink AR, van der Sanden WAB, Gardner SL, Bianucci R (2013) Parasitism of the Zweeloo woman: dicrocoeliasis evidenced in a Roman period bog mummy. Int J Paleopathol 3(3):224–228 Strabo (ca. 10 CE/1977) Loeb classical library, vol 2. Harvard University Press, Cambridge, MA Tacitus (98CE/1981) The Agricola and the Germania (trans: Mattingly H and Hanford SA. Penguin Books, New York Turner RC (1995) Recent research into British bog bodies. In: Turner RC, Scaife RG (eds) Bog bodies: new discoveries and new perspectives. British Museum Press, London, pp 108–122 Turner RC, Briggs CS (1986) The bog burials of Britain and Ireland. In: Stead IM, Bourke JB, Brothwell D (eds) Lindow man: the body in the bog. Cornell University Press, Ithaca, pp 141– 161 van der Plicht J, van der Sanden WAB, Aerts AT, Streurman HJ (2004) Dating bog bodies by means of 14C-AMS. J Archaeol Sci 31(4):471–491 van der Sanden WAB (ed) (1996) Through nature to eternity. The bog bodies of Western Europe. Batavian Lion International, Amsterdam van der Sanden WAB (2013) Bog bodies: underwater burials, sacrifices, and executions. In: Menotti F, O’Sullivan A (eds) The Oxford handbook of wetland archaeology. Oxford University Press, London, p 403 van Vilsteren VT (2004) The mysterious bog people. PalArch Newsletter 1(2):21–26 Wills B, Ward C, Sáiz Gómez V (2014) Conservation of human remains from archaeological contexts. In: Fletcher A, Antoine D, Hill JD (eds) Regarding the dead: human remains in the British museum. The British Museum Press, London, pp 49–74

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Cossack Mummy from the Peat Bog Sergey Mikhailovich Slepchenko, Igor Konstantinovich Novikov, Jong Ha Hong, Do Seon Lim, Chang Seok Oh, Jieun Kim, Hye Jin Lee, and Dong Hoon Shin Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 834 Archaeological Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 836 Historical Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 837 S. M. Slepchenko (*) Institute of the Problems of Northern Development, Tyumen Scientific Center of the Siberian Branch of the Russian Academy of Sciences, Tyumen, Russia Surgut State University, Surgut, Russian Federation I. K. Novikov Kurgan State University, Kurgan, Russian Federation J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea D. S. Lim Department of Dental Hygiene, College of Health Science, Eulji University, Seongnam, Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea J. Kim Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul, Korea H. J. Lee Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Republic of Korea Ministry of National Defense Agency KIA Recovery and Identification, Seoul, Republic of Korea D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_48

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Revisiting the Cossack Mummy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Radiocarbon Dating . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . CT Radiography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dental Prosthesis in 3D Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Histology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Stable Isotope Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mitochondrial DNA Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A Bog Mummy of West Siberia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preservation Status of Cossack Mummy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Archaeological surveys have been carried out on mummies found in peat bogs. This chapter is focused on one such mummy unearthed in Western Siberia, one of the largest wetland regions in the world. In the 1960s, a forensic expert examined a Cossack mummy discovered at a peat bog in the Kurtamysh district of the Kurganskaya Oblast. Despite this being a rare non-European “bog body” case, the investigations carried out to date have been insufficient. The data presented in this chapter, albeit still inadequate to reveal the complete nature of Siberian bog mummies, nevertheless, does present important insight into a preservation pattern not previously reported. Our report can provide fundamental information for future studies on bog bodies discovered in Western Siberian peat bogs. Keywords

Bog body · Mummy · Europe · Siberia · Cossack · Soldier · Histology · Radiology · DNA · Stable isotope · Peat bog

Introduction Many kinds of mummies have been subjected to scientific research. Mummies are generally classified into natural and artificial categories according to the mummification process. Egyptian mummies are an example of a type of artificial mummification induced by skilled embalmers. As for natural mummification, there are many ways in which it can occur. The permafrost condition is known to induce mummification, examples being the Greenland mummies (Hart-Hansen et al. 1991; HartHansen and Nordqvist 1996), the Tylorean Iceman (Hess et al. 1998), and the pre-Columbian Andean mummies (Reinhard 1996). The extremely dry condition, meanwhile, is known to result in cases such as the mummies of the Taklamakan Desert (Wang 1996). Another type of natural mummification is represented by peat bog mummies. Actually, studies on what are known in the vernacular as “bog bodies” have a long history. As the name suggests, bog bodies are mummies that are discovered in peat bogs of partly decomposed plants when peat is mined as a fossil fuel (Aufderheide

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2003). The first reports on bog bodies are as early as 1640, 1747, and 1781 CE (Aufderheide 2003). According to van der Sanden (1996), 122 bog bodies, mostly from the European Iron Age, have been identified and studied so far. Archaeologists have dated bog bodies to eras ranging from 8,000 BCE to about 1945 CE (Fischer 1998; Aufderheide 2003). Bog bodies heretofore were regarded as an historical phenomenon unique to ancient Danish, German, Dutch, British, and Irish societies. Over several centuries, data on ancient peoples’ life, religion, disease, and even fashions in dress and hair styles, etc., have been gleaned from studies on bog mummies (Fischer 1998). Lifelike displays of bog mummies in museums have deeply inspired the public as well as academics all over the world. Therefore, how bog bodies came to be mummified has been a hot topic among researchers. Besides the conditions generally accepted as causes of mummification (rapid exclusion of air or cold temperatures in peat bogs, etc.), the flora of bogs also has attracted attention as a material responsible for bog mummies’ near-perfect preservation status (Fischer 1998). In fact, Sphagnum (peat moss) is regarded as an inducing factor for mummification in peat bogs. This plant is known to include a substance sphagnan, which, when moss dies, is released into bog water, whereupon it is chemically converted into humic acid. The intermediate (sphagnan) and final compound (humic acid) definitively suppress bacterial proliferation, thus accelerating mummification (Painter 1991; Fischer 1998). However, peat bogs are present not only in Western Europe but are native also to countries such as Russia. Western Siberia is the world’s largest wetland, which covers almost two-thirds of that region. At least one-half of Western Siberia, in fact, consists of peatlands that include different latitudinal bog zones (Neustadt and Selikson 1971; Ivanov and Novikov 1976; Zhulidov et al. 1997; Kremenetski et al. 2003). The development of the peatlands has had a large impact on land-use in Siberia from the Mesolithic period (Praslov 1984; Kremenetski et al. 2003). Nevertheless, Western Siberia remains relatively little known to many academics in various research fields. The same is also true for mummy research. Except for a very brief mention on “Second World War victims found in the Russian wetland” (Fischer 1998) and the 10 “Russian cases” listed by Dieck in 1972 (Aufderheide 2003), there have been very few reports on bog bodies from Western Siberian peat bogs. Other, unreported bog bodies almost certainly have been found in that area, but the full extent of the presence of mummies there remains unknown. Meanwhile, as early as 1966–1968, a mummified Cossack individual was recovered from a peat bog in Western Siberia. The forensic specialist and archaeologists estimated, based on the typology of accompanying artifacts, that the person was likely to have lived during the seventeenth to eighteenth centuries. However, except for a brief forensic record, little information on the true nature of the Siberian bog mummy has been reported. Recently, we were fortunate enough to have an opportunity to examine this mummy by multiple scientific techniques. In this chapter, we will describe the overall preservation status of the Siberian bog mummy and show the extent of the similarity of this case to bog bodies reported from other Western European countries.

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Archaeological Information The first mention of the discovery of the Cossack mummy dates back to the late 1960s. It was in 1966–1968 that peat mining was productively carried out in Kurtamysh, a town located on the periphery of the Western Siberian lowlands of the Kurganskaya Oblast (Fig. 1). During the mining, well-preserved mummies of an adult man and his horse were discovered in a peat-rich swamp near Kurtamysh. Surprised at the discovery, the miners urgently called the police. Vasily Efimovich Kazantsev (1944–2005), a forensic expert, first examined the mummified man and horse in an on-site autopsy. In the autopsy, the man’s lungs

Fig. 1 Russian Federation and magnified image of Kurgan region. The Cossack mummy was found at the peat bog in the Kurtamysh district of Kurganskaya Oblast (Credit: Igor Novikov)

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were clogged with peat and silt, testifying that he might have drowned in the swamp. Kazantsev speculated, based on the thickness of the precipitated peat layer, that the tragedy had occurred at least 200 years ago. The man’s purse or belt types, moreover, as alluded to above with respect to cultural artifacts, suggested a Cossack horseman of the seventeenth to eighteenth centuries. According to Kazantsev’s description, the mummified man was dressed in a caftan with leather-covered wooden buttons. Whip and rusty weapons were discovered nearby, which seemed to confirm the man to have been a Cossack. Since Kazantsev ruled this case to be archaeological, not forensic, the investigation was closed immediately after completion of the on-site inspection and autopsy. The mummified Cossack and his horse thereafter were buried nearby. None of the clothing or accompanying inventory was preserved. Only the Cossack’s head was taken, which Kazantsev stored in the Bureau of Forensic Medical Examination as part of its anthropological collection. When Kazantsev himself died in 2005, memories of this case mostly faded away.

Historical Considerations In 2006, a new report on the Cossack mummy was presented at an academic conference organized by Kurgan State University and the Ministry of Education and Science of the Russian Federation. There, Maslyuzhenko and Shilov (2006) presented new speculations and conclusions. According to them, the mummy’s head represented a mustached man aged 40–45 years. The preservation status was particularly good, except for a skin defect made at the occipital part of head. Propagated downward from the lip corners were cracks that, they averred, might have been formed during improper treatment with preservatives in the 1970s. The mummy’s eyes and mouth were half-open. His hair was short, wavy, and dark-brown colored. The eyebrows and a long mustache were reddish in color. Maslyuzhenko and Shilov (2006) next discussed the date estimation. In their presentation, they estimated the date by considering the extant archaeological and historical knowledge. First, concerning the Cossack’s military caftan (a long suit with tight sleeves) described in Kazantsev’s original report in the 1960s, Maslyuzhenko and Shilov (2006) questioned its academic value in dating, given that, confusingly, many different types of military caftans had been used at that time. Instead, the leather-covered wooden buttons attracted their attention. They presumed that the buttons had been made by handicraft folk production that continued until being replaced by metal-button manufacturing in the mid-eighteenth century. To support the date estimation, it is necessary to consider the chronology of the Russian people’s eastern advancement into Siberian territory. Maslyuzhenko and Shilov (2006) noted the involvement of Cossacks in military conflicts with Siberian nomadic peoples. Actually, Kurtamysh is located on the border of the Steppe zone. In the seventeenth century, this territory was the political center of the nomadic Tatars of the Kuchumovichi, Nogaev, Kalmyks, and Kazakhs. The nomads occupied this territory and fiercely resisted Russian colonization. During this period, Russian

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patrols advanced into the southern area of the Kurtamysh River, though such reconnaissance units did not exceed two to three people in number (Maslyuzhenko and Shilov 2006). In the 1680s, when the first Russian settlements (Utyatskaya) appeared in the north of Kurtamysh, new conflicts were opened with the nomadic peoples. In 1698, a Siberian Dragoon Regiment was created to deal with this resistance. In 1745, the Siberian dragoons were replaced by the Yaits Cossacks unit after the construction of a fortress in Kurtamysh (Maslyuzhenko and Shilov 2006). Considering this chronology, the mummy found in the peat bog might have belonged to the one of the Cossack units that had operated in this region between the 1680s and 1740s (Maslyuzhenko and Shilov 2006). However, it should be noted that their presentation was a simple deduction based on Kazantsev’s incomplete description and partial historical knowledge accumulated to that time. Thus, Maslyuzhenko and Shilov’s (2006) argument was bound to be hypothetical in nature.

Revisiting the Cossack Mummy In 2019, the authors of this chapter revisited the Cossack mummy, this time with more sophisticated research tools. In our anatomical examination, even though it has been more than 50 years since the mummy’s discovery, the preservation status remained particularly good. As described in the report of Maslyuzhenko and Shilov (2006), the mummy sported a long mustache (Fig. 2). The skin defect at the occipital part of the head could also be identified. As for the “cracks” extending downward from the mouth (Fig. 2a), Maslyuzhenko and Shilov (2006) conjectured (as noted above) that they might have been formed during preservative treatment in the 1970s. These “cracks” remained identifiable to us (Fig. 2b and C). Another interesting finding was a dental prosthesis reinforcing the mandibular incisors (Fig. 2c). In our view, the prosthesis looked too modern to be from the Cossack’s time. However, it did not appear to be a twentieth-century prosthesis either. In terms of dental prosthesis typology, it most likely had been used at some point between the eighteenth and twentieth centuries, quite possibly the nineteenth. When we examined the foramen magnum, the mummified brain could be identified in the cranial cavity (Fig. 3). Since amplifiable DNA can be preserved very well in mummified brain tissue (Oh et al. 2013), we collected a sample (17.15 grams) with a sterilized knife. The tissue was hardened but very fragile (Fig. 3).

Radiocarbon Dating To estimate the date in a more scientific way, AMS radiocarbon dating was done on this brain sample and muscle samples as well. The calculated radiocarbon age of the Cossack soldier was 141.9 +/
0.4 pMC, and the conventional radiocarbon age was 140.8 +/
0.4 pMC. This means that the AMS radiocarbon dating of the

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Fig. 2 A Cossack mummy head. (a) ‘Crack’ on mummy’s right cheek (indicated by arrow). (b) and (c) Dental prosthesis (asterisk) in mandibular incisors. ‘Cracks’ (indicated by arrows) are found from nostrils to the lateral canthi of the eyes (Credit: Igor Novikov) Fig. 3 Mummified brain (asterisk) identified inside the cranial cavity (Credit: Igor Novikov)

Cossack brain and muscle could not be considered to be reliable, because this “extra” 14C likely originated from modern contamination (e.g., postbomb carbon). Meanwhile, in our additional testing with a cervical vertebra (the atlas), the date estimate of this case was revised to the years 1882–1915 CE (44.9%) in 68.2% probability and 1867–1919 CE (53.3%) in 95.4% probability. This means that the Cossack mummy was likely to have lived in the late nineteenth to early twentieth century.

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CT Radiography Multislice CT scanning was performed in the City of Kurgan to determine the preservation status and any atypical findings in the Cossack mummy’s head. A standard scanning technique with the slice thickness of 0.75 mm was used. Image analysis was performed using multiplanar reconstruction (MPR) in the axial, frontal, and sagittal planes (3D). The mummified brain remained within the cranial cavity on CT axial images (Fig. 4a). Although the brain had been reduced in size, it still occupied almost half of

Fig. 4 Cossack mummy head on CT axial images. (a) Mummified brain (Br) in cranial cavity. (b) The mummy’s eyeballs (Ey) inside the orbits. (c) Axial view at cervical vertebrae level. Neck muscles (MM) are visible. (d) Dental prosthesis (Pr) of mandibular incisors (Credit: Sergey Slepchenko)

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the cranial cavity. Specific parts of the hemispheres or lobes could not be differentiated (Fig. 4a). The mummy’s eyeballs were found inside their orbits. The size had been reduced by drying and contraction, but their appearance was well maintained (Fig. 4b). In the axial view at the cervical vertebrae level, the neck muscles were well preserved. The cartilage in the nose remained, thus contributing to the Cossack’s near-perfect countenance (Fig. 4c). A dental prosthesis for the mandibular incisors also could be identified on CT axial images (Fig. 4d). In the first molar (right side) of the mandible, we also found a radiological sign of dental treatment (Fig. 5). The Cossack might have had dental-filling treatment, not with a metal alloy mixture such as an amalgam, but with the same kind of metal used for his dental prosthesis. In the coronal view of the CT, the first-molar cavity was not completely filled (Fig. 5a). This means that the filling technique used for the Cossack was different from that of the twentieth century. In the coronal (Fig. 6a) and sagittal (Fig. 6b) views, the wellpreserved brain and tongue could be seen.

Dental Prosthesis in 3D Model Our CT-based 3D model exhibited the appearance of the dental prosthesis more successfully (Fig. 7). As seen in Fig. 4d, it had been set to reinforce the mandibular incisors (Fig. 7). It was clear that this type of prosthesis was not used among twentieth-century Russians. The cavity-filling technique used for the first molar of the mandible, furthermore, showed a similar, pre-twentieth-century pattern (Fig. 7b). However, as mentioned above, we are not sure whether the prosthesis belongs to either the seventeenth or eighteenth century. In fact, its precise date estimation remains elusive. Actually, the wooden buttons, dental prosthesis, cavity-filling technique, and information on historical artifacts clearly indicated that the Cossack mummy died at some point before the twentieth century. Unfortunately, knowledge of Russian peoples’ lives on the Siberian frontier is not yet sufficient for any closer estimation of the Cossack mummy’s chronology. Our carbon dating result (i.e., late nineteenth to early twentieth century) also was confusing. That he lived on the Siberian frontier at least 100 years ago is our best guess for now.

Histology Microscopic examinations were performed on Cossack mummy samples following the method of Slepchenko et al. (2019). First, a section of mummified skin was rehydrated in Ruffer’s solution (1921). Frozen skin tissue was sectioned on a cryostat in 12–20 μm thicknesses. The slides were stained in Masson’s trichrome solution (Sheehan and Hrapchak 1980). We observed the histological patterns under light microscopy. The keratinized layer could be seen, but the epidermis could not be identified. The dermis, being composed mostly of collagen fibers (see the next paragraph), was intact (Fig. 8a). We also discovered sebaceous glands in the dermis

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Fig. 5 Dental treatment sign in the first molar (right) of mandible (indicated by arrows) on CT images. The first molar cavity was not completely filled (Credit: Sergey Slepchenko)

Fig. 6 CT images of (a) coronal and (b) and sagittal views. Br, Brain; To, tongue (Credit: Sergey Slepchenko)

(Fig. 8b and c). In the layer below the dermis, muscle fibers were often found (Fig. 8d). The histology also was examined by scanning electron microscope (SEM) (Hayat 1970; Bozzola and Russell 1992). A sample was treated in fixatives, then postfixed

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Fig. 7 CT based 3D model. (a) and (b) Skull. (c) and (d) Teeth. Dental prosthesis is marked by arrows (Credit: Igor Novikov)

in osmic acid solution, and finally subjected to Pt-Pd coating. In the dermis, we found thread-like structures that most likely were collagen fibers (Fig. 9a). In highermagnification images, collagen fibers were seen to be aligned loosely in connective tissue (Fig. 9b).

Stable Isotope Analysis We used the Cossack mummy’s hair for stable isotope analysis by isotope ratio mass spectrometry linked with an elemental analyzer. Carbon and nitrogen isotope compositions (δ13C and δ15N) were calculated as: δ (‰) ¼ [(Rsample/Rstandard)
1]  1000 (R being the ratio of 13C/12C or 15N/14N). The analysis was carried out twice.

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Fig. 8 Histological patterns of Cossack mummy skin. Masson’s trichrome staining. (a) Most part remained in mummified skin is dermis. Collagen fibers are stained blue. (b) Sebaceous glands in dermis. (c) Magnified image of sebaceous gland in mummy skin. (d) Muscle fibers (stained red) observed in the deepest layer (Credit: Ji Eun Kim)

Fig. 9 SEM images of Cossack mummy skin. (a) Collagen fibers in the mummified dermis. (b) Magnified image of collagen fibers (Credit: Do Seon Lim)

The averages of the stable isotope analyses on the Cossack mummy hair were -20.03‰ for δ13C and 11.20‰ for δ15N. The C/N ratios (4.0 and 4.2) fell within the acceptable range for successful analysis (Harbeck et al. 2006; Meier-Augenstein

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2017). The δ13C value indicated that the Cossack mummy might have consumed more C3-based foods (soy, rice, wheat, potato, etc.) than C4-based foods (millet, maize, corn, sugarcane, etc.). The mummy’s δ15N value reflected the high consumption of nitrogen-rich products (meat, fish, milk, etc.) and is similar to that of modern Russians (9.61 to 11.90) (Lehn et al. 2015).

Mitochondrial DNA Analysis The Institutional Review Board (IRB) of Seoul National University Hospital confirmed that performance of aDNA analysis on the Cossack mummy was exempt from board review (IRB No. 2017-001). The setup of our DNA lab facility followed the protocol of Hofreiter et al. (2001). Researchers wore sterilized protection gear to reduce the chance of modern DNA contamination. After DNA was extracted from the Cossack mummy’s brain, it was mixed with PCR reagents and primers (Table 1). After PCR reaction, specific bands were observed in agarose gel electrophoresis (only 2 amplicons among 9 primer sets) (Fig. 10). Transformed bacteria were grown for cloning and sequencing. Purified DNA was sequenced on an automatic sequencer. The Revised Cambridge Reference Sequence (rCRS; accession number: NC_012920) was used for this analysis. Cloned sequences were the same except for a few clones with single nucleotide substitutions. The consensus mtDNA mitotypes are summarized in Table 2. The researchers’ mtDNA profiles were obtained under IRB permission (H-0909-049-295). The authenticity of the aDNA analysis could be confirmed by the finding that no identical sequences were detected between the Cossack mummy and a researcher (Table 2). We could not calculate Cossack mummy’s exact haplogroup, as the sequences obtained were too short for such determination.

A Bog Mummy of West Siberia Bog bodies commonly have been reported from European countries such as Denmark, Germany, Great Britain, Ireland, and the Netherlands. The socio-cultural aspects of those bog mummies have been studied by historians, archaeologists, and anthropologists. Many hypotheses as to why and how those individuals had been buried and mummified in peat bogs have been put forward. Among the bog bodies of Denmark, academic reports on Borremose Men, Grauballe Man, Elling Woman, and Tollund Man, among others, have been reported (Fischer 1998). Researchers noticed that one of the Borremose Men had been hanged or strangled to death before being abandoned in the peat bog. In the case of Tollund Man of Jutland, a rope made of braided leather strap was discovered around his neck. Similar evidence was also found in the case of Elling Woman, who might have been hanged (Fischer 1998). Grauballe Man was unique in that he had suffered a long cut

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Table 1 PCR primer sets used in this study Region HVI (15991–16390)

Set MPS1A

Primer F15989 R16158

MPS1B

F16112 R16237

MPS2A

F16190 R16322

MPS2B

F16268 R16410

HVII (034–369)

MPS3A

F34 R159

MPS3B

F109 R240

MPS4A

F151 R292

MPS4B

F220 R377

HVIII (423–548)

MVR2

F403 R569

50 to 30 CCC AAA GCT AAG ATT CTA AT TAC TAC AGG TGG TCA AGT AT CAC CAT GAA TAT TGT ACG GT TGT GTG ATA GTT GAG GGT TG CCC CAT GCT TAC AAG CAA GT TGG CTT TAT GTA CTA TGT AC CAC TAG GAT ACC AAC AAA CC GAG GAT GGT GGT CAA GGG AC GGG AGC TCT CCA TGC ATT TGG TA AAA TAA TAG GAT GAG GCA GGA ATC GCA CCC TAT GTC GCA GTA TCT GTC TAT TAT TAT GTC CTA CAA GCA CTA TTA TTT ATC GCA CCT ATT TTT TGT TAT GAT GTC T TGC TTG TAG GAC ATA ATA AT GTG TTA GGG TTC TTT GTT TT TCT TTT GGC GGT ATG CAC TTT GGT GTA TTT GGG GTT TGG TTG

Annealing temp. (°C) 50

Length (bp) 170

50

126

46

133

48

143

54

126

46

132

45

142

46

158

56

167

to the neck that was deep enough to reach his esophagus (Fischer 1998; Aufderheide 2003). In the Netherlands, there have also been reports of bog mummies such as Yde Girl and Weerdinge Couple. Yde girl had been strangled by a hairband found around her neck (Fischer 1998). As for the Weerdinge Couple, a deep abdominal stab wound was found in one of them (Fischer 1998). In the UK and Ireland, the discoveries of Worsley Man, Lindow Man II, and Meenybraddan Woman have been reported. Violent blows may have crushed Worsley Man’s skull, and a thin cord had been

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Fig. 10 Agarose gel electrophoresis of DNA amplicons (MPS2A and MPS3B) from Cossack mummy. Negative controls (NC) not exhibiting any amplified bands (Credit: Jong Ha Hong)

Table 2 Comparison of mtDNA D-loop sequences between Cossack mummy and researcher Sample Cossack Researcher

D-loop region HV1(15991–16390) 16223T 16093C, 16176T, 16223T, 16362C

HV2(034–369) 195C 73G, 94A, 194T, 263G, 309.1C, 315.1C

tied tightly around his neck. Lindow Man II also received two blows to his head and a blow to his back, a string also having been tied around his neck (Fischer 1998). As for Rendswuhren Man and Windeby Man and Woman from Germany, researchers discovered that their death might have been due to a triangular wound to the forehead (Rendswuhren Man) or strangulation by constriction with a hazel twig (Windeby Man) (Fischer 1998). All of this evidence indicates that bog mummies in Europe often show traces of murder. The actual purpose of placing dead bodies in peat bogs remains a subject of debate. The writing of Tacitus has often been referenced to learn about the sociocultural background of what was then known as Germania. According to his book, “On the Origin and Situation of the Germans,” traitors, homosexuals, and those accused of battle cowardice, among others, were drowned in swampy bogs as regular punishment. This description is corroborated by the physical signs of violence commonly observed of bog mummies (Fischer 1998; Aufderheide 2003). In Germania, human sacrifices also were made by certain cult groups hoping to appease their gods. In these cases, the victims were not culprits or criminals but had been chosen from among social elites (Fischer 1998; Aufderheide 2003). Tollund Man or Grauballe Man might be such cases. They were sacrificed for rituals and buried very respectfully in peat bogs. Actually, we note that, as far as the historical literature is concerned, Siberian peoples did not have any traditions of sacrifice or punishment at peat bogs. Indeed, there are as yet no archaeological traces or historical evidence of sacrifice or

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punishment at Siberian peat bogs. It should also be noted that, correspondingly, reports of Siberian bog bodies have not been nearly as common as reports from Western Europe, where sacrifice or punishment at peat bogs was not especially rare. Nevertheless, echoing Aufderheide (2003), we should also emphasize that bog bodies have been formed as the results not only of sacrifice or punishment but of accidents or murder. In this respect, the record of the forensic specialist Kazantsev in the 1960s did not describe any signs of trauma to the Cossack mummy. This suggests that he drowned in a state of unconsciousness after accidentally falling while riding a horse. Actually, drowning accidents can happen at any time and place in history. In any case, it is difficult to exclude the possibility that bog mummies caused by accidents or murders will be reported in the future, for Western Siberia and other regions as well.

Preservation Status of Cossack Mummy Aufderheide (2003) described a unique sign of peat bog mummies’ preservation status: decalcification of skeletons and teeth. Typically the bog mummy’s skull and limbs were deformed due to serious decalcification and soil pressure upon them. Radiological examinations of bog mummies thus provide little information about them. In histology, Aufderheide (2003) also mentioned that collagen fibers remained uncompromised in bog mummy’s skin and internal organs. Facial skin integrity was often intact, preserving the mummy’s lifetime countenance. Despite shrinkage, collagen-rich structures (e.g., the pericardial sac) have maintained their original size and morphology. The liver, lung, and kidney have been the most common organs found in bog bodies (Aufderheide 2003). Superbly preserved bog mummy cases such as Grauballe Man have even shown fingerprints or toe prints (Fischer 1998). The preservation patterns of the Cossack mummy, as revealed by anatomical, radiological, and histological analyses, were not seriously distinct from those of other cases of natural mummification. The preservation of the Cossack mummy’s head was perfect from a gross anatomical perspective. His brain was dehydrated, deformed, and displaced to the dorsal part of the cranial cavity, possibly due to the long-term action of gravity. The microscopic patterns of the Cossack mummy can be summarized as follows: disappearance of cells; atrophy of epidermis but preservation of dermis; and presence of collagen fibers in dermis, and maintenance of microscopic structures such as sebaceous glands. The Siberian peat bogs generally ensure excellent preservation status in general. Recently, aDNA analysis has become a useful tool to reveal the genetic affiliations of ancient humans or animal specimens discovered at archaeological sites. As discussed in this chapter, aDNA analysis also was performed on the Cossack mummy’s brain specimens. Judging from the preservation status of the mummy, we originally anticipated that genetic testing would show excellent-quality results. However, the outcome fell short of our expectations: only limited parts of the PCR amplicons could be obtained. We could not rule out the possibility that the poor aDNA results were due to the treatment by preservatives in the 1970s.

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Finally, we would like to point out some peculiarities of the Cossack mummy compared with other bog bodies. Unlike European bog mummies, in which serious decalcification was identified, the bone of the Cossack mummy was perfectly preserved. In short, decalcification did not occur in any parts of his cranium. With regard to why decalcification in this case was not as serious as seen in European bog mummies, we considered that there might be a fine difference between the peat bog environments of Western Siberia and Northern Europe.

Conclusion This chapter presents biological features of a bog body discovered in a Western Siberian peat bog. The Cossack mummy had been investigated in the 1960s, only the head having been collected and stored in a Russian institute. Since no detailed scientific investigation had been performed previously, in 2019, our multidisciplinary research team revisited the Cossack mummy case. Anatomical examination confirmed the excellent preservation status. Upon histological examination, many collagen fibers were observed in mummy tissue. Stable isotope analysis revealed the Cossack mummy’s diet. In a radiological examination, the Cossack mummy’s bone exhibited a different pattern from other bog mummies in that there was no decalcification evident on CT images. We speculate that the process of mummification in Siberian peat bogs might differ from European bog body cases in terms of the distinct respective environments. Since the scientific value of Siberian bog mummies is great, the academic community will certainly welcome more attention being paid to them in the future.

Cross-References ▶ Bog Bodies Acknowledgments This research was supported by Basic Science Research Program through the National Research Foundation of Korea by the Ministry of Education (2020R1A2C1010708).

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lenin’s Official Autopsy Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . External Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Internal Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Lenin’s Diseases During Lifetime . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Final Course of Lenin’s Malady . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Official Version of Lenin’s Cause of Death . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Fate of Lenin’s Brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Funeral That Never Took Place . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Immortalization of the Leader: A “Living Sculpture” . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Embodiment of the Leader . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Embalmers of the Communist Leaders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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R. Bianucci (*) New Mexico Health Enhancement and Marathon Clinics Research Foundation, Albuquerque, NM, USA Warwick Medical School, Biomedical Sciences, University of Warwick, Coventry, UK Legal Medicine Section, Department of Public Health and Paediatric Sciences, University of Turin, Torino, Italy e-mail: [email protected]; [email protected] F. M. Galassi Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia FAPAB Research Center, Avola, Italy e-mail: francescom.galassi@flinders.edu.au A. G. Nerlich Institute of Pathology, Academic Hospital Munich-Bogenhausen, Munich, Germany e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_52

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Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 875 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 875

Abstract

This chapter focuses on the fate of the body of Vladimir Il’ič Ul’janov (1870–1924), aka Lenin, from the day after his death, January 22, 1924, till the present day. It is aimed at considering the process of a conservation of the nonliving body of the founder of the Soviet Union over the last 96 years and to elucidate its sociocultural and political significance. Keywords

USSR · Lenin’s illness · Autopsy · Brain histology · Body conservation/ cultivation · Cultural and social anthropology

Introduction Vladimir Il’ič Ul’janov, (1870–1924), also known as Lenin (Fig. 1), the Bolshevik leader and the founder of Soviet Union, died on the 21st January 1924 at 6:50 pm at the Gorki estate near Moscow. Since no long-term preservation of the corpse had been planned, Lenin underwent autopsy in Gorki on the 22 January 1924 under the supervision of the chief pathologist, Professor Aleksei Ivanovich Abrisokov (1875–1955), in the presence of the Commissar of Public Health (between 1918 and 1930), Dr. Nikolai Semashko (1874–1949) (Lerner et al. 2004). In the former USSR, it was expected that all physicians who had treated Lenin had to participate in his autopsy. However, of the twenty-seven medical doctors who had assisted Lenin (including the foreigners Professors Nonne, Bumke, Struempell, Henschen, Vogt, and Foerster), only eight signed the final document. Therefore, it remains unclear how many of those physicians attended Lenin’s autopsy as several of his doctors were Germans and not present in the USSR at that time. Seven were Russians and one was the renowned German neurosurgeon, Professor Otfrid Foerster (1873–1941), who did not know Russian (Lerner et al. 2004). Vladimir Bekhterev (1857–1927), the Director of the Brain Institute in Petrograd, who had examined Lenin at least once during his malady, was not invited (Arutyunov 1999). During Lenin’s autopsy, the brain and the heart were removed, fixed in formalin and transferred to Moscow. While the brain was extensively investigated (see below), the fate of Lenin’s heart is unknown. It is thought that all remaining inner organs were removed during autopsy and, then, put back in the cadaver; in the end, they were removed by Professor Vorobiev during Lenin’s experimental embalming (Yurchak 2019).

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Fig. 1 Vladimir Il’ič Ul’janov, also known as Vladimir Il’ič Lenin, in July 1920 (Image freely available at https://en.wikipedia.org/wiki/ Vladimir_Lenin#/media/File: Bundesarchiv_Bild_18371043-0003,_Wladimir_ Iljitsch_Lenin.jpg)

Lenin’s Official Autopsy Report External Examination Body of an elderly man of normal build, adequately nourished. Small pigmented spots are noted on the interior aspect of the chest (acne). Obvious signs of cadaveric hypostasis are noticeable on the posterior aspect of the trunk and the extremities. A linear cicatrice 2 cm in length is noted on the skin in the area of the anterior end of the clavicle. Another cicatrice having an irregular outline and measuring 2
1 cm is located on the external surface of the left shoulder area. A round cicatrice about 1 cm in diameter is found on the skin of the spine above the ridge of the scapula. The outlines of the skeletal muscles are quite prominent.

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In the left clavicle at the border of the lower and middle third there is a slight thickening of the bone (bone callus). Above this area in the posterior part of the deltoid muscle, a solid roundish body can be palpated. Upon incision of this area, a deformed bullet is found, enclosed in a capsule of connective tissue at the border between the subcutaneous fatty tissue and the deltoid muscle.

Internal Examination The cranial bones are unchanged. Upon removing the skull cover, a solid fusion of the dura mater with the inner surface of the cranium is noted, primarily along the course of the longitudinal sinus. The outer surface of the dura is dull, pale; pigmentation of a yellowish hue is noted in its left temporal and partly frontal area. The anterior part of the left hemisphere seems slightly collapsed in comparison with the corresponding part of the right hemisphere. The longitudinal sinus contains a small quantity of liquid blood. The internal surface of the dura mater is smooth, shiny-moist, easily separating from the underlying arachnoid membrane except in areas bordering the sagittal suture where there are areas of fusion in the region of the pacchionian granulations. The dura of the base of the brain is normal; the basal sinuses contain liquid blood. Brain. The weight of the brain immediately after removal, freed of the dura mater, is 1,340 g. In the left hemisphere: (1). in the anterior central gyrus; (2). in the area of the temporal and occipital pole; (3). in the area of the fissura paracentralis, and (4). in the area of the high gyri, there are noticeable signs of pronounced collapse of the cerebral surface. In the right hemisphere at the border between the temporal and occipital poles there are also two adjacent spots of collapse of the brain surface. Above the described areas of collapse, the arachnoid membrane is dull, whitish, in places yellowish. In some areas overlying the fissures, including even some parts where there is no collapse, whitish regions are noted in which the arachnoid is hard and appears thickened upon section. Vessels of the base of the brain. Both arteriae vertebralis, and also the arteria basilaris are thickened, do not collapse; their walls are hard, irregularly thickened, of a whitish and in places yellowish color. Upon section, their lumen is seen to be extremely narrowed in places down to the dimensions of a tiny slit. Identical changes are also found in branches of the arteries in question (aa. cerebri posteriores). The internal carotid arteries, and also the anterior cerebral arteries, are similarly hardened with an irregularly thickened wall and in parts greatly narrowed lumen. The left internal carotid artery in its intracranial course has a completely obliterated lumen and upon section appears merely as a homogeneous solid whitish band. The left Sylvian (i.e. middle cerebral) artery is very thin, hardened, but upon section still shows a thin small slit. Incision into the

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vermiform process of the cerebellar convolutions reveals no change of the brain tissue. The fourth ventricle is free of any pathological contents. Resection of the brain according to Flessing shows the brain ventricles, particularly on the left side, to be widened and containing a transparent fluid. In the above noted areas of collapse of the brain there are areas of softening of the tissue, having a yellowish color and accompanied by formation of cyst-like structures filled with a turbid liquid. The areas of malacia involve the white as well as the gray brain matter. In other parts of the brain the tissue is moist, pale. The vascular plexus overlying the corpora quadrigemina is well irrigated with blood and there are signs of fresh hemorrhage in this area. Upon removing the skin of the trunk the good development of the subcutaneous and fatty tissue is noted. The muscular system is adequately developed. The muscular tissue is of the usual maroon color. The positions of the organs of the abdominal cavity are regular with the exception of the caecum which lies somewhat higher than is the norm. The omentum and the mesentery are rich in fat. The diaphragm runs from the level of the fourth rib on the right hand side to the fourth intercostal space on the left. At the region of the pulmonary apex, fibrous synechiae are visible in the right pleura. The left pleura also forms synechiae with the diaphragm in its lower part. No pathological conditions are noted in the region of the heart sac; the mediastinum shows no particular changes. Heart. Dimensions: transverse 11 cm.; longitudinal 9 cm.; thickness 7 cm. The epicardial surface is smooth and shiny; under the epicardium, mainly in the area of the left ventricle, spotty accumulations of fat are noted. The semilunar valves of the aorta are somewhat thickened at their bases. The mitral valve shows some thickenings at its margins and whitish opaque spots of the anterior cusp. The valves of the right half of the heart are without special changes. The interior of the ascending aorta shows a small number of convex yellowish plaques. The wall thickness of the left ventricle is 1¾ cm. and of the right one ½ cm. The coronary arteries gape upon section; their walls are very hard and thickened; their lumen definitely constricted. The inner surface of the descending aorta and also the inner surfaces of the large arteries in general show numerous very prominent yellowish plaques, partly undergoing ulceration and calcification. Lungs. The right lung is of the normal size and configuration; soft throughout, feels spongy. Resection of pulmonary tissue reveals it full of blood, a foamy liquid appears. The pulmonary apex shows a small retracted scar. The left lung is of the usual size and shape, its consistency being soft throughout. The posterior inferior part of the upper lobe has a scar reaching from the surface to a depth of 1 cm. into the pulmonary tissue. At the apex of the lung there is a small fibrous thickening of the pleura. The spleen is slightly enlarged and moderately filled with blood upon section. The form and size of the liver are normal. The border of the left lobe is somewhat sharp. The surface is smooth. Section reveals a moderate degree of so-called grapiness. The gall bladder and the bile ducts reveal no special changes.

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The stomach is empty. Its walls are collapsed. The mucosa shows clearly visible and normally arranged creases. No special conditions to be noted regarding the intestines. The kidneys are of normal size. Their tissues are clearly identifiable and the substance of the cortex can be easily distinguished from the medullary part. The capsule comes off easily. The surface of the kidney is smooth with exception of some small areas where small depressions of the surface are present. The lumina of the renal arteries gape upon section. The pancreas is of normal size. No special changes are noted following its section. Glands of Internal Secretion. No special changes are noted in the pituitary gland. The adrenals are somewhat smaller than the norm, especially the left one; the cortical substance is rich in stipples; the medulla is pigmented and brownish in color. Anatomical Diagnosis: Generalized arteriosclerosis with pronounced degree of affection of the cerebral arteries. Arteriosclerosis of the descending aorta. Hypertrophy of the right ventricle of the heart. Multiple foci of yellow cerebromalacia (based on vascular sclerosis) of the left cerebral hemisphere in a stage of resolution and of cystic change. Fresh hemorrhage into the vascular plexus overlying the corpora quadrigemina. Bone callus of the left clavicle. Incapsulated bullet in the soft tissue of the left shoulder. Conclusion. The basic disease of the deceased was disseminated vascular arteriosclerosis based on premature wearing out of the vessels (Abnutzungssklerose). The narrowing of the lumen of the cerebral arteries and the disturbances of the cerebral blood supply brought about focal softening of the brain tissue which can obtain all symptoms of the disease (paralysis, disturbance of speech). The immediate cause of death was: (1). The aggravation of the circulatory disturbance of the brain, and (2). Hemorrhage into the arachnoid pia mater in the area of the corpora quadrigemina. Gorki, 22 January 1924 PROF. A. I. ABRIKOSOV PROF. FOERSTER PROF. V. OSIPOV PROF. V. BUNAK PROF. A. DESHIN PROF. B. WEISSBROD DR. V. OBUKH DR. ELISTRATOV DR. V. ROZANOV N. SEMASHKO The “official” autopsy report indicates that the ultimate death was caused by the aggravation of the circulatory disturbance of the brain, and a hemorrhage into the arachnoid pia mater in the area of the corpora quadrigemina (Payne 1964; Service 2000). The etiology of Lenin’s illness has been discussed for decades and it is still a

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matter of debate. The “official” autopsy report, of which there have been at least eight different unofficial versions, was released in Gorki on January 22 1924 (Payne 1964; White 2009). The body of Lenin was transferred from Gorki to Moscow via train and exposed in the House of Trade Unions.

Lenin’s Diseases During Lifetime According to the ideology of the former USSR, the icon of the Soviet Union could not be affected by internal diseases, such as a venereal disease, e.g., syphilis. This would have produced a negative image of Lenin as a leader with a deviant morality (Hesse 1975). Therefore, it is extremely difficult to obtain robust information on Lenin’s illnesses. However, one event, which severely affected Lenin’s health, has been well documented. On the 30th August 1918, during an assassination attempt, the leader was injured by two pistol bullets (Boris Yeltsin Presidential Library (2018). The bullet close to the clavicle was removed on the 23rd April 1922 by Vladimir Nikolaevich Rozanov (1872–1934), the head of the surgical division at Soldatenkov Hospital in Moscow (now the S.P. Botkin Hospital) (Ochkin 1934). This bullet must have caused the callus on the clavicle. Three shots were fired. The autopsy report states that another was removed from the posterior deltoid, i.e., behind the shoulder. There were three scars. The final one being over the spine of the scapula. No bullet is mentioned related to this one. There is no other wound at the front of the chest to indicate a bullet passed through it. The description of the assassination states that the assassin, Fanja Kaplan (1890–1918), called out and Lenin turned and then she fired (Boris Yeltsin Presidential Library 2018). However, given two bullets became embedded subcutaneously, it must have been low velocity, and the posterior wounds indicate that he was shot at from behind, as well as in front. Perhaps he turned away after the first shot. The third shot hit the elbow of a nearby person. At that time, medical examination indicated that Lenin had the following ailments; eye problems, stomach ache, recurrent headache and sleeplessness. Furthermore, he complained of obsessive ideas, which were not further specified (Service 2000). Beyond those identified diagnoses, there is an on-going debate whether Lenin was suffering from tertiary neurosyphilis. This dispute, which is highly contentious, can only be resolved when the histopathological results of Lenin’s brain investigation are independently examined. (Lerner et al. 2004; Kreutzberg et al. 1992; Hesse 1975). A precise compilation of Lenin’s medical history (Hesse 1975) only adds evidence for pneumonia at the age of 25 and recurrent gastro-intestinal problems throughout his life. Otherwise, only indirect evidence exists on Lenin’s diseases prior to his final course. According to the autopsy record, scarring on both sides of the lungs with synechia of the pleura suggests the residues of an old-healed, inactive tuberculosis; a very frequent observation at that time without further clinical relevance. The adrenal glands are reported to show in “the cortical substance . . . rich in stipples” which may indicate multinodular adrenal gland hyperplasia, suggesting either clinically relevant or, much more likely, nonrelevant increase in adrenal

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function. In the first setting, enhanced corticosteroid or adreno-corticotropin hormone (ACTH) production may cause Cushing’s syndrome (Vassiliadi and Tsagarakis 2019), which is frequently associated with hypertonia. However, all other symptoms of Cushing’s syndrome are missing in Lenin’s anamnesis so that clinically nonrelevant conditions should be assumed. In their substantial review of Lenin’s malady, Lerner et al. (2004) reports on unpublished archival documents from the Soviet Ministry of Health indicating that Lenin had suffered for more than 10 years from “his illness.” Furthermore, letters from Lenin’s widow indicate that, at the end of 1902, her husband had already been affected by a severe nervous illness with a cutaneous eruption leading to a hospitalization in Zürich for 2 weeks (Lerner et al. 2004). Subsequently, Lenin was reported to have been irritable, ill-tempered, and suffering from headaches, that occasionally prevented him from working. It has not been reported whether these symptoms stopped or persisted until the final stages of his life.

Final Course of Lenin’s Malady The final events of Lenin’s life have been officially documented by physicians, many of whom were Germans. Political and cultural relationships and exchanges between the USSR and Germany dated back to the General Erich Ludendorff’s (1865–1933) support of Lenin’s return to Russia that ultimately lead to the collapse of Imperial Russia and the Treaty of Brest-Litovsk which put an end to WW1 on the Eastern Front. According to the records of Vladimir Petrovich Osipov (18?-1947), the President of the Petrograd Society of Psychiatrists and Neurologists, who attended the ailing Lenin, the leader’s malady can be divided into three main stages. The first began in March 1922 when Lenin, a passionate hunter, was forced to sit down frequently to rub his right foot such as in intermittent claudication (Kaplan and Petrikovsky 1992; Hachinski 1992). Further complaints of numbness of the right side and several episodes of loss of consciousness were associated with disturbance of speech. Lenin recovered by August 1922. The second stage began in December 1922 when Lenin developed new symptoms; an acute right hemiparesis, followed by a slow recovery in February 1923. The final stage was in March 1923 when his terminal illness began. It manifested as paralysis of the right arm and leg and an abrupt deterioration in speech (Kaplan and Petrikovsky 1992). Following this, there was no complete recovery, but several further episodes occurred. He was very agitated, had hallucinations, lost appetite, was sleepless, and had headaches. He was confined to a wheelchair (Fig. 2). Lenin’s conditions further worsened such that by mid-October 1923 (Fig. 3), his neurological symptoms deteriorated; with brief loss of consciousness. He became more and more apathetic until on January 21, 1924. He lost consciousness coupled with convulsive activity of upper and lower limbs and developing a pathologic breathing rhythm (Cheyne-Stokes respiration) suggesting damage to the respiratory centers in the brain or congestive cardiac failure. Attempts at resuscitation failed (Kaplan and Petrikovsky 1992).

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Fig. 2 Lenin in a wheelchair in the summer 1923 in Gorki. The right hand is clenched due to the strokes he suffered.© (Reproduced with permission of akg-images)

The Official Version of Lenin’s Cause of Death The difficulties with this document have already been discussed. This report states that generalized systemic arteriosclerosis was the major underlying disease with mainly the brain vessels affected. The coronary arteries, the aorta, and the large arteries of the trunk also showed evidence of arteriosclerosis. These observations support the official diagnosis. Interestingly, there does not seem to have been evidence for acute or old-healed myocardial infarction despite the coronary arteriosclerosis. There was a strong family history of arteriosclerosis and early death suggesting a genetic cause. As a consequence of the severe arteriosclerosis, Lenin’s brain showed typical signs of previous and recurrent insults. The collapse of the cerebral surface with cystic transformation suggests older lesions; the “softened areas” may indicate fresher ones. The yellowish transformation of the arachnoid membrane also suggests old bleeding deposits. Finally, the fresh bleedings of the plexus area overlying the corpora quadrigemina points to terminal hemorrhage. However, this does not indicate the

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Fig. 3 One of the last pictures showing the ailing Lenin at the end of 1923 with his sister Maria Ul’yanova and Professor Foerster. © (Reproduced with permission of akg-images)

diagnosis of a lethal intra-cerebral bleed (Lerner et al. 2004) Surprisingly, although Lenin’s brain was preserved for substantial histological examination, no report deals with the findings of these studies with respect to the clear circulatory changes.

The Fate of Lenin’s Brain Following Lenin’s death, his brain and body were immediately separated and two entire buildings, The Brain Institute (Instituta Mozga) and Lenin’s Mausoleum, were built to house each of them (Richter 2006). In 1925, the brain was transferred to the newly created Brain Institute. The prominent German neurologist and neuropathologist, Professor Oskar Vogt (1870–1955), who had examined Lenin during his illness, and his wife Cécile were called to Moscow to examine his brain (Kreutzberg

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et al. 1992). The Vogts at that time were deeply involved in the study of the cytoarchitecture of the cortex in relation to higher-level brain functions (White 2009; Bentivoglio 1998). Between 1926 and 1930, Oskar and Cécile Vogt remained in Moscow for long periods and supervised the work on Lenin’s brain by the Russian scientists who had been trained at Vogt’s Kaiser-Wilhelm-Institut für Hirnforschung (KWI for Brain Research) in Berlin. Professor Vogt’s influence on the Soviet authorities was so significant that an entire building, named The Institute of the Brain, was erected around the Bolshevik leader’s brain. The Institute was built with the aim of investigating the cyto- and myeloarchitecture of Lenin’s brain, and to eventually prove neuro-histologically that Lenin was a genius and an eminent leader. Professor Vogt employed a giant microtome to obtain full coronal sections of Lenin’s brain fixed in formalin. Reports of the number of slides vary from 1,108 to more than 31,000, each 20 microns thick (Kreutzberg et al. 1992). Even though Professor Vogt had exceptional abilities, it appears very unlikely that Lenin’s brain, as described at autopsy, would have retained its anatomical shape per section and that 30,000 full coronal sections were obtained (White 2009). In 1927, Oskar Vogt gave a preliminary report on his findings which read as follows: “In the third cortical layer (of several brain areas) particularly in the deep portions, I found pyramidal neurons of extraordinary size and number never previous observed by myself. . . These anatomical findings allow us to identify a brain athlete and an association giant” [Assoziationsathlet ¼ an athlete in associative thinking] (Vogt 1929; White 2009; Kreutzberg et al. 1992; Richter 2007; Max Planck Institute (MPI) (https://brain.mpg.de/institute/history/leninsbrain.html Accessed on 1 Jun 2020) (Fig. 4). Later in 1927, Professor Vogt showed his findings to Professor Otfrid Foerster (1873–1941) and Dr. Wilder Penfield (1891–1976), both distinguished neurosurgeons and neuroanatomists, who had come to Moscow to visit him. Although both scientists recognized that the pyramidal neurons in the third cortical layer of Lenin’s brain were enlarged in size, they did not agree that these findings could be interpreted as evidence of superior intelligence (Penfield 1977). In his later years, Professor Vogt regretted to have overexposed himself in stating that Lenin’s brain was that of a genius (Kreutzberg et al. 1992; Richter 2007). Between 1932 and 1936, further discoveries were made by the excellent scholars Semyon A. Sarkisov (1895–1971) and Ivan Nikolaevich Filimonov (1890–1966). They had been Vogt’s former Russian students. Both would later become highly respected members of the USSR Academy of Medical Sciences (Richter 2006). While Vogt’s had identified the vertical cytoarchitecture of Lenin’s brain, Sarkisov and his team were able to add more details about the horizontal cytoarchitectonics of Lenin’s cerebral cortex (Richter 2006). They compared Lenin’s brain cytoarchitecture with that of other “élite brains,” while no reference was made to the “average” man’s brain. The investigations on Lenin’s brain included comparisons with the brains of the Russian poets Vladimir Vladimirovich Maiakovski (1893–1930), Eduard Bagritsky (1895–1934), and Andrei Bely (1880–1934), the French writer Henri Barbusse (1873–1935), the Russian politician Anatoly

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Fig. 4 The material substrate of his genius: microphotographs of slices of the brain of Lenin (right) and an ordinary person (left). (From Prozorovskii 1929. Image freely accessible at: https:// jhiblog.org/2016/10/31/the-brain-for-itself-soviet-psychoneurologists-debate-the-psychophysicalproblem/)

Lunacharsky (1875–1933), the German politician Clara Zetkin (1857–1933), the Russian novelist Maksim Gor’ki, (1868–1936), and the Leningrad physiologist and Nobel prize winner Ivan Petrovich Pavlov (1849–1936), among others (Spivak 2001; Richter 2007). The findings of Sarkisov’s team were later published by Adrianov et al. (1993) and contained the following statements: 1. The measurement of the furrows of the frontal lobes and the furrows of the other lobes yielded the highest percentual value for V. I. Lenin’s brain in comparison with other [. . .] brains investigated along these lines. [. . .] Likewise, the lower parietal lobe is extremely convoluted and deviates greatly from the average brain in this respect. [. . .] The temporal lobes also exhibit a large number of furrows. 2. In V. I. Lenin’s brain, we found very large cells, especially in the III layer, in the frontal region, notably in areas 10 and 46, in the lower parietal region, in the upper temporal region, in the temporal-parietal-occipital region adjacent to area 19, and in the postcentral region.

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3. We came up with important results in the evaluation of the organisational level of the brain of V. I. Lenin by comparing the size of the frontal and temporal regions and the areas they form with the corresponding square measure of other brains. The frontal region in the brain of Vladimir Ilyich occupies 25.5% of the entire surface, in the brain of Skvortsov-Stepanov 24.0%, and in the brain of Maiakovski 23.5%. What is of particular interest here is that the dominance of the frontal region in Vladimir Ilyich’s brain mainly benefits the size of the areas connected with the particularly high-developed functions of this region. Area 10 in Lenin’s brain cortex p.e. occupies 6.3 % of the entire brain surface, in Bogdanov’s brain 6 %, in Skvortsov-Stepanov’s 5.8 % and in Maiakovski’s brain 4.2 %.

These results from Vogt’s Russian colleagues confirmed his initial findings (1927–1929). In the end, as pointed out by Richter (2007), they made a step forward “as they located the origins of the associative systems mostly in the cells of layer III in the cortex, the layer which plays a major role in higher nervous activity and in extremely highly developed functions of the intellect.” The correlation of anatomical results with Lenin’s personality characteristics, which Vogt had intended, was left out. The true nature of Lenin’s disease, however, remained undescribed. The Moscow Brain Institute (Instituta mozga) is still active today and has become the repository of the brains of the most important personalities of the nineteenth and the twentieth centuries (i.e., Armenian composer Aleksandr Afanasevich Spendiarov (1871–1928), the physician and politician Aleksandr Aleksandrovich Bogdanov (1873–1928), the publicist Ivan Ivanovich Skvortsov-Stepanov (1870–1928), the neurologist Grigori Ivanovich Rossolimo (1860–1928), and the neurologist Bekhterev, who had he himself created the term “Pantheon of Brains”) (Vein and Maat-Schieman 2008; White 2009; Richter 2007).

The Funeral That Never Took Place Since no long-term preservation of the body had been planned, Professor Abrikosov did not take care when cutting and opening the major blood vessels at autopsy. This would have been avoided prior to a planned artificial embalming. Professor Abrikosov resorted to the most popular short-term embalming procedure of that time. He injected 30 measures of formalin, 20 measures of alcohol, 20 of glycerine, 10 of chloride, and 100 of water in Lenin’s aorta (Zbarsky and Hutchinson 1997). Such provisional embalming was intended to maintain the body stable, in an open coffin, for the 4 days (23 to 26 January) of the public farewell in the Hall of Pillars in the House of Trade Unions. The estimate temperature inside the building was between +5 °C and +6 °C) (Yurchak 2019). The state funeral, which had been first scheduled for the 26th Saturday was, then, postponed to 27 Sunday at 4 pm (Zbarsky and Hutchinson 1997). On January 24, the Kremlin decided that a provisional wooden mausoleum would be built by the Red Army in 3 days. By January 27, the “temporary mausoleum” was ready. Iosef Stalin (1878–1953), Nikolai Ivanovich Bukharin (1888–1938), Lev Borisovich Kamev (1883–1936), and Grigory Yevseyevich Zinoviev (1883–1936),

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the most important communist leaders of the time, carried Lenin’s coffin into the provisional mausoleum. Leon Trotsky (1879–1940), who was not in Moscow at that time, was intentionally left out of the ceremony. The coffin was left open in a grave 3 metres deep (Zbarsky and Hutchinson 1997). The public farewell was extended because of the enormous crowds that gathered to bid farewell to their leader. The funeral, which should have been on the January 27, was never performed. From January to the end of March 1924, while people were still lining in front of their leader’s corpse, lengthy discussions went on among the prominent members of the Communist Party and the scientists. The very first idea of preserving Lenin’s body had already emerged at a secret meeting of the Politburo in late October 1923 when it was obvious that Lenin would not live for a long period. Now the idea, which had been aired by Stalin, had to be turned into reality (Zbarsky and Hutchinson 1997). If the archives of the Russian Centre for Preservation and Study of Contemporary Historical Documents (CRCEDHC) are to be credited, Feliks Dzerzhinsky (1877–1926) was the person responsible for the launch of the project that would have allowed to preserve Lenin’s body (Zbarsky and Hutchinson 1997). Two commissions, the “Commission for the Organisation of Lenin’s Funeral” and the “Commission for the Preservation of Lenin’s Body,” were created. The majority of the members of the party were part of both Commissions. (Yurchak 2015b, 2019) Dzerzhinsky was the powerful chairman of the Funeral Commission and the head of the first Soviet police organization then called OGPU. OGPU is the acronym of the Russian initials for United State Political Administration. It was named NKVD in 1934, and changed several acronyms over the years until it became the KGB in 1954, after Stalin’s death. On the evening of the January 23, during a meeting of the “Commission for the Organisation of Lenin’s Funeral,” Dzerzhinsky stated: “Kings are embalmed because they are kings. In my opinion the question is not so much if we should preserve Vladimir Ilyich’s body, but how” (Zbarsky and Hutchinson 1997). Not all the members of the party were in favor of preserving Lenin’s body. On the one hand, Leon Trotsky (1879–1940), Nicolai Bukharin (1888–1938), Kliment Voroshilov (1881–1969), and Lenin’s widow, Nadezhda Krupskaya (1869–1939), were strongly against it. They claimed that this action was counterrevolutionary and thought that it would have turned Lenin into a “religious” relic. Voroshilov emphasized that this action would have violated the principles of Marxism-Leninism (Tumarkin 1997). On the other hand, Vladimir D. Bonch-Brueyvich (1873–1955), who had been Lenin’s personal secretary, was in favor of building a public memorial to Lenin, but stressed that the corpse had to be placed in a closed burial (Tumarkin 1997). However, the majority of the party members, pushed by Iosef Besarionis dzе Jugashvili, aka Joseph Stalin (1878–1953), were in favor of keeping Lenin’s corpse exposed for a longer time period, potentially forever (Zbarsky and Hutchinson 1997). A “committee of three,” composed of the old Bolsheviks Avel Yekunidze, Vyacheslav Mikhailovich Molotov (1890–1986), and Leonid B. Krasin (1870–1926), was entrusted with the search for a “rapid” method to save that Lenin’s body from decay.

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Professor Vladimir Vorobiev (1876–1937), the Ukrainian Head of the Anatomy Department at Kharkov University, an expert in long-term conservation of anatomical specimens, received a telegram from Dzerzhinsky. He was requested to head to Moscow immediately. On the 28th February he met both Krasin and Professor Abrikosov. Meanwhile, Krasin had contacted the Ukrainian biochemist Dr. Boris Ilyich Zbarsky (1885–1954) (Zbarsky and Hutchinson 1997). On March 5, 1924, an important meeting of the Central Executive Committee took place. Several different proposals were discussed. The engineer Leonid B. Krasin (1870–1926), a member of the Central Committee, proposed that the sarcophagus containing Lenin’s body to be placed in a metal box with a glass lid. The box would have then been filled with a transparent embalming liquid invisible from the outside. However, this proposal did not meet the approval of Feliks Dzerzhinsky (1877–1926). Instead of submerging Lenin’s body in an embalming liquid like as though it were some kind of dead meat, Dzerzhinsky proposed freezing it. Other members of the Commission disagreed. Freezing a body was risky, as it would have preserved both the body, and its post mortal defects. As matter of fact, when Lenin’s body was transported in an open coffin from Gorki estate to the railway station (a few kilometres away), the eyelids, ears, hands, and feet were damaged by the extremely low temperatures. Furthermore, Lenin bore the stigmata of his debilitating illness (Zbarsky and Hutchinson 1997; Yurchak 2019). In reply to Dzerzhinsky, Grigory Belen’ki pointed out that freezing was not a reliable method. A series of recent experiments performed on human bodies had shown that, even when a slight change in temperature occurred, frozen bodies turned black (as soon as the temperature changed just a little, the body turned black. So, if something breaks down – say the freezer is not working for half of an hour- it will all turn black and everything will be lost) (Zbarsky and Hutchinson 1997; Yurchak 2019). Dr. Maksimilian A. Savel’ev (1884–1939), also known as with pseudonym I. Petrov, an Old Bolshevik and economist, suggested preserving Lenin’s body in a transparent capsule filled with pure nitrogen. This would stop the decomposition process. But Krasin opposed this idea. The meeting was closed by Yenukidze’s (1877–1937) and Voroshlikov’s statements (Yurchak 2019). Yenukidze, who was one of the most prominent Bolsheviks and member of the Executive Central Committee, stated that Lenin’s body had to be preserved the way it was (We should certainly understand that we will not be able to preserve Vladimir Il’ič for a long time. . . We will freeze the body without promising that this is done for posterity. If disaster strikes and it continues changing even when it’s frozen, we will have to enclose it) (Zbarsky and Hutchinson 1997; Yurchak 2017a). Freezing equipment was bought in Germany. It was sent to Moscow, but it was never transferred to Lenin’s Mausoleum (Yurchak 2017a). Hence, Voroshlikov made his final remarks were I propose doing nothing. If the body holds up for another year, without change, this is already good enough (Zbarsky and Hutchinson 1997; Yurchak 2015b). Even if the weather was extremely cold (the temperatures registered in those days were between 15 °C and 27 °C) (Yurchak 2019), by late February to the

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beginning of March, the body already showed early signs of decomposition (Zbarsky and Hutchinson 1997; Yurchak 2017a). On March 26, 1924, the examination commission in charge of constantly checking the integrity of Lenin’s body noticed that the skin of the face and hands had darkened, wrinkles could be seen on various part of the body. . . the lips had become slightly parted (Zbarsky and Hutchinson 1997). On the same day, in a cellar underneath the temporary Mausoleum Professor Vorobiev, Dr. Zbarsky, the Anatomy Professor Piotr Karuzin, and their assistants (Arnold Shabadach, Alexander Juravlev, Yakow Zamkovsky) started to set up the embalming procedure. The body was partially decomposed and smelling. Major signs of decomposition could be detected. More brown patches had appeared on Lenin’s lower legs, the left hand had turned to a green-gray color, the ears had crumpled up, the eyelashes were lost, and the lips had retracted and the mouth had dried up (Zbarsky and Hutchinson 1997). Professor Vorobiev started the procedure by removing all the sutures that had been used to sew up the head and the chest following the autopsy. He removed the lungs, liver, spleen, kidneys, intestines. No information is available as to the preservation of the internal organs. Then, the inner side of the rib cage was washed with distilled water and then the tissues were fixed with formalin. The face, the hands, and the body were covered with cotton wool wraps impregnated with 1% solution of formaldehyde (Zbarsky and Hutchinson 1997). All body cavities were cleaned with acetic acid and, then, formalin was injected into all tissues which had shown signs of softening. Since in late March the outside temperature was still under 0 °C, a series of stoves were installed in the cellar. A temperature of 16 °C had to be reached to allow Professor Vorobiev to carry out the work of anatomical conservation. The body was then immersed in a 3% solution of formaldehyde, avoiding the use of an ordinary metal bath; the surface might have chemically interacted with the chemical components of the embalming liquid. A rubber bath was needed and was quickly found (Zbarsky and Hutchinson 1997). The results of the first bath in the viscous liquid were not as satisfactory as hoped. The tissues had not completely absorbed the fluid. As a result, Professor Vorobiev made several incisions into the skin, muscles, abdomen, shoulders, lower legs, back, in the palms of the hands and feet and in the space between the fingers (Zbarsky and Hutchinson 1997). Meanwhile, the chemical composition of the initial bath was modified. A 20% solution of alcohol, which is known to improve the color of the skin and makes it more permeable, was added. Six days later, the percentage of alcohol was increased to 30% and also a 20% of glycerine was added. The body was left immersed in the bath for 15 days and was later immersed in a mixture of glycerine and water so that tissues could slowly recovered their initial elasticity. Large jars of potassium acetate were later poured into the bath (Zbarsky and Hutchinson 1997). By the end of June 1924, the bath contained 240 litres of glycerine, 110 kg of potassium acetate, 150 litres of water, and, as disinfectant between 1% and 2% quinine chloride (Zbarsky and Hutchinson 1997).

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Ilya Borisovitch Zbarsky (1913–2007), the Director of Vladimir Lenin’s Mausoleum (from 1956 to 1989), stated that this was the formula adopted for all subsequent treatments of the body, which even now [i.e. 1997 – authors’ note] still take place beneath the mausoleum every 18 months (Zbarsky and Hutchinson 1997; Quigley 1998). As member of the Museum Laboratory, the young Ilya, a renowned biochemist, had worked with his father on Lenin’s body from January 1934. It is worth mentioning that Professor Vorobiev resorted to a procedure for tissue preservation that another Russian academic, Professor Nikolay Fedotovic Melnikov-Razvedenkov (1866–1937) had already used for the first time in 1895 (Zbarsky and Hutchinson 1997). Professor Melnikov-Razvedenkov was a highly reputed anatomical pathologist from Moscow University who headed the pathology-anatomy department in Kharkiv (Ukraine) for 18 years (from 1902 to 1919) (Zbarsky and Hutchinson 1997). During his experiments, he used a similar highly hygroscopic solution containing glycerine, potassium acetate, and alcohol. Tissues immersed in this type of solution were able to absorb and to retain water, and held the moisture for a longer period. Professor Melnikov-Razvedenkov also noticed that glycerine preserved the elasticity of tissues and allowed the original color of the skin to be retained (Zbarsky and Hutchinson 1997). Dr. Ilya Zbarsky stated that: it may be said, therefore, that Professor Vorobiev’s real distinction lies in his adopting and improving upon Melnikov-Razvedenkov’s method (Zbarsky and Hutchinson 1997). One of the main problems Professor Vorobiev, Dr. Boris Zbarsky, and colleagues had to face was the appearance of dark spots on Lenin’s face and hands. These problems were solved by Professor Vorobiev by adding different chemical reagents between baths. Where an area of the skin was wrinkled or discolored, acetic acid diluted with water was applied. The original tissue coloring was restored by using hydrogen peroxide. Disinfectants like quinine or carbolic acid were used to eliminate damp spots (Zbarsky and Hutchinson 1997). Once all the visible defects had been eliminated, Lenin’s embalmers started to restore the eyes and the mouth. The eyes were removed and false eyeballs were inserted into the orbits. The eyelids, which tended to openwere closed and sewn in place. Stitches were inserted under Lenin’s moustache to keep the mouth closed (Zbarsky and Hutchinson 1997). Once the restoration was concluded, Lenin’s body could be dressed up. His widow provided Dr. Boris Zbarsky with the khaki-coloured tunic that the Leader had worn during his illness at Gorki (Fig. 3) which resembled a uniform. In mid-June 1924 Dzerzhinsky ordered that Lenin’s body to be dressed and to be lain in a cone shaped glass coffin. Lenin’s wife Nadezhda and his brothers and sisters were then invited to see the preserved body. Dimitry Ul’yanov was astonished on seeing his brother’s body and said I am very moved. . .it takes my breath away. He looks as he did when we saw him a few hours after he died- perhaps even better (Zbarsky and Hutchinson 1997) A second wooden Mausoleum designed by the famous architect Aleksej Ščusev (1873–1949) was completed in July 1924 and Lenin’s body was moved into it. Professor Vorobiev and Dr. Zbarsky had worked constantly on the body for 4 months. They reported the results of their experimental embalming to the

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Commission for the Immortalisation of Lenin Memory in late July 1924. They claimed that there was a chance of preserving Lenin’s body for a long period without specifying for how long. However, they underlined that regular treatment and reembalming would be needed to preserve the body’s shape (Yurchak 2019). The Commission for the Immortalisation of Lenin Memory was satisfied with their results. In 1929, the second Mausoleum was replaced by a third one (the pyramidal shape was similar but the wood was substituted with red and black granite) again designed by Ščusev (1873–1949) and Lenin’s body was moved in. Professor Vorobiev died on October 31, 1937 and Dr. Boris Zbarsky became the new director of the Mausoleum laboratory (Zbarsky and Hutchinson 1997). Repeated procedures were performed in the following years and several conservation problems arose. On January 19, 1939, the members of the Commission of the People’s Commissariat of Health for the Examination of Lenin’s Body reported that Lenin’s nose, whose original shape was lost due to exposition of extreme cold, had been successfully reconstructed. The eyelids were shown to have preserved their elasticity and the face makes a complete impression of a sleeping person rather than a corpse (Zbarsky and Hutchinson 1997; Yurchak 2019). Professor Nikolai Burdenko (1876–1946) noticed that there were new spots on the outer side of the left forearm and in the lower part of the body, especially in the pelvic area. Dr. Alexei Busalov, Director of Medical Administration of the Kremlin added that on the soles and toes there are some signs of mummification. In the pelvic area, there are hints of wrinkling and thinning. The scientists were then called to solve these problems that were due to oxidation and hydrolysis of the subcutaneous fat. An examination carried out 3 years later confirmed that these problems had been resolved, and the defects eliminated (Yurchak 2019). The problem of fat hydrolysis was solved with the development of an artificial filling material, which had the same characteristics of subcutaneous fat (consistency, suppleness, and firmness); this material was chemically neutral and did not liquefy at room temperature (Yurchak 2019). In November 1943, Dr. Zbarsky explained to the leadership how Lenin’s body had been preserved so far resorting to an artificial material composed of a mix of paraffin, glycerin, and carotene with the melting point of 56° Celsius. This mix in liquefied form can be injected under the skin, where it quickly hardens into a solid mass that can be easily shaped. After experiments in the lab it became possible to substitute hydrolyzed fats with this new mass. From a chemical point of view, this mass is inert and can be preserved without change. . .Two years of experiments in this area produced such good results that they could be applied to fixing defects in Lenin’s body (Yurchak 2019). This “re-sculpting” mix was inserted via microinjections in various parts of the bodies where defects were seen. A major accident occurred on February 14–15, 1945, when a piece of Lenin’s skin from the back of his right foot was lost during a reembalming procedure; specifically, during a foot bath aimed at restoring the elasticity of the skin. This was immediately reported by Lavrentiy Beria (1899–1953), the director of NKVD (later KGB) and supervisor in chief of the laboratory for the treatment of Lenin’s

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body, to Vyacheslav Mikhailovich Molotov (1890–1986), who was Chair of the Council of People’s Commissars in a letter dated March 9, 1945. Beria drew the missing piece on a one-to-one scale and ordered Dr. Zbarsky and colleagues to look for it (Zbarsky and Hutchinson 1997). The piece was never found. It had dissolved in the solution of glycerine. Hence, the members of the Laboratory, which had been created in 1939, 15 years after Lenin’s death and the first long-lasting embalming performed by Professor Vorobiev and Dr. Boris Zbarsky had to develop new materials to substitute the lacking piece. The material had to show the same characteristic of Lenin’s skin and reapplied in its original position. Of course, it was stressed that further loss of Lenin’s skin must be avoided (Zbarsky and Hutchinson 1997). For this reason, Beria ordered Dr. Zbarsky’s team to conduct experimental work on other bodies that he called “the objects” before applying any new method to the Lenin’s body. About two dozen bodies of unknown people were then used as “experimental objects.” All were embalmed using the same procedure used to preserve Lenin’s body. The aim was to ensure no further mistakes and loss of anatomical material (Zbarsky and Hutchinson 1997; Yurchak 2015a, b, 2017a, 2019). In the 1940s and 1950s, further methods were developed to allow a careful conservation of Lenin’s body. The members of the Mausoleum laboratory, created in 1939, and now called Centre for Scientific Research and Teaching Methods in Biochemical Technologies, progressively increased their studies on bodies of deceased people (Yurchak 2015a, b). Daily, weekly, and monthly experiments were performed. Since then, Lenin’s body is subjected to the so-called big procedures every 18 months, each lasting 2 months. The body is thoroughly examined, tested, embalmed, remodeled, and resculpted, to eliminate any defects. All embalming fluid is drained out of the body and subjected to analyses to avoid the presence of unwanted organisms (bacteria, fungi, etc.) that might affect the body. With the help of formaldehyde, which binds to fungal proteins, all infestation problems have been solved (Zbarsky and Hutchinson 1997; Yurchak 2015a). A series of microphotographs of the body are taken and compared to those from the previous reembalming procedure so if any changes had occurred. Microinjections are performed to reshape all parts of the body; reembalming liquids are introduced and then the body is submerged in various solutions (different solutions are designed for the different parts of the body) (Zbarsky and Hutchinson 1997; Yurchak 2015b). The first solution is a bath of glycerin, acetate potassium, and water. The body stands in the first solution for 1 month or more, depending on the condition of Lenin’s tissues. Other baths follow. According to Zbarsky, on average, the body has to be immersed in each solution for 2–3 weeks; some contain embalming materials, and others calcium, which is used to counteract the process of decalcification of the bones. As Zbarsky stated in 1939, once this procedure is completed, the body is as if he were living (Zbarsky and Hutchinson 1997; Yurchak 2015a). Two tasks were/are of vital importance for the preservation of Lenin’s body; the “calcium balance” and the “hydrochloric balance.” These procedures, which had started in 1924, became more and more refined over time.

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The Immortalization of the Leader: A “Living Sculpture” Professor Iurii Mikhailovich Lopukhin, the highly reputed anatomist and veteran of the Mausoleum Laboratory, describes Lenin’s body as a “living sculpture” (zhivaia skul’ptura), the sculpture of a body that is constructed out of the body itself (Yurchak 2019). The body exposed in the Mausoleum obviously belongs to Lenin. However, it is not a mere external representation of the body such an effigy, a painting, or a sculpture. The body of Lenin is that of a nonliving person that does not decompose, it is a body without organs whose perpetual form is obtained through a constant process of modification of his biomatter (Yurchak 2019). The aim of the embalmers has been to preserve the physical dynamic appearance of each single part of Lenin’s body (skin, bones, muscles, and tendons). Lesser importance was given to the preservation of the original biological material, which has been substituted with artificial materials when needed (Yurchak 2015b). The body has to keep its original shape, color, as well as his dynamic characteristics – its overall suppleness, elasticity of the skin, flexibility of the joints, internal pressure in muscle tissues and so on . . .scientists have maintained not only the features of Lenin’s face but also the shape of his heels, the pigmentation around his armpits, the strength of hair attachment on his chest and the flexibility of his knee joints (Yurchak 2015b) (Figs. 5a, b and 6). More and more organic material has been replaced with artificial material. The best example is the missing eyelashes, which have been replaced by commercial ones (Yurchak 2015a). Hence, Lenin’s body has been remodeled, reshaped, and resculpted over the past 96 years (Yurchak 2015b) (Fig. 5a, b). Professor Vladizlav Kozel’tev, one of the members of the conservation team, claimed that every wrinkle, cavity, or protrusion must be fixed. We are talking about tiny dimensions. Some amounts of artificial substitutes had to be introduced, which is quite difficult (Yurchak 2015b). As a result, all the members of the Mausoleum Laboratory (which consists of several laboratories and different scientific groups) agree in stating that Lenin’s body

Fig. 5 Lenin’s body in 1968 (a) and in 1993 (b). The leader lies in his sarcophagus beneath a glass lid. Only the head and the hands are exposed. Note that the left hand is flattened whereas the right hand is clenched into a fist. Following Todorov (1995), the clenched fist is the symbol of the revolutionary salute. However, it should also be noted that, from summer 1923, Lenin’s right hand was also clenched due to the strokes he suffered. © (Reproduced with permission of akg-images)

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Fig. 6 Lenin is exposed to public display in the basement of the Mausoleum bearing his name on the edge of the Red Square (Zbarsky and Hutchinson 1997). During the WWII, Lenin’s body was moved to Tyumen in Western Siberia. The body arrived in Tyumen on the 7th July 1941 and was taken back to Moscow in March 1945. While in Tyumen the body underwent careful checks (Zbarsky and Hutchinson 1997). © (Reproduced with permission of akg-images)

cannot be classified as a mummy. His body is soft and flexible in every single part. He does not dehydrate, darken, or distort. This makes him unique and different from naturally mummified, embalmed or plastinated bodies. In mummified and embalmed bodies, the original shape changes as the corpse undergoes dehydration and shrinking. Similarly, the original pigmentation of the skin and its elasticity are lost. In the natural process, the cadaver loses its life-like appearance and becomes unrecognisable (Yurchak 2015a). In the case of plastinated bodies, while the external appearance may be preserved, the flexibility and elasticity are lost. The plastinated bodies, which may apparently show an “in motion” posture, are stuck forever in the position they were given by the anatomist (i.e., Gunther von Hagens’ plastinated bodies) (Bianucci et al. 2015). Lenin’s body cannot be considered as a wax effigy as has occasionally been supposed and discussed (Aufderheide 1998; Yurchak 2015a, b). On several occasions, foreigners have had the opportunity to look at Lenin’s body in detail. An American journalist, who had been invited in the 1930s to see Lenin’s Mausoleum (Fig. 6), saw that Boris Zbarsky . . . tweaked Lenin’s nose and turned his head to the right and to the left (Yurchak 2015b). Similarly, the German criminal biologist Mark Beneke had access to Lenin’s body during a TV production in the 2010s (Wendorf and Benecke 2015). There, he confirmed that Lenin’s body was flexible and contained biological material.

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In 2008, the science writer Tiziana Lanza wrote a fictional science-based mummy novel, Rosalia Per Sempre, about the famous child mummy, Rosalia Lombardo, housed in the Capuchin Catacombs of Palermo (Lanza 2008; Sineo et al. 2008).The plot includes the invented idea of an interaction between the Bolshevik government and an Italian embalmer, Alfredo Salafia of Palermo (1869–1933) (Lanza 2008). A similar idea was proposed to the newspapers and media 5 years later (Arcovio 2013; Il Messaggero 2013; Russia Beyond 2013) by a mummy investigator working at the Capuchin Catacombs of Palermo. The researcher hypothesised that the Soviet embalmers may have been influenced by the techniques developed in Sicily by Alfredo Salafia (Corriere del Mezzogiorno 2013). However, no documents attesting to the existence of correspondence between the Bolsheviks and Salafia have been published. Any role for Salafia’s technique in the preservation of Lenin’s body remains to be scientifically proved. Similarly, the existence of scientific contacts between the Pre-Communist Russia and Salafia is not proved. Salafia received a letter from the Russian Embassy dated May 16, 1909 in response to his own letter to the Russian Queen Mother Maria Federovna (1847–1928). She had had the opportunity to witness Salafia’s work during a visit to Palermo. In this correspondence, the Queen Mother praised Salafia’s work (Piombino-Mascali 2009; third edition 2012). However, a letter of thanks does not imply the existence of a scientific collaboration. With the assassination of the Romanovs (Nikolai II and his family) and the Russian Revolution, the political and sociocultural environment changed dramatically. Marxism-Leninism was against bodily preservation after death. An exception was made with Lenin for ideological reasons that occurred after his death and his embalming occurred resorting to Melnikov-Razvedenkov’s method (Zbarsky and Hutchinson 1997).

The Embodiment of the Leader The reason for the exception against body preservation made for Lenin stems from the problem faced by the members of the Communist Party because his body had not been buried, and had started to decompose. It could have been buried in Spring 1924 but it was not. The historian Nina Tumarkin (1999) has suggested that Joseph Stalin built the cult of Lenin because bodily preservation of a “sacred relic” could continue to legitimize Soviet power and mobilize the population. At that time, the vast majority of the Russian population was living in the rural remote areas of the country. Religious orthodoxy included the cult of Saints (and the Tsar) and was deeply rooted in the minds of the ordinary people. The cult of Lenin, the “sacred symbol of the heroic revolutionary past,” was quickly assimilated (Tumarkin 1999; Yurchak 2015b). While it is more than plausible that the devotion to Lenin’s “sacred body” would become firmly established by the largely illiterate population, Yurchak (2015b) considers that this explanation alone does not completely justify the lengthy discussions that occurred between Lenin’s death in January and March 1924.

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During Lenin’s final illness, the leader was progressively removed from the political scene becoming completely isolated 1923. Meanwhile, during Lenin’s illness and in the immediate days after his death, Joseph Stalin, who had arranged to become Lenin’s heir, and the members of the Politburo siding him “constructed” a new ideology, Leninism (Zbarsky and Hutchinson 1997). In the Leninist organization, the Communist Party was characterized by a “charismatic impersonalism,” which was always right and legitimate (Jowitt 1992; Yurchak 2015b). However, the Communist party was not only a “charismatic collective impersonal agent” but was also the sovereign of the Soviet system, since it represented the “locus of Truth” (Yurchak 2015b). On July 24, 1924, the Commission for the Immortalization of Lenin’s Memory released a public statement, which was read by Avel Yenukidze. The statement was aimed at explaining to the Soviets why the Communist Party decided to preserve the body of Lenin: We did not want to turn the body of Vladimir Ilyich into some kind of “relic”, by means of which we could popularise and preserve his memory. He had already immortalised himself enough with his brilliant teaching and revolutionary activities . . .We wanted to preserve the body of Vladimir Ilyich. . . [because] it is of great importance to preserve the “fizicheskii oblik” (the physical appearance) of this remarkable leader for the next generation and all the future generations (Yurchak 2015b).

Based on Yenukidze’s statement, Yurchak (2019) introduced the concept of the dual bodies of Lenin, a concept created by the Communist party leaders. The dual bodies of Lenin represented both the Leader’s teaching to the present and future generations and, at the same time, represented the “truth” of Leninism transcending the man (Yurchak 2019). From the one hand, there was the body of Vladimir Ilyich, the individual who had died, and, on the other hand, there was the embodiment of the foundational Truth of the Communist ideology, Leninism, which was far bigger than Vladimir Ilyich himself (Yurchak 2015b, 2017b). The existence of a “dual” body of Lenin profoundly influenced the way the corpse has been preserved over the past 96 years. It explains why Lenin’s body is not considered as a mummy but “a living sculpture” whose physical appearance (“fizicheskii oblik”) has to remain unmodified through a continuous process of renovation (Yurchak 2017b). The anatomical image of Lenin (anatomischeski obraz) immortalizes the body of the Communist Party. From this point-of-view, the never-ending process of modification, resculpting and remodeling results in a preservation that does not have to be considered synonymous with conservation but with cultivation of the sovereign body. This concept transcends Lenin himself (Yurchak 2015b). This model of perpetual sovereignty did not develop from the previous tsarist monarchy nor from the religious orthodoxy. It came from the Soviet mentality, which found a similarity only in the ideals of the French Revolution. As a matter of fact, following the Russian Revolution, a new concept of “democracy” was reached (Yurchak 2019). The body of Lenin became the “locus of Truth” and the body of the Communist Party. It represented the political transformation both for its contemporaries and for the future generations, as explained by Yekunidze during his statement on

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July 24th, 1924 (Yurchak 2019). The material form of this immortal body was the perpetually reconstructed body of Lenin. This was the cultivation of the immortal, infallible, perpetually renewed body of the sovereign-party- the body that transcended individual mortal bodies of each of his members and leader (Yurchak 2015b). Based on this concept, each Soviet leader, including Stalin, the “Great Continuer of Lenin’s cause” was legitimized only with reference to Leninism, but he could not supersede Lenin. Stalin, the man with the absolute power and with an extreme cult of the personality, underwent a process of denunciation. Nikita Sergeevič Chruščëv (1894–1971), the Soviet leader who led the de-Stalinization process (in 1956), attacked Stalin’s cult of the personality and delegitimized him. Although Stalin’s figure was wiped out, the Soviet Party did not collapse, because Stalin was not the “locus of Truth.” The same happened to Chruščëv in 1964 when he was accused of violating the Leninist principle of collective leadership and was deposed by the Politburo (Yurchak 2015b). Neither Stalin nor any other following leader of the USSR would have ever been able to occupy “the locus of the sovereign power.”

Embalmers of the Communist Leaders The Mausoleum laboratory, which has guaranteed the conservation of Lenin’s body for over 90 years, was also involved in the permanent embalming of several other bodies. The most renowned are the bodies of nine state leaders; Georgii Dimitrov (Bulgaria, 1949), Khorloogiin Choibalsan (Mongolia, 1952), Joseph Stalin (USSR, 1953), Klement Gotwald (Czechoslovakia, embalmed 1953), Ho Chi Minh (Vietnam, 1969), Agostinho Neto (Angola, 1979), Linden Forbes Burnham (Guyana, 1985), Kim Il-sung (North Korea, 1995), and Kim Jong-il (North Korea, 2012) (Zbarsky and Hutchinson 1997). Since the 1970s, the political relations between the USSR and China have been extremely difficult; China being opposed to the political changes in Soviet Russia policies and considering itself to be the true descendant of the real Leninism. The Soviet specialists were not involved in the permanent embalming of Mao Zedong’s body in 1977. Chinese embalmers took care of this (Yurchak 2017b, 2019; The Guardian 2016).

Conclusions From a scientific point of view, the successful reproduction of the embalming procedures on both human beings used as Lenin’s templates (“experimental objects”) and other Communist leaders testifies to the high quality of the Melnikov-Razvedenkov method. Due to secrecy concerning Lenin’s health, and the paucity of medical reports available in English, the reconstruction of the leader’s medical history in his younger years is challenging. At autopsy, the brain was weighed and later histological investigations were performed. No information about the size of the heart, its weight, and the weight of the other inner organs were reported in the official autopsy report. Apparently, no

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histology was performed on them. Although many hypotheses have been proposed over the decades, significant pieces of information are still lacking. It is hoped that in the years to come more details on Lenin’s brain pathology. It is also hoped that access to X-rays and CT scanning will be granted to confirm or infirm the presence of pathognomonic skeletal lesions possibly linked to tertiary syphilis. When all these pieces of information are revealed, scholars will be able to fully appreciate the state of health and the ultimate cause of death of the founder of the USSR.

Cross-References ▶ Mummies in Crypts and Catacombs

References Adrianov OS, Bogolepova IN, Blinkov SM, Kukuey IA (1993) Issledovanie mozga V.I. Lenina (The Study of V.I. Lenin). Usp Fiziol Nauk 24(3):40–52 Arcovio V (2013) L’imbalsamatore siciliano che aiutò i russi. [L’intervista]. Il Messaggero, 13 gennaio 2013, p 18 Arutyunov A (ed) (1999) Lenin’s record without retouch. Veche, Moscow Aufderheide AC (ed) (1998) The mummy of Vladimir Il’iych Lenin. In: Aufderheide AC (ed) The scientific study of the mummies. Cambridge University Press, Cambridge, pp 210–211 Bentivoglio M (1998) Cortical structures and mental skills: Oskar Vogt and the legacy of Lenin brain. Brain Res Bull 47:291–296 Bianucci R, Soldini M, Di Vella G, Verzé L, Day JL (2015) The “Body Worlds” exhibits and the understanding of death: do we educate children to science or to voyeurism? Clin Ter 166(4): e264–e268 Boris Yeltsin Presidential Library (2018) Presidential library declassified materials, which cast light on assassination attempt on Lenin’s life. https://www.prlib.ru/en/news/1163306. Accessed 15 July 2020 Corriere del Mezzogiorno (2013) La mummia di Lenin? Tutta opera del palermitano Salafia, May 22, 2013. https://corrieredelmezzogiorno.corriere.it/palermo/notizie/cronaca/2013/22-maggio2013/mummia-lenin-tutta-opera-palermitano-salafia-2221273336298.shtml. Accessed 20 June 2020 Hachinski D (1992) Lenin’s foot. Neurology 42:1849 Hesse G (1975) Patient Lenin – ein Übermensch? Dtsch Ärztebl 72:683–686, 755–760, 835–839 Il Messaggero (13 gennaio 2013) L’imbalsamatore siciliano che aiutò i russi, p 18 of the printed edition Jowitt K (1992) New world disaster: the Lenisinist extinction. University of California Press, Berkeley, pp 1–6 Kaplan GP, Petrikovsky BM (1992) Advanced cerebrovascular disease and the death of Vladimir Ilyich Lenin. Neurology 41:241–145 Kreutzberg GW, Klatzo I, Kleihues P (1992) Oskar and Cécile Vogt, Lenin’s brain and the BumbleBees of the Black Forest. Brain Pathol 2:363–371 Lanza T (ed) (2008) Rosalia per sempre. Lulu.com, pp 40–44 Lerner V, Finkelstein Y, Witztum E (2004) The Enigma of Lenin’s (1870–1924) malady. Eur J Neurol 11:371–376 Max Planck Institute (MPI) Brain Research Lenin’s brain. https://brain.mpg.de/institute/history/ lenins-brain.html. Accessed on 1 June 2020 Ochkin A (1934) Pamiati Vladimira Nikolaevicha Rozanova. Sovetskaia klinika 20:117–118 Payne R (ed) (1964) The life and death of Lenin. Simon and Schuster, New York, pp 637–640

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Penfield WG (ed) (1977) No man alone. A neurosurgeon’s life. Little Brown, Boston, pp 172–173 Piombino-Mascali D (2009; third edition: 2012) Il Maestro del Sonno Eterno. La Zisa, Palermo, pp 58, 86 Prozorovskii NI (1929) Problema anatomicheskoi osnovy odarennosti i mozg V. I. Lenina. Chelovek i priroda 2:14–17 Quigley C (ed) (1998) Modern mummies: the preservation of the human body in the twentieth century. McFarland, Jefferson, pp 28–36 Richter (2006) Chapter 8: Castor and Pollux in Brain Research: the Berlin and The Moscow Brain Research Institutes. In: Gross Salomon S (ed) Doing medicine together: Germany and Russia between two wars. University of Toronto Press, Toronto, pp 325–367 Richter J (2007) Pantheon of brain: the Moscow Brain Research Institute 1925–1936. J Hist Neurosci 1:138–149 Russia Beyond (15 maggio 2013) Il mistero dell’imbalsamatore siciliano di Lenin. Accessed 27 June 2020. https://it.rbth.com/storie/2013/05/15/il_mistero_dellimbalsamatore_siciliano_di_ lenin_24137. Accessed 7 July 2020 Service R (ed) (2000) Lenin: Eine Biographie. Beck, München, pp 566–572 Sineo L, Manachini B, Carotenuto G, Piombino-Mascali D, Zink A, Palla F (2008) The Palermo Capuchin Catacombs Project: a multidisciplinary approach to the study of a modern Mummy Collection (CA 1600–1900). Conserv Sci Cult Herit 8:155–165 Spivak M (ed) (2001) Posmertnaia diagnostika genial’nosti. Eduard Bagritski, Andrei Bely, Vladimir Maiakovski v kollekcii Instituta mozga. Agraf, Moscow The Guardian (2016) Preserving Chairman Mao: embalming a body to maintain a legacy. https:// www.theguardian.com/world/2016/sep/11/preserving-chairman-mao-embalming-a-body-tomaintain-a-legacy. Accessed 27 June 2020 Todorov V (ed) (1995) Black Square Red Square. Organon for revolutionary imagination. State University of New York Press, Albany, p 127 Tumarkin N (ed) (1999) Lenin lives! The Lenin cult in soviet Russia. Harvard University Press, Boston Vassiliadi DA, Tsagarakis S (2019) Diagnosis and management of primary bilateral macronodular adrenal hyperplasia. Endocr Relat Cancer 26:R567–R581 Vein AA, Maat-Schieman MLC (2008) Famous Russian brains: historical attempts to understand intelligence. Brain 131(2):583–590 Vogt O (1929) 1. Bericht über die Arbeiten des Moskauer Staatsinstituts für Hirnforschung. J Psychol Neurol 40(3–4):108–117 Wendorf R, Benecke M (2015) Lenins Leichenzustand. www.osteuropakanal.uni-freiburg.de/ Textinterview/leninsleiche. Accessed 1 Nov 2015 White R (2009) Lenin’s brain. J Neurosurg 110:1327–1328 Yurchak A (2015a) Form versus matter: miraculous relics and Lenin’ scientific body. In: Hakola O, Heinämaa S, Pihlström A (eds) From individual to communal perspectives. Collegium: studies across disciplines in the humanities and social sciences, vol 19. Helsinki Collegium for Advanced Studies, Helsinki, pp 61–81 Yurchak A (2015b) Bodies of Lenin: the hidden science of communist sovereignty. Representations 129:116–157 Yurchak A (2017a) Re-touching the sovereign: biochemistry of perpetual Leninism. In: Ben-Dor Benite Z, Geroulanos S, Jerr N (eds) The scaffold of sovereignty: global and aesthetic perspectives on the history of a concept. Columbia University Press, New York, pp 246–274. https:// www.youtube.com/watch?v¼HqLauUxJpS0 Yurchak A (2017b) The canon and the mushroom: Lenin, sacredness and Soviet collapse. HAU: J Ethnogr Theory 7(2):165–198 Yurchak A (2019) Communist proteins: Lenin’s skin, astrobiology and the origin of life. Kritika 20(4):683–715. Slavica Publisher Zbarsky I, Hutchinson S (eds) (1997) Lenin’s Embalmers. Harvill Press, London, pp 15–32, 77–92, 116–125, 139, 172–190

Ancient Greece and Mummies: The Primacy of the Soul over the Body

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Francesco Maria Galassi and Elena Varotto

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Did Ancient Greeks Know About the Egyptian Practice of Embalming Cadavers? . . . . . . . . . . What Was the Ancient Greek Perception of Egyptian Mummies? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Where There Ever Ancient Greek Mummies? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Ancient Greece did not contemplate embalming practices meant to preserve bodies for eternity. Reports of a mummy found by Schliemann are not to be trusted, while there is some literary evidence of temporary forms of honeymediated embalming practices adopted for Spartan kings. Greeks rejected the importance of preserving bodies for eternity because, as it can be found in Plato’s work Phaedo, the soul was considered more important than the body and the only truly immortal element. Keywords

Ancient Greece · Archaeology · Mummies · Schliemann · Plato · Classics

F. M. Galassi (*) · E. Varotto Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia FAPAB Research Center, Avola, Italy e-mail: francescom.galassi@flinders.edu.au; [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_54

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Introduction There are striking differences between how ancient societies and empires viewed life after death, which are reflected on their funerary practices. While the embalming art was held in high esteem in ancient Egypt to the point of being uninterruptedly performed over thousands of years and originating in Prehistory (Jones et al. 2018), a radically different approach characterized most other civilizations in the Mediterranean Basin. One clear example is that of ancient Greece, whose societies attributed great value to the preservation of historical knowledge as exemplified by the Thucydidean motto κτῆμα εἰς α᾿ εί, “a possession for ever,” yet at the same time did not endorse such preservative customs. Indeed, the ancient idea of immortality did not include the preservation of bodies in the form of mummies, and other practices, namely cremation and in-ground burials, were favored. With particular reference to famous historical characters, this is highlighted by the case of Philip II (382–336 BC) of Macedon, cremated according to the traditional Macedonian-Greek custom, and his son Alexander the Great (356–323 BC), succumbed in Babylon (in modern-day Iraq), and later subjected to the embalming process in a foreign land (Habicht et al. 2017). This chapter briefly examines the ancient Greeks’ knowledge and perception of embalming practices and evaluates the existence of mummies on Greek soil (hence excluding Hellenized Egypt) by synthetically answering some key questions by means of re-analyzing published literature on the topic and reassessing ancient written sources (Rühli et al. 2016).

Did Ancient Greeks Know About the Egyptian Practice of Embalming Cadavers? The ancient Greeks were certainly acquainted with the Egyptian practice of preserving corpses for the afterlife through a complex sequence of embalming procedures. Their knowledge must have come from direct contact with Egyptians, in particular as a result of travelers’ tales. The first substantial piece of evidence is offered by Herodotus of Halicarnassus (484?–ca. 430–420 BC), also popularly known as “the Father of History,” in the second book of his monumental work Histories (II, 86–89) (Andrews 2004; Brier and Wade 2001). As far as the surgical procedures are concerned, Herodotus left only a brief description, but did mention some aspects, namely, brain removal with an iron hook and evisceration by means of a cut in a corpse’s flank, which would constitute a relevant part of the academic discussion on Egyptian mummies in later centuries. Contemporary studies have demonstrated these two assertions to be essentially correct although not all mummies were subjected to the excerebration procedure and still exhibit cerebral remnants within their cranial cavities, hence suggesting that varied embalming approaches existed in those days and were applied (Brier and Wade 2001; Wade et al. 2011). While it is unlikely that Herodotus was ever allowed to see the making of a mummy, hence

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achieving his very own idea of seeing countries, peoples, and their customs for himself (αὐτoψία) – some authors even questioning altogether the fact that he truly visited Egypt (Armayor 1978) – modern scholarship tends to believe that he must have collected his information through intermediaries, possibly fellow Greeks who were in Egypt as he visited it (Thomas 2002). The second famous description of the Egyptian embalming practice would be furnished much later by Diodorus Siculus (fl. first century BC) in the first book of his Library of History (I, 91) (Andrews 2004; Brier and Wade 2001). Among the additional details provided by the writer from Agyrium is the mention that the heart, unlike other visceral organs, was left in situ, which contemporary mummy research has shown not to be always the case, since, as Wade and Nelson write (2010), “[t]he stereotype of universal heart retention, or replacement on accidental removal, is far from the truth. The heart was uncommonly retained in situ, and rarely returned or replaced by a heart scarab.” From a scientific standpoint, these two ancient references are considered the most important ones – later, Imperial-era, accounts by Plutarch (AD 46–119) and Porphyry (ca. AD 234–ca. AD 305) being less reliable sources – since they provide their readers with a number of details which, with the help of contemporary technology and knowledge, can help shed light on the ancient Egyptian embalming procedures. Although what we read is unquestionably two accounts mediated by a Greek mindset and worldview, it is interesting to note that both Herodotus and Diodorus choose to focus on the technical details of the ancient Egyptian art of preserving bodies rather than expressing any personal thought on it or overly weighing a foreign practice against the customs of their countrymen.

What Was the Ancient Greek Perception of Egyptian Mummies? While travelers’ accounts from the land of the Pharaohs must have been read and listened to with a mix of awe and curiosity, in order to understand what cultivated ancient Greeks might have thought of Egyptian mummies (and of preserved bodies in general), we need to go back to the fifth century BC, shifting our attention from the words of a traveler-historian to those of a philosopher, Plato (ca. 470–399 BC), particularly what he presented in his work on the soul, the dialogue Phaedo (Centini 2012). In it [80c-d] Plato has Socrates (his mentor and main character of his philosophical dialogues) say: “[. . .] when a man dies, the visible part of him, the body, which lies in the visible world and which we call the corpse, which is naturally subject to dissolution and decomposition, does not undergo these processes at once, but remains for a considerable time, and even for a very long time, if death takes place when the body is in good condition, and at a favorable time of the year. For when the body is shrunk and embalmed, as is done in Egypt it remains almost entire for an incalculable time [ὥσπερ oἱ ἐν Αἰγύπτῳ ταριχευθEντες, ὀλίγoυ ὅλoν μE νει α᾿ μήχανoν ὅσoν χρóνoν]. And even if the body decay, some parts of it, such as the bones and sinews and all that, are, so to speak, indestructible.” (Plato 1903, 1966)

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In this passage, Plato seems to be aware of the influence of climate conditions on the decomposition process and of the Egyptian practice of embalming corpses. As pointed out by Forsey (Forsey 1926), Plato stressed the fact that emaciated bodies tend to preserve themselves more than fat bodies, which putrefy faster, and he finds the example of Egyptian mummies particularly suitable given the fact that these bodies are eminently thin (understandably after evisceration and dehydration). Nonetheless, in Plato’s view it is not important that the body is preserved or can avoid the effects of postmortem deterioration. What matters to him is the survival of the soul, the only true immortal component of a human being due to its divine nature. This view would later influence other philosophical schools and religions.

Where There Ever Ancient Greek Mummies? Although a high number of mummies found in Egypt dating to the Ptolemaic and Roman eras and one case of embalming from Greece dated around AD 300 (Papageorgopoulou et al. 2009) may erroneously induce one to think of a natural acceptance of this practice by the ancient Greeks, this is not the case. No Greek case of intentional long-lasting embalming has ever been found nor is attested in the ancient sources and in the archaeological record. One famous case of pseudo-mummy is that of the Mycenaean mummy allegedly found by Heinrich Schliemann (1822–1890) during his excavations at Mycenae, a topic that in recent years has once again become the matter of much attention (Graziadio and Pezzi 2006; Schofield 2007; Killgrove 2012). In his book Mycenae Schliemann (1878) wrote that in Shaft Grave V he had found a body in a very good state of preservation (“the round face, with all its flesh, had been wonderfully preserved under its ponderous golden mask: there was no vestige of hair, but both eyes were perfectly visible, also the mouth . . . the colour of the body very much resembled that of an Egyptian mummy”). However, as demonstrated through a reassessment of the material found by Schliemann, this appears to be a misnomer and probably Schliemann himself never actually meant a mummified body (Papazoglou-Manioudaki et al. 2010). While Mycenean mummies can be ruled out as a serious possibility, there is an instance of embalming that is worth mentioning. That is the case of Spartan kings, who, unlike their lower class comrades, happened to be temporarily preserved in honey or wax, due to these substances’ natural antimicrobial properties, in order to be collected from the field of battle and taken home so that solemn funerals may be celebrated. Thus, one may not speak of bodies meant to be preserved for eternity but for a very limited period of time. This destiny may have also been envisioned for king Leonidas who died at the battle of the Thermopylae against the Persians in 480 BC; however, it proved impossible to do so because he had already been beheaded by his enemies and likely had already decomposed (Cartledge 2011, 2012; Millender 2017).

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Conclusion In conclusion, it can be summarized that the ancient Greeks were well aware of the Egyptian practice of embalming corpses but they looked down upon it particularly due to their different approach to death and, influenced by Platonic philosophy, were more concerned on the survival of the soul than that of the body. In addition, no evidence of mummies exists for the most ancient and classical phases of ancient Greek history, with the sole exception of forms of temporary preservation using honey or wax, as in the case of Spartan kings, meant to transport the preserved bodies of famed deceased in order for them to receive the expected funerary rites. Acknowledgments We are grateful to Professor Simon T. Donell who generously proofread and edited this chapter.

References Andrews C (2004) Egyptian mummies. Harvard University Press, Cambridge, MA, pp 20–22 Armayor OK (1978) Did Herodotus ever go to Egypt. J Am Res Cent Egypt 15:59–73 Brier B, Wade RS (2001) Surgical procedures during ancient Egyptian mummification. Chungará (Arica) 33(1):117–123 Cartledge P (2011) Thermopylae: the battle that changed the world. New York, Pan Macmillan, Kindle Edition, p 255 Cartledge P (2012) The Spartans: an epic history. London, Pan Books, Kindle Edition, pp 441–442 Centini M (2012) Le mummie. Un viaggio alla scoperta della più affascinante sfida alla mortalità. Xenia, Milano, p 98 Forsey GF (1926) Plato, Phaedo, 80 c. Class Q 20(3-4):177–178 Graziadio G, Pezzi E (2006) Schliemann and the so called ‘Agamemnon’s mask’. SMEA 48:113–131 Habicht ME et al (2017) The lost mummy of Alexander the Great: theoretical considerations and hypothetical scenarios. In: “Disease and the Ancient World”, Green Templeton College, University of Oxford, 21–23 September. [Conference paper] Jones J et al (2018) A prehistoric Egyptian mummy: evidence for an ‘embalming recipe’ and the evolution of early formative funerary treatments. J Archaeol Sci 100:191–200 Killgrove K (2012) Schliemann’s Mycenaean Mummy. Powered by osteons [blog], online at http:// w w w. p o w e r e d b y o s t e o n s . o rg / 2 0 1 2 / 0 7 / s c h l i e m a n n s - m y c e n a e a n - m u m m y. h t m l ? fbclid¼IwAR0xDaov5tdlUGbya9iizdoN96-FkMUGL6gIor4taLSEZfRslwbhxNQ6xsw. Last accessed 12 Oct 2020 Millender EG (2017) Kingship: the history, power, and prerogatives of the Spartans’ ‘Divine’ Dyarchy. In: Powell A (ed) A companion to Sparta, vol II. Wiley, Hoboken, pp 452–479 Papageorgopoulou C et al (2009) Indications of embalming in Roman Greece by physical, chemical and histological analysis. J Archaeol Sci 36(1):35–42 Papazoglou-Manioudaki L et al (2010) Mycenae revisited part 3. The human remains from Grave Circle A. Behind the masks: a study of the bones of Shaft Graves I–V. Annu Br Sch at Athens 105:157–224 Plato (1903) Platonis Opera (ed: Burnet J). Oxford University Press, Oxford Plato (1966) Plato in twelve volumes, vol 1. (trans: Fowler HN). Harvard University Press/William Heinemann Ltd, London/Cambridge, MA

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Rühli FJ et al (2016) Palaeopathology: current challenges and medical impact. Clin Anat 29(7):816–822 Schliemann H (1878) Mycenae. Scribner, Armstrong & Company, New York, p 296 Schofield L (2007) The Mycenaeans. Getty Publications, Los Angeles, p 43 Thomas R (2002) Herodotus in context: ethnography, science and the art of persuasion. Cambridge University Press, Cambridge, p 277 Wade AD, Nelson A (2010) Heart treatment in ancient Egyptian mummification. In: 38th annual meeting of the Canadian Association of Physical Anthropology. Saskatoon, SK, 27-30 October. (Conference paper) Wade AD et al (2011) A synthetic radiological study of brain treatment in ancient Egyptian mummies. Homo 62(4):248–269

Guanche Mummies

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Conrado Rodríguez-Martín

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Problems in the Scientific Study of Mummified Remains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Problems in the Biological Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pseudopathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Problems in the Cultural Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Tenerife and the Canary Islands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A History of the Guanche Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Spoliation of Guanche Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Spain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . France . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Germany . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . England (United Kingdom) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Russia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Netherlands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Argentina . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Canada . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The United States . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Repatriation of Guanche Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Repatriation of Two Guanche Mummies from Necochea (Province of Buenos Aires, Argentina) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Repatriation of Three Guanche Mummies from Museum of Medical-Forensic Anthropology, Paleopathology, and Criminalistics “Reverte Coma” (School of Legal Medicine, Complutense University, Madrid) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Research on Guanche Mummies: CRONOS AND Athanatos Projects . . . . . . . . . . . . . . . . . . . . . . . . CRONOS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Athanatos . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Guanche Types of Burial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Types of Mummification Among the Guanches . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Natural or Natural-Intentional Mummification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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C. Rodríguez-Martín (*) Instituto Canario de Bioantropología and Museo Arqueológico de Tenerife, OAMC-Cabildo de Tenerife, Santa Cruz de Tenerife, Spain e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_24

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Anthropogenic Mummification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Comparative Studies Between Mummified and Skeletal Populations . . . . . . . . . . . . . . . . . . . . . . . . . Demography . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Diet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Markers of Physical Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Metabolic Stress Markers and Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Dental Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Skeletal Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Soft Tissue Pathology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Paleoparasitology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Guanche mummies from Tenerife have been a permanent focus of attraction for scholars from many countries, especially Europeans, and for the population of the island since the moment of the conquest (1496) when they were known by the Spaniards. Chroniclers of the conquest and ulterior historians of the Canaries wrote on these valuable specimens, on the mummification methods used by the Guanches and what the mummies represented for that society. However, until very recently the research lacked of direct observation and analyses using modern scientific methods and techniques. We will discuss in this chapter the history of these mummies, the analysis of the problems of human colonization of islands, and the comparative study between the mummified population (the elite of Guanche society) and the skeletal population represented by a larger number of individuals, in order to check differences and similarities from the demographic, dietary, nutritional, and pathological perspectives. Keywords

Guanche · Mummies · Canaries · Tenerife · Bioanthropology

Introduction Mummies are not mere objects expressing some partial aspect of a culture. They constitute an important link between human biology and the cultural practices of different societies. The amalgam between human biology and culture, synthetized in the mummies, makes their study a relevant field of research transcending their fundamental patrimonial value. The soft tissue conservation and, in some cases, the wonderful persistence of the external shape of the corpse are the true causes that the general interest for mummies and for their scientific study show a long tradition in different scientific disciplines. In general, mummies can be natural or anthropogenic. Natural mummification is due to the desiccation of the corpse by dehydration that delays or avoids its decomposition. In general, it occurs in those sites with low relative humidity, constant temperature, and abundant circulating air. The fast desiccation of the corpse makes impossible the bacterial growth. There are many

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examples: Egyptian desert, Atacama desert (northern region of Chile), areas of the southwest of the United States, coastal region of Peru, some places in Mexico, and the Canary Islands (Tenerife, La Palma, and Gran Canaria) (Rodríguez-Martín and González-Antón 1994). There are factors conditioning the degree of preservation: length of the interval between death and burial, presence of bacteremia in the moment of death, and climatic fluctuations in the region or in the burial place. Mummification can be complete or partial, and it always begin in the most exposed parts of the body (face, hands, and feet), and the process lasts normally between a month and a year, although it is very variable depending upon the environment and other factors. Artificial mummification is carried out by humans with the goal to keep a similar body shape to that of the person while was alive. Independently of the place where this practice was performed, the main goal was to get a fast dehydration of the body to avoid its decomposition. Although it is universal, the process has many variants depending of the place: • Extraction of all the internal organs, including muscles and subcutaneous tissue (sometimes even the bones) and substitution by mud and wood, like the case of the Chinchorro mummies in Chile • Extraction of internal organs prone to decomposition (spleen, pancreas, kidneys, brain) and superficial treatment with desiccating salts (like natron) like ancient Egypt • External treatment of the corpse using desiccating substances (grass, aromatic herbs, heather dust, resin, pumice stone) along with sun and smoke exposure like the mummies from Tenerife (Canary Islands) (Aufderheide et al. 1992a; Rodríguez-Martín and González-Antón 1994) (Fig. 1) Natural-intentional mummification consists in the use of favorable climatic and environmental conditions without any type of internal or external treatment (mummies of the Basket-Makers Indians of the southwest of the United States) (El-Najjar et al. 1985).

Problems in the Scientific Study of Mummified Remains Problems in the Biological Study Histopathological and biological analyses and, in a lesser degree, macroscopic and radiographic studies are hampered by four events that occur during the conservation processes, especially mummification (Aufderheide 1981): protein decay, loss of epithelial cells and nuclei, enzymatic action of lysosomes, and slow destruction of the soft tissues by bacterial and fungal action. It is evident that not all the body tissues suffer the same degree of decay. Epithelial cells are quickly deteriorated in contrast to supporting cells, and organs like the heart, kidney, or adrenal glands, to cite just a few examples, suffer a much

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Fig. 1 Subadult Guanche mummy. Barranco del Infierno (Adeje). Southwest of Tenerife.1216 AD–1527 AD. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros – Cabildo de Tenerife)

fastener and greater postmortem destruction than muscles that are usually well preserved. Tissues that are more active in life from a metabolic point of view suffer a faster and almost complete decay after death. An added problem is the histological analysis because those tissues that are worst preserved will disintegrate in the process of rehydration or suffer important structural changes that makes difficult not only the pathological diagnosis but even their identification (Aufderheide 1981; Zimmerman and Kelley 1982). On the other hand, the insertion loss of the viscera hampers their macroscopic identification due to the abnormal topographic location. Other complications may appear by overlapping of radiographic images, contamination of aDNA, or diagenesis appearing in many burial places when trying to carry out dietary chemical reconstruction.

Pseudopathology Pseudopathological changes are those produced in the normal tissue structure mimicking an antemortem lesion (Aufderheide and Rodríguez-Martín 1998). Pseudopathology can be related to funerary practices (substitution of body parts by

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others belonging to other corpses, resin molds, hair of other mummies, subdermal stuffing with sand or other conserving substances, tied tissues, obliteration of the body holes, fire action on the corpse producing abnormal postures like that of the “boxer,” and the effects produced by the burial site or the burial type.

Problems in the Cultural Study The absence of cultural artifacts along with lack, in most cases, of written sources that can guide to perform the research, the scarcity of secure ethnographic data and the paucity of radiocarbon dating in many specimens make difficult the assessment of the anthropogenic nature of many mummies, their adscription to a certain cultural group, the relation of mummification with other religious and cultural practices, etc. Other imponderables are added to the abovementioned: the constant plundering of burial sites, the intentional destruction of the specimens due to political or religious reasons, the mummy trading, reburial, etc. imply an enormous limitation for mummy studies (Aufderheide 1981; Rodríguez-Martín 1992a).

Tenerife and the Canary Islands The Canary Islands, also called Fortunate Islands in Antiquity, conform an archipelago located in the Atlantic Ocean, 100 km west of Morocco (northwestern Africa). The seven main islands are Tenerife, La Palma, La Gomera, and El Hierro (western Canary Islands) and Gran Canaria, Fuerteventura, and Lanzarote (eastern Canary Islands). All of them were inhabited at the moment of the European conquest. Tenerife is the largest island of the archipelago with a surface of 2035 km2. The pre-Hispanic inhabitants of the island of Tenerife were known as Guanches. Their culture is related to that of the Berber population in North Africa, and their technology was rather poor by the scarcity of raw material, especially metals. There are different theories about the islands settlement by human groups although the most plausible is that it was probably the final result of an extensive colonization process of the Western Mediterranean (during the 1st Millennium BCE) carried out by Phoenician-Punics (Strait Circle or Carthage Circle). After the battle of Zama (202 BCE), when the Roman Legions of Scipio “Africanus” defeated the Carthaginian Army of Hannibal, the final part of the process was done by Romans. As a consequence, Roman presence in the archipelago began to be notorious as it has been checked in Lanzarote (Atoche and Ramírez 2016) and in the islet of Lobos (in front of the northern shore of Fuerteventura) where a Roman factory of purple was discovered by chance and is being excavated and studied at present (Arco et al. 2016) (Fig. 2). Human colonization of islands can lead to their isolation, and this implies cultural answers to adapt to a new and different environment, usually with less natural resources than the mainland (Rodríguez-Martín et al. 2009). Different phenomena appear shortly after the settlement of the population. Changes in relationships, group cohesion system, funerary practices, etc. appear at a cultural level (Rodríguez-Martín

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Fig. 2 Archaeological excavation of the Roman factory of purple in the islet of Lobos (Fuerteventura). Picture C. del Arco

and González-Antón 2005). From the biological point of view, the existence of biological adaptation, including genetic phenomena – appearing specific markers – has been proved in different archipelagos around the world (Martín-Rodríguez 2008). As a scientific consequence, the islands are true laboratories for the study of human adaptability to different environments and situations like those created by the contact between islanders and Europeans bringing new diseases, especially epidemics (Crosby 1986; Rodríguez-Martín and Martín-Oval 2014). Recent radiocarbon dating on human remains belonging to the ancient inhabitants of Tenerife fluctuates between first and fourth centuries AD, although there are many cultural artifacts, animal bones, and charcoal dated as early as tenth century BCE in Lanzarote (Atoche and Ramírez 2016), Tenerife, and other islands, demonstrating that the archipelago was known by the Mediterranean cultures long before the permanent human settlement. The ulterior isolation from the Mediterranean cultures took place around third to fourth centuries AD lasting until the beginning of the European conquest in the fifteenth century AD.

A History of the Guanche Mummies The main problem of mummy research in this archipelago has been that the only basis for the study was the written sources until the decade of 1950, lacking direct observation, analysis of the internal content, use of modern techniques and methods of research, and, even, experimentation.

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Fig. 3 Eighteenth-century print showing one of the mummy caves in Tenerife containing hundreds of specimens inside. Credit: Centro de Documentación Canarias América (Organismo Autónomo de Museos y Centros – Cabildo de Tenerife)

Pierre Bontier and Jean Le Verrier, chaplains in the expedition of Jean de Béthencourt and Gadifer de la Salle to the Canary Islands at the beginning of the fifteenth century, did not mention the practice of mummification in Le Canarien, written during the first third of the fifteenth century. There are chronicles of the conquest and descriptions of the islands, written between the sixteenth and seventeenth centuries, describing the different mummification methods. Of all these volumes, the most known were those of Leonardo Torriani (1980), Alonso de Espinosa (1980), and Juan de Abreu Galindo (1978). Another author was the English physician and merchant Thomas Nichols who wrote A pleasant description of the Fortunate Islands (1526), being one of the first to mention the presence in Tenerife of funerary caves with thousand mummies. The most important source during the eighteenth century is José de Viera y Clavijo (1982) with his Noticias de la historia general de las Islas Canarias who visited one of these caves (Fig. 3). A common fact to the authors mentioned above is the eternal comparison between Egyptian and Guanche mummification, and, as occurs in the case of Egypt, most of them agreed that only the elite of the society was mummified.

Spoliation of Guanche Mummies Mummies from the Canary Islands, and especially those from Tenerife, have been objects of admiration and curiosity and, at the same time, have suffered plundering, spoliation, and speculation. There are many references on the presence of

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Guanche mummies out of Tenerife. We include here the most relevant and known examples.

Spain Museo Nacional de Antropología (Madrid): although this museum had five Guanche mummies at the beginning of the last century (Barras de Aragón 1929), until recently only one was exhibited there. This specimen, an adult male, is the best preserved of all Guanche mummies known at present and was sent to Madrid as a gift to the king Carlos III in the eighteenth century. The mummy was located first, 1776, in the Real Gabinete de Historia Natural (Mora-Postigo 1992) and, most probably, came from Barranco de Erques, between Arico and Güimar (southern Tenerife). It is believed that it belonged to the big funerary cave in which hundreds of mummies were found during the final third of the eighteenth century (Viera y Clavijo 1982). In December 2015 the mummy was moved to the Museo Arqueológico Nacional, also located in Madrid. The Cabildo de Tenerife (Government of the island) asked for the return of the mummy to the island in 1976, for the first time, without success. Thirty years later, the Cabildo began with the parliamentarian procedures in Madrid to get the repatriation of the specimen, but, once again, the Government rejected the idea arguing the risks of the trip and the environmental differences between Madrid and Tenerife.

France Musée de l’Homme (Paris): six Guanche mummies were moved to Paris by the French physical anthropologist René Verneau during his trips to the Canary Islands in the last third of the nineteenth century and the first third of the twentieth, along with hundreds of pre-Hispanic Canarian skulls, bones, and diverse archaeological material. Other part of that collection was donated by Gregorio Chil y Naranjo, physician and Director of El Museo Canario in Las Palmas de Gran Canaria, and by Sabin Berthelot, consul of France in the islands (Verneau 1879).

Germany Johann-Friedrich-Blumenbach Institut für Zoologie und Anthropologie (“GeorgAugust-Universität,” Göttingen): this university has a small collection on physical anthropology and paleopathology where a Guanche mummy is exhibited (one of the best preserved Guanche mummies). The mummy was brought there by JohannFriedrich-Blumenbach in 1803, who carried out the first examination noticing that the internal organs had not been removed. After studying the mummy with high resolution CT imaging, Gauert and Grosskopf (2018) affirm that the mummy belongs to a woman who died at the age of 30 or, at most, 40 years. Tissues are present in the abdominal and thoracic cavities, although most organs cannot be differentiated.

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Other places in Germany had Guanche mummies in some moment of their history. Mummified and skeletal remains were stored in the ancient “Königlichen Museum für Völkerkunde” (Ethnologisches Museum today) and in the “Berliner Gesellschaft für Anthropologie, Ethnologie, und Urgeschichte,” both in Berlin, but were destroyed by the bombings suffered during World War II (Rodríguez-Martín 1992a). It is probable that these mummies were those commented by Hooton (1970 [1925]) who affirmed that their state of preservation was excellent and that the specimens came from the caves of San Andrés (Santa Cruz de Tenerife).

England (United Kingdom) Museum of Archaeology and Anthropology (Cambridge University): a Guanche mummy of unknown origin and very well preserved that was brought by a certain captain Young of the vessel H.M. Sloop at the end of the eighteenth century is curated there. Brothwell et al. (1969) observed that it is a male between 40 and 44 years, robust, and with very short stature (155 cm), suffering lung anthracosis (this finding has been observed in other specimens from Tenerife) (Aufderheide et al. 1992a). Likewise, the mummy shows a very important perimortem or postmortem fracture on the head, involving the facial skull, the frontal, and the parietal bones, spondylosis at the thoracic level, and a healed fracture of the anterior third of the ninth right rib. An important datum is that the mummy shows an incision on the right side of the trunk for the introduction of sand and other absorbing substances. This type of incision is also seen in the buttocks and the two superior-posterior thirds of the thighs. These substances were also observed in other mummies at the collection of the Museo Arqueológico de Tenerife, although surrounding the mummy instead to be introduced through incisions (Sánchez-Pinto and Ortega 1992). British Museum (Natural History) (London): according to Diego-Cuscoy (1971), this institution curated human remains from Guanche origin until World War II. However, nothing appears in other sources regarding these alleged specimens.

Russia Kunstkamera Museum (St. Petersburg, Russia): in October 1803, arrived to Tenerife the vessels Nadejda and Neva of the Russian Imperial Navy for a scientific and diplomatic expedition. A complete Guanche mummy and two legs in a perfect state of preservation whose archaeological dating is unknown were given to the members of the expedition entering in the Imperial Academy of Sciences in St. Petersburg in 1808 (Langsdorff 1813–14).

The Netherlands Utrecht (unknown place): Viera y Clavijo (1982) affirms that a mummy from Tenerife showing an excellent state of preservation could be admired in this Dutch city. This is the only reference on the existence of Guanche remains in The Netherlands.

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Argentina Museo de La Plata (La Plata, Province of Buenos Aires): a very important part of the collection of archaeology and natural history of the Canaries belonging to the old Museo Casilda in Tacoronte (city in the northern slope of Tenerife) arrived to that institution when it was sold to several Argentine citizens at the end of the nineteenth century. Of the four mummies of that collection, only one (from Gran Canaria) remains in the museum (Rodríguez-Martín et al. 2005). Two were sent to Necochea (also in the Province of Buenos Aires) and since 2003 are exhibited in Museo de Naturaleza y Arqueología (MUNA) (OAMC-Cabildo de Tenerife) in Santa Cruz de Tenerife. The fourth mummy disappeared without traces many years ago.

Canada Redpath Museum (Faculty of Science, McGill University, Montreal): a Guanche mummy in a very bad state of preservation is curated in the collections of this museum and, according to several documents, was brought by the British physician and antiquarian E. I. Lambert in 1892 along with almost a dozen of mummified skulls and several archaeological objects (Horne et al. 1992). The mummy, registered as RED-1, is a male around 35–39 of age and 162–165 cm of stature, came from Barranco de Santos (Santa Cruz de Tenerife), and was dated 1380. A.P. Horne (1990) and Horne and Ireland (1991) affirm that most of the body is skeletonized with the exception of the skull and no viscera are present. Chemical reconstruction of diet demonstrates a high consumption of meat and dairy products and, in a lesser degree, vegetables. Elements of marine origin were almost absent. The most striking finding in this mummy is the presence of the moss Neckera intermedia, as an antiseptic used in the process of mummification. This analysis of Horne and Ireland was the first to demonstrate the presence of the moss, observed later in other two specimens from the Museo Arqueológico de Tenerife.

The United States Peabody Museum of Archaeology and Ethnology (Harvard University, Cambridge, Massachusetts): in 1915, after his scientific trip to Tenerife, Earnest A. Hooton ([1970]1925) illegally transported an osteological collection to the Peabody Museum, even when the permission he asked was denied, as he recognized in his book of 1925. The majority of these remains came from Barranco de la Baranda (Hoya Grande, municipality of Adeje in the southern part of Tenerife) and some others from the proximity of Santa Cruz de Tenerife, the capital of the island. The important number of plundered elements is an example of the events regarding Tenerife’s heritage that happened during and before that moment.

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Repatriation of Guanche Mummies All the stories told above implied that the research in this fascinating field of science, especially important in the Protohistory of the Canary Islands, has not been so brilliant as could be in a land where mummies have been found by hundreds during the last five centuries. The attempt to rescue this and other archaeological material in Tenerife began at the end of the nineteenth century with the creation of the Gabinete Científico in Santa Cruz de Tenerife and a little bit later with the opening of the Archaeology Section in the Museo Municipal of Tenerife’s capital. More recently, in 1958, the Cabildo de Tenerife created and funded the current Museo Arqueológico (Rodríguez-Martín et al. 2005). Two repatriations have succeeded until present.

Repatriation of Two Guanche Mummies from Necochea (Province of Buenos Aires, Argentina) These two mummies belonged to the Casilda Cabinet, collected by Sebastián Pérez Yanes. This man was a rich owner in Tacoronte that devoted most of his life to collect different natural and archaeological objects from the islands. After his death, the mummies were sold to Argentine citizens for increasing the collections of La Plata Museum at the end of the nineteenth century. What happens between the arrival of the mummies to La Plata and their presence in Necochea is unknown. The only clear datum is that the mummies appeared in Necochea in the decade of 1920 and were located firstly in the National School remaining there for almost 50 years until Necochea’s Municipal Museum of Natural Science (Necochea’s Municipality) and Colegio Nacional agreed to bring both specimens to the museum for public exhibition. The contacts for the repatriation began in 2000 to know the opinion of the Argentine scientific community, and the next step was to carry out the administrative and political work (Rodríguez-Martín et al. 2005). The final agreement was ready at the middle of 2003, and the Guanches of Necochea could finally make the trip back home, after 105 years, on September 2003 (Figs. 4 and 5).

Repatriation of Three Guanche Mummies from Museum of MedicalForensic Anthropology, Paleopathology, and Criminalistics “Reverte Coma” (School of Legal Medicine, Complutense University, Madrid) These mummies probably come from Araya (Candelaria, in the south of the island) and La Orotava, in the northern slope, and were discovered during the second half of the nineteenth century and sent to Madrid some years later (Mora-Postigo 1992). The fragmented remains of the three mummies – along with a large collection of American and Egyptian specimens – were transported from the Museo Nacional de Antropología to the University of Madrid in 1975 and they never returned there. In 2009, the Organismo Autónomo de Museos y Centros signed a Collaboration

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Fig. 4 NEC 1. Young female Guanche mummy from Tenerife showing the typical funerary bundle made of animal leather. The mummy returned from Necochea (Argentina) to Tenerife’s Archaeological Museum in 2003. Southeastern Tenerife. 1148 AD–1277 AD. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros – Cabildo de Tenerife)

Fig. 5 Male mummy (25–29 years) sold to Argentina at the end of the nineteenth century and returned to Tenerife’s Archaeological Museum in 2003. North of the island. 1019 AD–1183 AD. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros – Cabildo de Tenerife)

Agreement with the Universidad Complutense de Madrid for the scientific study of these remains and their further exhibition in the island where they are curated since 2011 (Fig. 6).

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Research on Guanche Mummies: CRONOS AND Athanatos Projects CRONOS The so-called CRONOS Project. Bioanthropology of Guanche Mummies (1989– 1992) was the first interdisciplinary research project carried out on Guanche mummies and skeletal remains by a team of Spanish and North American scientists in order to get as much as possible information on these valuable specimens. Besides the research project, the entire program consists in the holding of the I World Congress on Mummy Studies, under the Presidency of Arthur C. Aufderheide from the University of Minnesota-Duluth, and an international exhibition entitled “Momias, los secretos del pasado” (Mummies. Secrets of the Past), both organized by the Museo Arqueológico de Tenerife with the collaboration of more than a dozen institutions (national and international) that took place on February 1992.

Fig. 6 Head of a Guanche mummy. It belonged to Museo Reverte (School of Legal Medicine, Universidad Complutense de Madrid) until 2011 when was returned to Tenerife’s Archaeological Museum. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros - Cabildo de Tenerife)

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Athanatos Athanatos. Death and Immortality in Past Populations (2016–2018) was organized by the Instituto Canario de Bioantropología and Tenerife’s Archaeological Museum (both belonging to the OAMC-Cabildo de Tenerife) to commemorate the 25th anniversary of CRONOS Project. It consisted in an homonymous international exhibition on mummies in the Museo de Naturaleza y Arqueología and the Extraordinary World Congress on Mummy Studies, both held in Santa Cruz de Tenerife during the first half of 2018, as well as a multi-annual (2016–2021) research program entitled “Guanches, una visión integradora” (“Guanches, an integrating view”). The goals of this study are the following: • Correct location of the archaeological sites. • Increase the chronological data of mummies and other archaeological material. • Integration of the archaeological data through the complete documentation of every site. • Chemical dietary reconstruction of the whole island. • Review the demographic data of the selected areas. • Continuation of the analysis of metabolic stress and physical activity markers. • Continuation of the paleopathological studies until complete the whole pre-Hispanic population of the island. Another goal of these studies is the inclusion of the contact period between Guanche and European (with special attention to the contact diseases introduced in the island) because Tenerife was an important port of connection between the Old and New World and Africa and many epidemic diseases affected the island severely (Rodríguez-Martín 1994; Rodríguez-Martín and Martín-Oval 2014) (Fig. 7).

Guanche Types of Burial The Guanches buried their deceased in natural caves normally located in the slopes of the island ravines. There were different types of burial in Tenerife prior to the conquest (Arco 1976; Rodríguez-Martín and Martín-Oval 2009): • Burials with a single individual (mummified or not). This is the most uncommon type. • Caves with mummified individuals only. These caves can contain a variable number of specimens, from a few to dozens or hundreds. According to some historical sources (Nichols 1526; Viera y Clavijo 1982), the number of these large necropolises account, at least, for 20 caves distributed through the island. Nichols affirmed that the “old Guanches” showed him a cave of this nature in acknowledgment for his medical services and Viera y Clavijo mentions the discovery of a cave with hundreds mummies inside, in the Barranco de Erques, between Arico and Güimar, in the eighteenth century. An important number of specimens were sent to different museums, cabinets, and universities around Europe.

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Fig. 7 Partial aspect of the exhibition Athanatos. Death and Immortality in Past Populations (December–June 2018, Museo de Naturaleza y Arqueología, Santa Cruz de Tenerife). Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros - Cabildo de Tenerife)

• Burials with skeletonized individuals. Most of them are ossuaries. As in the case of mummies, the skeletonized remains may account from several individuals to hundreds like the cases of Uchova (San Miguel de Abona in the southern part of Tenerife) and Los Guanches (Tegueste, in the northeast), to mention only two of them (Rodríguez-Martín 1992a). • Caves with mummified and non-mummified individuals. Natural mummification cannot be excluded in this case because the rule was to bury mummies in specific places.

Types of Mummification Among the Guanches A long debate on the different types of mummification and, in general, on Guanche mummies has existed since the moment of the conquest up to the half of the twentieth century (and even later). In general, the corpses show two types of mummification:

Natural or Natural-Intentional Mummification It is difficult to distinguish between both cases. One of the few examples of natural mummification from Tenerife is a female mummy, aged 25–29 years, coming from

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the Museo Reverte at the School of Legal Medicine in Complutense University, Madrid. Most of the caves (more than 50 are known at present) in the island have the parameters to produce partial or complete mummification. Criado and Clavijo (1992) were the first to develop these studies.

Anthropogenic Mummification Most Guanche mummies are anthropogenic as it is proved by the presence of conserving, absorbing, and antiseptic materials and the typical external funerary bundle of animal leathers, mainly goat. The extraction of organs that according to some historical sources, like Abreu Galindo (1978) or, much later, Chil y Naranjo (1876), practiced the Guanche population has not been proved in the specimens that have been studied in Tenerife or other parts of the world. The most typical method is based in the external treatment of the corpse with dehydrating absorbent and antiseptic substances like pumice stone, heather bark, and pine powder and, sometimes, the use of a moss, Neckera intermedia, with antiseptic properties (Horne and Ireland 1991; Rodríguez-Martín and González-Antón 1994; Aufderheide et al. 1992a; Rodríguez-Martín 1998; Rodríguez-Martín and Martín-Oval 2018). At the same time, the corpse was exposed to the sun’s rays during the day and to smoke at night (Fig. 8). This treatment lasted around 15 days, and then the corpse was wrapped in animal leathers being the number of layers dependent of the individual’s social status (some specimens show up to 10 and 12 layers). The final step was the introduction of the mummy upon a funerary board (“chajasco”) inside “inaccessible” caves with dry environment that were closed with stones walls to avoid plundering and predators attack. According to the written sources, the mummification process was carried out by specialized persons that did not take contact with the rest of the population. The work was divided by sex according to the sex of the deceased.

Comparative Studies Between Mummified and Skeletal Populations Demography The demographic data of both populations show significant differences probably based in the type of diet consumed (Rodríguez-Martín 2000a; Rodríguez-Martín and Martín-Oval 2009, 2018). Mummies had a life expectancy at birth around 34.5 years and a crude mortality rate of 28 per thousand, while the skeletal population shows 30.9 years of life expectancy at birth with a crude mortality rate of 33 per thousand. It is important to mention that these parameters offer a better perspective in Tenerife than in European and Northern African human groups, in general, at that time (Rodríguez-Martín 2000a).

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Fig. 8 Guanche mummy showing bowels and genitalia in a good state of preservation. El Chorrillo cave. Southeastern Tenerife. 1117 AD–1453 AD. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros - Cabildo de Tenerife)

Diet The analysis of the skeletal content of trace elements (Sr, Ca, and Zn) and stable isotopes (C and N) demonstrated that Guanche diet was rich in meat and dairy products (50–55%), while vegetables showed a lesser ratio (30–35%). Marine resources (30%). Mummies show less than 30% of attrition and around 15% of periodontal disease (Langsjoen 1992; Rodríguez-Martín and Martín-Oval 2009). Caries is rather uncommon in both populations accounting around 5% in the skeletons (it is more common in the north as a consequence of the higher intake of sugar) (Langsjoen 1992) and is almost absent in mummies with a frequency of 1.5% as much.

Skeletal Pathology The high prevalence of conditions like postcranial trauma, degenerative joint disease, and osteochondritis dissecans demonstrates that physical stress was constant and intensive among the Guanches. A significant difference in the topographic distribution of these lesions exists: the lower limbs are more affected in the south what may be indicative of intense shepherding in comparison to the north. On the contrary, in the northern slope of the island, the lesions involve upper limbs and spine, and this points out to farming and gathering activities.

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Congenital malformations are present with very high prevalence in all the demarcations of pre-Hispanic Tenerife, especially transitional anomalies at the lumbar-sacral level (spina bifida occulta with prevalence between 25% and 40%, sacralization of L5 – 10–15% – and lumbarization of the first sacral segment – 8– 10%). This high prevalence points out to inbreeding by the isolation of the Guanches from the mainland during 11 centuries before the conquest (Fig. 9). Cranial trauma caused by violence (fights and ambushes) shows a very high frequency. Many cranial fractures have a more or less rounded shape corresponding to stone objects made for their use as mazes and others show an irregular contour suggesting stone shooting (Rodríguez-Martín 2000b). More than 80% of the cases show good healing. There is a significant difference between the prevalence in the south (10–18%) and the north (2–4%) indicating battles between both sides that were motivated by cattle robbery, invasion of the territory, etc. provoked by the inhabitants of the north in searching for resources in the south (Fig. 10). Other bone diseases like tumors, infections, and hematologic (anemia), endocrine, or metabolic disturbances had scarce incidence in the island (RodríguezMartín 1992a; Rodríguez-Martín and Martín-Oval 2009). In general, skeletal pathology, including trauma and congenital malformations, show no differences between mummified and skeletal populations. Fig. 9 Guanche mummy showing talipes equinovarus (clubfoot). Araya (Candelaria). Southeastern Tenerife. 1031 AD–1416 AD. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros - Cabildo de Tenerife)

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Fig. 10 Partially mummified Guanche head with a perimortem fracture on the right frontal. Barranco de Hendía (El Escobonal, Güímar). Southeastern Tenerife. Credit: The Museo Arqueológico de Tenerife (Organismo Autónomo de Museos y Centros - Cabildo de Tenerife)

Soft Tissue Pathology Lung anthracosis is the most common disease of the soft tissues producing fibrosis and respiratory distress (Brothwell et al. 1969; Aufderheide et al. 1992a). The cause of this condition is the practice of cooking inside or in the entrance of the dwelling caves with scarce ventilation.

Paleoparasitology Paleoparasitological studies have been carried out in historical and pre-Hispanic mummies, and multidisciplinary analyses were conducted in coprolites and abdominal-sacral sediments. These samples are usually rehydrated, and paleoparasitological research is conducted by spontaneous sedimentation and microscopic examination. aDNA is also extracted to elucidate and confirm parasite infection using different molecular targets. The results have demonstrated the presence of helminth eggs of Ascaris sp., Trichuris trichiura, Enterobius vermicularis, and hookworms (Mayo-Iñiguez et al. 2018).

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Conclusions Anthropogenic mummification was only applied to the elite of Guanche society. Natural and, probably, natural-intentional mummies have been also observed. Evisceration has not been proved among the Guanches, despite references like those of Abreu Galindo (1978) or Chil y Naranjo (1876). The treatment of the corpse was external and based in the use of dehydrating and absorbing material (pumice stone, heather bark, and pine powder) as well as antiseptics (Neckera intermedia) in some cases. In some instances, as the case of the Guanche mummy curated at the Museum of Archaeology and Anthropology (Cambridge University, England), subcutaneous incisions for stuffing with sand and other absorbing substances were performed too, although it was not the rule. The identical frequency of congenital malformations implies that mummies were not more affected by inbreeding than the rest of the island’s population. The elite carried out more or less similar daily activities than the skeletal population as indicated by the similar frequencies of physical activity markers in both groups. However, as dietary chemical reconstruction demonstrates, mummies show a higher consumption of proteins, and this is also corroborated by the lower frequency of metabolic stress markers; the lower prevalence of dental diseases related to plant consumption, especially caries; the higher life expectancy at birth; and the lower crude mortality rate observed in the group of mummified individuals. No pathological differences between populations are shown in the studies performed up to present.

References Abreu Galindo J (1978) Historia de la conquista de las siete islas de Canaria. Goya, Santa Cruz de Tenerife Arco C (1976) El enterramiento canario prehispánico. Anuario de Estudios Atlánticos 22:13–124 Arco C et al (2016) Un taller romano de púrpura en los límites de la Ecúmene. Lobos 1 (Fuerteventura-Islas Canarias). Primeros resultados. Museo Arqueológico de TenerifeInstituto Canario de Bioantropología (Monografías Canarias Arqueológica, 6). Santa Cruz de Tenerife Atoche P, Ramírez MA (2016) C14 references and cultural sequence in the Proto-history of Lanzarote (Canary Islands). Paper presented at Congreso de Cronometría para la Historia de la Península Ibérica, Barcelona 17–19 October, 2016 Aufderheide AC (1981) Soft tissue paleopathology – an emerging subspecialty. Hum Pathol 12(10):865–867 Aufderheide AC (2005) Chemical dietary reconstruction of Guanche NEC-2. ERES Arqueología/ Bioantropología 13:31–35 Aufderheide AC, Rodríguez-Martín C (1998) The Cambridge encyclopedia of human paleopathology. Cambridge University Press, New York Aufderheide AC, Rodríguez-Martín C, Estévez F, Torbenson M (1992a) Anatomic findings in studies of Guanche mummified human remains from Tenerife, Canary Islands. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992

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Aufderheide AC, Rodríguez-Martín C, Estévez F, Torbenson M (1992b) Chemical dietary reconstruction on Tenerife Guanche diet using skeletal trace elements content. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Barras de Aragón F (1929) Estudio de los cráneos antiguos de Canarias existentes en el Museo Antropológico Nacional. Actas y Memorias de la Sociedad Española de Antropología, Etnografía y Prehistoria VIII:3–153 Bontier P, Le Verrier J (2003) Le Canarien. Instituto de Estudios Canarios, San Cristóbal de La Laguna Brothwell D, Sandison AT, Gray PHK (1969) Human biological observations on a Guanche mummy with anthracosis. Am J Phys Anthropol 30:333–347 Chil-Naranjo G (1876) Estudios históricos, climatológicos y patológicos de las Islas Canarias. Miranda, Las Palmas de Gran Canaria Criado C, Clavijo M (1992) Características geográficas de los enterramientos con momias de la isla de Tenerife. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Crosby A (1986) Ecological imperialism: the biological expansion of Europe, 900–1900. Cambridge University Press, New York Diego-Cuscoy L (1971) De la inacabada historia del hombre canario. El Día 8 Diciembre 1971: 4 El-Najjar M, Aufderheide AC, Ortner DJ (1985) Preserved human remains of the southern region of the North American continent: report of autopsy findings. Hum Pathol 16(3):273–276 Espinosa A (1980) Historia de Nuestra Señora de Candelaria. Goya, Santa Cruz de Tenerife Gauert O, Grosskopf B (2018) The Göttingen Guanche mummy – recent research and findings. Paper presented at the extraordinary world congress on mummy studies, Instituto Canario de Bioantropología y Museo Arqueológico de Tenerife (OAMC-Cabildo de Tenerife), Santa Cruz de Tenerife, 21–25 May 2018 Hooton EA (1970 [1925]) The ancient inhabitants of the Canary Islands. Kraus Reprint Co, New York Horne P (1990) From the canaries, via Canada. Paleopathol Newsl 71:4 Horne P, Ireland R (1991) Moss and a Guanche mummy: an unusual utilization. Bryologist 94(4):407–408 Horne P, Lawson B, Aufderheide AC (1992) Examination of the Guanche mummy RED-1. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Kelley MA, Boom K (1992) Harris lines and environment: the early inhabitants of Tenerife. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Langsdorff GH (1813–14) Voyages and travels in various parts of the wolrd: during the years 1803, 1804, 1805, 1806 and 1807. H. Colburn, London Langsjoen OM (1992) Dental pathology among the prehispanic Guanches of the island of Tenerife. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Martín-Rodríguez A (2008) La necrópolis de la Hoya del Villano (La Montañeta, Garachico): estudio bioantropológico. Canarias Arqueológica (Arqueología / Bioantropología) 16:131–139 Mayo-Iñiguez A, Gijón-Botella H, López-Gijón, R et al (2018) Paleoparasitological and paleogenetic studies: a decade of collaborative project Brazil-Spain. Paper presented at the extraordinary world congress on mummy studies, Instituto Canario de Bioantropología y Museo Arqueológico de Tenerife (OAMC-Cabildo de Tenerife), Santa Cruz de Tenerife, 21–25 May 2018 Mora-Postigo C (1992) Momias guanches en el Museo Nacional de Etnología. Paper presented at Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Nichols T (1526) A pleasant description of the Fortunate Islands. Thomas East, London

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Rodríguez-Martín C (1992a) Una historia de las momias Guanches. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Rodríguez-Martín C (1992b) Osteopatología del habitante prehispánico de Tenerife. Islas Canarias. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Rodríguez-Martín C (1995) Biología esquelética de la población prehispánica de la comarca IsoraDaute. In: González Antón R et al (eds) La piedra Zanata. Museo Aqueológico de TenerifeOrganismo Autónomo de Museos y Centros-Cabildo de Tenerife, Santa Cruz de Tenerife Rodríguez-Martín C (1998) The Guanche mummies. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, disease and ancient cultures, 2nd edn. Cambridge University Press, Cambridge Rodríguez-Martín C (2000a) Estudio Demográfico de la Población Guanche de Tenerife. Chungará. Revista de Antropología Chilena 32(1):27–32 Rodríguez-Martín C (2000b) Cranial fractures in the Guanche population of Tenerife (500 BC – 1500 AD). In: González-Feria L (ed) Quality management in head injuries care. Critical approaches to health economics, patient selection and acute treatment. Euroacademy for Multidisciplinary Neurotraumatology – Servicio Canario de Salud, Santa Cruz de Tenerife Rodríguez-Martín C, González-Antón R (1994) Momias y momificación en las Islas Canarias prehispánicas. ERES-Serie de Arqueología 5:117–131 Rodríguez-Martín C, González-Antón R (2005) Colonización y asentamiento en islas por grupos humanos: aspectos biogeográficos y bioantropológicos. ERES-Serie de ArqueologíaBioantropología 11:115–133 Rodríguez-Martín C, Martín-Oval M (2009) Guanches. Una historia bioantropológica. Museo Arqueológico de Tenerife-Instituto Canario de Bioantropología (Monografías Canarias Arqueológica, 4), Santa Cruz de Tenerife Rodríguez-Martín C, Martín-Oval M (2014) La peste. El cuarto jinete. Epidemias históricas y su repercusión en Tenerife. Instituto Canario de Bioantropología (Organismo Autónomo de Museos y Centros del Cabildo de Tenerife), Santa Cruz de Tenerife Rodríguez-Martín C, Martín-Oval M (2018) Skeletal vs mummified populations in the protohistory of Tenerife. Paper presented at the extraordinary world congress on mummy studies, Instituto Canario de Bioantropología y Museo Arqueológico de Tenerife (OAMC-Cabildo de Tenerife), Santa Cruz de Tenerife, 21–25 May 2018 Rodríguez-Martín C, Guichón, R, Flegenheimer N et al (2005) The restitution of two Guanche mummies from Tenerife by Necochea, Argentina. J Biol Res. Paper presented at V World Congress on Mummy Studies LXXX:268–271 Rodríguez-Martín C, González-Antón R, Arco C (2009) La colonización humana de islas en la prehistoria. Un modelo teórico para el estudio de poblamientos insulares. Instituto de Estudios Canarios, San Cristóbal de La Laguna Sánchez-Pinto L, Ortega G (1992) Análisis del material localizado en la cavidad abdominal de dos momias guanches. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Tieszen LL, Matzner S, Buseman SK (1995) Dietary reconstruction based on stable isotopes (13C and 15C) of the Guanches of Prehispanic Tenerife, Canary Islands. Paper presented at I Congreso Internacional de Estudios sobre Momias, Museo Arqueológico y Etnográfico de Tenerife, Santa Cruz de Tenerife, 3–6 February, 1992 Torriani L (1980) Descripción e historia del Reino de las Islas Canarias, antes Afortunadas, con el parecer de sus fortificaciones. Goya, Santa Cruz de Tenerife Verneau R (1879) De la pluralité des races anciennes de l’Archipel Canarien. Bulletin et Mémoires de la Societé d’Anthropologie de Paris I:420–436 Viera-Clavijo J (1982) Noticias de la historia general de las Islas Canarias. Goya, Santa Cruz de Tenerife Zimmerman MR, Kelley MA (1982) Atlas of human paleopathology. Praeger, New York

Embalming in France (from Twelfth to Nineteenth Century): Principle and Development of Techniques

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Origin of Embalming in France . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Embalming Techniques: Modus Operandi . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Embalming Techniques from the Sixth to the Fifteenth Century . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Agnès Sorel (1450) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . King Louis XI and Charlotte de Savoie (1483) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Embalming Techniques from the Sixteenth to Nineteenth Century . . . . . . . . . . . . . . . . . . . . . . . . . . . . King Henri IV of France (1610) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Louis XIII of France (1643) and Louis XIV of France (1715) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Louis XVIII of France (1824) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions or Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Why and how does one embalm a dead body in France? Beyond the simple material aspects of conserving a corpse in the different time periods, the choice of the type of embalming, the products, the fragmentation, and the burial sites are important. In the present research, we are interested in both the practical (operational) and the symbolic (speculative) aspects of embalming, over a period of 14 centuries (from sixth to nineteenth century).

P. Charlier (*) Laboratoire Anthropologie, Archéologie, Biologie (LAAB), Université Paris-Saclay (UVSQ), UFR des Sciences de la Santé, Montigny-Le-Bretonneux, France Fondation Anthropologie, Archéologie, Biologie (FAAB) – Institut de France, Paris, France Département de la Recherche et de l’Enseignement, Musée du quai Branly – Jacques Chirac, Paris, France e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_55

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Keywords

Embalming · France · Fragmentation · Conservation · Palaeopathology · Chemistry · Reliquary · Religion

Introduction Why in the West, and specifically in France, were the bodies of the elite embalmed? Why were the corpses of the powerful, be they kings, queens, aristocrats, or ecclesiastics, preserved? How does the dead body become an object of power, a foil in the service of a family, a clan, a dynasty? What modus operandi allows this partial or complete conservation of the corpse? The aim of this chapter is to answer these questions using descriptions of the embalming techniques and the conservation of famous people and the subsequent examination of their corpses over the course of nine centuries.

The Origin of Embalming in France Embalming was begun with a practical goal in mind – to make the body of a dead individual presentable to the public at a distance from their burial place (Kantorowicz 1989). Examples are the pilgrim or crusader who fell in the Holy Land, a traveler or ambassador. In these circumstances, the first stage of embalming was performed by the butchers and cooks since they were used to opening animal carcasses, knew how to bleed meat, and had access to aromatics. Thus, they temporarily delayed the decomposition and putrefaction by removing the internal cadaveric blood deposits, eviscerating the trunk (and sometimes by removing the brain from the cranial cavity), and then filling the empty cavities with drying and odoriferous substances such as spices, salt, incense, and aromatic herbs. Extraculinary elements could be added to this “kitchen of the dead,” e.g., copper, mercury, and quicklime. Stuffed poultry or a calf’s heads are the same as an embalmed corpse, except that the deceased is presented publicly for an almost medico-legal purpose: to ensure that the one whose death is declared is indeed the “good” person. Recognition of the individual’s physical traits or peculiarities is essential (Charlier 2006a, b). When it is impossible to bring back the entire body for practical reasons such as when postmortem changes begin with the impossibility of ensuring the correct embalming technique, or for economic reasons such as to pay the embalmer and/ or for the raw materials, which were particularly expensive, then only “symbolic” elements can be repatriated to the widow or to the beneficiaries: the head, upper limbs, hands, or heart. This practice was called mos teutonicus, inspired, it is said, but without a definitive source, by the Teutonic knights who died in the exercise of their duties in the Baltic States, the Danube, and the Middle East. This dividing of the body was not intended to disperse the corpse, since it was brought back whole to its native land and then buried (Le Breton 1993).

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However, at this time, practice changed with the intention of distributing the parts of the dead body over a larger area. This was called the dilaceratio corporis, “the breaking up of the corpse.” Starting from a single deceased person, two or three burial sites could be consecrated. In 1199, Richard the Lionheart (Richard I of England) was mortally wounded in Châlus (Limousin, France). In the 10 days that his agony lasted, he had time to organize his funeral. His heart was to be entombed in Rouen cathedral (then English territory), his entrails in the church at Châlus (the viscera were left where the embalming took place), and his body in Fontevraud Abbey (dynastic necropolis of the Plantagenet family) (Charlier et al. 2013). Another example is Louis IX of France, whose body was dispersed along the route from his place of death in 1270 to the royal necropolis that he had established at the Basilica of Saint-Denis. His skin was buried in Tunis, his entrails in Monreale (near Palermo, in Sicily, in the kingdom held by his own brother Charles d’Anjou), his heart at Saint-Chapelle (in Paris, near the relics of the Passion of Christ; this point is still disputed by some specialists), and his body in Saint-Denis (Erlande-Brandenburg 1975). At the time of his canonization in 1297, his body was furthered dispersed (Fig. 1). This cadaveric dispersal left nothing to chance and indeed constituted a postmortem territorial marking, a way of carrying out a demarcation of the royal territory by the cadavers’ parts, such that they defined the ownership of the land. Louis IX, like his successors, will never stop renewing these ties of blood and soil, having his relatives (including his children) buried in religious establishments far from each other. Paraphrasing the French philosopher Blaise Pascal, one could say: “Pleasant territory bounded by corpses.” Later, since physicians decided to stick to antemortem care, being reluctant to practice embalming, it was mainly apothecaries and barber-surgeons who dealt with the dead body, the habit of offering one’s remains posthumously began to abate (Giesey 1987). In 1643 and 1715, Louis XIII and Louis XIV affirmed their support and gratitude toward the Society of Jesus by having their hearts placed in vermeil reliquaries in the church of Saint-Paul-Saint-Louis (Paris), as an offering to the General of the Jesuits. In a private setting, at his death (1788), the Comte de Buffon

Fig. 1 The reliquary of Saint Louis’ jaw bone in the cathedral Notre-Dame-deParis. (Credits: Philippe Charlier)

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offered his fixed brain to Madame Necker (his last love) and his embalmed heart to the National Museum of Natural History in Paris (Charlier 2006a).

Embalming Techniques: Modus Operandi The recent proliferation of osteo-archaeological studies has made it possible to put better into perspective the treatises on surgery (whether medieval or later) describing the modus operandi of embalming care. Several treatises on medieval surgery provide precise descriptions of the embalming process: Chirurgia magna (Great Surgery) by Guy de Chauliac in 1363 (Nicaise 1890), La Chirurgie (Surgery) by Master Henri de Mondeville composed from 1306 to 1320 (Nicaise 1893), and Chirurgia (Surgery) by Guglielmo da Saliceto (Guillaume de Salicet) in 1275. According to these various treatises, dating from the thirteenth and fourteenth century, but still widely used during the next two centuries, embalming consisted of seven independent stages: 1. 2. 3. 4. 5. 6. 7.

The opening of the cavities and withdrawal of the viscera. The preparation of the viscera. The incisions. The (internal) treatment of cavities and incisions. The external application of the liniment. The wrapping of the deceased. The deposit of the body.

The preparation of the deceased consisted of extracting the viscera and incising certain parts of the body to infiltrate odorous substances (traces of these longitudinal incisions are still sometimes visible in the form of a cut by definition without signs of cicatrization) along the diaphyses of long bones, particularly on the anterior and posterior sides of the femora. Henri de Mondeville then recommended approaching the heart by passing through the abdomen, i.e., without opening the thoracic cavity. This process was indeed of great practical ease, since it avoided not only sectioning the sternum and the ribs, but also the need to splint them during closure. Sometimes, a sternotomy was used, i.e., a more or less median vertical cut of the entire sternum, splinting being relatively easy when the cadaver was closed (Nicaise 1893). The paleopathological examination of the sternum must be particularly carefully done in the context of embalming, because besides the original sternotomy cut, there may be a deep laceration on the internal (visceral) face of the manubrium caused by the embalmer’s blade when the mediastinal fat was removed. In the same way, when pulling the aorta in front of the spine, from top to bottom with the index finger inserted in the vascular lumen, the other hand cuts the costal arteries one by one, creating anterior marks on the vertebral bodies. It was not until the seventeenth century that embalmers cut the costo-sternal cartilages, thus extracting not only the sternum but also the entire ribcage (thoracotomy), leaving free access to the entire

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thoracic cavity. This was described by Jacques Guillemeau (1612) in the seventeenth century and Pierre Dionis (1716) in the eighteenth century, respectively: “Various ways of embalming dead bodies. (. . .) First at the end of not showing so many incisions and openings, and the way of embalming is so misshapen and hideous to see, you will make a long incision, which will start at the neck, internal part above the chin, as high as you can, as above the knot in the throat. You will make it reign all along the sternum by its middle, continuing it on the belly, following the white line, going down to the pubic bone. (. . .) Such a section being thus conducted, it will be necessary to separate the leather and the muscles, which are located under the chest, and this joining the ribs, as well on the dextral side as on the sinister one. This makes the cartilages of the sternum and the ribs appear, which will be cut on the one hand and the other, then the sternum will be lifted by cutting the mediastinum, which separates the lungs partly right and left. (. . .) And should be noted that if the parents or friends want to keep the heart apart, it will be separated. And to do this, you will make an incision in the pericardium in which it is enclosed, then will be pulled and set apart” (Guillemeau 1612). “In order to be able to penetrate the chest, you have to be able to separate the muscular parts that cover it from the breastbone, and with a strong scalpel cut the cartilages that are at the end of each rib, both on the right and on the left side. Then separating the first bone of the sternum from the two ends of the clavicles, with which it is strongly attached, it is necessary to lift the entire sternum, as I said in my anatomy, in order to see the parts contained more easily” (Dionis 1716).

One last maneuver is often described in medieval embalming treatises: the opening of the skull with a saw in order to remove the brain. In fact, the skulls of embalmed subjects frequently show signs of the technique with an opening and incisions, generally of small size, required by the reflection of the scalp. Archaeological cases have shown the marks on the cranium (see Louis XI and Charlotte de Savoie cases, just after). In medieval times, the reality was more complex than this, as there are different levels of preparation of the body depending, not only on social status, but also on the time that the corpse had to be kept and presented to the people or the family: “If the corpses must be kept for more than four nights and there is a special privilege of the Roman Church, we will incise the anterior wall of the belly from the middle of the chest to the pubis if it is a question of a man, in women, one will incise of the fork or opening of the stomach while descending following the form of an inverted shield until the two sides or iliums. Then the entire wall between the two incisions will be overturned on the sexual parts and all the viscera will be extracted up to the anus” (Nicaise 1893). Surfaces were covered and cavities were filled with a powder of myrrh, mummy (mumie, generally Egyptian mummies reduced to dust, or possibly a fruit with the smell of camphor and turpentine), aloes, and other spices. The abdominal and thoracic cavities were filled with fragrant herbs (camomile, pennyroyal, mint, costmary chrysanthemum, lemon balm, etc.), and the body was stitched up until it returned to its “usual and decent shape.” Balms could then be applied to the integuments and hair, or even bandages, Egyptian style, limb by limb. Guy de Chauliac summed this up thus laconically as “And another kind are prepared the

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dead bodies, which suddenly one opens them by the belly, and one pulls out all the entrails. And let the belly be filled with the aforesaid powder, and a large quantity of salt and cumin. And either sewn and wrapped as said is” (Nicaise 1890). Sometimes, one of the liniments used was of metallic origin, mercury (or quicksilver), the use of which is corroborated by medieval surgeons and technicians. For Henri de Mondeville, “if you have to keep your body with your face uncovered, you will not sew your mouth or close your nostrils, but you will put quicksilver in each of them, about 6 ???; immediately afterwards we will introduce silk so that it does not appear outside the nostrils” (Nicaise 1893). Quicksilver was also used by the same author, but in half the quantity “for the bodies of the rich which must necessarily be kept and prepared,” without specifying the exact place where it was deposited in the body (Nicaise 1893). For Guy de Chauliac also, in embalming of the external type without evisceration, quicksilver was used to fill the nostrils, the mouth, and the ears “because with that one keeps that his brain does not flow” (Nicaise 1890). For embalming with evisceration, he clearly did not use mercury. Archaeology provides further evidence of the use of this metal in certain cases of bodily preservation, notably for aristocratic burials. When, shortly after 1860 in the cathedral of Rouen, the tomb of John of Lancaster, Duke of Bedford (who died in 1435, the same year Joan of Arc was burnt), was discovered and examined, and the black coloration of the bones was immediately linked to body preservation practices (Fig. 2): “The head (. . .) was wrapped in a crust of dough that was a good two centimetres thick. In this paste were perfectly visible black hair and a little rolled up. (. . .) In this embalming must have entered a lot of mercury, because drops of this liquid filled the cavities of the paste. Such a quantity escaped that small streams of mercury formed at the bottom of the coffin.” An analysis was performed on this paste, the results of which were as follows: metallic mercury – 11.25%, watersoluble matter which does not contain chlorine or sulfuric acid – 11.33%, balsamic resin having the character of benzoin – 8.20%, insoluble organic matter, nitrogen – 59.00%, water and loss – 10.22%, (Total – 100%) (Charlier et al. 2016). Fig. 2 General view of the modern reliquary of the remains of the Duke of Bedford. (Credits: Philippe Charlier)

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It was noted above that Henri de Mondeville described the use of aromatics deposited in the abdominal cavity: powder of myrrh, mum, aloe, other spices, chamomile, pennyroyal, mint, costmary chrysanthemum, lemon balm, etc. The analysis of historical texts confirms the use of other plants, salt, and bean flour. Wine was also used, as mentioned in The Death of Roland (Charlier 2006a). Later, works were written on the use of plants and seeds in embalming, such as the Treaty of Embalming according to the Ancients and Moderns, with a description of some balsamic and odoriferous compositions by L. Pénicher in Paris (Pénicher 1699). Not only are openings made to extract the viscera (sometimes brain, or heart and often the intestines), but holes are also made, that is, notches carried out in the flesh so that the blood was evacuated and replaced by aromatics slipped into the interstices. These berries and plants also filled the natural cavities: the mouth, nose, ears, anus, and vagina. The tongue and testicles were sometimes removed (Pénicher 1699).

Embalming Techniques from the Sixth to the Fifteenth Century The cases of the Merovingian queen Arégonde (died circa 574–580) (Gallien et al. 2009) and the early Christian bodies of Saint-Victor de Marseille were not, for certain, embalmed. It may be that some plants or odoriferous substances were placed on the remains without being dispersed within a previously opened corpse. King Charles the Bald is the first Western individual for whom the definitive textual evidence mentions embalming. He died in Savoy in 877and underwent evisceration (corpus more regio curatum). The corpse was then preserved using wine and aromatics. Unfortunately, the technique was not efficient, and the rotting corpse was buried in Nantua (200 kilometers from his place of death), and secondarily transported to the definitive place of inhumation some years later, according to the Annales Bertiniani (877). In 1040, Foulque Nerra III, the count of Anjou, known for his atrocities, died in Lorraine after returning from one of many pilgrimages to the Holy Land. The body was opened by a physician. The entrails were left near the cathedral of Metz, while the rest of the corpse, coated externally with aromatics, was buried in Beaulieu-lèsLoches near a relic of the Holy Sepulchre (Charlier et al. 2009a). The recent reopening of the grave of Philippe I of France (died 1108) found a corpse covered with a mass of foliage and branches (mint, walnut, angelica stems, and walnut leaves), the head resting on a pillow of iridaceous leaves, a bouquet thrown over the body, and clothes made of wool, hemp, velvet, linen, and silk. Unfortunately, it was not possible to specify whether the dead body had undergone any anatomical preparation, since there was significant fragmentation of the skeletal elements coupled with the noninvasive nature of the archaeological inspection (Georges 2006). More recently, the interdisciplinary study of the embalmed heart of Richard I of England revealed an extremely complex process associated with myrtle, daisy, mint, pine, mercury, copper, quicklime, birch pitch, and frankincense (Fig. 3). The latter

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Fig. 3 General view of the modern reliquary of the remains of Richard the Lionheart. (Credits: Philippe Charlier)

aromatic, generally absent from the embalming process during this period, must be considered as an exceptional product intended to promote the apotheosis of the English sovereign particularly decried on his return from the Third Crusade. Indeed, the Bishop of Rochester did say, soon after his death, that Richard had not yet reached Heaven but still had to atone for his crimes during 33 years in Purgatory (Charlier et al. 2013). Particularly expensive spices were used on Jean I Duke of Berry in 1416. The internal embalming was based on bean flour, olibanum, myrrh, frankincense, mastic, mummy, “militillas” (a nonidentified product), Armenian bowl, dragon’s blood, cypress nut, fragrant herbs, mercury, camphor, musk, rosin (Greek pitch), black pitch, and cotton. The corpse was wrapped in a canvas of Reims, tied with ropes, and placed in a lead chest without a lid which, in turn, was placed in a wooden coffin with iron rings. Three burials were consecrated, comparable to royal use: the body in a tomb in Bourges Cathedral, the heart in a grave in Saint-Denis Basilica, and the entrails in a tomb in Saint-André-des-Arts church (Paris) (Lehoux 1956). In 1435, John of Lancaster, Duke of Bedford, died in Rouen. The recent study of his remains revealed an embalming (internal and/or external) based on mercury, myrtle, mint, frankincense, lime, and copper. Historical records attest that the body was placed in a lead container inside an oak coffin (Charlier et al. 2016). Other bodies have been described as covered with mercury or “drowned” in a coffin filled with mercury, for example, King Charles VII of France (died 1461) and Queen Anne of Brittany (died 1514), at least as attested during the revolutionary desecration of their bodies and profanation of their graves in the Royal Basilica of Saint-Denis (Dom Druon 1793). In 1450, Agnès Sorel, official favorite of King Charles VII, died in Jumièges (Normandy). Her embalming was performed on-site. Some of the viscera were buried in the abbey, while the embalmed body was buried 500 kilometers away in Loches (next to the River Loire). Her interdisciplinary study revealed the use of

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white mulberry fruits and seeds, rhizomes (ginger?), and grey “maniguette” pepper from West Africa. This embalming is worth considering in more detail (Charlier et al. 2009b).

Agne`s Sorel (1450) The Lady of Beauty, official mistress of King Charles VII, died on February 3, 1450, in Jumièges in Normandy, from a “belly flow.” A complete paleopathological examination of her remains highlighted the exact cause of death (Fig. 4): acute mercury poisoning from a de-worming (antiascarid) treatment. However, it is not known whether this overdose was accidental (therapeutic error) or intentional (murder by poisoning). The demonstration of this high concentration of mercury in the proximal ends of the integuments (for lack of prolonged survival of the subject) confirmed that it was not a question of metallic body preservation care, especially since the skull showed no sign of sawing or perforation of the toothed blades and therefore no sign of excerebration. On the other hand, we know that Agnès Sorel’s heart was left in Jumièges, as the contemporary historian Jean Chartier (Chartier 1858) recounts: “Which has since been opened and her heart carried and buried in the abbey, why had made very great donations.” The black funerary slab of this heart burial is still visible in the abbey, after the tomb was looted and the funeral slab reused as a... cutting table for a butcher from Rouen! More poetically, Charles VII would have said, burying his lover’s heart there “It is a little of myself that I am abandoning you. . . .” In all cases, there was a need for care in conservation of the body, it having to be transported from Jumièges (Normandy) to Loches (Indre-etLoire). Admittedly, it was in February, but the trip was long (nearly 10–15 days due to the many festivals and ceremonies punctuating the transport of the remains), and decomposition would take its inevitable course. It is probable that the entrails were extracted along with the heart, but we have no evidence of this. Fig. 4 General view of the conserved mummified remains of Agnès Sorel. (Credits: Philippe Charlier)

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The multidisciplinary study carried out on the remains of the Lady of Beauty has made it possible to find traces of her embalming method. Indeed, very many seeds and fragments of small plants were isolated from the funeral urn, mixed with the bones. We have seen above that a common practice in medieval times was to extract all of the viscera (lungs, intestines, kidneys, etc.) at the same time as the heart, and to fill the abdominal cavity and/or the thorax with berries, seeds, and aromatics. These are the ones that were found in the funeral urn, testifying to the practices of the preservation of the body. This is how “maniguette” pepper from West Africa and fragments of rhizomes and seeds of white mulberry from China have been identified. However, the latter’s cultivation for silkworm breeding was not introduced into France until the end of the fifteenth century. This choice is not by chance, and it is not from flowers being placed on the body of the deceased some time before the coffin was closed. In fact, these are the remains of the products of her embalming, carried out according to the prescriptions of the time, following the precepts of the surgeons Guy de Chauliac and Henri de Mondeville (Charlier et al. 2009b). On the other hand, it is certain that only a few elements of Agnès Sorel’s embalming have been identified, and that the complete list of plants and herbs remains unknown. The great variety of botanical species used in embalming in the middle of the fifteenth century can be obtained from the funeral accounts of Philippe le Bon, Duke of Burgundy, who died in 1467. The substances listed for the preservation of his corpse (internal and external embalming) were black pitch, resin, rosin, frankincense, mastic, styrax calamite, arabic gum, tragacanth, aloe, myrrh, Gallia muscata (medicinal preparation), Alipta muscata (officinal preparation), musk chervil, cypress nuts, turpentine, canvas, pepper, salt, camphor, cumin, bowl of Armenia, sigillate earth, henna, pomegranate bark, galbanum, aloe wood, alum, zedoa (spread on the body after embalming and bandaging), and artificial balm (on the face). The corpse was then placed in an iron and then a lead coffin (Baveye 2011).

King Louis XI and Charlotte de Savoie (1483) At the request of paleo-anthropologist Patrice Georges, access was granted to examine the cranial remains of Louis XI in order to carry out a complete paleopathological study (Fig. 5). The inventory of bone remains will not be listed here, but rather the details of the lesions related to the embalming observed during this examination. Using modern forensic techniques it was possible to work out the procedures used to access the brain. Of note, the cranial vault was not cut in a horizontal plane but in an oblique backward, downward, and to the right manner. The embalmer probably wanted to use the corpses’ rigor mortis as a “third hand,” the head of the deceased being turned, in this case, to the left. Rigor mortis, by paralyzing the muscles of the neck and immobilizing the head in relation to the trunk, facilitated the cutting of the cranium by giving a better grip to the saw and avoiding the forward and backward movements of this extremity of the corpse. Rigor mortis typically ceases within 48–72 h following death, indicating the timing of opening of the skull and therefore the embalming (Charlier and Georges 2009).

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Fig. 5 Details of saw lesions on the skull vault of King Louis XI. (Credits: Philippe Charlier)

Numerous traces of cuts were visible on the surface of the exocranium, on all sides of the dome, suggesting some technical difficulties and several attempts. There were three striations on the left parietal bone, two old defects (chips) to the left hemifrontal bone, a splinter and striae on the right hemi-frontal bone, a right temporal bone peri-mortem fracture, and finally, a stria on the right occipital bone. The embalmer, probably inexperienced, therefore, began to saw the skull at the left parietal level, after having previously separated the temporal muscles from the bone. Nevertheless, the aponeuroses, obviously still in place at the start of the sawing procedure, hampered it to begin with. It required three attempts, but maybe the embalmer was not helped by the position of the head. He did not start on this side so perhaps he was left-handed. In all the attempts, the last few mid-occipital centimeters of bone were not sawn through. On the contrary, the practitioner leaned on the forehead three times in a row (twice on the left, and once on the right) to rotate the skull backward, finally cracking off the uncut occipital segment (Charlier and Georges 2009). Another example of this kind of practice (use of rigor mortis as a third hand) has been noted in the skeleton of vault F of the Basilica of Notre-Dame de Cléry. These were fragments of a skull from an immature individual (less than 16 years old) that had been subjected to a postmortem craniotomy, and the exact procedure could be precisely reconstructed. Unlike normal practice, the sawing plane was strictly vertical to the anatomical plane. The corpse was therefore placed in the dorsal decubitus position, head turned to the left. The right temporal and the right parietal bones were therefore uppermost and available for cutting. Rigor mortis was probably still present and holding the head still. Locked in this position, the skull could only be approached by the right temporal and the right parietal bones. The saw was used in the horizontal plane (being more convenient for the practitioner), but it was a horizontal plane relative to the autopsy table and not with respect to the skull. On the other hand, the absence of maceration of the bone can be explained by the young age of the deceased. The extraction of the brain was facilitated by dismantling the cranium via the still widely open sutures. Since rigor mortis ceases naturally after

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Fig. 6 Right lateral view of the sawed skull of Queen Charlotte de Savoie. (Credits: Philippe Charlier)

48–72 h from the time of death, it implies that the skull was sectioned within 72 h of his death (Charlier and Georges 2009). Permission to gain access to the cranial remains of Queen Charlotte de Savoie in order to carry out a complete paleopathological study was also obtained as part of the same scientific study (Fig. 6). Here too, the skull was not cut in a horizontal plane but slightly obliquely from above, forward, and to the right. Again, evidence of the cuts was still visible on the surface of the exocranium. There were several striations on the right temporal bone (corresponding to the cutting of the temporal muscle), a score mark and a chip at the level of the right frontal bone (corresponding to the point of impact of the lever during the separation of the cranial vault from the rest of the skull), a stria at the level of the left hemi-frontal bone, a stria at the level of the left parietal bone, and, finally, a left occipital peri-mortem fracture when the dome was removed from the rest of the skull (Charlier and Georges 2009).

Embalming Techniques from the Sixteenth to Nineteenth Century The almost complete skeletonization of the remains of Diane de Poitiers (died 1566) was identified by a forensic process in a mass grave after her exhumation by revolutionaries in 1795. The only embalming product found was bituminous residues (Fig. 7). The author of the text on the body’s conservation must have been Ambroise Paré, the appointed surgeon, since one of the duties of a surgeon was opening the dead body as a part of continuing care of the patient, logical although not therapeutic (Charlier et al. 2009c). Since the second part of the sixteenth century, embalming was particularly well documented because some French surgeons did not hesitate to use famous corpses for self publicity. For example, Jacques Guillemeau published in his Les Oeuvres de chirurgie (Works of surgery) in 1598 his autopsy reports of Charles IX (died 1574), Henri III (died 1589), and Henri IV (died 1610). Meanwhile, chroniclers, servants, and chaplains also gave detailed accounts of this.

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Fig. 7 CT-scan 3D-rendering of the whole conserved skeleton of Diane de Poitiers. (Credits: Philippe Charlier)

King Henri IV of France (1610) King Henri IV of France was assassinated on May 14, 1610, by François Ravaillac with two stab wounds to the heart. He was transported to the Louvre Palace (Paris) that day, dying shortly after. His body was autopsied and embalmed on May 15, 1610, and then buried on June 30. During the desecration of the Basilica of Saint Denis by revolutionaries, his remains were removed. On Saturday, the 12th of October 1793, his coffin was opened where it was noted that “his body was found well preserved with well recognisable facial features, he remained in the passage of the low chapels wrapped in his shroud also well preserved. Everyone had the freedom to see it until Monday morning, the 14th October. We brought him into the choir at the edge of the steps of the sanctuary, where he remained until two hours afternoon. We laid him in the cemetery (known as Valois) in a large pit in the bottom right of the north” (Dom Druon 1793). His head (Fig. 8), separated from the body during this time and passed from hand to hand, was definitively identified by a group of interdisciplinary researchers (Charlier et al. 2010, 2014a). Although no official embalming report has been kept for Henri IV, several texts document the funerary practice, including the final phase where strips of material are used completely to wrap the body. In his Les Oeuvres de chirurgie (1598), Jacques Guillemeau outlines this practice during the preparation and embalming of King Charles IX’s body. He said that he performed the same techniques when preparing Henri III and Henri IV’s corpses for burial. During the final stage of embalming, Guillemeau describes two different methods, one for the body and the other for the head. Initially, the body is “ put in a shroud and wrapped in oilcloth [. . .] twisted to either side with rope untied enough, and above, all will be wrapped again with a good and strong oilcloth and firmly tied with rope and wrapped, to be placed in a lead coffin [. . .] the said coffin will be exactly welded and put in another one that will be made of good wood” (Guillemeau 1649). Second, the head, like the whole body, was bandaged with clean strips soaked in a melted mixture of liquor made of fresh wax, dust iris flowers, turpentine of Venice, Elemi gum, benzoin and styrax,

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Fig. 8 Right lateral view of the mummified head of King Henri IV. (Credits: Philippe Charlier)

natterjack, myrrh, aloes, wormwood oil, and balsam of Peru. So that the bandage bonded well, it had to be placed in a coal basin filled with the ingredients listed to soften them. The bandaged head was provided with a tissue headdress. The body was then covered with oilcloth and firmly tied with a large cord and wrapped in a shroud (Guillemeau 1649). In his Chirurgia, Pierre Pigray (1610) offers little detail about the embalming and bandaging. He only tells us that after having sprinkled the body with a powdery mixture, “we bandage the legs to contain it, and we will wrap all with a well oilcloth and sprinkled said powders.” The use of bandaging for embalming is also attested by engravings printed during and some years after the exhumation, clearly showing the bandages around the cadaver; the one around the detached head has the face exposed when the royal tombs at Saint-Denis were desecrated on October 23, 1793 (Augias et al. 2017). In addition to these written sources, the presence of embalming bandages is confirmed by the analysis of the head of Henri IV through different biomedical techniques, a medical scan performed at the University Hospital of the Pitié Salpêtrière in Paris, as well as observations on the head itself, including the help of binocular microscopic observations. These allow movement of the soft parts and

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distinguish the dynamic in the actions of the embalmer guided by the head position in the struggle to reopen the jaw during rigor mortis, from the subsequent filling of the orifices (nostrils, mouth, etc.). Initially, forensic analyses have highlighted four main areas of folds of the skin: these areas are the neck (anterior and posterior part), nose, and right ear. Indeed, several crushing lesions by bandages are observed on the nape and upper anterior surface of the neck, just below the root of the chin. The nevoid lesion on the right nostril is crushed and transversally elongated by two contiguous bandages. There is an anterior fold of 2 cm of the pinna with respect to the right external auditory meatus caused by an overlapping bandage directed from back to front and left. In addition, the soft tissue mass on the posterior surface of the skull looks like superimposed folds at the nape of the neck. Skin folds are visible at the top of the skull between the vertex and the lambda; the convolutions at the back of the skull indicate that they were created starting at the nape of the neck and finished at the top of the skull. The orientation of these folds suggests the bandage placement was directed from left to right on the posterior surface, and right to left on the anterior side (Fig. 9) (Augias et al. 2017). Several zones of pulled-out skin at the top level of the parietal and temporal bones, and the left mandible, are also visible which may correspond to adhesion surfaces of the waxed bandages mentioned by Guillemeau (Guillemeau 1649). The open position of the mouth and these pulled-out areas are then the result of the removal of these waxed bandages during the desecration of the tomb, when the Fig. 9 Restitution of the position and movements of the strips during the embalming of King Henri IV head. (Credits: Philippe Froesch, Anaïs Augias & Philippe Charlier)

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king’s face was uncovered to extract the teeth and dentures with gold dental work, as stated in the Revolutionary texts (Augias et al. 2017). Such forensic analyses have highlighted the application of the embalming bandages on Henri IV’s mummy and that they were directed from the bottom up and to the left. Crosschecking the historical and technological data allowed reconstruction of the embalmer’s actions in one of the final uses of this practice reserved for the elite. The involvement of forensic and anthropological perspectives helps interpret postmortem changes (anatomical displacements) and traces of human actions (Augias et al. 2017). In some cases, archaeo-anthropology allows textual and scientific data to be compared, for example, in the case of Anne d’Alègre, Countess of Laval (Brittany). She died in 1619 and underwent complete evisceration and craniotomy. The viscera and brain were replaced by a compact light brown powdery odoriferous material: a mixture of wood chips, segments of stems and rootlets, floral calyces, seeds, and some leaves (90% thyme, plus oregano and juniper). The cavities of the calvarium and rib cage were also filled. The body was then taken in a canvas shroud held in place with hemp cords, placed in an anthropomorphic lead coffin enclosed in a wooden coffin on which the lead “reliquary” of the heart was placed (Colleter et al. 2011). Twenty years later, when a young English nobleman, Thomas Craven, died of the plague, his body was placed in a lead coffin, a bunch of dried plants with long stems placed between his legs, and the inside of his corpse was “stuffed” with a substance composed of Artemisia sp., Lamiaceae, and Apiaceae (Hadjouis and Corbineau 2009). The latter case is indicative of the danger of embalming a dead body; some practitioners did not hesitate to open dead bodies in the context of a particularly contagious (and dangerous) epidemic disease, while others, as with the smallpox of Louis XV of France (died 1774) simply refused to perform this treatment because of the extreme danger, as reported in a letter from Doctor Lemonnier to Dean Lethieullier: “Mr. Dean, Mr. Bordeu, our colleague, spoke to me yesterday about the custom that I was ignorant of calling the Dean of the Faculty and an assistant to the opening of the body of kings of France. We would not have failed to maintain the rights of the Faculty, if the sad ceremony had taken place; but given the kind of illness from which His Majesty died, we will simply bury the body in a waxed taffeta well garnished with aromatic powders; we will finish filling the coffin with lead. This is how the king’s apothecary has just told me that it will be practiced this evening at five o’clock. “ I have the honour, etc. Lemonnier” (BIUM 1764–1777). Some surgeons asked for a risk premium. Because embalming involved decomposition and putrefaction which, in themselves, were considered a great source of miasma, then they correctly understood that working with dead bodies put one’s life in danger. Funeral financial accounts confirm that, far from being a thankless task, embalming the bodies of the well-to-do was a particularly profitable business. A final type of embalming involves those who were recognized as holy. It constitutes a field of research on its own. Our inter-disciplinary team examined the mummified heart of Blessed Anne-Madeleine Remuzat (1696–1730). This organ was studied in the context of an official canonization process. The analysis was

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related to important aspects of the early history of anatomy in Europe – that of “Holy Autopsies” – and to the relationship between anatomical investigations, Catholic theology, and religious or medical customs. According to the anatomical, genetic, toxicological, and palynological analyses, it was shown that this heart had not been naturally (i.e., miraculously) conserved but embalmed using myrtle, honey, and lime. After its surgical opening, and following extraction of the bloody fluids, all four cavities were filled with botanical elements, then sewn up with linen fibers. Moreover, right ventricular dilatation was diagnosed that may represent a posttuberculosis condition and may have played a role in the cause of her death (Charlier et al. 2014b).

Louis XIII of France (1643) and Louis XIV of France (1715) Since the desecration of the royal burials in the Basilica of Saint-Denis in October 1793, and of the vaults of the Notre-Dame-de-Paris Cathedral, all that remains of the royal corpses of Louis XIII and Louis XIV are the hearts (Figs. 10 and 11). These are now deposited in the Chapelle des Princes, within the crypt of the Basilica of SaintDenis (but, initially placed in the church of Saint-Paul-Saint-Louis, in Paris, following a postmortem donation of these very symbolic organs to the general of the Jesuits, for services rendered). As a source of information, therefore, we only have the autopsy reports, by order of organ appearance, on the general organization of the practice of opening the bodies. On the other hand, no details are known concerning the secondary embalming time itself – a posteriori conservation of the remains. Fig. 10 General view of the mummified heart of King Louis XIII. (Credits: Philippe Charlier)

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Fig. 11 The original copper plaque of the King’s Louis XIV heart reliquary. (Credits: Philippe Charlier) “The following day, at the sixth hour of the morning, the body of the deceased king was opened in the presence of the serene prince Monseigneur de Nemours, marshal-general of the camps, of M. de Vitry, of M. de Souvré, first chamberlain, of the ordinary chamberlains, premieres of the king and queen and ordinary physicians and surgeons on both sides. We found many purulent ulcers, sanieux, tabescens (meaning unknown), located in different places, in the mesocolon, in the small intestines. There was one at the end of the colon, and which had gnawed and perforated the intestine, whence a large purulent collection coming from the putrefied glands and vessels of the mesocolon had accumulated in the lower abdomen and could have been fill three demi-setiers, measure of Paris. In the right kidney, an abscess was found, but small, and which must have had no influence on the disease. At the bottom of the stomach was a somewhat larger abscess, and several other very small ones, brown, sooty, greenish, blackish, analogous to those observed throughout the intestinal canal. The gall vesicle, adhering to the liver, was almost empty. The liver was almost parched and shriveled, pressed against the abdominal walls and crushing into lumps. The lobe of the left lung was adhered to the pleura by a large and deep cavern, full of pus. This is what we scrupulously observed the dean of the Faculty of Medicine, Michel Delavigne and

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René Moreau, doctor and royal professor, who both, during the space of twenty-six days, with the aforementioned doctors gave their care to the very Christian king, called from Paris as consultants on Monday April 20 of the year of the Lord 1643” (BIUM 1636–1652).

This means that the order of Louis XIII’s organ dissection, then precise examination, was as follows: the intestines, kidneys, stomach, gallbladder, liver, and lungs. It is astonishing that no mention was given of the heart (maybe because it was not examined carefully but extracted for a further embalming process) nor of the brain (maybe because no craniotomy was performed). “Today, the second of September in the year 1715, we assembled at 9 a.m. in the Castle of Versailles to open the body of the king, where we found the following. On the outside, the entire left side looked gangrenous from the tip of the foot to the top of the head. The epidermis was usually removed from the whole body on both sides, the right side was gangrenous in several places, but much less than the left, and the belly appeared extremely puffy. When the lower abdomen opened, the intestines were found to be altered with some signs of inflammation; mainly those which were located on the left side, and the large intestines, prodigiously dilated. The kidneys were pretty much in their natural state. Only in the left was found a small stone of the same size as the one he passed in urine several times during his life, with no sign of pain. The liver, spleen, stomach, bladder were absolutely healthy and in their natural state, both inside and out. Upon opening the chest, we found the lungs healthy, as well as the heart, the ends of the vessels and some valves of which were bony; but all the muscles of the throat were gangrenous. When the head was opened, all the dura mater was found adhering to the skull, and the pia mater had two or three purulent spots along the scythe to the rest of the brain was in its natural state both at -in and out. The left thigh, in the interior, was found gangrenous, as well as the muscles of the lower abdomen and this gangrene went up to the throat. The blood and lymph were in complete dissolution, universally in the vessels.”

This means that the order of Louis XIV’s organ dissection, then precise examination, was as follows: the intestines, kidneys, liver, spleen, stomach, bladder, lungs, heart, and brain. Precise analysis of altered limbs was also carried out (Charlier 2006a).

Louis XVIII of France (1824) Although we have almost all the autopsy records of French monarchs, we rarely have the embalmer’s accounts. Particularly interesting is therefore the report on the embalming of the body of Louis XVIII (1824) carried out just after the autopsy. Pharmacist Labarraque first sprayed the corpse with a sodium solution to rid it of the horrible smell of putrefaction mainly due to gangrene (Charlier 2006). Other processes aimed at body preservation (including the heart which was separately conserved) were then used: “Today the 17th September 1824, immediately after the opening of the body of the late King Louis XVIII, and in accordance with the instructions given to us by the Marquis de Brézé,

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Grand Master of Ceremonies of France, we, the undersigned, proceeded to the embalming in the following manner. The King’s heart, after having been opened, emptied, washed and macerated for four or five hours in an alcoholic solution of deuto-chloride of mercury or corrosive sublimate, and having been filled and surrounded by selected aromatics, was enclosed in a lead box, bearing an inscription indicating the precious object it contains. The viscera of the three great cavities of the body, after having been similarly opened, incised, washed and macerated for six hours in the aforesaid solution, were penetrated, filled and surrounded with aromatics, and enclosed in a lead barrel, bearing an inscription indicative of the parts it contains. The entire surface of the body and that of the large cavities were washed successively with a solution of sodium oxide chloride and with an alcoholic solution of deuto-chloride of mercury. The fleshy parts, both of the trunk and of the limbs, have been cut widely and deeply. They were then washed with the above solutions. The surfaces of the body, those of its cavities and incisions, have been coated with alcohol varnish several times. All the cavities were filled with powders made up of aromatic species and various resinous substances. These cavities were closed by the application of their walls, supported by means of numerous sutures. The limbs, pelvis, stomach, chest, neck and head were successively surrounded by several methodically applied bands. The entire surface of the body thus enveloped was covered with several coats of varnish. On this varnish, bands of gummed diachilon were applied. On the strips of diachilon, other strips of glazed taffeta have been applied. Finally, a last layer of bands has been applied to the glazed taffeta. When the embalming was completed, the head of the late King was covered with a cap, his body with a shirt, his arms and chest with a sleeved white silk vest. The whole body of a Baptist shroud. It was in this state that the body of the King was handed over to M. de Brézé to be placed in the coffin which is to enclose his mortal remains in Saint-Denis. Signed: Portal, Alibert, Dupuytren, Fabre, Distel, Thévenot, Portal pour Ribes, Auvity, Breschet, Marx, Moreau, Bardenat, Vesques, Dalmas, Delagenevraye” (Charlier 2006b).

It should be noted that this is merely an improvement on preexisting techniques: evisceration, washing of the cavities, filling with aromatics, and mercurial imbibition. The development of chemistry is very clearly visible, with the use of many products that were once unimaginable; embalmers took advantage of the latest technical innovations, in large part due to the growing involvement of pharmacists and apothecaries in these macabre activities (Charlier 2006b). Finally, it would be theoretically possible to judge the effectiveness of these conservation treatments: if the body has not been abused by revolutionary actions or wars, it is therefore still present in the tomb, and therefore ready to be examined by paleopathologists.

Conclusions or Perspectives This juxtaposition and succession of both practical and theoretical cases clearly shows the development of the use of embalming. Initially, it was intended to keep the corpse from smelling while the funeral was organized. It was performed by cooks,

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who were used to opening carcasses and stuffing the cold meat to smell good. Subsequently, embalming became a more symbolic practice. The body was divided to cover a wider area postmortem, thus territorial demarcation was made by separate burial places for different parts of the body. At the same time, creating a pleasant smell was important (close to the smell of holiness) in order to exalt the memory of the deceased and facilitate their apotheosis (entry into Paradise). The technique was professionalized at the same time. Since the preparation of the body was always done in the same manner, it gradually moved away from being performed by barberssurgeons, and became the specialty of apothecaries and chemists. Acknowledgments I am grateful to Professor Simon T. Donell who generously proofread and edited this chapter.

References Annales Bertiniani Auctore Hincmaro, Pars Tertia Auctore Hincmaro, Remensi Archiepiscopo. a. 877, MGH, Scriptores, VI, 137 Augias A, Huynh-Charlier I, Froesch P, McMullan-Weitzel B, Charlier P (2017) Detection of an embalming strip on a French royal mummified head (Henri IV, 1610): virtual reconstruction of a technical gesture. In: Charlier P (ed) Actes du 6ème Colloque International de Pathographie (mai 2015). De Boccard, Paris, pp 7–16 Baveye L (2011) La mort de Philippe le Bon, duc de Bourgogne (15 juin 1467) d’après une lettre de son apothicaire Poly Bulland et les comptes des funérailles de ce prince. http://cour-de-france.fr/ article2123.html. Accessed on 14 Aug 2020 BIUM, Ms XIII (Com. Fac. Med.: 1636–1652), fol. 173 BIUM, Ms XXIII (Com. Fac. Med.: 1764–1777), fol. 548 Charlier P (2006a) Médecin des morts. Récits de paléopathologie. Fayard, Paris Charlier P (2006b) L’évolution des procédures d’embaumement aristocratique en France médiévale et moderne (Agnès Sorel, le Duc de Berry, Louis XI, Charlotte de Savoie, Louis XIII, Louis XIV et Louis XVIII). Med Secoli 18(3):777–798 Charlier P, Georges P (2009) Techniques de préparation du corps et d’embaumement à la fin du Moyen Age. In: Alduc-Le Bagousse A (ed) Inhumations de prestige ou prestige de l’inhumation ? CRAHM, Caen, pp 405–437 Charlier P, Embs A, Ubelmann Y, Patou-Mathis M, Huynh-Charlier I, Lo Gerfo L (2009a) Le tombeau dit « de Foulque Nerra III » : étude archéologique et anthropologique. In: Charlier P (ed) Actes du 2ème Colloque International de Pathographie (avril 2007). De Boccard, Paris, pp 73–120 Charlier P, Poupon J, Bouchet F et al (2009b) Etude ostéo-archéologique des restes d’Agnès Sorel (Loches, Indre-et-Loire). In: Charlier P (ed) Actes du 2ème Colloque International de Pathographie (Avril 2007). De Boccard, Paris, pp 419–527 Charlier P, Poupon J, Huynh-Charlier I, Saliège JF, Favier D, Keyser C, Ludes B (2009c) Fatal alchemy. Br Med J 339:1402–1403 Charlier P, Huynh-Charlier I, Poupon J, Keyser C, Lancelot E, Favier D, Vignal JN, Sorel P, Chaillot PF, Boano R, Grilletto R, Delacourte S, Duriez JM, Loublier Y, Campos P, Willerslev E, Gilbert MT, Eisenberg L, Ludes B (2010) Multidisciplinary medical identification of a French King’s head (Henri IV). Br Med J 341:c6805 Charlier P, Poupon J, Jeannel GF, Favier D, Popescu SM, Weil R, Moulherat C, Huynh-Charlier I, Dorion-Peyronnet C, Lazar AM, Hervé C (2013) The embalmed heart of Richard the Lionheart (1199 A.D.): a biological and anthropological analysis. Sci Rep 3:1296

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Charlier P, Froesch P, Huynh-Charlier I, Fort A, Hurel A, Jullien F (2014a) Use of 3D surface scanning to match facial shapes against altered exhumed remains in a context of forensic individual identification. Forensic Sci Med Pathol 10(4):654–661 Charlier P, Huynh-Charlier I, Poupon J, Lalueza-Fox C, Keyser C, Mougniot C, Popescu SM, Brun L, Piétri S, Thévenard F, Laquay L, Hurel A, Ellul JP, Hervé C (2014b) The heart of Blessed Anne-Madeleine Remuzat: a biomedical approach of a « miraculous » heart conservation. Cardiovasc Pathol 23(6):344–350 Charlier P, Poupon J, Jeannel GF, Favier D, Popescu SM, Huynh-Charlier I, Laquay L, Boudouma O, Dorion-Peyronnet C (2016) The embalming of John of Lancaster, first Duke of Bedford (1435 AD): a forensic analysis. Med Sci Law 56(2):107–115 Chartier J (1858) Chronique de Charles VII, roi de France: nouvelle édition, revue sur les manuscrits, suivie de divers fragments inédits, publiée avec notes, notices et éclaircissements par Vallet de Viriville. Pierre Jannet, Paris Colleter R, Charlier P, Treguier J, Pruvot S, Poupon J (2011) Les derniers jours des Comtes de Laval. Etude ostéo-archéologique des restes de Guy XX et d’Anne d’Alègre. In: Charlier P (ed) Actes du 3ème Colloque International de Pathographie (Bourges, avril 2009). De Boccard, Paris, pp 449–500 Dionis P (1716) Cours d’opération de chirurgie démontrés au Jardin Royal. Laurent d’Houry, Paris Druon D (1793) Journal historique de l’extraction des cercueils de plomb des Rois, des Reines, Princes et Princesses, Abbés et autres Personnes qui avaient trouvé sépultures dans l’Eglise de l’Abbaye Royale de St Denis en France. Archives Nationales, Paris Erlande-Brandenburg A (1975) Le Roi est mort. Etude sur les funérailles, les sépultures et les tombeaux des rois de France jusqu’à la fin du XIIIe siècle. Droz, Genève Gallien V, Périn P, Rast-Eicher A, Darton Y, Rucker C (2009) La tombe d’Arégonde à Saint-Denis. Bilan des recherches menées sur les restes organiques humains, animaux et végétaux retrouvés en 2003. In: Alduc-Le Bagousse A (ed) Inhumations de prestige ou prestige de l’inhumation? CRAHM, Caen, pp 203–226 Georges P (2006) L’embaumement médiéval des nantis. Pour la Science 50:98–101. https://www. pourlascience.fr/sd/histoire-sciences/lembaumement-medieval-des-nantis-1886.php Giesey RE (1987) Le roi ne meurt jamais. Les obsèques royales dans la France de la Renaissance. Flammarion, Paris Guillemeau J (1612) Œuvres de chirurgie. Nicolas Buon, Paris Guillemeau J (1649) Œuvres de chirurgie. Jacques Viret, Rouen, pp 856–858 Hadjouis D, Corbineau R (2009) Analyses d’une momie d’un protestant anglais mort en 1636 (Saint-Maurice, Val-de-Marne). In: Bizot B, Signoli M (eds) Rencontres autour des sépultures habillées. GAAF, Paris, pp 127–135 Kantorowicz E (1989) Les deux corps du roi. Essai sur la théologie politique au Moyen Age. Gallimard, Paris Le Breton D (1993) La Chair à vif, Usages médicaux et mondains du corps humain. Éditions Métailié, Paris Lehoux G (1956) Mort et funérailles du duc de Berry (juin 1416). Bibliothèque de l’Ecole des Chartes 114:76–96 Nicaise E (ed) (1890) La grande chirurgie de Guy de Chauliac, chirurgien et maistre en médecine de l’université de Montpellier, composée en l’an 1363. Alcan, Paris Nicaise E (ed) (1893) Chirurgie de Maître Henri de Mondeville, chirurgien de Philippe le Bel, roi de France, composée de 1306 à 1320. Alcan, Paris Pénicher L (1699) Traité des embaumements selon les anciens et les modernes, avec une description de quelques compositions balsamiques et odorantes. Girin, Paris Pigray P (1610) Chirurgia, cum aliis medicine partibus juncta. Marc Orry, Paris

Part VII Mummies in South America and Pacific Region

South American Mummies

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Guido Lombardi and Bernardo Arriaza

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Bioarchaeological Review . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Environmental Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Pharmacological Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chronic and Metabolic Diseases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Regional Trends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Argentina . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Brazil . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Peru . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

South America’s climate primarily explains the existence of the largest set of mummies in the world, out of Egypt. Andean cultures understood these conditions and took advantage of them, developing technical strategies that benefited from natural dehydration in different ways – mostly to preserve food –, among which mummification was an ancient and paramount trait. Over the nineteenth century and the first half of the twentieth century, large bioarchaeological collections originating from this part of the world reached local and distant universities and museums. Studies on those collections have supported the development of important theoretical concepts of modern archaeology, such as the rise of agriculture, sedentarism, civilization, and disease. G. Lombardi Cátedra Pedro Weiss, Universidad Peruana Cayetano Heredia, Lima, Perú B. Arriaza (*) Instituto de Alta Investigación, Universidad de Tarapacá, Arica, Chile © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_25

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More recently, mummy studies have deepened into the description of population size problems that affect people today too, either locally, such as the case of arseniasis, or worldwide, as it is the case with atherosclerosis. Regarding arsenic pollution, its effect in people might have triggered the creation of the oldest mummy-making tradition in the world among the Chinchorro. In the case of atherosclerosis, its description among ancient peoples came as a revelation against the long-held notion of being a “disease of modernity.” Finally, soul searching still affects everyone involved in mummy studies due to the tragic loss of valuable collections over the National Museum of Brazil fire. New and known ways to deal with these risks should be considered in all collections, including the display of replicas. Keywords

Bioarchaeology · Paleopathology · Atacama Desert · Paleoenvironment

Introduction South America holds, probably still mostly underground, the largest number of naturally dehydrated human remains in the world, as well as the oldest, artificially prepared, the Chinchorro. Roughly, a few thousand mummy packages of “bundles” are stored in local museums and, in Peru, appropriate storage facilities for all of them poises a problem for most museum keepers. The existence of mummies and their preservation over centuries and millennia are closely linked to the particular set of environmental conditions that created their setting: the Atacama Desert, the driest place on earth. These conditions run along the western coast of the continent, at the Pacific foothills of the Andes and, therefore, it is no wonder its peoples profited dehydration in other ways too. For instance, Andean cultures sundried their harvested corn, beans, chili peppers, etc. to both store them and to boost the exchange of goods across very early trade networks. Dry potatoes in several forms, even freeze-dried, dry fish and meats – jerky comes from the Quechua word charqui –, are still staple in the Andes today (Fig. 1). Nowadays, the majority of South American mummies are part of Peruvian and Chilean museums’ patrimony. Nevertheless, anthropological collections among the largest developed countries’ museums also keep mummies from this part of the world, usually through travelers and explorers’ acquisitions over the nineteenth and early twentieth centuries (Ordóñez 2019a). Since most local collections also come from a time when archaeology was still shaping up, it is a shame that the scientific value of them is limited due to, in many cases, the lack of appropriately recorded contextual data. Fortunately, an effort has been set up lately by some curators in North American and European museums to recontextualize these collections and, therefore, improve research possibilities among them (Ordóñez 2019b). Over the last few decades, both case and series’ studies based on mummies from this part of the world, provided clear information about the existence of infectious

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Fig. 1 The monumental archaeological site of Huaycán de Pariachi (ZAMHP) holds numerous underground storage wells, Lima, Perú (Credit: Guido Lombardi)

diseases such as tuberculosis and syphilis in the pre-Columbian Americas; later, the documentation of other infectious conditions followed, as well as the description of cases of cancer. Clearly, the South American mummy corpus has proved its utility as a time capsule for past human characteristics and medical conditions. In consequence, though South American mummies have been studied for over a 100 years now; a truly modern scientific approach – multidisciplinary and epidemiologically meaningful – is very recent and still, for different reasons, almost exceptional. This review presents recent and relevant contributions to the field of South American mummies’ studies from thematic and geographical standpoints, followed by a discussion considering general trends. Readers interested in learning more on the subject are directed to previous publications with a wider approach (Verano and Lombardi 1999; Auderheide 2003; Aufderheide and Rodriguez – Martín 2011; Gerszten et al. 2012).

Bioarchaeological Review Since its inception by Marc A. Ruffer over a century ago, paleopathology has traditionally been divided between – physical – anthropologists, dealing more with conditions seen in skeletal remains, and physicians, attempting to pinpoint relevant diagnoses on mummified cadavers. Naturally, both branches have been closely related to the setting up of archaeological research projects around the world; among which Andean South America has contributed some of the most numerous and important cases; for instance, on stress markers, cranial trepanations, infectious

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diseases, early tattooing, human sacrifice, and cranial remodeling (Verano 2016; Verano 2020). Despite the rich contributions by different authors over the decades, paleopathology, from the theoretical point of view, has kept itself mainly subordinated to archaeology driving themes (developmental processes, raise of agriculture, sedentarism and urbanization, warfare and imperial expansion, for instance) or to medicine – raise of new diseases, origin of atherosclerosis, etc. (Gerszten et al. 2012). Lately, there is an effort to grow beyond the level of standardized diagnoses, so to truly develop theoretical lines of research proper of an independent science. One of the best examples is the field of bioarchaeology of care, which synthesizes its observations from skeletal and mummified materials, as well as from contextual data, including coprolites (Nystrom 2018; Nystrom and Tilley 2019).

Environmental Studies Though ancient Andeans settled in diverse territories, their peopling along the Atacama Desert deserves special attention. There, the rivers that enliven coastal and inland areas present high levels of natural environmental contamination by different minerals (e.g., arsenic, lithium, etc.). It has been postulated that the ancient local people were severely contaminated by a polymetallic mix, as many toxic chemical elements appear diluted simultaneously in the different water resources available (Arriaza et al. 2018). Geographically, natural contamination of surface and groundwater shows a mosaic pattern and is associated with long-term volcanism of the Andes. That is, some geographic areas present good quality water sources, while others, for example, show high arsenic levels, up to 100 times above the 10 μg/L, acceptable limit set by the World Health Organization (2008). Continuous arsenic exposure causes an array of chronic health problems including dermatological lesions, different types of cancer, slow growth, and development in children, and differential mortality (Milton et al. 2005). Bioarchaeological analysis of Andean mummies shows that, chronologically, all populations, from Chinchorro to Inca, were affected by natural contamination, particularly from arseniasis, likely due to hydroarsenicism, in addition to consuming contaminated food irrigated by contaminated waters (Arriaza et al. 2018; Echeverría et al. 2018; Swift et al. 2015). In antiquity, this polycontamination may have contributed to or even triggered social change. It has been proposed that the high levels of arsenic in the Camarones Valley, was an initiating factor for the development of the oldest anthropogenic Chinchorro mummies (Arriaza 2005). The rational is that water at the Camarones river, with over 1,000 μg/L of arsenic, significantly affected Chinchorro pregnant women, producing high rates of miscarriages and premature births. As a cultural response to assuage the grief of losing many babies, the Chinchorro people began ornamenting their tiny dead with colored earth, clay, pigments, and reeds. The oldest Chinchorro mummified remains correspond to fetuses and newborns in the Camarones valley. Artificial mummification of children must have been an emotional and successful response to alleviate their grief.

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It is interesting to highlight that after hundreds of generations consuming arseniccontaminated waters, some current Andean populations, such as those from San Antonio de los Cobres (Argentina) and Camarones (Chile), present high tolerance to this element. These populations present a 68% frequency of the AS3MT gene protective variant. AS3MT is a key enzyme involved in arsenic methylation which allows quick elimination of this semimetal through urination (Apata et al. 2017; Eichstaedt et al. 2015; Schlebusch et al. 2013). In these areas, arsenic levels in water reach on average 300 and 1,000 μg/L, respectively. This gene frequency value is very high compared with populations living in nonarsenical areas. The discovery of similar adaptive mechanisms to chronic mercury in Huancavelica natives could lead to interesting paleopathological studies on pre-Columbian and Colonial period quicksilver miners (Lombardi et al. 2012).

Pharmacological Studies Documentation of psychoactive molecules in ancient human remains, particularly mummy hair, is not a novelty; nevertheless, this field has particularly been very active and creative on South American mummies. Trapped catabolites from local ancient plants with a magical or shamanic use, such as tobacco or coca, have been documented numerous times in the hair shafts, particularly since Larry Cartmell pioneered the field (Cartmell et al. 1991). Though population size studies are still lacking, in order to reconstruct the dynamics of, for instance, coca leaf use (age of first consumption, gender differential use, chronological origin, and territorial diffusion), interesting advances have occurred recently in that direction (Fig. 2). Passive tobacco smoking has been demonstrated in the hair of a formative period perinate from Northern Chile (Niemeyer et al. 2018). Moreover, this finding comes along with the demonstration of the mastery of up to three different psychotropic plants – yopo, coca, and ayahuasca – simultaneously in a 1,000-year-old shamanic bundle from the Bolivian highlands (Miller et al. 2019). No doubt, upcoming research will and should shed light onto the use of these and other plants among beyond the dominant, hallucinogenic field. For instance, potato and tomato alkaloids and their catabolites could be analyzed, as well as those from local medicinal plants.

Chronic and Metabolic Diseases As previously implied, South American mummies, due to their relative abundance, appear demographically appropriate for inferring valid paleoepidemiological information regarding important medical conditions, such as degenerative diseases (for instance, atherosclerosis, diabetes or high blood pressure), which are directly or not, linked to the leading causes of current morbimortality. As noted also, elucidating the existence of past conditions’ prevalence and evolution, could add to our understanding of their etiology, and eventually, such knowledge could lead to better controlling and even reducing their negative effects on mankind.

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Fig. 2 Coca plant, leaves and fruit (Credit: Guido Lombardi)

The Horus research team of paleocardiology pioneered studying atherosclerosis in past populations, through CT-scanning well preserved mummies from Egypt and the Americas. The results of their studies shattered the long-held notion that atherosclerosis is a modern condition directly linked to low nutritional value food consumption. Moreover, their work has also incorporated epidemiological studies of atherosclerosis among the Tsimane, an Amazon basin native group from Bolivia, who hold the lowest prevalence in the word for this ailment. Taken together, so far, these findings suggest not only that atherosclerosis existed in the past, but also that it particularly affected the most sedentary members of their societies, independently from their diets; and that the condition is reduced by continuous, but not exhausting, physical activity (Lombardi et al. 2017; Thompson et al. 2013). On the other hand, the application of new techniques is broadening the possibilities to track down earlier cases of atherosclerosis (Mohammad et al. 2019). While the Horus team method has focused on the quantification and distribution of calcified atheromatous plaques as observed and quantified on CT scans taken to mummies, it has been demonstrated that near-infrared spectroscopy allows to detect plaques in mummies prior to calcification (Madjid et al. 2019). This contribution will definitely improve our knowledge of the raise of this deadly condition among past coastal South American populations.

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Regional Trends Argentina The Llullaillaco mummies are, without a doubt, the best-preserved frozen bodies in the World. Just the same as with the celebrated Ötzi, the Tyrolean Iceman, in Bolzano, Italy, the bodies of these children have prompted the creation of a specialized research center and museum around them, in Salta (Ceruti 2018). Frozen mummies store cells, bacteria, DNA, and all sorts of molecules whose research potential is difficult to fathom. Devoted centers as those mentioned are absolutely necessary to protect their precious biomes and proteomes from external contamination and unnecessary handling. Moreover, these centers encompass all necessary tools to handle new discoveries of this sort, which lately, are more likely to happen. Frozen mummy studies have gotten a new twist under the current climate-change scope; ice cores are being studied all over the World to assess the periodicity and extension of this phenomenon. No place on earth is more affected today to ice melting and receding glaciers than the Tropical Andes. Therefore, the mediatic appeal of these mummies presents an opportunity to engage the public into the topic, otherwise difficult to comprehend or too distant, of global warming. Public education, which is at the center of any museum’s mission, gets an opportunity to provide insights into another developing and worrisome subject, through mummies (Veettil and Ulrich 2019).

Brazil Brazil’s National Museum (BNM) fire – Rio de Janeiro, September, 2018 – brought up the tragic loss of precious artistic and scientific heritage, including unique bioarchaeological specimens. Though a few elements of this collection were spared, most of its mummified human remains became unaccounted for in the aftermath, namely, for instance, the unique Mundurucu heads (Rodrigues Carvalho 2019). As it should happen upon tragedies, this one triggered solidarity from both Brazilian and international peer institutions to help rebuild and reassemble BNM collections. Nevertheless, in the scope of this book, it should also prompt all museums’ curators and stakeholders to evaluate safety measures to prevent and avoid events like this one by all means. It should also induce all of us to set up backed-up digital databases of all materials stored and to focus research on most vulnerable materials. We should also keep in mind that every bioarchaeological remain is ultimately perishable; despite all our best efforts, it will eventually disappear in an unforeseeable future. Besides digital imaging, museums dealing with unique materials should also consider displaying state-of-the-art replicas, while originals could be kept in risk proof conditions.

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Chile Northern Chile and neighboring Southern Peru host the ever-surprising Chinchorro mummies, the world’s oldest artificial ones. Though nature would have probably preserved their bodies without much intervention; early embalmers’ skill and centuries-long tradition created statue-like idols which, back in their time, could still partake from the lives of their surviving kin (Arriaza 2016). The mystery of the Chinchorro deepens into the past even more, as the material culture that has survived to our times is basically reduced to their bodies themselves and few artifacts. Lately, two important and promising avenues have sprung on Chinchorro studies: paleoenvironmental and sociological. The first one deals with the realization that most of these societies lived on, as aforementioned, a naturally arsenic-polluted land. Research on the geological distribution of this feature, its concentration and deleterious effect on the human remains, and perhaps most importantly, the effect of currently living communities occupying the same lands, has produced surprising and useful policy-leading results. The second avenue involves teaching local communities about the Chinchorro in a way to create both a sense of belonging as well as a sense of pride for their ancestors. The Chinchorro settlements are currently being nominated as Heritage of Humanity. This is a very important issue, particularly in Northern Chile, where previous disconnection between researchers and collection keepers, and native populations provoked the de facto closing of San Pedro de Atacama’s famous Le Paige museum. Another important element developed by Chilean scholars is the use of well-developed mummy replicas, both for teaching and displaying. This methodology not only protects the original specimen from unnecessary manipulation, but also helps to raise public consciousness on the adequate conditions that should be met to protect storage rooms from more dangerous threats, such as natural disasters, fire, etc. (Figs. 3 and 4).

Peru Every few years, important archaeological discoveries occur in this country; therefore, it comes almost naturally that recently, a large set of Chancay mummy bundles was excavated at the Cerro Colorado archaeological site (North of Lima), over a salvage program due to urban development. Around 300 hundred bodies were recovered, the largest archaeologically led mummy excavation made in Peru since the Puruchuco – Huaquerones find of over 2000 bundles, also in a salvage project. An important proportion of the observed bodies in Cerro Colorado have tattoos: over a hundred of them, whose comprehensive study would definitely allow for understanding the demographic distribution and, perhaps, elucidate the evolution of this cultural practice among the Chancay (Van Dalen and Majchrzak 2019). Similarly, a pre-Columbian cemetery devoted to over a hundred dogs was also identified in Lima’s Parque de las Leyendas zoo in 2002. The study of their partial mummies led to the identification of four breeds and the mostly sacrificial cause of death. These findings add up to the knowledge obtained from the celebrate Chiribaya dog cemetery (Venegas–Gutiérrez 2019) (Fig. 5). Nevertheless, not everything has been good news. Over the last few years, grave looting has obtained an added value thanks to an intricate partnership with publicity-

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Fig. 3 Female mummy adult. Black style. (Credit: Bernardo Arriaza)

driven ufologists. Though mostly innocuous, this liaison turned bitter when skilled scammers began to manipulate and even mutilate real Nazca mummies to mimic imaginary alien bodies, attempting to remove the human condition to our ancestors, a veritable crime against humanity (Heaney 2017). The unfortunately ongoing hoax has been denounced by mummy research experts in different forums, and today, the general public is aware of this ill-oriented business (see ▶ Chap. 49, “Fake and Alien Mummies”). Finally, there is great local expectation regarding the opening of the new National Museum – MUNA – in Pachacamac (South of Lima) in 2021, on occasion of the Peruvian independence bicentennial. Some of the best archaeological and anthropological collections in the country will be taken to new, state-of-the-art facilities, which should prove to become an asset on Sonia Guillén’s recent position as Minister of Culture of Peru. To our knowledge, this is the highest public position a mummy expert has ever held in the world.

Conclusion South American human remains have participated from numerous and very important studies that have shaped up archaeology and physical anthropology as modern fields of human knowledge. Though in the beginning, most publications dealt with skeletal series linked to processual archaeology, lately some shifts have occurred. The new paradigm is incorporating, from the standpoint of medicine, the prevalence

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Fig. 4 Replica of the previous case. (Credit: Paola Pimentel)

Fig. 5 Andean mummy being CT-scanned (Du Brulle 2017) (Credit: Thomas Wilski - KMKG / MRAH - CUSL)

of contemporary diseases that kill people in the developing world, such as cardiac infarcts secondary to atherosclerosis to the focus of mummies’ studies. Pollution as a worldwide problem is also driving analyses on human remains, such as it is the case of Chinchorro mummies. From the point of view of anthropology, social dynamics could also be read through careful evaluation of even individual bodies, such as it is

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the case of the bioarchaeology of care. The rise of new diseases and/or their reappearance is also another line for current and future surveys of large collections of mummies stored in our museums and universities, in South America and beyond (Fig. 5). The last Paleopathology Meeting in South America (PaMinSA 2019) held in Sao Paulo was particularly revealing of the fragility of our field, not only due to the loss of valuable human remains in Brazil’s National Museum, but for the reduced funding for scientific research is hitting hard several of our countries. Different reasons could be blamed for this negative tendency; nevertheless, it is important to stress that perhaps in part, our research has not reached the mainstream as it should have, so that public investment in primary research would be more valued by everyone. Science is at risk, in general, so our task should also encompass strategies to keep our field and work advancing and not receding. With very few exceptions, despite de aforementioned difficulties, important research is currently being held on South American mummies, basically fueled by North American and European universities’ funds. This symbiotic contribution, which historically drags from colonialism, still jags on the process of transference of knowledge back to the sources of information. This is a complex sociological problem in our countries too, since native populations, whose direct ancestors are the ones subject to study, have also been postponed by the local elites (Lombardi 2008). An effort is being made to bridge that gap, for instance, in Huaycan, East of Lima, to put together the locals with visiting scholars and the knowledge they obtained from studying their abuelos (grandparents, as they refer to the mummies of their ancestors). Free weekly lectures – online over the pandemic – are provided to the public so to teach them about archaeologists and related-field specialists’ work (Lombardi 2015). As mentioned, most all mummies in South America have been naturally produced. This means even today, most bodies get dehydrated by natural factors, without any human intervention. More importantly, this phenomenon, which depends fully on concurrent natural factors, could and should be taken advantage of, when dealing with human remains’ storage, particularly mummies. We should not fight nature but play along. Therefore, Andean countries could reduce budgets and save energy by using these factors appropriately. For instance, instead of fighting the high humidity of the Peruvian coast, some archaeological collections, including mummies, could benefit from low-cost storage in the highlands, were low temperatures and low humidity prevail naturally almost all year long. If applied, this procedure, based on native lore, would not only provide jobs to native people but also improve their self-esteem. Though, through time, we have learned a lot from South American ancestors’ bodies and conditions, it is not late for us to learn from their original wisdom.

Cross-References ▶ Fake and Alien Mummies

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References Apata M, Arriaza B, Llop E, Moraga M (2017) Human adaptation to arsenic in Andean populations of the Atacama Desert. Am J Phys Anthropol 163(1):192–199 Arriaza B (2005) Arseniasis as an environmental hypothetical explanation for the origin of the oldest artificial mummification practice in the world. Chungara Rev Antropol Chil 37:255–260 Arriaza B (2016) Cultura Chinchorro: Las momias artificiales más antiguas del mundo. Editorial Universitaria de Chile, Santiago Arriaza B, Amarasiriwardena D, Standen V, Yáñez J, Van Hoesen J, Figueroa L (2018) Living in poisoning environments: invisible risks and human adaptation. Evol Anthropol 27(5):188–196 Auderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge, MA Aufderheide AC, Rodriguez – Martín C (2011) The Cambridge encyclopedia of human paleopathology. Cambridge University Press, Cambridge, MA Cartmell L, Aufderheide A, Springfield A, Weems C, Arriaza B (1991) The frequency and antiquity of prehistoric coca-leaf-chewing practices in Northern Chile: a radioimmunoassay of a cocaine metabolite in human-mummy hair. Lat Am Antiquity 2(3):260–268 Ceruti C (2018) Inca mountaintop shrines and glaciers in the high Andes. J Glacial Archaeol 3:59–78 Du Brulle C (2017) Rascar Capac passe au scanner. Daily Science 23 March 2017 https:// dailyscience.be/23/03/2017/rascar-capac-passe-au-scanner. Accessed 16 Jan 2020 Echeverría J, Niemeyer H, Muñoz L, Uribe M (2018) Arsenic in the hair of mummies from agroceramic times of Northern Chile (500 BCE–1200 CE). J Archaeol Sci Rep 21:175–182. https:// doi.org/10.1016/j.jasrep.2018.07.008 Eichstaedt C, Antao T, Cardona A, Pagani L, Kivisild T, Mormina M (2015) Positive selection of AS3MT to arsenic water in Andean populations. Mut Res 780:97–102 Gerszten E, Allison M, Maguire B (2012) Paleopathology in south American mummies: a review and new findings. Pathobiology 79:247–256 Heaney C (2017) The racism behind alien mummy hoaxes: pre-Columbian bodies are once again being used as evidence for extraterrestrial life. The Atlantic, August 1 Lombardi G (2008) Indigenous health, South America. In: Kirch W (ed) Encyclopedia of public health. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-5614-7_1675 Lombardi G (2015) Revisiting PaMinSA: a perspective from Peru. Paleopathol Newsletter 172:37–39 Lombardi G, Lanzirotti A, Qualls C, Socola F, Ali AM, Appenzeller O (2012) Five hundred years of mercury exposure and adaptation. J Biomed Biotechnol 2012:472858. https://doi.org/10.1155/ 2012/472858 Lombardi G, Thompson R, Allam A, Frohlich B, Rowan C, Michalik D, Sutherland ML, Sutherland J, Narula JL, Wann LS, Finch C, Kaplan H, Thomas G (2017) Revisiting atherosclerosis: what have we learned from Paleopathology and Anthropology? In: 7th paleopathology meeting in South America, Arica, Chile Madjid M, Safavi–Naeini P, Lodder R (2019) High prevalence of cholesterol-rich atherosclerotic lesions in ancient mummies: a near-infrared spectroscopy study. Am Heart J 216:113–116 Miller M, Albarracin – Jordan J, Moore C, Capriles J (2019) Chemical evidence for the use of multiple psychotropic plants in a 1,000-year-old ritual bundle from South America. PNAS 116(23):11207–11212 Milton AH, Smith W, Rahman B, Hasan Z, Kulsum U, Dear K, Rakibuddin M, Ali A (2005) Chronic arsenic exposure and adverse pregnancy outcomes in Bangladesh. Epidemiology 16:82–86 Mohammad M, Safavi-Naeini P, Lodder R (2019) High prevalence of cholesterol-rich atherosclerotic lesions in ancient mummies: a near-infrared spectroscopy study. Am Heart J 216:113. https://doi.org/10.1016/j.ahj.2019.06.018 Niemeyer H, Souza d, Patricio CC, Echeverria J (2018) Chemical evidence of prehistoric passive tobacco consumption by a human perinate (early formative period, South-Central Andes). J Archaeol Sci 100:130–138 Nystrom K (2018) The bioarchaeology of mummies. Routledge, New York

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Nystrom K, Tilley L (2019) Mummy studies and the bioarchaeology of care. Int J Palaeopathol 25:64–71 Ordóñez M (2019a) Unbundled: European collecting of Andean mummies 1850–1930. Doctoral Thesis, Archaeology. Leiden University Ordóñez M (2019b) Bundling objects, documents, and practices: collecting Andean mummies from 1850 to 1930. Museum Hist J 22(1):75–92 Rodrigues - Carvalho C (2019) Dealing with human osteological collections: questions and reflections before and after the fire at Museu Nacional. 8th paleopathology meeting in South America. Sao Paulo, Brazil Schlebusch C, Lewis C Jr, Vahter M, Engström K, Tito R, Obregón-Tito A, Huerta D, Polo S, Medina A, Brutsaert T, Concha G, Jakobsson M, Broberg K (2013) Possible positive selection for an arsenic-protective haplotype in humans. Environ Health Perspect 121:53–58 Swift J, Cupper M, Greig A, Westaway M, Carter C, Santoro C, Wood R, Jacobsen G, Bertuch F (2015) Skeletal arsenic of the Pre-Columbian population of Caleta Vitor, northern Chile. J Archaeol Sci 58:31–45 Thompson RC, Allam AH, Lombardi GP, Wann LS, Sutherland ML, Sutherland JD, Soliman MA, Frohlich B, Mininberg DT, Monge JM, Vallodolid CM, Cox SL, Abd el-Maksoud G, Badr I, Miyamoto MI, el-Halim Nur el-Din A, Narula J, Finch CE, Thomas GS (2013) Atherosclerosis across 4000 years of human history: the Horus study of four ancient populations. Lancet 381(9873):1211–1222. https://doi.org/10.1016/S0140-6736(13)60598-X Van Dalen P, Majchrzak L (2019) ¿Quién era quién en la cultura Chancay? Los resultados del estudio estratigráfico, antropológico y arqueométrico de los fardos y restos humanos encontrados en los sitios de Cerro Colorado y Macatón. 8th Paleopathology Meeting in South America. Sao Paulo, Brazil Veettil B, Ulrich K (2019) Global disappearance of tropical mountain glaciers: observations, causes, and challenges. Geosciences 9(5):196. https://doi.org/10.3390/geosciences9050196 Venegas–Gutiérrez KE (2019) Análisis de la relación entre cánidos y humanos en el Complejo Maranga Lima, período Intermedio Tardío: Estudio de cánidos del cementerio Huaca 33. M.A. Thesis in Archaeology. Pontificia Universidad Católica del Perú, Lima Verano J (2016) Holes in the head: the art and archaeology of trepanation in ancient Peru, Dumbarton Oaks Pre-Columbian Art and Archaeology Studies Series 38 Verano J (2020) Looking back, looking forward: paleopathology in Andean South America. Int J Paleopathol 29:150–152. https://doi.org/10.1016/j.ijpp.2019.10.001 Verano J, Lombardi G (1999) Paleopatología en Sudamérica Andina. Bull Inst Français d’Études Andines 28(1):91–121 World Health Organization (2008) Guidelines for drinking-water quality. Incorporating the first and second addenda. Genoa. Edition. Vol 1. Recommendations www.who.int/water_sanitation_ health/dwq/fulltext.pdf. Accessed 22 Jan 2018

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Archaeological Site Lapa Do Boquete: Peruaçu Valley . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Church of Santo Antonio Aparecido: Municipality of Itacambira . . . . . . . . . . . . . . . . . . . . . . . . . Archaeological Site Gruta Do Gentio II: Municipality of Unaí . . . . . . . . . . . . . . . . . . . . . . . . . . . . Archaeological Site Caverna da Babilônia: Municipality of Goianá . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Mummified bodies are found in many areas due to processes that interfere with the influence of external and internal agents on corpse. Factors such as climate, temperature, and soil are external agents that directly influence the mummification process. Brazil, despite being a tropical country, has in some regions very local climate, temperature, and soil that favor the process of total or partial conservation of bodies. However, it is possible to find a large number of skeletons that, as well as mummies, are relevant for studies by professionals from different areas, such as anthropologists, archaeologists, and paleoparasitologists. Certain locations in Brazil gained prominence due to the findings. This chapter presents some of the important findings in very peculiar regions of our country. Although there are relatively large numbers of skeletons in Brazil, scholars are surprised by the findings of fully or partially conserved bodies found in a country with such peculiar and unique climate characteristics. Keywords

Brazil · Mummies · States · Climate · Temperature · Soil S. P. Corrêa Novo (*) Escola Nacional de Saúde Pública Sergio Arouca – ENSP, Fundação Oswaldo Cruz – Fiocruz, Rio de Janeiro, Brazil © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_26

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Introduction Brazil is a country of many peculiarities and singularities. Hence, this is no exception to the climate and the different regions we find in our country. Thus, it would have no point to speak of Brazil’s mummies without first saying about the climatic and regional conditions of this country. Another important topic to expose, even if it is brief and without going into through details, is the question of mummification. The association of natural or spontaneous mummification with local climatology is of essential importance, as it exerts great influence on the process of preserving bodies. In most of the countries located in the Americas in the prehistoric period, intentional mummification did not occur due to cultural issues. Naturally mummified bodies are mainly found in dry climate regions. These bodies are recovered from areas such as shelters and caves where they are spontaneously desiccated and mummified (Cárdenas-Arroyo 1998). Thus, some questions are always raised when talking about mummification in Brazil. So, why not take advantage of this chapter to answer some questions and what exactly is mummification and how does it happen? Why does Brazil have more skeletons than absolutely or partially preserved mummies? Mummification is the process of preserving bodies when basically a corpse or its tissues remain so physically preserved that they resemble their morphology when they are alive, but resists further deterioration for a long postmortem break (Zimmerman and Tedford 1976). But how does this happen? Immediately after death, decomposition begins. A complex chemical process develops in the body tissues and the physical effect is initially observed with the softening of the tissues and finally with the liquefaction or gasification of the tissues involved. The products generated by the body in this process can react with environmental substances such as air, soil, composts, etc. This process is known as autolysis and begins immediately after the individual’s death (Aufderheide 2003). According to Aufderheide (2003), initiated by intracellular enzymes normally already present in structures named lysosomes, the enzymatic process of autolysis (self-destruction) begins immediately after death. During the lifetime, these enzymes are highly regulated and activated only when their effects are useful for the wellbeing of the cell. After death, such regulation ceases and the released enzymes soon self-digest and liquefy the cells. The second stage begins with the arrival of bacteria from the body itself, such as the oral and fecal microbiota or from the external environment. These bacterias secrete more enzymes, resulting in greater tissue liquefaction to generate additional molecules that the bacterias absorb for their self-nutrition (Aufderheide 2003). Some flies and beetles are attracted to them and, in the body already in decomposing process, lay their eggs and the larvae, when they hatch, they feed on the decomposing tissue (Haskell et al. 1997). Afterwards, the odor of gases released by this process attracts other insects that voraciously ingest part of the liquefied tissue and the rest of the decomposed tissue. Thus, the process eventually finishes with environmental effects involving the dissolution of skeletal tissue by the interaction of bone mineral with ions

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in groundwater, basically functioning as a “bone cleansing” (Evans 1963). So that, it is possible to understand that the mummification mechanisms are related to some factors which influence the action of chemical substances in this enzyme-dominated decomposition process. The factors which influence this process slowing down and even blocking the action of these decomposition enzymes are the aqueous medium, soil pH acidity, temperature, substrate specificity, and inhibitors (Aufderheide 2003). As mentioned before, a simple and fast explanation of the mummification process has been presented in the paragraphs above, basically to start this chapter. The mummification process goes beyond what has been explained above. However, the explanation is sufficient to understand why we found more skeletons than mummies in Brazil. Desiccation of body tissues is undoubtedly the most common form of spontaneous mummification. Not surprisingly, spontaneous mummification occurs most commonly in the hyperarid deserts of the world, including regions of Egypt, China, Peru, northern of Chile, and some southwestern regions of the United States (Dall 1880; Hrdlicka 1941; el Najjar et al. 1985; Schobinger 1991; Aufderheide 1996; Aturalyia and Lukasewycz 1999). It is important to highlight that man-made microclimates, usually due to cultural habits and where desiccation would not be expected, such as stone baskets, caves, and urns, can also lead to desiccation in humid climate areas (el-Najjar et al. 1998). These cultural habits have also been documented in Brazil. Brazil is a country with a tropical climate and a moist and acidic soil, which speeds up the decomposition of bodies. Taking into account the great influence that climate has on the soil, and characteristic of both in our country, it is important to mention for each region where mummies were found in our country, the characteristic of their respective climate and soil. Numerous archaeological sites have been discovered in different regions of Brazil. Some of them showed only the human presence due to excavated artifacts that proved that presence at the time in the region (Laming-Emperaire 1979; Guidon 1984; Da-Gloria et al. 2017). However, elsewhere, spontaneous or fully mummified bodies and many skeletons have also been excavated. Some of them articulated, but others not. Our chapter focuses on the mummies found in Brazil and was a satisfactory survey related to some of these mummies which we will mention after. There are many archaeological sites found in our country, so that it would take many pages to present if we would mention them at all. Although the natural preservation of the human body is not frequent in Brazil, it has been described in other known cases in dry regions, both hot and cold (Beltrão and Lima 1986) and, since we consider all the findings, not only in Brazil, but worldwide, of utmost importance, will be presented to you, as a reader, some archaeological sites of our country and the mummies found in them. Some Brazilian sites with precious archaeological finds have been found in the state of Minas Gerais.

Archaeological Site Lapa Do Boquete: Peruac¸u Valley It is located in the Peruaçú River Valley, in the northern region of Minas Gerais State, central part of Brazil. The region is characterized by savannah vegetation composed

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of stunted and twisted trees and gallery forests along the rivers. The site is located in an environmentally protected area of the Peruaçú Caverns, covering 1.440 km2 (IBAMA 2003a). The site is a multipurpose rock shelter occupied in the past by an extinct society that practiced mixed farming, gathering wild plants and hunting. The occupation dates from 600 to 1200 years ago. The sediment in the region is powdery, ash-rich and of basic pH, and has subsurface limestone blocks (Prous and Schlobach 1997). Such characteristics are believed to contribute to the preservation of organic remains. Although there are gallery forests along the rivers, the vegetation is characteristic of the cerrado. This explains the finding of some partially mummified bodies (Prous and Schlobach 1997). One of the bodies found is male, aged between 35 and 40 years old, dated 540
40 BP (Before Present), buried in an oval grave inside a stone shelter near the Peruaçú river (Kipnis 2008). The body was found in a fetal position, protected by leaves and covered with baskets. His head was wrapped in leaves and his body was partially mummified. The arms and legs, abdominal skin, and musculature of some parts are well preserved. Many personal objects were found in the tomb. Some consolidated fractures were identified in the bones of the feet, as well as oral lesions, tooth wear, tooth decay, and dental abscess. An observed fact, which is no less relevant, was an accumulation of feces forming a mass in the abdominal cavity, which was later confirmed as chagasic megacolon (Fernandes et al. 2008). In addition, paleoparasitological analyses showed infection by Echinostoma sp. and hookworms (Sianto et al. 2005). The other body found is a female, approximately 35–39 years old, dating from 7000–4500 years AP (After Present) and belonging to a hunter-gatherer population. In this body was also observed Trypanosoma cruzi infection (Lima et al. 2008).

Church of Santo Antonio Aparecido: Municipality of Itacambira The municipality of Itacambira is located in the north of Minas Gerais, a transition region between the semiarid and the cerrado, at the coordinates 8138407.717 N and 727907.697 E, with a short and humid summer period and long dry winter periods. At an altitude of 1048 m, it currently has a total of 4988 inhabitants in an area of 1.788, 445 Km2 (IBGE 2010). The settlement of the village of Itacambira took place in 1674, after the increase in diamond mining, the local population was definitively established. The mummies originating in that locality consist of 3 partially preserved bodies, 2 adult males approximately 40 years old, and 1 female child approximately 6 months old. Found along with other skeletonized bodies in the basement of the church built at the end of the seventeenth century, these mummies were provided by Professor Simeão Ribeiro Pires (in memory), from Montes Claros, Minas Gerais, to Dr. Adauto Araújo (in memory) and Dr. Luiz Fernando Ferreira (in memory) of the National School of Public Health Sergio Arouca, ENSP, Oswaldo Cruz Foundation, Fiocruz, Rio de Janeiro, Brazil. Today, they are part of the Collection of Coprolites and Materials of Paleoparasitological Interest Luiz Fernando Ferreira/ENSP, Fiocruz.

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The burial of these bodies is supposed to have occurred as early as 1700 AD (Anno Domini), according to the death certificate files and the oral tradition of the city, although it is not possible to precisely date them. However, over 100 years ago, these bodies were removed from the church yard and deposited in the basement after church repairment. Paleoparasitological analyzes on these bodies revealed infections by Trichostrongylideo, Trichuris trichiura, and Leishmania tarentolae (Araújo et al. 1988; Novo et al. 2015).

Archaeological Site Gruta Do Gentio II: Municipality of Unaí This archaeological site is located in northwest of the state of Minas Gerais, at a height of approximately 600 m and geographic coordinates 16 00 22 '4500 S and 46 00 53 '4500 W. The cave where the mummified body was found is located at a 2.5 km long limestone bank (Ferreira et al. 1980). The region has a tropical climate with dry season in winter and savannah vegetation. However, the cave that housed the mummy has a semi-arid climate with vegetation very similar to savannah (Ferreira et al. 1980). The site shows evidence that it was used by prehistoric humans as housing and also served as a burial site. Analyzes by the Carbon 14 (C14) method showed that there were two occupations at this site (Ferreira et al. 1980; Ferreira et al. 1983). The mummified body was found naturally and it is from a child aged 8–9 years. The body has been found to be fully encased in a network of fragile plant material and it is dating from 3490
120 to 430
70 BP (Before Present). Next to it, there was the presence of some ornaments and the head was separated from the body, also presenting small fragments of soft tissues in the occipital region. The radiographic analysis showed that the legs were flexed with the knees pulled up. It is assumed by the characteristics of the mummy, including clothing and ornaments found near the body, that it is from an indigenous origin (Ferreira et al. 1983). The paleoparasitological analyzes diagnosed in this mummy the presence of hookworm and Trichuris trichiura eggs (Ferreira et al. 1983). Nowadays, the mummy we refer to is curated by the Brazilian Institute of Archaeology (IAB) in Rio de Janeiro, Brazil. In the same cave were also found a partially mummified human chest and bones, with mummified skin and muscle fragments (Ferreira et al. 1980).

Archaeological Site Caverna da Babiloˆnia: Municipality of Goiana´ The municipality of Goianá, located in the state of Minas Gerais, is characterized by its tropical altitude climate, with mild summers and average temperatures of 23 °C, with higher temperatures in January and February. In winter, it reaches its peak in July and August (PMGIRS 2006). In the region, there are basically two types of soils: oxisols and alluvial soils. Oxisols are from a very ancient origin, deep and well-drained soil. They form wavy

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and soft undulating reliefs. They have low natural fertility and high acidity, requiring fertilization and limestone correction. Once there was an unfinished forest with original vegetation and now it has been replaced by extensive pasture. Alluvial soils occur in the floodplains of Rio Novo, formed by material deposited by floods. In some places they have excessively high humidity and soaking areas occurring over extended periods such as 1 year (PMGIRS 2006). At this archaeological site were found a 25-year-old adult female and two funeral burdens: one at her feet, closed, containing a small child, and another, near the head, open, exposing a newborn. The adult individual was only partially preserved, while the abdominal region and the left upper limb have been damaged, leaving only the exposed bones. The head, neck, part of the chest, right arm and hand, and lower limbs were in good preservation condition. In the head, some suggestive elements were observed, such as an excessive amount of skin in the lip region. A small dark brown, almost black tuft of hair above the left ear was also recorded and the neck muscles was perfectly preserved (Beltrão and Lima 1986). Although the bodies were found together in the same cave with the remains of other individuals, it was not possible to claim which of them is the mother or the children. The bodies were found tied to bones, fiber-strung bags, a hammock, a thick bead, and a cross of threads. Such objects and the location of the findings indicate that at least the woman would be from the Botocudo’s group, from the Maxakali, Kanacam, or Makuni ethnic group (Beltrão and Lima 1986). As you can notice, most archaeological sites where some mummies were found, mostly in partial conservation, are located in the state of Minas Gerais. The first thing that leads us to an explanation for the existence of mummified bodies in this region is related to the climate issue. However, the other regions from our country, Brazil, have provided us with the same important archaeological evidence. Thus, we cannot fail to mention some other archaeological sites where skeletal bodies of extreme importance were found within archaeology, anthropology, paleoparasitology, etc. We can mention here the region of Lagoa Santa (Minas Gerais) where some important archaeological sites are located; some of the archaeological sites are Lapa Vermelha IV, Lapa do Braga, and Lapa do Santo. Reverence we do to the archaeological sites located in the Serra da Capivara National Park (Piauí) and the archaeological site Furna do Estrago (Pernambuco). Throughout the Lagoa Santa region, the climate is warm, with the coldest month temperature exceeding 18 °C, and two alternate seasons, one rainy (summer) and one dry (winter), with annual thermal amplitudes below 5 °C (Ribeiro 1995). The soil of the region has a high calcium carbonate content of the limestone rock that composes its structure and, together with the high volume of water and the characteristic climate of the region, form the main corrosion factors of the reliefs. Alongside these chemical corrosion processes, there are also physical processes of underground slumps and the collapse of stone blocks and walls (Kohler 1989). The soil is characterized as dystrophic red-yellow podzolic and the morphoclimatic domain is the Cerrados (Ab’saber 1977), which flora is composed by savannas and general fields (Romariz 1972).

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The entire region of Lagoa Santa is a karst area, topography of which includes hundreds of caves and rock shelters in limestone outcrops, which have favorable geomorphological characteristics for the preservation of ancient bone material. Despite having a relatively acidic soil, cave soil chemistry works as a fossilizing agent, retarding the degradation of the inorganic part of the bones (Auler and Piló 2015). The findings of abundant human bone material and fauna led the first researchers in the region to soon realize the potential of the area to study the first occupations of the continent. In fact, the human skeletons found in the area, in the order of 200–500 individuals (Mello and Alvim 1977), form the largest initial Holocene osteological collection in the Americas and are today crucial for the study of the first human groups on this continent (Neves and Piló 2003). Until nowadays, the oldest dating from human remnants has been recorded on an exhumed skeleton at the Lapa Vermelha IV. We are talking about our famous Luzia, whose Carbon 14 in carbon dating indicated a deposition of human bones between 11,000 and 11,500 years BP (Before Present) (uncalibrated) (Prous and Fogaça 1999; Prous 2002). Luzia’s finding helped to rekindle an old debate about the origins of the American man. The morphology of Luzia’s skull would bring it closer to the current aborigines of Australia and natives of Africa. A slightly more recent date in another human bone fragment exhumed by Lund at another archaeological site in the region – Lapa do Braga – was 9780
70 years BP (Before Present) (uncalibrated) (Lund 1844). In another archaeological site in the region, Lapa do Santo, 26 burials were found with 36 individuals, for which seven dates were obtained directly from the human bones, defining an occupation period between 8730 and 7400 AP (After Present) (Neves and Piló 2003; Strauss 2010). Serra da Capivara National Park (PNSC), located in the State of Piauí, is based between coordinates 08°260 500 and 08°540 2300 south latitude and 42°190 5700 and 42°450 5100 west longitude, occupying an area of 129,953 hectares and a perimeter of 214.23 km, comprising part of the municipalities of São Raimundo Nonato, Coronel José Dias, and Brejo do Piauí (IBGE 2010). The local geographical position, the geomorphological, climatic, and vegetation aspects gave it its own characteristics that influenced the human settlement during pre-history (Mützenberg 2010). The climate of the region is part of the general conditions of the eastern sector of the Brazilian subequatorial semiarid region. It is currently semi-arid, corresponding to the Köppen Bshw type, with an annual average temperature of 28 °C with maximums 47 °C and minimums 10 °C. Due to low rainfall, the dry season is extended (about 8 months). The rainy season runs from October to May, with little capacity to supply the region, as it has a potential annual evapotranspiration of the order of 1.400 mm (IBGE 2010). The vegetation coverage belongs to Caatinga’s domain, so that the soil is very sandy and stony, with high water scarcity. It is a typical dystrophic red-yellow Latosol (low fertility soil); this soil is associated with dystrophic quartz sands, lithic and eutrophic soils, and noncalcic bruno soils (IBGE 2010). For a better description of this type of soil, we can say, according to the source of the IBGE’s Library, they are deep soils, little susceptible to erosion, low natural

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fertility due to the absence of easily weathered, highly leached minerals. However, litholithoseutrophic soils contain high levels of well-drained iron oxides with shallow to medium-deep soils, with possibilities for agricultural use due to physical and chemical characteristics, and soils with high nutrient levels for plants (red latosol) (yellow + red-yellow podzol + noncalcic bruno). However, the presence of quartz sands with litholites makes the soil acidic and with low water retention (IBGE 2010). It is noteworthy that, according to Oliveira et al. (1999), the caatinga vegetation, although less remarkable in antiquity, was already in the landscape, increasing, however, after 4240 years AP (After Present) (Oliveira et al. 1999). Archaeologists from many areas of the worldwide which have performed studies on archaeological sites located in the PNSC have proven that in the sites of Boqueirão da Pedra Furada Burrow, Sítio do Meio Toca, and Toca do Caldeirão do Rodrigues I provided the earliest evidence for the presence of human groups in this region. However, not all of these sites showed human traces. In some of them were found only traces that proved the presence of man in the region (Mützenberg 2010). The archaeological site Furna do Estrago is located in the Municipality of Brejo da Madre de Deus, in Pernambuco state. It consists of a granite leftover shelter located at the foot of the Serra da Boa Vista at an altitude of 650 m. The shelter has an opening of 19 m, facing northeast, height of 4.80 m and depth of 8.80 m. The studies carried out with the traces and bone materials of Furna do Estrago have biologically and culturally characterized the human population buried in this site and were of paramount importance to meet the objective of identifying these individuals (Mello e Alvin and Mendonça de Souza 1983–1984; Lima 1984, 1985, 2001; Carvalho et al. 2007; Lima 2012; Schmitz et al. 2012; Alencar 2013, 2015; Silva 2016). All Furna do Estrago burials are considered simple, only one individual per grave. Regarding the gender of the individuals, the proportions are 15.1% female (corresponding to 11 individuals); 35.6% male (corresponding to 26 individuals); in 49.3% of the cases, archaeologists were able to identify gender (corresponding to 36 individuals). However, from the total of undetermined individuals regarding gender, twenty (20) are infants and children, three (3) are adolescents. The thirteen (13) remaining individuals are adults. We can notice that they were also found along skeletal bodies, adornments, and mats, characterizing the tradition of peoples for a long time (Lima 2001). According to Lima (2001), the burials were organized in sets, and overlapping. The skeletons have been relocated in some places to make way for new burials. In these, the deposition was not the result of secondary burials, but rather an “arrangement” of old skeletons. As a result of this kind of action, many were partially destroyed, mutilated, and friable. In some cases, only the head was rescued. This also explains the presence of decontextualized bones in the upper levels of the shelter (Lima 2001). As the parameters of pit depth and stratigraphic record it was hypothetically defined at the time of the studies, for the occupation of the site as a cemetery, that the depth of the skeletons were a chronological indicator. Three levels of burials were established by Lima (1985, 2001), the former being the burials

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positioned below 80 cm deep (23%); intermediate occupation, with burial pits between 50 and 80 cm deep (51%), and recent occupation, those skeletons positioned up to 50 cm deep (26%). The chronological positioning of the site occupation as a cemetery was between 1000 and 2000 BP (Before Present) (Lima 2001). A dating of 1040
50 was initially used as a reference to burials, which would be older than this date.

Conclusion The survey conducted for the preparation of this chapter, through analysis by professionals from different areas, in a multidisciplinary interaction, showed that Brazil, a country with such peculiar and characteristic climate, temperature and soil, is capable of, in certain regions, partially or totally conserve the bodies. The truth is that the vast majority of the findings are skeletonized. However, regions of the states of Minas Gerais, Piauí, and Pernambuco revealed bodies in a state of spectacular preservation. The findings in Brazil shed light on important historical facts such as the migration of some peoples and even the emergence of the first human groups on our continent.

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Fire Mummies of the Kabayan Region of Benguet Province, Luzon, the Philippines

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Context and Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Symbols of Status -The Practice of Tattooing as Seen on Fire Mummies . . . . . . . . . . . . . . . . . Headhunting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cultural Aspects of Mummification Practice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . When It All Began . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . When It All Ended . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Funerary Rituals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cave Burials and Coffins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Death Rituals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientific Perspectives . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification Science . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Fire Mummies in Today’s Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Current Death Ritual Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Current Attitudes Toward the Mummified Ancestors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

This chapter describes the Fire Mummy tradition of the Ibaloy in the Kabayan region of Benguet province, Luzon, the Philippines. The reader is provided with a background regarding the physical context of the Kabayan region as well as the oral history context as related to the Fire Mummy tradition. Social stratification of the ancient society is discussed and placed in the context of the broader Igorot culture. This discussion includes descriptions of classes and the symbols of status R. G. Beckett (*) Biomedical Sciences, Bioanthropology Research Institute, Quinnipiac University, Hamden, CT, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_33

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including the practice of tattooing. The chapter discusses the alleged beginnings of the Fire Mummy tradition including the Apu Anno legend. Several possible reasons for the decline of the practice are also debated. The chapter explores the cave burial practices of the Ibaloy including a description of coffin preparation and coffin styles. Death rituals are discussed looking at the ancient cultural practices and identifying those that persist into modern times. The chapter presents the Fire Mummification method as handed down through oral histories and applies scientific reasoning as to the efficacy of the techniques employed. The chapter finishes with a discussion of the attitudes of living descendants as they relate to the Fire Mummy tradition. Keywords

Fire Mummies · Mummification · Kabayan · Ibaloy · Philippine culture

Introduction Context and Culture The focus of this chapter is the “Fire Mummies” of the Kabayan region of Benguet province within the Cordillera administrative region on the island of Luzon, Philippines. The Republic of the Philippines is an archipelagic country in Southeast Asia. Situated in the western Pacific Ocean, it consists of about 7,641 islands. These islands are categorized broadly under three main geographical divisions from north to south: Luzon, Visayas, and Mindanao. The Philippines are often considered to be in the broader context of Oceania. Oceania is a collective name for the islands scattered throughout most of the Pacific Ocean. The term includes the entire island regions of the Pacific ocean between Asia and the Americas (Lewis and Wigen 1997). There are some exclusions, however. The Ryukyu, Kuril, and Aleutian islands and the Japan archipelago are not considered a part of Oceania, nor is Indonesia, Taiwan, and the Philippines. The exclusion is based on the premise that the peoples and cultures of those islands are more closely related historically to the Asian mainland. This distinction becomes important as we consider cultural aspects as they relate to the mummification practices of the Philippines. The climate of the Philippines is tropical maritime. Thus, it is usually hot and humid. Annual rainfall averages around 200 in with frequent typhoons, called Bagyo. The coordinates for the island of Luzon are 6
310 0800 N, 126
050 3800 E, about 451 miles (726 km) north of the equator. Given that the island has generally a wet, moist environment and climate, mummification would necessarily be an active process. According to the Provincial Summary of Regions (2016), the Cordillera administrative region is subdivided into six provinces, one of which is Benguet. Benguet has only one major city, Baguio, with around 140 villages called barangays (Fig. 1). The Kabayan region of Benguet is a rugged mountainous terrain and consists

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Fig. 1 Detailed map showing Luzon, the Philippines. The Benguet province (circled) and the Kabayan region (arrow) (Map used with permission from www.freeworldmaps.net)

of thirteen barangays where the Ibaloy culture has practiced terraced farming for centuries (Fig. 2). The crops include tomatoes, carrots, beans, potatoes, cabbages, and upland rice (Anonymous 1985, 2003). The Ibaloy hold the tradition of prestige rites know as a peshit. A peshit is a tradition where a community member enhances their social position through a series of ceremonies. These ceremonies include butchering pigs and other animals and sharing them with kin and neighbors (Bagamaspad and Pawid 1985; Ballard and Afable 2011). The Ibaloy are part of an indigenous culture collectively known as the Igorot. Reportedly, the Ibaloy have practiced a form of mummification for centuries spanning from the 1700s through the 1900s (Picpican 2003). While it is impossible to know for certain, other sources suggest that the mummification practice began many centuries prior to this suggesting that it began around 1200 AD or even as early as 2000 BC (Merino 1989; Morimoto 1993). The Igorot of the Benguet province are a collection of indigenous peoples with the Ibaloy and the Kankanaey being the two dominant groups. The Ibaloy occupy the

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Fig. 2 The rugged Cordillera region. Note the terraced farming that has been practiced for centuries (Credit: Ronald Beckett)

southern portions of Benguet, and though distinct in many ways, they share the beliefs of animism and ancestor worship with the broader Igorot groups. The concept of ancestor worship is often misunderstood. The actual practice of maintaining the ancestors through mummification is more aligned with ancestor appreciation and ancestor guidance in that the mummies continue to be an active part of the daily culture. The mummies are consulted regarding hunting and agriculture efforts. The associated death rituals of the Ibaloy including the creation of mummies were carried out by the priests known as mambunong. Many of these rituals, especially those related to the fire mummies, are still conducted today. In addition to terrace farming, the prehistoric Igorot also mined for gold. This mining resulted in social class formation among the Igorot which included an upper class (mine owners/discoverers) called the baknang, the gold workers, or abiteg, and the slave class known as the bagaen (Picpican 2003). This social stratification helps explain how individuals, those who made an impact on their culture through wealth cultural contributions, were selected for mummification.

Symbols of Status -The Practice of Tattooing as Seen on Fire Mummies It is important to place the Fire Mummies in the larger context of the Ibaloy culture. The stratification of local societies within villages helps explain not only the why and

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how of mummification but also demonstrates the impact of social status regarding who was mummified. To understand the social structure and status, it is important that we examine the practice of tattooing as a social statement. The Fire Mummies of the Benguet region display a wide array of symbolic tattooed art that remains on the bodies through today. Many of the Fire mummies present with elaborate tattoos. The tattoos are called batuk or burik among the Ibaloy and throughout the region. Since there is no written history of Ibaloy culture, it is difficult to say when tattooing became commonplace among the Igorot. It is thought that tattooing may have become a common practice that was influenced by a man named Baglao who was the half-brother of Apu Anno, a great leader and demigod. Apu Anno, thought to have been the first mummy of the region, had elaborate tattoos all over his body. It is speculated that Baglao assisted with the mummification of Apu Anno and helped spread that tradition throughout the region. It would hold then that the practice of tattooing also may have been spread by the same man. This would have been in the twelfth century, the time of Apu Anno’s reign. Apu Anno is discussed in some detail in a later section of this chapter. Batuk is a pre-Hispanic practice that flourished from its inception as a cultural practice through the sixteenth century. Considering the patterns seen, there were likely various purposes for the use of tattoos. Given the variety and recurring motifs associated with the tattoos remaining on the fire mummies, it is surmised that the batuk served as a visual and material record of one’s life, a record of experiences and of deeds while alive (Salvador-Amores 2013). Tattoos were not only considered a rite of passage into adulthood but were also seen as bodily adornment. Often the tattoos presented a record of an individual’s personal history, a life story of sorts. Still other tattoos were intended to be symbols used to ward off evil forces. Others served as marks of bravery or of prowess in activities such as hunting. As mentioned, tattoos were seen as symbols of status and of influence. The greater the degree of tattooing and the symbology used would indicate one’s place in the societal structure. The arms, legs, ankles, waist, chest, abdomen, pectorals, and back may all bear tattoos. The face, ears, chin, and eyes may also be tattooed yet these locations were reserved for the fiercest warriors of the village. The practice of batuk was not limited to the males of the society (Fig. 3a–b). Females also had elaborate tattoos. At the Timbac rock-shelters in Kabayan, one can observe mummies of adult females who were tattooed on the forearms and upper arms, while the adult male mummies tended to have whole body tattoos including tattoos on their fingertips. Some of the Fire Mummies showed tattoos that were incomplete, a work in progress when they died. The tattoos were of varied thematic designs. Geometric designs including circles, triangles, chevrons, various lines, etc., were common. In addition to the geometric patterns, there were also tattoos that represented animals, plants, and humans. Among the Fire Mummies, patterns depicting deer, snakes, lizards, centipedes, indigenous flora, and human shapes can still be seen. Other aspects of the environment are also depicted. The sun, mountains, water, and lightening are common tattoo designs seen on the Fire mummies.

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Fig. 3 Examples of tattoo patterns: (a) Showing a variety of geometric patterns on the hand and arm, and (b) Showing horizontal lines on lower extremities (Credit: Ronald Beckett)

The significance of the tattoos likely varied from one location to another. Some tattoos were aesthetic in nature which served to identify social status and was perhaps a tradition within a familial group. Still other symbols used as tattoos held some specific significance. Generally tattooing was thought to assure a long life and serve as individualistic ornamentation. Tattooing was also believed to have the power to cure and ward off disease. The main motif indicated a self- sense as a prime element within the context of the individual’s existence and reality. The snake motif in particular indicated a belief in the spirit, the soul, and life after death. The snake, as well as other tattoo designs, was a recurring themes seen in coffin carvings associated with the Fire Mummies and is discussed later. Thus, tattooing seemed to be both practical in identifying the status or significant life events of an individual for others to see, and they also held some magico-spiritual significance as well. There were specific individuals who created the tattoos. These practitioners were called manbatok. Depending on the size of a given village, the individual may have been a resident with plenty of work to keep them busy, or they could have been an artist who traveled between and among regional locations. There are several oral reports regarding the actual procedure employed to create the tattoos. Likely there were several methods which varied from practitioner to practitioner. Needless to say it was a time-consuming process that required a variety of resources. As with all tattooing procedures, the integument of the surface is disrupted by some sort of puncturing process. This is an important consideration since the skin is a major defense mechanism against infections for the human body. The process then had to have been carried out skillfully and the correct substances had to have been used. There appeared to be two major approaches to creating

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the tattoo, one being an already inked puncture “instrument,” the other being an initial design produced on the surface followed by the spread of the “ink” over the design puncture sites. The puncture instrument was a thorn from a local plant called maguey (Furcraea foetida) or the thorn of citrus trees. The thorn was then dipped into a slurry made from ash and crushed native tomato plant leaves. This mixture served as the “ink.” The design was then produced on the skin with the preinked thorn. An alternate description was that the thorn was used first to puncture the skin creating the entire pattern or sections of a pattern. Then the “ink” was spread across the puncture sites with the excess being washed away leaving the design. The “ink” served as both the coloring agent as well as a healing solution at the puncture sites. If the ink slurry was made of the juice of fruits, tomatoes or their leaves, the citric acid present in these products could kill some of the surface bacteria thus serving not only as a coloring ingredient but also as an antiseptic agent warding off potential infections at the puncture sites. The resulting color on these “inks” was a bluish (indigo blue being the most prominent) or greenish black hue. The process was time-consuming, labor-intensive, and painful. Tattooing a single arm could take an entire day, while a whole body could take months or years depending on the pain tolerance of the individual. Even with the antiseptic characteristics of the citric acid based “ink,” infections were common prolonging the process. Traditional tattooing continues in parts of the Cordillera region, specifically in Kalinga. However, it has all but disappeared among the Ibaloy communities in Benguet. The decline in the tradition of tattooing is due in part to the loss of artisans and due to a decline in the significance of tattoos brought on by Christianity and education. The pain of the tattooing process may have decreased the number of individuals willing to endure the procedure. Social structures change over time and perhaps the significance of the tattoo was no longer in vogue. Tattooing is an expensive product and the expense may have exceeded the reach of middle and lower classes with precious resources being used for the most necessary needs for sustenance. With that said, today there is an increase in contemporary tattoo artists who create these traditional tattoo patterns as seen on their mummified ancestors (Salvador-Amores 2012, 2013). Tattooing is again on the rise as people rekindle their connection to the traditional ways of the past and perhaps as a result of improving local economies.

Headhunting In the past the Ibaloy had been headhunters. Much of what is known is gathered from oral histories handed down through the generations. Using heads made from the trunk of a giant fern called tibangdan ritualistic headhunts are still practiced. The tibangdan is used in a dance ceremony called the bendiyan. The tibangdan is carved to look like a human head and is mounted on a pole (Pungayan 1985). The bengdiyan is considered by the Ibaloy to be a thanksgiving ritual while along with the dance, traditional songs are sung.

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Cultural Aspects of Mummification Practice When It All Began It is difficult to say when mummification began with any degree of accuracy. As mentioned previously, it could have been as early as 2000 BC or later around 1200AD. It is most commonly thought that mummification began sometime in the 1700s. As with many other cultures around the world, the practice of mummification began by accident. People observed that under certain naturally occurring conditions, the body of a loved one did not decay. In these situations, development of mummification as a process was a bit trial and error. In the case of Ibaloy mummification, it appears to have been a deliberate attempt to preserve the body as there were no accidental or spontaneous mummies to lead the way. The first mummy that remains in the collective memory of the Igorot is that of Apo Anno. Apu Anno’s story is the stuff of legends. Consider this excerpt from the province of Benguet website regarding Apu Anno’s origin and life: (https://www.benguet.gov.ph/index.php/11-municipalities/398-the-legend-of-apoanno-buguias): A Long time ago, Tugtugaka, a brave chieftain of Nabalicong set out to hunt a white deer, which had eluded many hunting trips and had posed a big challenge to many hunters. Tugtugaka got obsessed with the deer and spent many countless days and nights, which greatly exhausted him. Then one day, his dog sensed the white deer and the hunting ensued. After covering a great distance of running and tracking, the deer was finally trapped against a waterfall. Spear in hand and ready, Tugtugaka poised for a good strike when he heard a soft and powerful woman’s voice “Don’t hurt my pet.” Bewildered, Tugtugaka saw a young woman bathing by the crystalline waterfall. Tugtugaka was so awed with her beauty and nakedness, as he did not understand why he didn’t notice her earlier by the waterfall. She continued: “My name is Cuyapon, and that deer is my pet. Don’t hurt it lest you get the ire of the folks. This place is our home.” Tugtugaka was so puzzled. He saw neither sign of dwelling nor of human activity. As she was dressing, “I see that you are a brave man, and I suppose, you are a good leader of your village,” she commented. “Come inside, you are hungry and in need of rest. You are exhausted. We can also talk so that you will understand my folks,” she said as she led him to a small cave opening. Tugtugaka was even more surprised when, as they reached the cave opening, it became a spacious entrance, and inside, a room only meant for a princess. All around are serenity and a paradise. Soon, he realized what he entered is no ordinary world and Kuyapon is no ordinary mortal! She is a fairy who becomes visible when she likes. At the end of the meeting, Tugtugaka left for home with a sackful of meat more than what he could have obtained from the white deer. “Keep our encounter a secret,” he was advised. From then on, Tugtugaka visited the fairy regularly and they fell in love with each other. Realizing that their love is to be blessed with a child, Kuyapon instructed Tugtugaka: “Don’t come back until eight moons and a half from now.” After laboring wait, Tugtugaka returned to find Kuyapon delivering a child. It was a healthy baby boy. He was advised: “Go home and come back only after eight moons and a half from now. You have to take out our child since he cannot live here. His mortal blood destroys the air of peace of our world. But take good care of him as I do in my spirit way. He will grow to be a good hunter and a worthy father of your village. Give him the name Anno, for he will shadow his generation with abundance and good will.”

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Anno was then taken by Tugtugaka and raised to be a brave man and a good hunter. As a leader, his saga includes repealing the “buso” (headhunters from another place and other enemies), yet he was a peaceful man and abhorred waging war even against the “buso” which earned him the respect of his villagers and other tribes. He used to hunt along the river now named after him, the Agno River. As Anno was in his advanced age and sensing death is near, he asked to be buried in a place now called Nabalikong. The people protested about his coffin; a large hollowed log would be too heavy to be carried over a mountain. “Just float it on the river, it will be carried by the river to the site,” he instructed. The people were puzzled because the burial cave is higher than the river and the river does not pass through the burial site. “You will find people to help you there and animals and food to eat,” he further said. Nevertheless they followed his instructions and indeed, the coffin floated all the way to the burial site. They also met people to assist, the food and animals as he said. Since Anno has a high status and is a regarded man in the village, he has to be mummified; mummification is a long process. Right after his last breath, they opened his mouth and forced him with strong brine solution. They even used their mouth to pump the solution into his stomach. After three days, his body was bathed with different kinds of herbs alternately every day. After the bath, the body was sundried. The process went for at least three months. Every day, his animals were butchered for food of the people doing the work. After it had dried and hardened, his body was put in the coffin to be interned in a cave. His generation flourished and peopled many villages far and wide from Benguet to Ifugao and Vizcaya.

Apu Anno’s legend is the first reference to postmortem body preparation resulting in mummification. If this is in fact the case, mummification among the Igorot was first seen in the twelfth century AD. The story also explains how ancestor respect and worship became a part of the Igorot culture. Yet his story did not end with a peaceful coffin burial within a cave. Sometime in 1918, the mummified remains of Apu Anno were stolen. Apu Anno was seen in 1922 in Manila as a carnival attraction. The mummy was exhibited in a museum in Seattle, Washington, in the 1940s. The mummy was not seen for some time until it was re-discovered in an antique shop in 1984 where it was reclaimed by the National Museum in Manila. It is also reported that the mummy was donated to the museum by a Ms. Conception Cortes who stated that she purchased the mummy in 1973 from the estate of Don Antonio Jiminez de Vigan, Llocos Sur. From 1995 through 1998, several attempts were made by Benguet officials to have Apu Anno returned. Then in May of 1999 Benguet elders and officials prevailed and returned Apu Anno to his original burial cave where they affixed a security gate to the cave opening. Apu Anno’s original coffin was long gone so a new one was fashioned. The mummy became a tourist attraction and in 2016 the caretakers were ordered to stop public showings of Apu Anno due to deterioration of the mummy. The coffin can still be viewed from the gate. As of this writing, Apu Anno’s mummified body is invaded with fungal spores and there are active efforts being made to arrest the spread of the fungal invasion (Lapniten 2019). If Apu Anno is not restored, not only will the mummified remains of Apu Anno continue to deteriorate, but the locals fear that disaster may befall them as told in tales of old. It is thought that a man named Baglao was Apu Anno’s half sibling and was directly involved with the mummification process. Since Baglao knew the process and technique of mummification, it is believed that he is the person who began to

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spread the tradition of mummifying revered members of villages when they died. The economic conditions had to be right for this to occur since the process is resource heavy both in time commitment, which would have taken villagers away from their usual tasks, and in materials. As the culture advanced, it was seen as a practice set aside for the wealthy since they would likely have the labor and resources available to them or have access to trade goods needed for the process. While the wealthy were measured in property, pigs, or the ability to trade having gold in one’s possession, it is thought that it was the poorer of the village who actually carried out the mummification process. Given that the process was so timeconsuming, the relatives of the wealthy likely oversaw the process while their abiteg (workers) or bagaen (slaves) carried out the actual mummification. There is speculation that the poor indeed where at times honored by being mummified and placed in the caves with their deceased benefactor.

When It All Ended Just as it is difficult to pinpoint when the practice of mummification began, so is it elusive as to when the tradition ended. It is likely that the practice slowly disappeared over a period of several hundred years. The mummification ritual may have died out in one area while it continued in another. Conventional thinking states that the practice persisted until the late nineteenth century. With that said, there are still reports that aspects of the death ritual continue to the present without seeing the process through to complete mummification. There may be several reasons why the practice of mummification began to decline and eventually cease. It is important to remember that the ritual of mummification was costly in terms or resources and was primarily a procedure reserved for the wealthy. As time passed, development of the region progressed creating more opportunity for mobility among the members of a village. Thus, the workers and slaves began to migrate away from their villages. As the poor moved away, families no longer had the means to conduct body preservation of their family members. Since it was the task of the poorer members of a village to conduct the mummification process, the wealthy who were left behind did not have the skills to continue the associated tasks. The wealthy also migrated away which added additional difficulty to maintaining their traditional practices. Another possibility is that, like the travelling tattoo specialists (manbatok), there may have been individuals who would travel among villages who served as mummification specialists. When these persons died, the practice and procedures may have eventually died off with them. Also, mummification may have been a family tradition handed down through the generations. Each generation would have been challenged to continue the practice yet family fortunes may have devolved over time to the point where the family could no longer afford to continue the tradition. The practice would then eventually die out. A final possibility is the phenomenon known in the developing culture of the region as a pejew. A pejew is a socio-cultural attempt to purge a given practice or

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tradition from the memories of the people. The tradition may have become something that brings up past feuds or painful memories. Or the practice may have caused the community disgrace or loss of prestige. The tradition in question may have been that of mummification. Through cultural pressures, it would never be talked about and eventually it would be removed from oral traditions until it would be forgotten completely. While this is a possibility, oral histories and stories of the mummification practice persist to the present day (Picpican 2003). External pressures from missionary efforts in the early twentieth century may have also impacted the continuation of the practice. Missionaries would have instilled their funerary and burial beliefs and encouraged the indigenous to adopt in ground burials instead of body preservation. More than likely, the end of the mummification practice was brought on by a combination of the various factors presented here. A combination of economics, increased mobility of the populace, enhanced opportunities, and social pressures combined to reduce and eventually halt the tradition of mummification.

Funerary Rituals Cave Burials and Coffins To the Ibaloy, the mountain is the home of the ancestral spirits (kaapuan) and placing the mummified bodies within the mountain rock-shelters or facing the mountain symbolized returning the deceased to Kabunyan, the mountain God. Kabunyan, also known as Lumawig, is the primary god of the Igorot (Fig. 4). The belief is that Kabunyan came down from the sky and married Bangan. They had three children. Once each month, a ceremony is held in his honor in a sacred grove where the trees are believed to have sprung from the graves of his children. Kabunyan was believed to have created the Igorot and taught them how to plant, reap, and overcome the variances of nature itself. The Igorot have many other deities, like the spirits of things and places, in keeping with animistic beliefs. In keeping with the belief in the ancestral spirits, the mummies were placed in wooden coffins being entombed high in the mountains surrounding the Kabayan region (Fig. 5). The coffins were of various shapes and sizes depending on the intended burial practice. Burial locations were typically natural rock-shelters or caves. While many of these caves were naturally occurring, some were increased in size or honed out to accommodate additional coffins (Panganiban and Omana 1998). The labor involved was intense and the resources that were spent further attest to the dedication to the process and the appreciation of the ancestors. There seems to be little consistency in the design of the coffins as various coffin styles were used to entomb their dead. These coffins were wooden and often carved from hollowed pine trees (Pinus benguetensis). In addition to the carved coffins, others were constructed from planks made from similar wood. There appeared to be at least three basic coffin shapes or designs associated with the Ibaloy mummies. These shapes fell into the broad design of ellipsoidal, parallelepiped, or in the shape

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Fig. 4 An example of a cave used for internment among the Ibaloy of the Kabayan region, Benguet Province, Luzon, the Philippines. The natural cave opennings were sometimes made larger to accommodate the burial coffins (Credit: Ronald Beckett)

Fig. 5 A cave used for internment. Note the wooden coffin (dungon) placed into or constructed within the cave (Credit: Ronald Beckett)

of a carabao (Bubalus bubalis) (Merino 1989). Some of the coffins had carvings adorning the tops and sides. As with the tattoos, there were geometric, anthropomorphic, and/or zoomorphic patterns. One coffin pattern common also as a tattoo

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present on the fire mummies was that of the snake. The snake appears in many cultures and has various meanings. Often the snake symbolizes the circle of life and the afterlife. Perhaps it held a similar meaning for the Igorot/Ibaloy culture. The general bioanthropological features of the mummies throughout the region include mummies of all ages at the time of death, with both male and female sexes represented. There are large coffins (dungon) in some caves representing familial burials. Given that it was usually the wealthy or the person who had distinguished himself, it would seem that not all members of that familial line would have contributed to the village in the same manner. Even if family members of a deceased wealthy person were not noteworthy members of the village, they would be mummified and placed with the individual in an attempt to assure that the honored ancestor was kept happy in the afterlife. In this way the living could avoid the wrath of the “spirit” after death. Not only does oral history support the family burial construct in both coffin burials and specific family tombs, radiographic evidence has also supported this concept with images showing inheritable features such as wormian (Inca) bones. As globalization has become more prevalent with travelers seeking to see facets of cultures unlike their own, souvenir seeking and vandalism have plagued the coffins, caves, and the mummies themselves (Fig. 6). Touching the coffins or the mummies increases the rate of deterioration of these priceless ancestors. The plague of “I was here” carvings can be seen within the caves themselves. Parts of mummies or ancestral bones have been taken as a keepsake by unthinking and uncaring adventurers. Indeed entire mummies and coffins have been removed and stolen in the past. The local municipalities have developed efforts and mechanisms to protect the ancestors within the known burials in the Kabayan region. Locked iron or steel gates placed over the mouth of the caves or rock shelter are commonly seen (Fig. 7). Often, a guard is placed at the most common or well-known locations of the burials. Gates on pathways and trails leading to the caves are the norm. Any ladders that once led up to caves on cliff fronts have been removed. Even with these efforts, looting and vandalism persists. The area is vast and the resources to protect the many burial sites are less than would be Fig. 6 Looted cave of an Ibaloy burial in the Kabayan region, Benguet Province, Luzon, the Philippines (Credit: Ronald Beckett)

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Fig. 7 Locked iron gate at the opening to a burial cave on Timbac (Credit: Ronald Beckett)

required to fully assure safety of the ancestral sites. Still there are sites whose locations are unknown. The potential for continued desecration is high. One thing that is in favor of continued preservation and protection of these sites is that of nature itself. Consider the burial caves on Mount Timbac (160 3900 3 N, 1200 4700 43 E), at 2,715 m above sea level (8,907 ft) it is difficult to get to. The rain is ever present in the region and travel to these remote areas is challenging even with good weather conditions. When it rains, as it often does, getting to these locations can be treacherous with many mud and rock slides obscuring the makeshift roads and paths. The province of Benguet receives an annual rainfall of 4,500 mm (177 in), which can at times render the burial sites inaccessible.

Death Rituals In the present, the Ibaloy continue to practice and demonstrate various traditional rituals associated with the ancestors as well as with their interactions with the dead. When there is a death and some of these traditional rituals are conducted, there is initially a dance (tayaw) and prayers or songs that are chanted (bad-iw) for the dead. These rituals are performed by the villagers. They are communicating their support for the deceased in their journey to the next level of existence, an expression of appreciation and caring. Traditional clothing is worn and the participants are often adorned with a white headband. There is specific symbolism within the dance moves. One includes a soaring motion with the arms outstretched as if the spirit of the deceased is being carried on wings to the mountain. When a visit to the burial rock shelters is planned, additional rituals are performed to determine if the tombs can be visited. Traditional clothing is worn while the villagers chant or sing a song to welcome visitors. Local red-rice wine is then shared with all who are present in a wooden bowl which is passed through the group. Forest

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pigs are sacrificed and their livers and gall- bladders are removed. The livers and gallbladders are then “read” by a mambunong (native priest or shaman) to determine if the ancestors approve of a visit to the burial sites. Before traveling to the tomb, other live animals are collected, usually chickens, for rituals to be conducted at the cave entrance. A song for the mummies and the dead called the angba is chanted at the destination. Rice wine is again shared among the participants with some given to the ancestors. At the entrance to the tomb, a small amount of wine is offered and poured at the threshold of the cave. Typically, the chicken or rooster that was brought up to the mountains from the village is sacrificed at the rock shelter entrance by hitting or beating the animal with the blood being poured nearby the cave. Food, betel nut, and matches are also offered at the tomb entrance. Only after the rituals at the village and the burial site have been completed, can the visit with the ancestors take place (Beckett et al. 2017). There is an ensemble of villagers and officials who accompany anyone who wishes to visit the burial sites. The entourage is an attempt to assure that respect is maintained, and it is likely that the large group is intended to dissuade would-be thieves. This is of course the case with organized sanctioned excursions. There is still the potential of uninvited visitors to the cave burial sites. It is considered taboo to bring the mummies out of the tomb with the fear of displeasing the spirits of the dead and the resulting trouble that may bring. If viewing or research is to be conducted the mummies can be moved toward the entrance of the tomb but no farther (Fig. 8a–b). Exceptions certainly have been made. The mummies have come out of the rock shelters for photography, but most of the work associated with research projects at the sites takes place within the tombs themselves. While viewing and examining the mummies, there is someone from the village, often the elder, who is talking to the mummies as if they were living. This person is explaining to the ancestors who was there and why they have come. In the case of research expeditions, any scientific procedures being conducted are explained to the ancestors. These various rituals as well as the interaction with the mummies seem to confirm a strong sense of ancestor respect among the living Ibaloy. These practices are also an attempt to keep the traditions of their culture intact for future generations as the village consort is made up of old and young alike.

Scientific Perspectives Mummification Procedure The actual mummification method or procedure is illusive in that there are no living individuals who have actually practiced or produced a mummy in the region. However, although the tradition is no longer practiced, there are oral accounts of the process handed down through generations (Baucas 2003; Cardin 2014). According to these oral testimonies of the living Ibaloy of the region regarding the funerary and burial traditions, a reasonable presentation of the mummification

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Fig. 8 (a) Research being conducted within the cave as a village elder, Baban Berong (1922–2005) looks on often speaking directly to the ancestor. Note the many coffins stacked in the background. (b) Radiographic analysis of a skull being conducted near the mouth of the cave respecting the tradition of keeping the ancestors with in the cave or rock shelter (Credit: Ronald Beckett)

method is derived. In addition, radiographic images and visual inspection provide evidence that supports several of the oral claims (Beckett and Conlogue 2010). It is important to recall that the process of body preservation among the Igorot was an active process. The rainfall and humidity are in no way conducive to preservation of organic material. In fact, the conditions enhance the decomposition process. Human intervention and ingenuity attest to the desire to preserve the respected deceased. Also, according to the Apu Anno legend, the method was handed down from Apu Anno’s mother, Kuyapon. Thus, there was no trial and error. Another possibility is that the ancient populations had devised a way to preserve deer and other animal meats and then transferred this knowledge to the deceased humans. Regardless, as you will see there is scientific support for the methods described. The mummification method was as time-consuming as it was complex. Beginning at or around the time of death, near to the final breath taken by the individual, the deceased was infused through the mouth with a salt water solution. There are some accounts that this initial step took place just prior to the final breath. From a scientific perspective, if this salt water solution was forced prior to death, severe

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diarrhea, and vomiting would ensue eliminating body water and thus enhancing the potential for dehydration. Further, any residual salt in the intestinal tract would draw body water towards it and may be eliminated as well. The less body water remaining, the greater is the chance of dehydration slowing decomposition while other aspects of the mummification method are employed. Once the individual was dead, he/she was washed in cold water. Following the cold water bath a funerary or death blanket called kolebao or pinagpagan was wrapped around the individual. Then, along with a head scarf called sinalibudo, the body was secured to a sangachil (death chair). The practice of wrapping the deceased with a death blanket is supported as many of the fire mummies have textile impressions on various parts of the body likely made by a death blanket used during the process. The sangachil played an important role in the mummification process with regards to eliminating body fluids through gravity. The individual was placed in a seated position in the death chair with his or her head held in position by the scarf or band. The head scarf/band often held the head in a hyper-extended position. This next step, exposing the deceased to fire and smoke, is reported to begin at different times in the process by different accounts. Yet clearly this procedure was employed at a time that helped to maximize the depletion of body fluids. The chair with the seated deceased was placed and secured to a ladder facing the front of a traditional stilt house. A low fire was lit below and near to the mummy, which likely enhanced dehydration of the individual. Thus, the Ibaloy mummies became known as “the fire mummies.” The next step was to enhance the removal of body fluids. These fluids were expressed manually and through gravity dependence in the death chair were collected in a jar. When the dripping had slowed or stopped, the body was removed from the death chair and laid out in the sun. Here the relatives or elders performed the duduan which is the peeling of the epidermis further enhancing fluid removal. The body was again washed with cold water and once body fluids no longer seeped out, the individual was wrapped again in a blanket and returned to the death chair. Any worms that became apparent on the skin were removed by hand. Various juices were then applied to the body. Juices from the pounded leaves of local plants, including guava (Psidium guajava), diwdiw (Ficus septica), patani (Phaseolus lunatus), duming (Dolichos lablab), and besodak/sopedak (Eubelia philippinensis) (Madulid 2001) were used and it was thought that these rubbings kept the body dry. Oral histories report that the ears and nose were plugged using leaves of the same plants in an attempt to keep insects out of the body orifices. The fire was maintained underneath the body in the death chair. The heat and smoke was continued for 40–60 days. In some cases it has been reported that the fire and smoke process could be maintained much longer, up to two years. Oral reports further state that during the process, tobacco smoke was blown into the mouth and nose intended to assist with the mummification of the internal organs. The time required to achieve mummification must have varied depending on many factors such as time of year and location. It is suggested that the process required several months of constant tending to the body (Morimoto 1993). In the final stages, the body may have been laid out in the sun during the day and returned to the smoky

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fire at night. Once the process was complete, the body was placed into a flexed position, wrapped with layers of funerary blankets and taken to the mountains for entombment within the appropriate coffin and cave or rock shelter (Merino 1989; Picpican 2003). Although the climate patterns of the region are wet and humid, the burial caves were relatively dry and cool. Coupled with the protection provided by the coffin, the fire mummies were provided with an environment that was conducive to preservation and minimized deterioration over hundreds of years.

Mummification Science Much of the information related to the fire mummy process is anecdotal. The efficacy of some of the steps in the process can be called in to question, such as the use of an expensive substance like salt and the possibility of ingesting the solution prior to death. However, many features reported in the accounts of the process would enhance the potential for mummification. The seated position produces a gravity dependence impacting the fluid movement. This would enhance desiccation, as the enzyme-laden fluids of the small intestine would have drained out through the perineum (Aufderheide 2003). The textile blanket used would have served to wick fluids from the body. This is apparent in the burial practices of many other ancient cultures that used textile blankets which helped to draw water out of the body through wicking. Such textile blanket wrappings were used in many mummies of South America including the Chiribaya and the Inca cultures. In the case of the fire mummies, the blanket used to cover the cadaver would have increased the rate of water evaporation by creating such a wicking. Even in such a humid environment, the heat created by the associated fire would have played an additional role in the desiccation process due to the decreased humidity and the greater elevation in temperature (Aufderheide and Aufderheide 1991). The warmth and lower humidity would enhance evaporation of fluids from the death blanket allowing for additional fluids to wick into the blanket from the corpse and then out to the environment. The practice of peeling of the epidermis would also greatly enhance the expression of fluids from the body. Any additional manipulations such as rubbing or massaging the body surface would assist in body fluid elimination. In wet and humid environments, special conditions need to be created if mummification is to be successful. Since the death chair was placed near the entrance to the traditional house, the fire and associated smoke would have created a microenvironment increasing the potential for mummification. Microenvironments such as those associated with mummification practices have been demonstrated in other cultures within tropical forest regions. A good example is the smoked body practice among the Anga of Papua New Guinea (Beckett et al. 2011). Wood smoke has certain chemical characteristics that would enhance the potential for mummification. Wood smoke has phenolic compounds that are released and would have aided in the preservation of tissues. Their antioxidant properties would have inhibited decay of fats and their antimicrobial action would have prevented

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bacterial growth. Wood smoke often contains formaldehyde and acetic acid which create a hostile environment for bacteria. At the same time, it encourages the crosslinking of collagen fibers expelling water from tissues and, as a consequence, decreasing the enzymatic action which occurs during decomposition. The smoke also acts as deterrent to insect infestation (Beckett et al. 2011). While many of the procedures would have enhanced the desiccation process, others must have played a very limited part in the process. The introduction of the salt solution into the mouth seems to have had no impact on organ preservation. At death the cessation of peristalsis in a deceased individual would not move fluid beyond the stomach. The tobacco smoke blown into the mouth would have caused only limited exposure to the internal tissues. The role of the plant materials employed in the process is not known. There exists the potential that these plant juices create a hostile pH environment, either acidic or alkaline, limiting the growth of surface bacteria. In summary, mummification in the Ibaloy tradition was likely achieved primarily from enhanced dehydration accomplished by the heat from the fire, the position in the death chair, the potential microenvironment, the peeling of the epidermis, and wicking of body fluids into the death blanket. The forces of decomposition may have further been delayed by the smoke chemistry, the increased barrier to insects provided by the smoke, and the plant chemistry. Plugging the body orifices also would have kept flies and insects from invading the corpse. If a salt solution purge was successful, this may have contributed to enhanced dehydration. We do know that the process was effective and desiccation, along with other factors happened relatively quickly (Fig. 9a–b). Not to the point of complete mummification, but enough to avoid early stages of decomposition. Internal preservation including the internal organs is highly variable. It is not uncommon to find variations in preservation among cultures who practiced mummification. Some of the variances seen may be explained by a potential delay in initiating the process following death. The skill of the practitioner also may have impacted the quality of the initial mummification procedure. The antiquity of the individual and taphonomic changes over time are other variables that would influence what remained of the initial internal preservation. A number of the mummies presented with a hyperextension of the head, which was related to the Ibaloy mummification practices described above. An interesting case examined was that of an adult female mummy housed in a small one room museum in the village of Kabayan. The mummy had an inverted T-shaped incision on the anterior abdominal wall, which was interpreted as a possible sign of disembowelment. An alternate explanation of the incision derived from oral histories and suggested by local museum personnel was that this particular mummy was subjected to a caesarean section using a sharpened bamboo knife. Local folklore further suggests that if the baby survived, they were fed with the juices from sugar cane. The internal preservation of this mummy was very good attesting to the efficacy of the process. Although enlarged anuses were observed on some subjects suggesting a form of organ removal, evisceration was not evident in these mummies as demonstrated by radiographic analysis (Fig. 10).

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Fig. 9 (a) Ibaloy adult mummy in Timbac mountain of the Kabayan region, Benguet Province, Luzon, Philippines. Note the excellent preservation on this mummy as well as the elaborate tattooing. The extended head position validates the oral history of the deceased placed in a death chair (sangachil) during the smoking process. Mummification also involved the application of a pastes made from various plant materials. Also pictured is Orlando Abinion of the National Museum of the Philippines, Manila, the Philippines. (b) Example of preservation variations: An Ibaloy infant mummy in the Kabayan region, Benguet Province, Luzon, the Philippines. Note the advanced deterioration on this mummy. It is “taboo” to bring the mummies outside the cave entrance (Credit: Ronald Beckett)

In an attempt to determine how long ago the mummies of the Timbac burial cave complex were placed, permissions were granted and permits were obtained which allowed for Radio Carbon Dating (RCD) of a single rib found in the cave. The RCD results from the sample indicated that the rib material was 185 plus or minus 43 years BP (1785–1871 AD) (Beckett et al. 2017). This finding in no way pinpoints the end of the fire mummy tradition yet if does confirm that the practice continued at least into the late eighteenth century and perhaps into the mid-nineteenth century.

The Fire Mummies in Today’s Culture Current Death Ritual Practices It is reported that the sangachil (death chair) is still used when a loved one passes away. This is a clear connection to the body preservation practices of their ancestors and also represents the level of respect afforded the loved deceased. Family and

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Fig. 10 Radiograph of the thoracic region of an Ibaloy mummy at the Kabayan Municipal Museum demonstrating the state of internal preservation. In addition, evisceration was not a part of the Fire Mummy process. Arrow indicates the well-preserved airways within the chest (Credit: Ronald Beckett)

friends gather around smoky fires sharing meals and stories. Songs are sung and rice wine is consumed. The celebration serves as a “wake” for the individual. There are many regional funerary customs spread across modern day Luzon. Specific to the Fire Mummies are the current burial and funerary practices in Benguet province. The dead are blindfolded (similar to the death scarf) and sat on a chair (sangachil). The arms and legs are tied in a manner that will allow the seated posture to be attained. This chair is placed next to the main entrance of the house of the deceased. The deceased is there in the chair for eight days and is tended to by the family and friends. On the eve of the funeral, the elders perform a chanted narration known as a bangil rite. This chanted narration is a biography of the life of the individual. Bamboo sticks are struck together as the body is lowered into the place of final rest. The bamboo stick process is intended to direct the dead toward heaven (Nobel 2013). There are cemeteries in the remote villages of the region where many of the deceased are interred, while others are not.

Current Attitudes Toward the Mummified Ancestors Recent ethnographic research (Piombinio-Mascali et al. 2013) was conducted in an attempt to ascertain the attitudes of the living toward the Fire Mummies and the associated death rituals. Researchers interviewed individuals willing to discuss their

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current beliefs regarding the mummified ancestors and their relationship to the mummified dead. One of these individuals was Baban Berong (1922–2005) (Fig. 11). Baban Berong was the village elder of the Kabayan municipality. Berong’s beliefs are in line with those of his ancestors. As with other reports (Picpican 2003) he believes that God is a mountain and the burials of the revered ancestors should be within or facing these sacred mountains. The Fire Mummy is considered to be sacred and thus contains the soul. His belief is that the spirit of the individual remains with the body, and therefore, the mummified remains must be treated with respect. The village and burial rituals expressed, as well as the practice of not removing the mummies from the tombs, are all representative of this respect. Berong and the Ibaloy of Kabayan speak to the mummies as if living. Researchers and other visitors are introduced to the mummies as if they were living. Baban Berong clearly holds the more traditional view of the relationship of the Ibaloy to their mummified ancestors. However, others within the community hold dichotomous views. Camilio Alumet (45 at the time of the study) is from the village of Poblacion. He believes that the spirits of the mummies exist and that these spirits are responsible for periodic calamities when they are angry with the living for some reason. In contrast, a weaver, Marcela Torren (79 at the time of the study), believes

Fig. 11 Former village elder Baban Berong (1922–2005) showing the location of a cave burial in the Kabayan region. Berong held a traditional view of ancestor respect and protection (Credit: Ronald Beckett)

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that the old ways are a dead tradition. Torren feels that if there is a calamity, it has nothing to do with the mummies or their spirits. It is interesting to note that one elder, Berong, has an entirely different view than another elder, while a villager of middle age leans toward the older beliefs. Clearly there are opposing views regarding the mummified ancestors and their spirits. An interesting fact is that while there is a cemetery in the villages, villagers tend not to use it. Gertrude Mangusan (50 at the time of the study), of Poblacion explained that most would rather bury their deceased loved ones in a garden of their homes in soil graves or in nearby rock tombs. It appears that the sense of a spirit of the deceased still exists, as they prefer to have their loved ones close by. Another possible explanation may be that the family members do not want the bodies of their loved ones stolen, as has been the case in the past with many mummies from the region (De Leon 1976). Ana Labrador is a social anthropologist and the assistant director of the National Museum in Manila. Based on her interactions with the Ibaloy culture, Labrador has developed some useful insights into the mummification practices of the ancients. Following the mummification process, the tightly flexed position resembles the fetal position. Labrador sees this as fitting in with the Ibaloy belief that life is preparing one for a transition to another life beyond death. Thus, the flexed or fetal position is possibly symbolic of a return to the beginning, a new life that follows this one. This concept is not dissimilar to Baban Berong’s view that the God is a mountain and that the mummies are being returned to where they came from, the womb of the mountain. Mummy tourism has grown in the region. As more and more people learn of the Fire Mummies, more people find their way to these remote enclaves of ancient traditions. The mummies have always been a part of the local culture but they have become a symbol of regional identity. Labrador suggests that while mummy tourism helps the community providing jobs and income there are always trade-offs. Not all of these are positive. While mummy tourism may be fiscally beneficial, the trails, roads, and the mummies are showing signs of deterioration beyond the effects of climate change, tropical storms, typhoons, and earthquakes. The growth of tourism and interest in the Fire Mummies has out-paced the villages’ ability in terms of resources to adequately regulate the influx of world visitors.

Conclusion The Fire Mummies of the Benguet province are truly a treasure. The rich context and legends associated with the cultural practice of human body preservation informs us about who we are as a human species. The traditions paint a picture of care and compassion while including aspects of the socio-economic impacts on these very practices and rituals. As with many such practices, it is becoming increasingly more challenging for the Ibaloy to keep the traditions alive. In a sense, the ancestors are still looking out for their descendants by providing a means of monetary support in the form of mummy tourism. Clearly the Fire Mummies are still an important part of

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the living culture today. In this chapter we have explored the context, the meaning, the method, and the science of the Fire Mummy tradition.

Cross-References ▶ Smoked Bodies of Papua New Guinea

References Anonymous (1985) Kabayan ‘85. A socio-economic profile. Kabayan Anonymous (2003) Comprehensive land use plan, Kabayan, Benguet – 2003–2012. Prepared by: The Municipality CLUP – MTWG of Kabayan, Benguet. Part I and II Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Aufderheide AC, Aufderheide ML (1991) Taphonomy of spontaneous (“natural”) mummification with applications to the mummies of Venzone, Italy. In: Ortner DJ, Aufderheide AC (eds) Human paleopathology: current syntheses and future options. Smithsonian Institution, Washington, DC, pp 79–86 Bagamaspad A, Pawid Z (1985) A people’s history of Benguet province. Baguio Printing and Publishing, Baguio City Ballard L, Afable P (2011) Ibaloy: dictionary, phonology, grammar, morphonemics and history. Dinteg Inc. and the Cordillera Studies Center, University of the Philippines Baguio, Baguio City Baucas BL (2003) Traditional beliefs and cultural practices in Benguet. New Baguio Offset Press, La Trinidad Beckett RG, Conlogue GJ (2010) Paleoimaging: field applications for cultural remains and artifacts. CRC Press, Boca Raton Beckett RG, Lohmann U, Bernstein J (2011) A field report on the mummification practices of the Anga of Koke village, Central Highlands, Papua New Guinea. Yearbook Mummy Stud 1:11–17 Beckett RG, Conlogue GJ, Abinion OV, Salvador-Amores A, Piombino-Mascali D (2017) Human mummification practices among the Ibaloy of Kabayan, North Luzon, the Philippines. Pap Anthropol XXVI(2):24–37 Benguet website – Apu Anno. https://www.benguet.gov.ph/index.php/11-municipalities/398-thelegend-of-apo-anno-buguias. Retrieved 3 Aug 2019 Cardin M (2014) Mummies around the world. An encyclopedia of mummies in history, religion, and popular culture. ABC Clio, Santa Barbara De Leon L (1976) The mummies of Benguet. Philippine Panorama 8:52 Lapniten K (2019) Benguet community races against time to save Apo Anno. https://newsinfo. inquirer.net/1081535/benguet-community-races-against-time-to-save-apo-anno. Accessed 20 July 2019 Lewis MW, Wigen KE (1997) The myth of continents: a critique of metageography. University of California Press, Berkeley. ISBN 978-0-520-20743-1 Madulid DA (2001) A dictionary of philippine plant names, vol 2. Bookmark, Makati City Merino FS (1989) The Kabayan mummies and the Bendiyan Canao. The Author, Kabayan Morimoto I (1993) Mummies of the Ibaloi (Benguet-Igorot), Luzon, Philippines. In: Group for research of Japanese Mummies (ed) Worship of mummies in Japan and China. Heibonsha Limited Publishers, Tokyo, pp 369–407 Nobel J (2013) Death rituals of the Philippines. Digital dying. Retrieved 2017-07-01 Panganiban MB, Omana LT (1998) Geological and geomorphological perspectives of mummy caves of Kabayan, Benguet. Natl Museum Pap 8:2, 1–2,22

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Picpican I (2003) The Igorot mummies. A socio-cultural and historical treatise. Rex Book Store, Quezon City Piombinio-Mascali D, Abinion O, Salvador-Amores A, Beckett RG (2013) Human mummification practices among the Igorot of North Luzon. Bull Swiss Soc Anthropol 19(2):45 Provincial Summary (2016) Number of provinces, cities, municipalities, baraguays, by region. Retrieved from http://nap.psa.gov.ph/activestats/psgc/SUMWEBPROV-SEPT2016-CODEDHUC-FINAL.pdf Pungayan EL (1985) The Bendianan unsavored reminder of a buried Ibaloi past. Saint Louis Res J 16(1):90–111 Salvador-Amores A (2012) The recontextualization of burik (traditional tattoos) of Kabayan mummies of Benguet to contemporary practices. Human Diliman 9(1):55–94 Salvador-Amores A (2013) Tapping ink, Tattooing identities: tradition and modernity in contemporary Kalinga society. University of the Philippines Press and the Cordillera Studies Center, University of the Philippines Baguio, Quezon City

Smoked Bodies of Papua New Guinea

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Context and Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Smoked Body Mummification – Cultural Aspects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . When the Smoked Body Method Began . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . When the Smoked Body Method Ended . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Purposes – Past and Present . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Burial and Funerary Practices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Warrior Culture of the Anga . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientific Perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Smoked Body Mummification Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Smoked Body Science – Impact of Procedural Steps on Mummification . . . . . . . . . . . . . . . . . Smoked Body Mummification Limiting Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ethnographic Aspects and Current Thinking Among the Anga . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Current Thought Regarding the Smoked Bodies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Inter-village Politics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

This chapter describes the smoked body tradition of the Anga in the Aseki region of Morobe province, Papua New Guinea. The Anga are the only known living culture to practice anthropogenic mummification to the present day. The reader is provided with a background regarding the physical context of the Aseki region as well as the oral history context as related to the smoked body tradition. The Anga culture, formerly known as the Kukukuku, is described including their warrior ways of the not so distant past. This discussion includes descriptions of the role of R. G. Beckett (*) Biomedical Sciences, Bioanthropology Research Institute, Quinnipiac University, Hamden, CT, USA e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_34

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the smoked bodies in past and current contexts. Anga spiritualism is presented as it relates to the context of the smoked body tradition and its origins. Several possible reasons for the decline of the practice are also debated including the caveat of a modern smoked body mummification of a village elder in 2015. The chapter explores the cliff and rock shelter burial practices of the Anga including a description of varied and transitional burials throughout time. The purposes of smoked body mummification are explored as is the impact of missionary efforts to halt the practice. The chapter presents the smoked body mummification method as handed down through oral histories and from recent research. The chapter applies scientific reasoning as to the efficacy of the techniques employed. Limitations to the method are described with an assessment of taphonomic impact on the smoked bodies in various burial contexts. The chapter finishes with a discussion of the attitudes of living descendants as they relate to the smoked body tradition. Keywords

Smoked bodies · Mummification · Aseki · Anga · Kukukuku · Papua New Guinea

Introduction Context and Culture The focus of this chapter is on the smoked body mummification practice of the Anga culture in the Aseki region of Morobe province of Papua New Guinea (Fig. 1). The Independent State of Papua New Guinea is the eastern half of the Island of New Guinea. New Guinea is considered a part of Oceania which is a collective name for the islands scattered throughout most of the Pacific Ocean. The term includes the entire island regions of the Pacific Ocean between Asia and the Americas with some exceptions. New Guinea is in the southwestern part of the Pacific Ocean north of Australia and is divided into the eastern and western halves. The western half is a part of the Indonesian archipelagic and includes the provinces of Papua and West Papua. Papua New Guinea is considered a nation of tropical climate. However, there are variances determined by region. In the lowlands, the maximum high temperatures is around 90  F (32 ) with the minimum around 75  F (24  C) with little seasonal variation given its proximity to the equator. The coordinates for Papua New Guinea are between 0 and 12 south and 140 and 160 east. The topography and the wind systems (the trade winds and the monsoon) dictate the average annual rain fall. The highlands receive rains throughout the year totaling around 130 in. (plus or minus 30 in.) with the southward facing highlands receiving around 300 in. In contrast, the capital of Port Moresby may only receive around 50 in. per year which is usually in the months of December through March. The Aseki region receives about 100 in. of

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Fig. 1 Map of Papua New Guinea. Circle represents Aseki region in Morobe province (Credit: retrieved from the Open Database www.openstreetmap.org_copyright)

rain falling throughout the year. The humidity is between 70% and 90% on average at all times. Clearly these conditions are not conducive to preserving human remains. The area is home to a vast variety of trees, including garamut, nutmeg and fig trees, magnificent Araucarias, rosewoods, mahoganies, walnuts, and Pandanus pines, adorned with vines and creepers. Between 8 and 10 thousand feet, just up from the Fringe Highlands, the forest changes to oak, beech, bamboo, red cedar, and pine covered in mosses, lichens, and luminous fungi, surrounded constantly by swirling clouds of rain. The smoked body mummification method is an intentional act and a very active process requiring special knowledge and skill to stave off the tropical climatic characteristics of the region. Papua New Guinea has unparalleled diversity of butterflies, birds, plants, strange marsupials, crocodiles, 90 species of snakes including the taipan, death adder, and Papuan black python, and at least 3,000 varieties of orchids. More species are being discovered as researchers find their way into the deeper parts of the interior.

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Papua New Guinea gained its independence from Australia in 1975. The geography of Papua New Guinea, along with its tropical climate and rainfall, make travel very difficult and while the economy is improving, infrastructure lags behind most developing countries. The highlands consist of dense mountainous rainforests with the mountains creating a “spine” running the entire length of New Guinea. There are 20 provinces and two autonomous regions in Papua New Guinea. Each province is broken down into districts or regions. Within these regions, there is a district center with unknown numbers of villages spread throughout (Fig. 2). There are many unknown villages in the deepest parts of Papua New Guinea who are not beholding to any administrative region or concerns. The deep forests between the more worldly coastal regions and the highlands remain wild with tribal social systems by nature. Papua New Guinea has over 820 indigenous languages, many with unknown origins. Most of the languages have fewer than 100 speakers making communication between and among populations in remote regions difficult if not impossible. The official language of Papua New Guinea is Tok Pisin (New Guinea pidgin) which often affords some means of common communication. The physical and language impediments to communication factor into one of the purposes of the smoked body tradition in that the mummies themselves mark territorial limits of a given clan. The smoked body tradition is practiced by the Anga culture. The Anga, previously known as the Kukukuku, were once the most feared tribe due to their violent raids and prowess as warriors. Tribal warfare was prevalent with both

Fig. 2 Koke village in Aseki region. Smoke Bodies mark their territory (Credit: Ronald Beckett)

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unprovoked attacks as well as revenge “wars” on neighboring villages. The Anga are a collection of peoples who speak a number of related languages and inhabit the high mountainous areas of south western Morobe province. This area is known as the Fringe Highlands. The Anga wore only grass skirts and cloaks made from the beaten inner bark of local trees. This bark cloth is called tapa and is still used today. The Anga of the Aseki region speak Hamtai, their traditional language, as well as Tok Pisin. Yet within a short walk from their own village, many Anga cannot communicate because of dialectic variation or a completely different language being spoken. They then resort to gestures and sign language. The Anga, as with other tribes in the past, practiced cannibalism. Cannibalism was both ritualistic and important in their warring culture. Captured enemy combatants would be cannibalized as a form of intimidation. There are reports that the practice has continued well into the 21st century (Raffaele 2006; New Zealand Herald 2012). For millennia, the Anga culture has been based on subsistence living, relying on what nature can provide. They are expert hunters but they also grow crops of taro and coffee. While the reliance on the rain forest has been offset by missionary efforts and the encroaching world, hunting and scavenging practices remain to this day. The Anga would mummify their dead using a smoking method which will be described in detail later in this chapter. Although their process was unique to the region, other cultures in Oceania used smoke to preserve their dead as well. The smoked body method has the most similarities to the methods used in the Queensland, Cape York, and Torres Strait regions. The similarities in methods are most likely the result of the geographic proximity (Beckett et al. 2011a; Aufderheide 2003; Cockburn et al. 1998).

Smoked Body Mummification – Cultural Aspects When the Smoked Body Method Began The Anga did not record the passing of time. This may be due to the fact that seasonal differences were not readily perceptible or that it did not occur to them to mark time. Since there is no written record with their past being handed down through oral traditions and stories, it is not possible to determine when the practice of smoked body mummification began with any certainty. Thus, when the Anga are asked about how long ago the mummification practice began, they simply reply, “It is our way,” implying that they have always mummified their dead. The practice is woven into their creation myths and belief that “spirits” are as real as the living. The Anga legend of origin includes many references to the “spirit” world. This origin legend was told to the famed ethnographer Beatrice Blackwood (1939, pp. 209–242) in the Amatchime dialect and helps explain the sense of afterlife behavior of Anga “spirits.” “Once, a group of spirits known as ye ’ama (in the Amatchime dialect) were crying out beside a river. They saw a man coming down to the water, they caught him and threw him away into the bush. Then they

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went back to the water. The man’s wife came to look for him. They caught her too and threw her away into the bush. Now all our people who die turn into spirits (ye ’ama). The spirits of the dead can return in the form which they had in life or in the form of some other person to deceive the living.” Blackwood (1939) further explained that the Kukukuku (Anga) believed that there were two types of spirits or supernatural beings. In the first sense, the spirits are the ghosts of people who have died. They are intended to live in the ground yet they can come up and make themselves visible to men. They are most often in the form that they were when they passed away. These spirits like to sit in the branches of trees (likely the Pandanus tree which served as early “burial” sites among the Anga). These spirits are called yakova. The second type of spirits, called yakova in the Manki dialect and pahanga in the Nauti dialect, were held in awe as it was believed that they had the power to harm the living. Blackwood could not find a clear or concise connection between the yakova/pahanga and the deceased people of the village. Blackwood suggests that in one sense they were just like the others, that is people who have died, and in another sense they were regarded as never having been men. Given the many references to the “spirits” of the dead being able to impact the living, it may be surmised that the smoked body practice of mummification was in some way an attempt to keep the spirit among them as something tangible. The only hint of an afterlife construct among the Anga rests in the sense of ghosts interacting with the villages. The Anga believe that if the deceased is not provided with a “home” as in a mummified body, the spirit of the deceased causes havoc. This may take the form of deaths among the villagers, poor hunting, and poor crops. Another critical point is that they must “see the face” of the deceased as this is key to keeping the spirit connected to his mummified body. If the spirit can “see” his mummified face, he will return there at night (Fig. 3). Even in today’s culture, the Anga consult with the mummies with regards to hunting and planting as well as other concerns that arise in their interactions with those around them. The living family members talk to the mummies and they also want to “see the face” to make a stronger connection with the departed (Lohmann et al. 2016a). The mummies are like photographs for the Anga. The mummies are treated with reverence and are included in village ceremonies. When someone first meets a smoked body, they are introduced as if the mummified remains are still alive and able to comprehend the interactions afforded them.

When the Smoked Body Method Ended As difficult as it is to determine when the smoked body mummification tradition began, when it ended is also an elusive question. The reason for this ambiguity is that as recently as 2015, a village elder named Gemtasu requested that at his death he become the most recently smoked body to be produced. This was accomplished by his relatives and the villagers of Koke village in the Aseki region. Prior to his own

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Fig. 3 Being able to “See the Face” allows the spirit of the smoked body to find its way home and allows communication between the living and the smoked body spirit (Credit: Ronald Beckett)

mummification, Gemtasu was present when the body of his father was preserved by the smoked body method. Gemtasu knew the method first hand. In order for Gemtasu to be mummified, he needed to be assured that it was not against Papua New Guinea Law. Gemtasu was concerned as the missionaries told him that it was against the law but they were referring to God’s law. With the assistance of Ulla Lohman, a photo journalist and ethnographer, Gemtasu traveled to the coast to meet with a legal advisor. The advisor assured Gemtasu that there was no law in Papua New Guinea that would prevent Gemtasu from practicing traditional burial practices of the Anga, specifically the smoked body mummification method. Armed with this new knowledge, Gemtasu set about a plan to rekindle interest in the smoked body tradition. His plan was two-fold. First he wanted to be sure that the method was clearly understood. To accomplish this, he had his sons, relatives, and villagers learn and practice the smoked body method on at least two forest pigs. Researchers were present to document the procedure (Beckett et al. 2011a) in an attempt to better understand the Smoke Body method described in detail later in this chapter. Secondly, Gemtasu wanted help restoring the smoked body of his father named Moimango. Moimango was a honored village elder who was a

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powerful warrior and shaman of Koke village. Recall that Gemtasu was present for his father’s mummification. Gemtasu asked for assistance and in 2008 a research team traveled to Koke to assess and conserve the smoked body of Moimango (Beckett et al. 2011b). The team examined Moimango and devised methods to conserve the mummy using only materials available to the villagers of Koke. The sap from the Kumaka tree served as an adhesive allowing the surface skin of the mummy to be repaired. The Kumaka sap was smoothed using a glowing ember held close to the surface enhancing the sealing and adhesive nature of the material. Tapa, a textile-like material made from the inner bark of a Mulberry tree (Barker 2008), was used to patch several openings in Moimango, one being made by vermin. Bilum twine and bush rope was used to secure those parts of Moimango needing support. Lichen had started to grow on the toes of the smoked body. The team tested many jungle materials and found that Suca, a substance made from lime used by the Anga to offset the bitter taste of betel nut when chewed, had a pH of ~12. The Suca eradicated the lichen without harming the underlying mummified tissues (Fig. 4). Restoration efforts were re-evaluated in 2010 finding that the 2008 efforts were effective in conserving Moimango. The areas treated for lichen showed no new invasions. Readers desiring greater details with regards to the restoration project are

Fig. 4 Lichen eradication during the restoration of Moimango. (4.1, 4.2) Lichen present, (4.3) Suca (pH~12) application, (4.4) lichen removal (Credit: Ronald Beckett)

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referred to Beckett et al. (2011b) and Beckett and Nelson (2015). Throughout the process, relatives and villagers were actively involved in the restoration procedures which helped Gemtasu restore interest in the smoked body tradition. It is important to remember that Papua New Guinea is one of the most remote places on earth. As of this writing, there are places that are yet to be seen by foreign eyes. The jungles are extremely dense and it is speculated that many of the old traditions, including the smoked body method of preserving the dead (or some variation), are still practiced. It is known that the tradition waned with the influx of Western missionaries who encouraged the adoption of inground burials stating that the smoked body method and the display of the mummies on cliff faces or rock shelters was “unclean” and against the teachings of the missionaries religious beliefs. Burial and funerary practices of the Anga will be discussed later in this chapter.

Purposes – Past and Present A spiritual connection is the major purpose for mummification of the honored deceased. As described previously, there is a great degree of spiritualism associated with the smoked body mummies. The traditional Anga have no afterlife construct yet they do believe in a “ghost” world after death. The living derive comfort in having their deceased around and they derive a power of sorts from the mummies. The spiritual connection is lost if the bodies deteriorate to the point where the person they once were can no longer be recognized. The second purpose assigned to the mummies is that they represent a physical marker of the territory occupied by the clan or village. While this seems to be an old tradition, it is still present today. Native Papuans are allowed to maintain their ancestral lands and the mummified remains, often overlooking a village, helping that village maintain their territory. The living are able to say, “that is my ancestor, we have been here, this is our place.” This is an important function of the smoked bodies as inter-tribal disputes regarding land boundaries are often settled by the presence of smoked body mummies. An emerging purpose of the mummies is one of economics. Several of the villages, even in the fringe regions, have started rudimentary businesses of mummy tourism. Adventure seekers are willing to travel to these out of the way places and pay to see or photograph the smoked body ancestors. There are several locations where rudimentary mummy tourism businesses have sprung up in the Aseki region. Of these, the most accessible is the village of Angapenga which has fourteen smoked body mummies. The mummies can be viewed and photographed for a fee. Yet the hike up to the traditional placement of these smoked bodies in a rock shelter above the village can discourage some who would like to see them. Another location is in the village of Oiwa. Here three mummies are kept in a locked tin shed along with other artifacts (Fig. 5). Again, there is a fee for entrance and a separate fee for photographs. The cliff face above Koke village holds the remains of 18 individuals in various states of deterioration. The gallery at Koke is the most difficult to get to.

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Fig. 5 Tin shed at Oiwa village. A fee is charged to see and photograph the smoked bodies held there (Credit: Ronald Beckett)

Burial and Funerary Practices There is considerable variability in the funerary rituals practiced by the Anga. Most ethnographic sources represent the Anga’s treatment of the dead as a monolithic practice by generalizing from single sites or events to the cultural group as a whole. However, it is clear that the practice of smoking the bodies and displaying them in galleries as demonstrated in the Aseki area is different from that described by Blackwood (1978) for the Upper Watut area and by Mbaginta’o (1976) for the Dunkwi Anga, where bodies were only partially smoked and were placed in trees. Variability was also observed within the Aseki area as some individuals were mummified, others were not, some were placed in galleries with several others, while others were placed on a cliff face by themselves and yet others were placed in a cave. The evolution of burial practices can be seen the many of the villages. The oldest known burials were those in which the smoked body was place in the basket-like branches of a Pandanus tree. At times, additional strappings made from strips of bamboo were used to secure the individual to the branch nest. Following the basket burials, traditions shifted to group presentations of smoked bodies. These groups were no longer in tree branch nests but placed in high places overlooking their native village. The groups were placed on benches fashioned from native trees with rows of benches seen at times. Following the bench burials individual “chairs” made of branches and tapa became the display style of choice.

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These chairs were placed high on the cliff and secured with other poles or branches. In a way the chair burial, being an individual presentation rather than a group display, may have been a return to the individual basket style burials. In all burial styles, it is common to see artifacts, such as bundles of arrows, a broken arrow, a bow, or spear, as inclusions with the individual. These items likely belonged to the individual and were representative of that person’s prowess in hunting or warfare. From the perspective of funerary ritual, one of the more interesting burial practices seen in Koke village is the demonstration of different burial styles that represent a culture in transition. There not only are the traditional basket and bench style smoked body burials and newer “chair” or seated burials found, but also wooden coffins representing the arrival and influence of the missionaries to the region (Fig. 6). The villagers say that the missionaries decreed that the smoked body practice was against God’s law and that they needed to place bodies in coffins and bury them in the ground. So the Anga of Koke smoked the bodies, placed them in coffins, but then placed the coffins there on the same cliff with the other smoked bodies. Now the Anga of Koke bury their deceased in coffins but in the consecrated ground of the village cemetery. Thus, the burial presentations on the cliff are an excellent example of funerary practice transition as influenced by external forces. The in-ground burials encouraged by the missionaries are in stark contrast to the traditional beliefs of the Anga. In some ways, this insistence on burying the dead in

Fig. 6 Variety of burial styles at the Koke smoked body gallery showing burial tradition in transition. Note the basket (a), bench (b), chair (c), and coffin burials (d) (Credit: Ronald Beckett)

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the ground has led to divisions among the Anga. While many have adopted the “new ways,” others have questioned the burials and are determined to live in a manner true to their traditions. For the traditionalist, an in ground burial creates an imbalance. They feel that once the earth tastes human blood, it will crave more. If the earth does not receive more blood, it will no longer provide for them in terms of the subsistence they require to survive. Related to the topic of transitions is that of a move toward a monetary system imposed by the missionaries and indeed the local municipalities. This is again in contrast to the subsistence hunting and farming practiced by the Anga. The transition has been difficult and it is not unusual to find a villager disappear for several days to hunt in the jungle while others gather local coffee beans to be sold at markets miles away.

The Warrior Culture of the Anga There is considerable evidence that the Anga (Kukukuku) were a violent group. In the early 1900s, near the time of first contact, the Anga were recognized as fierce warriors. A post was established “with the object. . .of pacifying the truculent little semi-nomads. . .name of Kukukuku, . . . (1908:17)” (in Bamford 1997). Into the early 1950s, the Anga maintained wars with bordering groups to the north and west as well as amongst themselves. Under Australian rule in this time period, efforts were made to contain the war-like stance of the Anga. Raids still persisted. Then around 1951, missionaries began their conversion works which by the 1960s had brought reasonable peace among the Anga with ever increasing numbers becoming Christians. The propensity toward warring is grounded in the Anga concept of Jerungdu which is the ultimate expression of what a male is to be in body and spirit (Herdt 1987). Initiation rites were harsh for boys between 9 and 13 years old often involving violence. This rite was used to instill the strength to do battle and to affirm the power of masculine traits. Status within the Anga warriors was established and maintained through repeated feats of courage and prowess in battle. If there was some minor insult from a neighboring village, a “little battle” would ensue. Little battles were a more ritualistic battle in that arrow volleys were exchanged at a great distance. These could grow into “big battles” where great numbers of warriors would mount a charge on their enemy using not only arrows but clubs and axes as well. Other than little battles, illness thought to be brought on by sorcery, assassination of a “leader,” theft of garden goods or pigs, could all escalate into a big battle. The weapons used included the bow and arrow, bamboo knives, and wooden or stone clubs. All were very effective in battle. One tribesman said that you cannot get out of the way of a Kukukuku arrow. If you hid behind a tree, the arrow would find you there. While the ethnographic evidence clearly paints a picture of a warring people, a study of smoked body mummies and skeletal remains of the Anga conducted by Nelson and Beckett (2011a) provides ample bioarchaeological evidence for violence and warfare. In the study, researchers examined 32 remains

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Fig. 7 Trauma was not uncommon among the warring Anga. This penetrating skull fracture shows signs of healing (arrow) indicating that the individual survived the initial insult. (Koke gallery K-8) (Credit: Ronald Beckett)

from three different sites in the Aseki region. The Koke village smoked body mummy gallery, the tin shed mummies of Oiwa, and the remains in the Imauka cave. Of those 32, 11 had enough exposed skull or remaining crania to make an assessment with regards to cranial trauma. Five of the 11 had depressed or penetrating skull fractures (Fig. 7). Some showed signs of healing indicating that the individual survived the insult. In addition, two mummies from the Oiwa tin shed group showed extensive perimortem slash wounds to the skin of the face and torso (Fig. 8). Associated artifacts with these burials included arrows, bows, and spears. In the village of Koke, researchers were allowed to examine stone axes of the type that could inflict such cranial trauma. Given that this bioarchaeoological evidence was not restricted to a single site, researchers further concluded that the weapons used (stone clubs) were more consistent with descriptions of “big battles” and that some individuals did survive as evidenced by new bone growth and bone “healing” at the injury site. The bioarchaeological evidence from this study was consistent with the ethnographic reports regarding the violent nature of the Anga. Violence was clearly a part of life and death for the Anga. The concept of Jerungdu, the continual need to demonstrate one’s maleness, was a driving force for the life of the Anga. Those warriors who distinguished themselves in battle or who were a renowned hunter or otherwise served the village in a meaningful way were treated with honor and respect after death by being mummified with the smoked body method. Recalling the spirituality of the Anga, there was also likely a fear that if the body was not preserved in this manner, that a “ghost” of this powerful warrior would cause

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Fig. 8 Trauma: In addition to skull fractures, the Oiwa tin shed smoked bodies have multiple slash wounds that were stitched using bamboo. Arrows indicate slash wounds on face (a), torso (b), and on top of the head (c), each with bamboo stiches (Credit: Ronald Beckett)

trouble for the living. It was not only men who were mummified but the families of those men may also have become smoked bodies and placed on the cliff face above the village. Women and children are seen among the smoked bodies in the mummy galleries. Perhaps this was an attempt to appease the deceased powerful warrior/ hunter or it may have been intended to further strengthen a clan claim to the territory. Regardless of the reasons, the smoked body method was effective and ingenious in that preserving a human in this wet and humid environment required a great expenditure of time and resource. The smoked body method is an active method and although we cannot know when it began, we do know that the tradition is continuing to a small degree. The mummy makers knew how to outrun the intent of nature which is to decompose all that dies.

Scientific Perspective Smoked Body Mummification Procedure Several studies have gathered considerable data regarding the smoked body method (Beckett et al. 2011a; Beckett and Nelson 2011; Lohmann et al. 2016b). These studies involved direct interaction with villagers and the oral history as related by a village

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elder named Gemtasu who helped mummify his father and hopes to inspire the clan to continue the smoked body tradition by being mummified himself. Additionally, the researchers learned along with the villagers the smoked body method through experimentation on two occasions using forest pigs. The village elder, Gemtasu, passed away in 2015 and was mummified according to the smoked body tradition. Gemtasu had lost his wife some months earlier and did not eat or drink many days before he passed away. It is thought by the villagers that this was an attempt to make it easier for everyone in the village. The method was documented and what follows is an accurate reflection of the procedure. It is important to note that this is perhaps one of the only witnessed mummifications within the natural cultural setting. The experimentation and the mummification of Gemtasu occurred in Koke village.

Step 1 The body is viewed for three to five days prior to active intervention. During this time, the body begins to swell as is seen in late stage one and early stage two decomposition. During this period, a location is chosen and a ceremonial smoking hut is constructed using local materials including palm leaves and bamboo (Fig. 9). The three to five day period also serves as a time for mourning. In the case of Gemtasu, seven men had been selected to perform the mummification ritual. A vegetable garden is planted to provide food for the seven men. The men are not allowed to drink except juice from sugarcane and may only eat food prepared on the

Fig. 9 Special smoking hut is constructed for the smoked body mummification method (Credit: Ronald Beckett)

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smoking fire. They may not leave the area. A chair in which the body will sit is constructed. This is one of two that will be used.

Step 2 A broken arrow ceremony is conducted. The body is undressed and secured to a “chair” in seated posture and a smoky fire made from Tomunga wood is started and maintained in the hut. The chair with the body is suspended above the fire and secured with bush rope made from strips of bark (Fig. 10). After one week, the epidermal layer and hair are scraped with a Kuyeka bristle plant and edged bamboo. The Kuyeka plant serves to both express and absorb body fluids. Body fluids continue to be expressed through the skin and ritualistically rubbed onto preparers’ own skin. This practice represents intimacy with the dead and continued mourning. The skin is punctured with sharpened bamboo in the gravity dependent portions of the body enhancing body fluid removal. The body fluids are collected. Step 3 The smoky fire is continued for around 30–60 days with constant attention and is maintained until the body dries. The seven men live in the smoking hut throughout the process, take their meals there, and cannot wash. An occasional change in body location within the hut ensures all areas of the body are exposed to the smoke and heat. Once the body is dry enough, a new chair, a Hangowa, is made. Ceremonial red ochre clay is applied to the smoked body as well as to the faces and bodies of the preparers. The red ocher clay applied to the deceased acts as a desiccant and draws more fluids up and out of body. The men continue to manually express (massaging and milking) remaining fluid from the body and apply it to their own skin. If needed, Kuyeka plant is again used as well as tapa to absorb any remaining fluid. Villagers report that the body fluids may be drunk and used to garnish taro root that are heated by the stones of the fire ring prior to eating. A stick is passed through Fig. 10 A forest pig is suspended over the smoky fire in a “chair” similar to what a human would sit in. The position helps bring the body fluids to the gravity dependent areas for removal (Credit: Ronald Beckett)

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the rectum to assure that the intestines are not retaining any fluids. The fluids collected from the intestines are collected in a bag.

Step 4 When the body is completely desiccated, preparation for its placement among the smoked body ancestors is begun. The mummified smoked body in its seated position is further secured to the chair made from local branches and bush rope. The body is dressed in traditional style including a grass skirt and tapa head cape. A ceremonial application of red ocher clay is the final step (Fig. 11). The red ocher clay may serve to further dehydrate the body and offers some protection from the environment. Step 5 A ceremonial procession of the smoked body is conducted as it is placed on a limestone cliff ledge among other ancestors overlooking the village.

Smoked Body Science – Impact of Procedural Steps on Mummification In order for a body to decompose, natural postmortem conditions need to be met. The body’s natural intracellular process of autolysis, the enzymatic activity of indigenous and exogenous bacteria, and larval insect infestation all contribute to the decomposition of organic remains. Each of these processes requires an aqueous environment. From a scientific perspective, the smoked body method is successful in that it controls three major variables associated that would halt

Fig. 11 Ochre clay application is both ritualistic and may serve to further draw fluids from the body. The clay offers some protection to the smoked body as a physical barrier (Credit: Ronald Beckett)

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decomposition. Those three include the creation of a micro environment (hut), elimination of body fluids (dehydration), and chemical impediment to bacterial activity (smoke chemistry).

The Smoking Hut Construction of a ceremonial smoking hut: As described, the environment in all of Papua New Guinea is tropical. The wet and humid forests are not conducive to body preservation. The smoking hut produces a concentrated microenvironment protecting the remains from ambient moisture and temperature fluctuations while serving to enhance the preservation characteristics of the smoke and fire. The impacts of smoke and fire are discussed below. During the initial 3–5 day waiting period, the normal forces of decomposition begin as the body is exposed to the ambient conditions. This is a vulnerable period for the mummy as preservation will be challenged. Elimination of Body Fluids Beyond this waiting period, many of the procedural steps enhance dehydration of the body. The slightly abrasive Kuyeka plant helps to open the pores of the skin reducing the impediment to fluid escape from the body. If initiated early enough, as in the late stages of stage 1 or early stage 2 decomposition, fluid removal is enhanced. The body being placed in a seated position helps body fluids migrate to the gravity dependent areas. Here the sharpened bamboo punctures along with the manual expression accelerates body fluid removal (Fig. 12). The stick passed through the rectum helps to remove fluids that had been retained in the gastrointestinal tract. The use of tapa and Kuyeka plant also helps absorb the fluid leaving the body. These procedures occur prior to as well as after the body has been placed into the smoking hut. All of these procedures enhance and expedite dehydration. In doing this, reduction in the aqueous environment decreases the activity of the enzymes of Fig. 12 Manual expression of fluids (massaging and milking) from the large muscle groups of a forest pig (Credit: Ronald Beckett)

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decomposition. Additionally, the Kuyeka plant has a slightly alkaline pH (like soap). When applied to the body surface, it produces a hostile environment for bacterial enzymes.

Impact of Smoke and Fire Within Smoking Hut The fire and smoke are key factors in the smoked body process. Four factors associated with the fire and smoke within the smoking hut are critical to body preservation and ultimate mummification. These factors include temperature, humidity, smoke chemistry, and physical barrier. Dehydration and bacteriostasis are greatly enhanced by the increased temperature within the smoking hut. According to researchers (Beckett and Nelson 2011) the measured temperatures inside and outside of the smoking hut varied dramatically with temperatures at the body surface within the hut of 51.7  C (125  F) while outside of the hut the temperature was 22  C (71.6  F). The elevated temperature effected rapid evaporation of fluids while at the same time creating a temperature that was not conducive to bacterial survival reducing the enzyme activity. Beckett and Nelson (2011) further report that the humidity within the smoking hut is reduced as well decreasing the amount of moisture available for decomposition. In their study the inside of the smoking hut exhibited a relative humidity of 55% while outside of the hut the relative humidity was 83% (these variable of course depend upon the ambient conditions of a given day). Essentially, the smoking hut is an artificial microclimate including heat and lower humidity, aided by smoke. The smoke chemistry plays a critical role in the smoked body method. The pH of the smoke is quite acidic at 5.5. This pH is very hostile to any bacterial activity. While working in the smoking hut, the acidity of the smoke impacts the preparers as the smoke affects the mucous membranes of eyes, nose, throat, and lungs which begin to inflame and burn on immediate contact (Fig. 13). This level of acidity, mixing with any body fluids, creates an environment which would kill or render Fig. 13 The interior of the smoking hut is very hostile environment for bacteria, insects, and for those who are preparing the smoked body (Credit: Ronald Beckett)

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nonfunctional any surface bacteria that may normally aid in decomposition. The bacteriostatic or bacteriocidal impact of the acidic smoke is likely an important factor enhancing the efficacy of the smoked body mummification procedure. The smoke also contains traces of formaldehyde which alters tissue and molecular structures essentially “tanning” the organic material which in turn expels water and resists rehydration (Vollhardt and Schore 2006; Aufderheide 2003). There is a reduction in the activity of the enzymes of decomposition as the enzymes are incapable of digesting the new substrate. Combined with the bacteriostatic nature of the smoke, the presence of formaldehyde greatly enhances the potential for mummification.

Physical Barrier Finally, the smoke acts as a physical barrier to insect infestation. The combined characteristics of the fire (elevated temperature, decreased humidity), smoke (pH and formaldehyde), expression of fluids (enhanced dehydration), and physical impediment to flies and other pests all disrupt any larval life cycles that may have begun during the waiting period. Once mummification is achieved, the application of red-ochre clay, while generally ceremonial, has desiccant characteristics. Further the clay serves as a physical barrier to the mummy’s constant environmental exposure on the cliff niche thereby aiding the continued preservation of the bodies.

Smoked Body Mummification Limiting Factors The smoked body method is an effective process that staves off decomposition and preserves the human body very well. Internal organs are found intact in the bodies with their size reduced through dehydration. Given that the environment of Papua New Guinea is one that is warm, hot, humid, and wet, it favors rapid decomposition making smoked body mummification an even greater challenge. Once the mummy is created, the major factor in determining its continued state of preservation is the location where it is displayed. Taphonomic forces play a major role in the preservation of the smoked bodies with taphonomy being influenced by how and where the bodies are displayed.

Taphonomic Change/Impact on Smoked Bodies In the Beckett and Nelson study, smoked bodies were examined at several sites in the Aseki region. Oiwa tin shed mummies were protected from the environment while the mummies from Koke village were exposed to the elements. Researchers have been able to examine and compare the external and internal state of preservation and consider the impact of taphonomic change in these and other varied environments (Beckett and Nelson 2011). In this study, comparisons of the taphonomic characteristics of remains in the Imauka cave and the Yeakunga cliff were also made. Extensive external observations as well as assessment of the internal state of preservation using the paleoimaging method of field endoscopy were employed. As one might expect,

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Fig. 14 Preservation variations can be seen among the various burial display sites. The Oiwa tin shed mummies are the best preserved (a), the Koke gallery shows a greater degree of deterioration due to environmental exposure (b), and complete skeletonization is found at the Imauka cave (c) (Credit: Ronald Beckett)

the mummies in the tin shed who have been protected from the environment for around 40 years had a superior state of external preservation (Fig. 14). Internal preservation including recognizable organ systems was superior among the Oiwa tin shed mummies when compared to the remnants of internal organs seen within the Koke mummies. Only one particular Koke mummy did have better internal preservation than the others of the same gallery but not to the degree found in the Oiwa tin shed mummies. While these data inform us regarding the taphonomic changes seen in the exposed mummy group, the data are also a testament to the exceptional preservation impact of the smoked body mummification method. At another site in the Imauka cave, the exposed burials were all skeletonized burials There are several possible explanations for this variance: (1) the Anga did not smoke these bodies and the Imauka cave represents an alternate burial practice that excluded mummification, (2) extreme deterioration of smoked bodies with no remaining soft tissue evidence, or (3) a rapid cliff burial in a time of war with no time to smoke the bodies and, upon returning to the cliffs, found the bodies either in an advanced state of decomposition or already skeletonized. There is clear evidence at the Imauka cave of organized placements with one burial including two individuals with cultural artifacts including two bundles of arrows, at least one spear and a bow. The Yeakunga cliff mummy showed evidence of having gone through the smoked body mummification process as some soft tissue is evident. Given the exposure and rather shallow cliff niche burial, deterioration is advanced demonstrating the taphonomic power of the harsh environment.

Miroclimates in the Burial Cliff Galleries Another factor related to the continued preservation status are the conditions specific to the burial display. At the smoked bodies mummy gallery of Koke, rainfall, wind, temperature, and humidity were analyzed and compared to the same variables in the

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village proper 1077 ft (328.2 m) below. The rock shelter or cave niche does in fact provide some protection from the elements. Researchers found that the cave niche stayed completely dry while 2 m out from the back of the niche, and in Koke village below, torrential rains fell (Beckett and Nelson 2011). At one moment there may be no wind that would have driven the rain into the cliff niche yet a major wind and rainstorm could certainly drive rain into the niche and onto the mummies. There are variances in the other parameters as well. Temperature, precipitation, and wind are all significantly less in the cave niche while relative humidity was about the same, only slightly less at the cave niche (Fig. 15). The researchers realize that a longitudinal study still needs to be conducted but state that the microclimate study was a “snapshot” that was measured and observed. It suggests that the cave niches did serve as a microclimate, thereby increasing the preservation potential for the smoked bodies. Elemental characteristics of the Koke and Imauka cave niches have been established using x-ray fluorescence. The Koke cave is a limestone matrix with embedded cobble. Limestone is hydrophilic and provides the cliff niche with a slight decrease in ambient humidity adding to the consistency of the mummy gallery environment. The Imauka cave is a sandstone matrix which also has some desiccant characteristics. More than likely, the Anga selected the burial sites with two things in mind, marking their territory and the convenience of the site. The burial site then determined the degree of deterioration of the smoked bodies over time due to the site

Fig. 15 Assessment of the weather conditions. Here a weather station is set up at the Koke smoked body gallery high above Koke village. Another unit was set up in the village proper below (Credit: Ronald Beckett)

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offering a reduction in taphonomic impact as well as the presence of a microclimate which would provide additional protection.

Ethnographic Aspects and Current Thinking Among the Anga The smoked bodies serve three major functions in the regional Anga culture. First, there is the spiritual connection with the mummified ancestor. The face of the individual holds their spirit. Recall that the Anga held no afterlife constructs prior to arrival of the missionaries in the 1950s. Their cultural custom was to preserve the body so the face could be seen and the living could continue to communicate with the ancestor and include them in village affairs. Second, the Smoked Body Galleries serve as a cemetery for particular lineages and as territorial markers for those lineages. Many of the villages and their boundaries are delineated by geological structures called “stones.” For example, there is a Koke stone and a Windgea stone identifying the village/clan boundaries. The smoked bodies are placed on or around these stones high up on cliff faces to further demonstrate that village’s land claims (Fig. 16). Third, the smoked bodies are more recently being seen as a source of revenue for the village. A mummy viewing, photographing, and studying industry is blossoming in the bush country of Papua New Guinea. Consider these examples: The tin shed mummies of Oiwa are kept locked up until fees for seeing, photographing, a specific time limit, and a fee for any interviews with the mummy keeper are agreed upon and paid. There is a vested interest in the restoration of the mummies in an attempt to develop and attract mummy

Fig. 16 The smoked body mummy gallery as timeless sentinels overlook Koke village below marking their claim to the territory (Credit: Ronald Beckett)

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tourism. Gemtasu of Koke was actually mummified using the smoked body method when he died in 2015. Prior to his death Gemtasu’s son Yangdeng presented a fee schedule for the upcoming “event.” Neatly drawn up, Yangdang developed an agreement with prices broken down for attending the mummification, for filming the mummification, and for services rendered by the villagers during the mummification. One might think that this enterprising concept was brought on by the missionaries and the advent of the change from subsistence living to a monetary value system. There is ample evidence that the indigenous populations of Papua New Guinea are in fact natural capitalists and have been prior to first contact. Blackwood (1978) states that in the 1930s after receiving permission to take a photograph in the Anga region, she was asked for compensation for the photo. Leopold Pospisil (1978) who lived among the Kapauku of West New Guinea in the 1950s reported that they had a carefully considered value system which included pigs, crops, wives, and shell currency. So it really is not surprising nor is it correct to assume that what you will experience with regards to negotiations is any different from what early ethnographers discovered. Indeed the current shift to a monetary system does have an influence but to the villagers themselves it is totally unnecessary as many villages still choose to live “off the jungle.”

Current Thought Regarding the Smoked Bodies Ethnographic data demonstrates that there appears to be generational variance regarding the cultural significance of the smoked bodies. The elders believe that their customs and rituals have spiritual importance. The next generation holds some residual respect for the older traditions but see the value in developing a mummy tourism enterprise. The younger generation, particularly those who have seen the modern world, feel that the tradition of smoked body mummification will necessarily fade. It appears that the younger generation’s interests are more in-line with the encroaching ways of the Western World, although they were also interested in the economic aspects of mummy tourism. With that said, there are many younger villagers who exhibit a great deal of respect for their elders and for the smoked bodies till this day (Nelson and Beckett 2011b).

Inter-village Politics It is beyond the scope of this chapter to delve into the complexities of the rivalries, concerns, and relationships among and between villages. However, it is very apparent that land disputes and territorial “maneuvering” continue to this day. Inter-village relationships in nearby regions are contentious at best. Local Aseki magistrates help a great deal in the ensuing negotiations between and among villages regarding a variety of issues including village boundaries, distribution of goods, “road” construction, taxation, and mummy tourism or research. As an

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example of these tense inter-village relationships, porters from Aseki may refuse to enter the disputed lands of nearby villages with the fear of being shot with an arrow. If negotiations were successful, porters are only willing to carry equipment to a certain point. Porters from the other village would then come out to that point a retrieve the gear. One has to keep in mind that the farther out one travels from municipal regions, the more intense inter-village relationships can become. Any intervening from local magistrates may place all involved at great risk.

Conclusion The Anga reside in the remote highland fringe of the Morobe province in Papua New Guinea, a region known to be rich in biological diversity yet still with little known about the culture of its inhabitants. While the jungle creates a natural geographic barrier to the outside world, the Anga have been challenged to question their cultural beliefs by the encroaching process of globalization and the influx of western missionaries. As the Anga struggle to shift from a subsistence economy to a monetary value system, their unique cultural, religious, and mythological traditions are beginning to diminish and are at risk of vanishing altogether. At the same time, the Anga are hoping to reaffirm their cultural identity. Of these cultural practices, their method and ritual of smoked body mummification may be in its final stages. This is the only place on Earth where a living culture is a single generation away from the practice of mummification and who have conducted the smoked body mummification method recently to one village elder, Gemtasu (Lohmann et al. 2016b). Other cultures that practiced mummification such as in Egypt and Peru are “dead” cultures in that there are no recent direct attachments to the preserved remains other than ancestry and cultural pride. In contrast, on the fringe of the Eastern Highlands of PNG the living people, the Anga, are the people who made the mummies that mark the territory of their clans. This chapter has offered the reader a glimpse into the Anga culture and their unique mummification method. The chapter also presented an assessment of the direct relationship between the smoked body preparation and ritual and the living culture of the Anga. The smoked body method is a very effective means of mummification in an environment that begs to decompose human remains. The Anga have practiced this method for as far back as the oral history reaches. As expressed in Gemtasu’s recent mummification, there is a clear desire among the Anga to carry on the tradition as they move forward in time.

Cross-References ▶ Fire Mummies of the Kabayan Region of Benguet Province, Luzon, the Philippines

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References Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Bamford SC (1997) The containment of gender: embodied sociality among a South Angan People. Dissertation, Department of Anthropology, University of Virginia Barker J (2008) Ancestral lines. Broadview Press, Peterborough Beckett RG, Nelson A (2011) Scientific exploration of the smoked body mummification practice of the Anga of Koke village, Papua New Guinea. Presentation at the 7th world congress on mummy studies, San Diego, June 12–16 Beckett RG, Nelson A (2015) Mummy restoration project among the Anga of Papua New Guinea. Anat Rec 298(6):1013–1025 Beckett RG, Lohmann U, Bernstein J (2011a) A field report on the mummification practices of the Anga of Koke Village, Central Highlands, Papua New Guinea. Yearbook of mummy studies, vol 1, Dario Piombino-Mascali (guest editor) – Accademia Europea di Bolzano, Bolzano. ISBN 978-3-89937-137-6. EURAC, Bolzano, June, pp 11–17 Beckett RG, Lohmann U, Bernstein J (2011b). A unique field mummy conservation project in Papua New Guinea. Yearbook of mummy studies, vol 1, Dario Piombino-Mascali (guest editor) – Accademia Eurpea di Bolzano, Bolzano. ISBN 978-3-89937-137-6. EURAC, Bolzano, June, pp 19–27 Blackwood B (1939) Life on the upper Watut. Geogr J 94(1):11–24. Blackwell Publishing, UK Blackwood B (1978) In: Hallpike CR (ed) The Kukukuku of the Upper Watut. The Pitt Rivers Museum, Oxford Cannibal cult members arrested in PNG (2012) New Zealand Herald. July 5th. ISSN 1170-0777. Retrieved Nov 2015 Cockburn TA, Cockburn E, Rayman T (1998) Mummies, disease, and ancient cultures, 2nd edn. Cambridge University Press, Cambridge UK Herdt G (1987) The Sambia: ritual and gender in New Guinea. Holt, Rinehart and Winston, New York Lohmann U, Beckett R, Nelson A, Piombino-Mascalli D, Lywood V (2016a) See the face: a living history of the cultural/spiritual implications of mummification practices among the Ibaloy of the Kabayan Region of the Philippines and the Anga of Papua New Guinea. Presentation at the 9th world congress on mummy studies, Lima Lohmann U, Beckett R, Nelson A (2016b) A modern cultural mummification. Gemtasu’s final wish and the smoked body tradition of the Anga of Papua New Guinea. Presentation at the 9th world congress on mummy studies, Lima Mbaginta’o IG (1976) Medical practices and funeral ceremonies of the Dunwki Anga. J Soc Ocean 32:300–305 Nelson A, Beckett R (2011a) The warrior ways of the Anga, trauma and artifact analysis. Presentation at the 7th world congress on mummy studies, San Diego, June 12–16 Nelson AJ, Beckett RG (2011b) The meaning of mummification among the Anga of the Aseki region of Papua New Guinea: A tradition in transition. Presentation at the 7th world congress on mummy studies, San Diego. June 12–16 Pospisil L (1978) The Kaapauku Papuans of West New Guinea. Holt, Rinehart, and Winston, Orlando Raffaele P (2006) Sleeping with cannibals. Smithsonian Magazine, September Vollhardt KPC, Schore NE (2006) Organic chemistry: structure and function, 5th edn. W.H. Freeman and Company, New York

Part VIII Mummies in Asia

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Mummies in Siberia Sergey Mikhailovich Slepchenko, Jong Ha Hong, and Dong Hoon Shin

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Frozen Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies of Ukok Plateau . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies of Oglakhta (Tashtyk Culture) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Yakut Frozen Bodies Found in East Siberia . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . West Siberian Arctic Graves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preservation of Siberian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientific Analyses of Siberian Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Despite the academic works of Siberian mummies over the past decades, except for the Pazyryk mummy (Siberian Ice Maiden) in Ukok Plateau, the academic achievements are not well known to the outside world. Nevertheless, the discovery and scientific research of mummies therefrom continue to be performed in the S. M. Slepchenko (*) Institute of the Problems of Northern Development, Tyumen Scientific Center of the Siberian Branch of the Russian Academy of Sciences, Tyumen, Russia Surgut State University, Surgut, Russian Federation e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_27

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artic or subarctic zones of Siberia. In this chapter, the authors summarize the brief history and recent research updates of Siberian mummies discovered under different geological and environmental conditions in the Russian Federation. This mummy research in Siberia, as in other countries, vividly restores the lives of ancient or medieval people at a place that could not be completely revealed by any historical, archaeological, and ethnographic studies. Keywords

Siberia · Mummy · Ukok Plateau · Pazyryk · Zeleny Yar · Histology · Preservation

Introduction Many important scientific advancements have been attained through the study of many famous mummies worldwide. As seen in the mummies of ancient Egypt, Peruvian Inca, or Europe, etc., such studies have given us important academic knowledge about the actual lives of those peoples who lived in the past. Over the past decades, however, mummy discovery and research has also been performed in the areas where mummy cases were rarely reported before. In fact, the more information about the mummies we accumulate, the deeper our understanding of ancient or medieval peoples and societies grows (Aufderheide 2003; Cockburn et al. 1998; Lynnerup 2007). Since these discoveries of mummies were novel and inspirational, the findings attracted much attention of relevant researchers in the world. Actually, Siberian mummies of the Russian Federation are less well known compared to those mummies of other countries and continents. Except for a famous Princess of Ukok or the Siberian Ice Maiden, a mummified female that made a huge impact all over the world, people still don’t know much about mummies discovered in Siberia. Nonetheless, the Russian academic tradition of mummy studies in Siberia is by no means less rigorous. Over the years, a number of researchers were involved in various studies of Siberian mummies, thus providing invaluable information about ancient or medieval peoples who lived in the area and buried at the nearby cemeteries (Fig. 1). In this chapter, we summarize the brief history and recent updates of the Siberian mummy research reported to date.

Frozen Mummies Most Siberian mummies reported so far are frozen mummies found in a permafrost area. In general, the permafrost mummies were naturally formed by the action of very low temperature and high speed wind, etc. The removal of water from the tissues induced by repeated freezing or evaporation, successfully protected the mummies from putrefaction or decomposition (Hart Hansen 1998). The Arctic and Antarctic zones or high-altitude mountain areas are places with ideal conditions for successful mummification (Hart Hansen 1998; Lynnerup 2007; Slepchenko et al. 2019; Zimmerman 1998).

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Fig. 1 Red dots 1–4: Geographic locations where Siberian mummies were reported. (1) Zeleny Yar; (2) Ukok plateau; (3) Oglakhta VI; and (4) Verhoyansk. Black dots indicate the locations of Greenland or Alaska where frozen mummies were discovered before (Credit: Dong Hoon Shin)

In previous reports, permafrost mummies were found in the graves of the Greenland or Alaskan Inuits. These were indigenous people who have lived in the Arctic region for a long time prior to European immigration (Hart Hansen and Nordqvist 1996; Lynnerup 2015). As early as 1972, a native Arctic woman’s frozen body (dated 200–500 CE) was discovered on the Saint Lawrence Island of Alaska. Scientific investigation revealed that the female mummy might have died of suffocation, presumably caused by an accident due to a landslide or an earthquake (Hart Hansen and Nordqvist 1996; Zimmerman 1998). In 1982, the indigenous people’s mummified bodies were found at Utqiagvik as well, the northernmost region of Alaska. They might have died of an “ice shove” (Inupiat: ivu), a powerful inland shift of drifting sea ice by winds or tides. In 1994, a young female’s mummified body, presumably belonged to the semi-nomadic society of the Thule culture (800– 1200 CE), was also reported from Ukkuqs in Alaska (Zimmerman 1998). Studies on the Arctic mummies have also been conducted in Greenland on a much larger scale. Briefly, sixteenth-century mummies with well-preserved garments were found in the Pisisarfik Mountains of the Nuuk/Godthaab district (Hart Hansen and Nordqvist 1996; Lynnerup 2015). In 1972, fifteenth-century mummies were also discovered in a rock crevice at Qilakitsoq, Northwestern Greenland. They wore skin garments representative of the Thule culture. The Qilakitsoq mummies, still one of the most important reports about mummies found in the Arctic region, provides comprehensive insight into the native Thule society who once prospered in

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Fig. 2 Anthropological study of Siberian mummy in the physical anthropology lab at the Institute of the problems of Northern development, Siberian Branch of the Russian Academy of Sciences (Credit: Irina Sharova)

the area (Cockburn et al. 1998; Hart Hansen and Nordqvist 1996; Lynnerup 2015; Slepchenko et al. 2019). Thus far, studies on frozen mummies have mostly concentrated on the Arctic countries of Western hemisphere (Greenland, Alaska, Canada). In the case of the Eastern hemisphere, like the territory of Russian Federation, there were very few reports on permafrost mummies in Arctic or subarctic regions. Nonetheless, it is not accurate to say that such frozen mummies have never been discovered in the Eastern hemisphere. Rather, these findings about permafrost mummies have been reported steadily from the ancient or medieval Siberian cemeteries which were investigated by Russian archaeologists. For example, frozen mummies were found at a high-altitude Ukok plateau of the Altay Mountains in the South-Central Siberia (Rudenko 1960; Polosmak 1994). In the arctic area, the aboriginal mummies were investigated and reported by Russian archaeologists (Slepchenko et al. 2019). Through well-organized interdisciplinary studies of archaeology, history, ethnography, anatomy, parasitology, medical science, and aDNA analysis, etc., researchers successfully unveiled cultural or biological facts of those prehistoric Siberian native peoples hitherto obscure (Fig. 2).

Mummies of Ukok Plateau As mentioned above, the Siberian mummy that was the most famous in the outside world was from the Ukok Plateau, the high-mountainous plain situated at the Altai Mountains. The plateau is close to the four corners between Russia, Kazakhstan, Mongolia, and China. This area, one of the highest plateau areas in Altai Mountains

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(an altitude of 7500 feet which is 2286 m above sea level), is covered by vast grasslands and a part of them was also surrounded by large glaciers (Polosmak 1994). In ancient times, nomadic Pazyryk people must have wandered around this plateau. In the fifth to third century BCE, the seminomadic Pazyryk culture prospered in the Ukok area. Since the plateau was the major crossroad in the Eurasian steppe, it became important research subject of Siberian archaeology for the past century (Kang 2015). In the twentieth century, Soviet Russian archaeologists began to investigate the Iron Age burial mounds (kurgans) scattered about the Ukok Plateau. The Pazyryk mounds were first explored by S. I. Rudenko. He published many reports on various aspects of the ancient Pazyryk people’s life. When Rudenko (1960) investigated the big Pazyryk mounds in the excavation of 1947–1948, the mummies (n ¼ 4) were found inside the mounds #2 and #5 in pairs (a man and a woman) for each. The mummified man discovered at mound #2 had a drawing tattooed on his skin. The mummy is currently displayed at the Hermitage Museum in Saint Petersburg, Russia (Barkova and Pankova 2005). In the 1990s, once again, there was a great academic discovery reported from the Ukok plateau kurgans by a group of archaeologists from the Russian Academy of Science (led by N. V. Polosmak). High-level investigations were conducted on the mummified or skeletonized remains found at the Pazyryk kurgans. This interdisciplinary study by different research fields produced invaluable scientific knowledge that could not have been known before. Using the ethnographic data accumulated in Siberia and Central Asia, they also proposed a novel perspective on the ancient Pazyryk people who once lived in the Ukok Plateau. This excavation and research has become a symbol of Russian field archaeology in Siberia. The archaeological excavation showed that the Ukok Plateau kurgans were covered with a pile of rocks. Under the rock heaps, there was a subterranean larch log chamber that resembled the Pazyrik people’s winter cabins (Rudenko 1953; Polosmak 1994). A larch log coffin was discovered inside of log chamber. When the log chamber and coffin were found as low as the permafrost table, the burial complex was frozen all year round (Kang 2015; Polosmak 1994; Rudenko 1953). Therefore, after hot water was poured into the log coffin to melt the ice, Russian archaeologists eventually found well-preserved cultural relics and Pazyrik people’s mummified dead bodies. During the excavation, a male mummy was discovered in the burial ground of Ak alakh #3; and a female mummy (nicknamed as princess) was found in Verkhny Kuldzhin #2. Actually, in the Soviet era and subsequent years of the Russian Federation, the studies of these mummies and accompanying cultural remains found in the Pazyryk kurgans have attracted great interest from archaeologists around the world (Kang 2015). In the Pazyryk kurgans, archaeologists also found well-preserved clothes, important cultural remains to the dress historians in Russia. Because silk clothing was discovered in the kurgan, dress historians presumed that it might have been used in the Ukok Plateau before the Silk Road was finally opened by the ancient Chinese explorers (Kang 2015; Polosmak 1994). This goes against the general theory of when silk was imported to the Ukok region. Other organic materials found inside the

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kurgan are also invaluable to understand the actual life of ancient Pazyryk people. In the votive pots, archaeologists found milk and meat dedicated to the dead individual, revealing the funeral custom of ancient Pazyryk people. Through an analysis of the rugs discovered, researchers revealed that Pazyryk people weaved rugs out of camel hair obtained locally. The botanical analysis of the plants collected from the Pazyryk kurgans also showed the herbs used for food or medicine at that time (Kang 2015). Nevertheless, the most remarkable findings Polosmak and Russian archaeologists got from the Pazyryk kurgan were the mummies. In the excavation, they found frozen mummies in different state of preservation. The mummies were lying on their sides inside the log coffin (Polosmak 1994). When the Pazyryk people died, they were not likely buried immediately. Rather, to prevent decay, artificial embalming was performed on the dead individuals (Kang 2015; Polosmak 1994; Rudenko 1960). According to anthropological survey, embalmers seem to have removed the mummy’s internal organs and then filled the empty cavities with various kinds of grasses, herbs, and wool, etc. The Pazyryk embalmers also treated the mummy’s skin with a mercury containing solution. In addition, we also note that the Pazyryk people dug to the permafrost table to place the embalmed dead body within it. The presence of the burial complex in permafrost table must have had a greater impact on the mummification in Ukok kurgans. When it rained or snowed after burial, the log coffin and chamber must have been quickly flooded by rainwater; and then, everything froze with the drop of temperature (Kang 2015; Polosmak 1994; Rudenko 1953). In such conditions, the Pazyryk mummy could be preserved inside over thousands of years. In Siberia, one of the most famous Pazyryk mummies discovered so far must be “The Siberian Ice Maiden” or “The Princess of Ukok” (Polosmak 1994). Although people nicknamed her princess, she does not seem to have been an actual princess, but, rather, likely to have been a female shaman (Kang 2015). Anthropological or paleopathological studies assumed that she might have suffered from various diseases like osteomyelitis or fractures, etc. A very interesting finding of the Siberian Ice Maiden was a tattoo. In fact, tattoos were not found only in the Siberian Ice Maiden, but were also reported from other mummies of the Ukok Plateau (Barkova and Pankova 2005). Generally speaking, the tattoos are archaeologically very important in that they give a glimpse of ancient people’s outlook on the world. In case of the female (shaman or princess) mummy, a vivid tattoo from the shoulder to the hand depicting a mythical creature, a motif also found among the Scythians (Kang 2015; Polosmak 1994). At the request of the indigenous people, the Siberian Ice Maiden mummy was moved from Novosibirsk to the place where she was discovered (Kang 2015) though some other Pazyryk mummies are still on display and under study in Novosibirsk.

Mummies of Oglakhta (Tashtyk Culture) The first mention about the discovery of mummies of the Tashtyk culture dates back to the beginning of the last century. When the burial ground of Oglakhta in the territory of modern Khakassia was excavated, Adrianov (1903) found mummies

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with various states of preservation. In one of the burials, the mummified remains of a man and a woman were found. Their hair, mummified faces, shoulder girdles, chests, etc., were well preserved. He found the osteological evidence of post-mortem craniotomy. These trepanations were located on the occipital bone, and the brain was presumably removed through these openings (Adrianov 1903). In 1969, Moscow State University under the leadership of L.R. Kyzlasov resumed the archaeological work at the burial ground of Oglakhta VI (Kyzlasov and Pankova 2004). In the excavation, a family grave of the Tashtyk culture was discovered. Inside the burial chamber, in addition to well-preserved objects made of wood, leather, and fur, the human remains of a mummified man and a woman and the bones of a child were also discovered. Funeral sculptural masks of gypsum mass were found on their faces (Kyzlasov and Pankova 2004). All the findings of the Oglakhtinsky tomb, including mummified remains, were transferred to the State Hermitage Museum for storage and restoration. In 2002, a study of the Oglakhta mummies was performed in the laboratory of the State Hermitage Museum for the purpose of scientific restoration of monuments of applied art. Researchers from the department of forensic medicine and the Military Medical Academy were also invited to the investigation. After the mummy’s clothing (fur jacket or leather pants) was removed, the scientific investigation began. In the gross examination, there were very few signs of violence and no traces of artificial mummification were noted. When the mummies’ skins were examined with infrared light, tattoos were noticed (Kyzlasov and Pankova 2004). No evidence of incisions or sutures were detected in the mummified body. The internal organs of the mummy were not removed as there was a lung visible through the hole at the shoulder. Therefore, the experts presumed that the mummies were formed naturally and not in an artificial way (Kyzlasov and Pankova 2004). During the investigation, the sign of trepanation was reconfirmed on the heads of the mummies (Kyzlasov and Pankova 2004) which was first reported by Adrianov (1903). In Russia, many studies have been done regarding the Oglakhta mummies. However, the complete story behind the mummy is not yet fully revealed. Further research on the mummies would provide novel data on the population of the Iron Age Tashtyk culture.

Yakut Frozen Bodies Found in East Siberia Siberian mummies examined through a series of scientific research are very important to the related Russian academia. Through the study of mummies, valuable information about ancient or medieval Siberian people and society can be attained. However, the mummies of the Ukok Plateau or Oglakhta are not all that have been found in Siberia. Siberia is an immensely vast area and there are many places with suitable conditions for natural mummification. Mummies of academic value have been continuously discovered in the region, mostly in permafrost areas like the Siberian arctic or subarctic regions. One such example is the frozen mummies discovered at the medieval burial grounds of the Sakha Republic (Central Yakutia). The republic is one of the largest

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autonomous regions in the Russian Federation (Dedouit et al. 2010). From 2002 to 2012, a Russo-French team excavated the sixteenth to nineteenth century graves in central Yakutia, Vilyuy, and Verkhoyansk. During the excavations, many frozen bodies (over 140 individual cases) were discovered in these locations. The preservation status was generally good due to inhumation in permafrost area (Dabernat et al. 2014). Nonetheless, the discovered remains were in different preservation states, ranging from skeletons to mummified frozen bodies (Dedouit et al. 2010). The authors claimed that their findings represented one of the largest pools of frozen mummies ever discovered in the entire world (Dabernat et al. 2014; Dedouit et al. 2010). The people buried in the Central Yakutia graves seem to be the Yakuts. In the eleventh to thirteenth century CE, persecuted by the expansion of Mongolian tribes, a group of Turkic-speaking cattle breeders moved to the Lena river basin. They further developed to become the Yakuts when contacting with Tungustic-speaking indigenous peoples. From that point, the Yakuts spread over the vast area of northeastern Siberia (Keyser et al. 2015). Through research on the Central Yakutia mummies, researchers acquired anthropological knowledge about Siberian indigenous people that was not found in any other historical works (Dabernat et al. 2014; Dedouit et al. 2010; Keyser et al. 2015). For example, in a case of a female mummy found at one of the Yakut graves archaeologists presumed that she was likely a legendary udagan, a female shaman politically superior to any male counterparts in the Central Yakutia society (Dedouit et al. 2010). By CT scan as well as autopsy, researchers deduced anthropological data from the udagan mummy: her age at death was likely around 19 years old; stature was 146 cm (57.4 inches); and body weight was 49 kg (108 lbs). The craniometric analysis also showed that she was very similar to the Buryat population (Dedouit et al. 2010). The study on the frozen mummy specimens of Central Yakutia also informed the infection history of Mycobacterium tuberculosis in East Siberia. To comprehend the M. tuberculosis infection among medieval Yakut population, multidisciplinary research was conducted for paleo-epidemiological data accumulated so far. According to the research, the first contact of Yakuts with European population occurred when Slavic Russians eventually reached Yakutia (Dabernat et al. 2014). The researchers argued that tuberculosis began to spread throughout Yakutia when the Yakut people contacted Russian immigrants in the early seventeenth century (Dabernat et al. 2014). The modern Yakut population is likely the final outcome of adaptation affected by selection pressure over the past several centuries after the initial outbreak of tuberculosis in East Siberia (Dabernat et al. 2014).

West Siberian Arctic Graves Similar studies have also been done on the permafrost burial grounds found in West Siberia (west of the Yenisei River). The burial grounds were located inside the Arctic Circle of the Russian Federation. For several decades, a collaborative team of

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archaeologists and anthropologists have undertaken excavations at the Arctic permafrost zone in Yamalo-Nenets Autonomous Okrug (YaNAO) (Fig. 3). The Okrug encompasses a vast area of the arctic or subarctic areas where many native Siberian peoples coexisted while sticking to different subsistence strategies (Lee et al. 2019). Of them, the Tatars, a Turkic-speaking ethnic group in West Siberia, were fishermen, hunters, and pastoralists around the Irtysh River basin. The Selkups, the presumptive descendants of the Kulayskaya culture (500 BCE– 500 CE), depended on fishing, deer farming, hunting, and wild berry and pine nut gathering. The Khanty, mostly fishermen and hunters, lived in the forest tundra and north taiga zones of the Ob regions. The Nenets were hunters, fishermen, and reindeer herders of the Near Arctic and Arctic Circle areas (Bagashev 2017; Lee et al. 2019; Perevalova 2004; Poshekhonova et al. 2018; Slepchenko and Ivanov 2015; Slepchenko et al. 2016; Slepchenko 2017). The archaeologists located many indigenous people’s burials during the investigation of the Arctic permafrost area in Yamalo-Nenets Autonomous Okrug (Figs. 4 and 5). One such investigation is an archaeological excavation conducted from 1999 to 2002. The Russian archaeologists led by N. V. Fedorova surveyed the twelfth to thirteenth century Zeleny Yar (Z-Y) site. The place is situated on a flood plain island (66 190 4.5400 С; 67 21013.5400 В), about 40 km (24.8 miles) away from Salekhard, the capital of Yamalo-Nenets Autonomous Okrug. A monograph about the 1999–2002 excavation of Z-Y site was published in 2005, giving the details about human remains and their related cultural remains discovered inside the graves

Fig. 3 Archaeologists need equipment to fight against mosquitoes in the forest tundra area of the Arctic region (Priuralsky district of Yamal-Nenets Autonomous Okrug, Russia) (Credit: Dong Hoon Shin)

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Fig. 4 An example of Siberian native (the Nenets) people’s burials found in subarctic region (the burial ground at Nyambouto 1, Tazovsky district of Yamal-Nenets Autonomous Okrug. Russia) (Credit: Sergey Slepchenko)

Fig. 5 Excavation of Siberian burials in subarctic zone (the nineteenth-century burial ground at Nyubrymposlor, Priuralsky district of Yamal-Nenets Autonomous Okrug, Russia) (Credit: Alexandr Gusev)

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(Aleksashenko et al. 2005; Slepchenko et al. 2019). During the excavation of the burials, archaeologists found very well-preserved mummies (Aleksashenko et al. 2005; Bagashev et al. 2017; Slepchenko and Ivanov 2015). The scientific analyses of these mummies yielded invaluable anthropological information about West Siberian indigenous peoples (Bagashev et al. 2017; Sato et al. 2011; Slepchenko and Ivanov 2015). In brief, using the mummy specimens, Sato et al. (2011) revealed the genetic features of native medieval people buried at Z-Y site. Slepchenko and Ivanov (2015) obtained a parasitological result from the samples of 1-year-old infant from the site. Bagashev et al. (2017) also conducted anthropological and radiological analyses (Fig. 6), from which they were successful in facial reconstruction of a male buried at the Z-Y site. Actually, this was the first report to show what the face of a local mummy looked like (Fig. 7). Recently, Slepchenko et al. (2019) also reported on another interdisciplinary research of the permafrost mummies that were newly discovered at the Z-Y site. In the continued excavation during the 2013–2016 season, 47 burials were investigated at the Z-Y site; and from them, 49 individuals were also identified. The grave was in trapezoid form, being oriented mainly on the north-south axis. A birch-bark cocoon was discovered on the bottom of each burial pit. The cocoon was the bundle in which human and cultural remains were packed together (Fig. 8). To accommodate additional scientific research, the cocoon was moved to the lab at Shemanovskiy Yamal-Nenets District Museum (Slepchenko et al. 2019).

Fig. 6 The Siberian male mummy currently displayed at Yamalo-Nenetskiy Okruzhnoy MuzeynoVystavochnyy Kompleks Imeni I.s. Shemanovskogo (Credit: Sergey Slepchenko)

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Fig. 7 Facial reconstruction of the Siberian mummy from the grave no. 27 of Zeleny Yar burial ground. The reconstruction was performed by Alekseeva E.A (Credit: Alexeeva Elena)

Fig. 8 Birch bark cocoon found in the West Siberian grave of arctic area (burial no. 53 in the Zeleny Yar burial ground) (Credit: Alexandr Gusev)

CT analysis was conducted to estimate the condition inside a birch bark cocoon before scientific investigation began (Fig. 9) (Slepchenko et al. 2019). Human remains were found inside the birch bark cocoon. The preservation status was very

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Fig. 9 CT scan of Siberian mummy. The mummy was transported from Zeleny Yar site to YamaloNenetskiy Okruzhnoy Muzeyno-Vystavochnyy Kompleks Imeni I.s. Shemanovskogo (Credit: Evgeniya Zhulanova)

diverse. Some of the Siberian people’s remains were almost skeletonized but the others were fortunately mummified (Figs. 10 and 11). The cocoon’s internal structure could be revealed during the investigation at the museum lab. First, the cocoon was formed by multiple layers of birch bark and bast fibers. When the barks and fibers were removed, a fur veil was then exposed. Various bronze or iron objects were also discovered inside the fur veil (Fig. 12). After the removal of fur veil, human mummified remains were noticed and anthropological examinations were performed on them (Slepchenko et al. 2019). Different from the mummies of Ukok Plateau, all of the mummies discovered in Siberian arctic zone are naturally formed ones.

Preservation of Siberian Mummies In general, considering Egyptian mummies or other countries’ mummies that have been studied for a long time, the preservation status of these mummies was already revealed to a great extent by many scientific research reports. However, regarding the Siberian mummies, it has not been known to date what the similarities and differences are in preservation status when compared to the other mummies. The mummy found at the West Siberian graves was therefore subject to anatomical, radiological, and histological analyses to discover its preservation pattern. Slepchenko et al. (2019) discovered that the Siberian mummies’ preservation pattern (Fig. 13) was generally similar to those of the other naturally mummified

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Fig. 10 CT scan of Siberian mummy from burial no. 78 in the Zeleny Yar burial ground (Credit: Sergey Slepchenko)

Fig. 11 A child mummy (burial no. 53) found in the Zeleny Yar burial ground. Siberia. The mummy’s body was exposed after the removal of fur veil (Credit: Sergey Slepchenko)

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Fig. 12 Cultural remains found inside the cocoon from burial no. 53 in the Zeleny Yar burial ground. Bronze artifacts were identified (Credit: Evgeniya Zhulanova)

cases. Briefly, the internal organs, on CT images, still remained inside the mummy’s body cavity. Mummy brains were observed in the cranial cavity. Eyeball tissues were detected inside the orbital cavities. The other mummified organs such as lung, liver, intestine, and muscle, etc., could be discovered in both thoracic and abdominal cavities (Slepchenko et al. 2019). In the previous studies on Korean mummies of Joseon period (1392–1910 CE), the internal organs were mostly shrunken, distorted, and dorsally displaced, presumably due to the long-term effect of gravity and/or dehydration. Internal organs were proven intact albeit seriously deformed, displaced, and dehydrated (Kim et al. 2014). Likewise, the internal organs (brain, lung, liver, etc.) of Siberian mummies were collapsed to the dorsal part of thoracic or abdominal cavity during their natural mummification process (Slepchenko et al. 2019). Regarding the histological analyses conducted on Siberian mummy samples (e.g., intestine, urinary bladder, skin, muscle, and hair), there was not much difference in the findings of light and electron microscopy when compared to other naturally formed mummies (Hess et al. 1998; Shin et al. 2003). In brief, the collagen fibers were intact though the cells were almost disintegrated (Fig. 14). In

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Fig. 13 A male mummy from burial no. 79 in the Zeleny Yar burial ground (Credit: Sergey Slepchenko)

Fig. 14 Histology of West Siberian mummy (Credit: Dong Hoon Shin)

the case of the skin, serious atrophy was observed in the epidermis due to the loss of cells while the dermis of Siberian mummies looked intact by the preservation of collagen fibers (Slepchenko et al. 2019). Histology of mummy muscles also exhibited collagens in perimysium remnants but the myocytes disappeared. In urinary bladder tissue, transitional epithelium and submucosal cells also became atrophic, leaving very thin film-like collagenous structure in the bladder wall. Collagen fibers were also profusely observed in mummified intestines (Slepchenko et al. 2019).

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In sum, the histological findings commonly observed in most naturally mummified cases (excellent preservation of collagen fibers but the absence of cells in mummified organs) could be evidently reported from Siberian mummies as well (Slepchenko et al. 2019). The permafrost arctic area in Siberia ensures the excellent preservation of mummified organs, due to local climate of year-round low humidity as well as low temperature (Cockburn et al. 1998; Hart Hansen and Nordqvist 1996). Notwithstanding the respective mummification processes having occurred under different geological and environmental conditions, the preservation pattern of the Siberian mummies was not so distinct from those of other naturally mummified cases in different countries.

Scientific Analyses of Siberian Mummies Considering that the mummified bodies were not subject to any artificial embalming treatment, the academic value of the Siberian mummies is very high indeed because the internal organs were not removed and naturally preserved even after death. Using the mummy specimens, scientific researchers have produced a lot of important information about the indigenous people who lived in Siberia. First, mummified organs were sampled and used for the aDNA analysis (Slepchenko et al. 2019). Since aDNA molecules are generally degraded into very short pieces, the test is very difficult when compared to modern DNA analysis in many respects (Ho and Gilbert 2010; Pääbo 1989). To guarantee the authenticity of aDNA work, researchers also followed the recommendation of strict lab maintenance (Ho and Gilbert 2010; Hofreiter et al. 2001). aDNA analysis was utilized as it is a good research technique to trace ethnic origins (Iwamura et al. 2004; Ricaut et al. 2006). The study on the mummies discovered at Qilakitsoq of Northwestern Greenland is one such example. In the study, aDNA analysis performed on the mummies’ hair and nail specimens revealed that their genetic kinship is more complex than originally expected (Lynnerup 2015; Slepchenko et al. 2019). Genetic features of West Siberian people were also studied by Sato et al. (2011). They analyzed mtDNA haplogroup of West Siberian native people by examining the bones excavated from two different archeological sites: Saigatinsky 6 (eighth to eleventh century) and Zeleny Yar (thirteenth century). Since the sequences of native West Siberians were mtDNA haplogroups B4, C4, G2, H and U, Sato et al. (2011) claimed that Saigatinsky and Zeleny Yar peoples might belong to the modern native Siberian populations of Mansi, Ket, and Nganasan. This means that some medieval West Siberians must have been in direct ancestral lineages of modern native populations at the same region (Sato et al. 2011). For native Siberian mummies, the consensus sequences were also determined by the studies of Slepchenko et al. (2019). Their studies revealed that mtDNA haplogroups of West Siberian mummies (of Z-Y site) belong to five different groups: D, D4j8, H3ao, U4b1b1, and U5a, those commonly observed among modern Siberian native peoples (Achilli et al. 2004; Derbeneva et al. 2002a, b; Derenko et al. 2010; Fedorova et al. 2013; Jin et al. 2009; Malyarchuk et al. 2010;

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Sato et al. 2011). Overall, aDNA analysis revealed that the mitogenome of West Siberian mummies is a unique combination of Eastern- (D and D4) and WesternSiberia-specific (H3, U4, and U5) mtDNA haplogroups (Slepchenko et al. 2019). Analysis of the samples obtained from the abdominal cavity of natural mummies is also highly informative. Actually, the specimens of mummy intestines can be a reliable source of informed scientific research concerning diet and health (Dean 2006; Gonc¸alves et al. 2003; Reinhard et al. 1986). By microscopic or molecular analyses of the intestinal contents obtained from the mummies, parasitologists revealed the parasite infection pattern prevalent among prehistoric peoples. In general, the skin of the mummy suprapubic area was incised; and for parasitological examination, the samples were taken from the abdominal cavity. One of the best examples of archaeoparasitological study in Siberia was a child mummy discovered at the Z-Y burial ground. The microscopic examination of a child mummy’s intestinal contents revealed the ancient parasite eggs of Opisthorchis felineus. This finding was corroborated by historical reports that West Siberian people of medieval times enjoyed raw fish in great amounts as early as the twelfth and thirteenth centuries CE (Nosilov 1937; Vizgalov et al. 2013). Ethnographic records also showed that Siberian people were fed with raw or undercooked fishes from early childhood (Vizgalov et al. 2013), which was also confirmed by the detection of O. felineus eggs in the archaeological specimens from the Z-Y child mummy (Slepchenko et al. 2019).

Conclusion Pazyryk mummies found in Ukok Plateau of the Altai Mountains are among the most famous ever reported having a great impact on archaeologists all over the world. However, in the Siberian arctic or subarctic zone, there were also many important findings about permafrost mummies. Through studies on these mummies, Russian anthropologists secured invaluable information that could not have been attained by archaeological or historical approaches alone. However, as a whole, we also admit that the extant reports about Siberian arctic or subarctic mummies cannot reveal detailed story in their collectivity. These reports can only be a preliminary step to fulfilling Siberian permafrost mummies’ academic potential. Nonetheless, this knowledge obtained from Siberian mummies to date could be the fundamental basis of forthcoming studies on the mummies discovered in Siberia, finally revealing the anthropological details of ancient indigenous peoples who inhabited in those regions (Slepchenko et al. 2019).

Cross-References ▶ Bog Bodies and Natural Mummification of Siberia ▶ Mummies and Skeletonized Individuals to Reveal the Relationship of Parasitism, Social Complexity, and Subsistence Strategy in Eurasian Continent

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Acknowledgments This work was supported by the Ministry of Education of the Republic of Korea and the National Research Foundation of Korea (NRF-2019S1A5C2A01083578). This study was also funded by State Task (program XII.186.4, project No. АААА-А17-117050400143-4 of Tyumen Scientific Center SB RAS).

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In Uk Kang, Jinseong Han, Jong Ha Hong, Jieun Kim, Dong Hoon Shin, and Victor H. Mair

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Xiaohe Cemetery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Investigations by Chinese Archeologists Since 1999 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Otani Collection in Seoul . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cultural Remains of the Xiaohe Cemetery in NMK . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . History of the Investigation of the Tarim Basin Graves and Cemeteries . . . . . . . . . . . . . . . . . . . . . People from the West . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Research Updates on the Identity of Tarim Basin Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Mummies in Xinjiang Museum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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I. U. Kang (*) · J. Han Department of History (Archaeology), Kyung Hee University, Seoul, South Korea e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea J. Kim Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul, Korea D. H. Shin Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea V. H. Mair Department of East Asian Languages and Civilizations, University of Pennsylvania, Philadelphia, PA, USA © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_28

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Abstract

In this chapter, through descriptions of the Xiaohe cemetery, related Silk Road archaeological sites, and the mummies discovered therein, we have reviewed the present situation of research on the Bronze Age cultures left by ancient IndoEuropeans who drifted into northwestern China as early as 2000–1500 BCE. The mummies and the early Bronze Age artifacts associated with them found in the Xinjiang area have been investigated in the last century, but professional analyses based on substantive evidence has been limited. But because of archaeological information that has recently emerged from the Xinjiang Silk Road reinvestigation, the significance of the mummies has been validated anew. The Xinjiang mummies currently maintained in the local museums have significance in two aspects: their contribution to the history of Silk Road researches and to mummy studies worldwide. Especially for the latter, the mummies and their culture could provide invaluable information about the first demic diffusion of Indo-Europeans to the east in the prehistoric era. The Xinjiang mummies are also significant because of their superb preservation, even after the lapse of several millennia. As is well-known, naturally mummified examples provide the best research subjects for scientific analysis for revealing the life and health status of ancient peoples. Since the mummies are very rare cases that have been preserved in the remote periphery of China during ancient times, sensitive and detailed scientific studies on them in the future could provide wonderful data to archaeologists, anthropologists, and paleopathologists worldwide. Keywords

Mummies · Xinjiang · Tarim basin · Silk Road · Xiaohe · Otani Collection · Loulan · Lop-Nor

Introduction The so-called Xinjiang mummies have attracted the attention of archaeologists and the public worldwide. The findings were already well described in the monumental publications by Cockburn et al. (1998), Mallory and Mair (2000) and Aufderheide (2003). According to these, Taklamakan Desert is one of the most arid places in the world, and it was there that China’s oldest naturally formed mummies were discovered (Map 1). There are two different routes of the Silk Road around the Tarim Basin. The southern route skirts the Taklamakan Desert along its edge at the slopes of the Kunlun Mountains. Meanwhile, the northern route connects various communities at Kongque (Konche or Konchi, and Kɵnqi dəryasi darya in the Uyghur language) and the Tarim River, leading to Loulan Kingdom (Lou-lan Kingdom) and Lop Nur (Lop Nor, nur meaning “lake” in Mongolian), where multiple Xinjiang mummies were discovered (Aufderheide 2003) (Figs. 1, 2, 3, 4, 5, and 6). In The Scientific Study of Mummies (2003), Aufderheide comments on several mummified bodies in graves near Loulan. Of these, Aufderheide describes the

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Map 1 Locations of sites with mummies around Tarim Basin, Xinjiang, China Fig. 1 Xiaohe mummy, male, Museum of Xinjiang Institute of Cultural Relics and Archaeology. (Photo taken by Kang, In Uk)

mummified female (the “Beauty of Loulan” or the “Loulan Beauty”) and other mummies found at the Lop Nur area as well as in Qiemo (Cherchen in the Uyghur language) county along the desert’s southern edge. These were pioneering achievements, but more detailed information is not well-known to mummy researchers, because most of the reports on these archaeological sites and associated mummies were published only in Chinese. In addition, with the rapid modernization of China,

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Fig. 2 Xiaohe mummy, child, Museum of Xinjiang Institute of Cultural Relics and Archaeology. (Photo taken by Kang, In Uk)

Fig. 3 Xiaohe mummy, female, Xinjiang Uyghur Autonomous Region Museum. (Photo taken by Kang, In Uk)

archaeological survey in the Tarim Basin has developed very fast, and the general knowledge concerning the mummies around this area has become much more evident than ever. In order to fill the communication gap, this chapter summarizes the most recent findings, based on reports from China, on the status of research on the Xinjiang mummies.

The Xiaohe Cemetery The Xiaohe (meaning “Little River”) cemetery is enormous. There are no contemporary sites comparable to the Xiaohe cemetery in the southern route of the Silk Road discovered so far. The Xiaohe cemetery is one of the prehistoric burial sites

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Fig. 4 Mummy from Niya cemetery 95 (M5), child, Museum of Xinjiang Institute of Cultural Relics and Archaeology. (Photo taken by Kang, In Uk)

Fig. 5 Mummy from Wupu cemetery, male, Turfan Museum. (Photo taken by Kang, In Uk)

Fig. 6 Mummy from Cherchen, female, Xinjiang Uyghur Autonomous Region Museum. (Photo taken by Kang, In Uk)

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that were constructed on a sand dune during the early Bronze Age (2000– 1500 BCE). From the earliest days of study of the Silk Road, the Xiaohe cemetery was already very famous among researchers and grave robbers for the discovery of invaluable artifacts and mummies in the vicinity, making the area a prime target for their academic investigations as well as grave plundering. The most remarkable investigations on the Xiaohe cemetery in the earliest days were those of Sven Hedin (1865–1952) and Folke Bergman (1902–1946). When they examined this area in the 1930s, the mummies and other artifacts already had been pillaged in repeated grave robberies, and finally the graves were themselves abandoned, their contents scattered around the cemetery site. At that time, Bergman’s investigation of the cemetery was assisted by Ördek (Ördeck), his Uyghur guide (the cemetery was thus named after him, originally being referred to as Ördek Necropolis). According to Ördek’s account, when he explored the Xiaohe cemetery site, news soon hit the area of an enormous amount of valuable treasures found during Sven Hedin’s excavation of Loulan in the 1910s. In 1906, the discovery of a prehistoric site very similar to the Xiaohe cemetery was reported by Ellsworth Huntington (1876–1947), the American geographer. The increase in interest in the ancient remains of this region made grave robbery frequent around the Xiaohe cemetery, where valuable artifacts could be acquired with ease. In fact, many records describe fierce competition between the inhabitants and the grave robbers in collecting artifacts from the Xiaohe cemetery.

Investigations by Chinese Archeologists Since 1999 The Xiaohe cemetery had not been reinvestigated after the survey conducted by Sven Hedin and Folke Bergman. Even the correct location of the cemetery site could not be accurately identified by any archaeologists. However, as late as 1999, through the efforts of the Xinjiang Archaeological Institute led by Binghua Wang, the location of Xiaohe cemetery was finally reconfirmed. Afterward, a Xinjiang archaeological expedition directed by Idris Abdursal (Yidilisi Abuduresule in the Chinese pinyin system) led to 167 graves being newly identified and thoroughly investigated in 2002–2005. Despite a full report about the reinvestigation of the site not being published yet, brief reports have been available (Mair 2006; Xinjiang Institute of Cultural Relics and Archaeology 2004, 2007). According to these Chinese reports, the Xiaohe cemetery is located in an oval sand dune in a branch of the Kongque River, 4 km east from the Xiaohe. The total area of the site is 2500 m2, measuring 7  74 m, and 7 m high. Owing to the many wooden pillars erected to mark the area, before regular archaeological excavation began, the cemetery was easily identifiable even at a considerable distance, attracting many grave robbers. The whole cemetery complex was delineated by wooden fences. Five different layers of archaeological stratigraphy were identifiable in the northern and southern areas. Graves at each layer were constructed at regular intervals. In most graves at the upper geological layers of the site, human skeletons and coffin wood had already

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been exposed by the depredations of robbers. In brief, when 13 graves in the first (uppermost) layer and 27 graves in the second layer were investigated, 21 graves had already been destroyed before the investigation began. Despite the destruction of these graves, those located in the third to fifth layers still were intact, possibly because the intruders might have had difficulty digging in the sliding sands. It is likely that each graveyard at each separate chronology was consistently occupied by specific groups.

Otani Collection in Seoul In the early twentieth century, Kozui Otani (1876–1948) of Japan conducted a major Silk Road exploration. The Otani expedition carried out research on the Silk Road remains, especially regarding the importation of Buddhism into East Asia, because the subject had not yet been studied sufficiently by academics at that time. Following in the footsteps of Hedin, Zuicho Tachibana, a member of the Otani expedition, in 1908–1912, surveyed Loulan and Lop Nur twice, collecting or buying artifacts from these regions. Some of the collection made by the Otani expedition is currently displayed in the National Museum of Korea (NMK), and it is very significant for researchers of Silk Road history. Bergman pointed out in the 1930s that the collection might not have been formed by regular archaeological excavation, but was purchased during the survey in Lop Nur, from local people or Chinese ambans, high officials dispatched to the periphery territory during the Qing dynasty. Owing to the fact that the artifacts were collected unskillfully during the early twentieth century, especially during the period of colonization, it has been difficult to analyze the Otani collection in the NMK, and such analysis has been conducted only very rarely. As a result, the meaning of these pieces has not been properly established. Nevertheless, the Lop Nur collection in the NMK is remarkable for incorporating artifacts of the Xiaohe cemetery, which reveal evidence of the prehistoric east–west cultural exchange along the Silk Road. In addition, during the reevaluation of the artifacts, the date estimated by the Tachibana explorations of some of the Xiaohe cemetery remains in the NMK was confirmed, making them possibly the earliest ever collected from the cemetery. Comparison of the Xiaohe artifacts in the NMK with recent archaeological updates in the Xinjiang region could lead to their reinterpretation and greatly advance our understanding of related mummies found in the region, and of the Silk Road generally.

Cultural Remains of the Xiaohe Cemetery in NMK Over all, the estimated dates of some Tarim Basin graves have been more certainly established. In the case of the Xiaohe cemetery, its southern part is dated to 1800– 1400 BCE, while the northern portion has been dated to 1950–1500 BCE. The Keliyahe northern cemetery is dated to 1880–1700 BCE. In fact, the Xiaohe

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cemetery has huge archaeological significance because it was used by an early Indo-European nomadic culture that arrived via the southern route of the Silk Road during the early Bronze Age (2000–1500 BCE). It was long thought that Xiaohe cemetery remains had not been collected by any scientific explorers prior to the Bergman’s excavation in the 1930s. It now appears that the Xiaohe cemetery collection formed by Tachibana’s expedition in 1908–1912 might have been the first ever of that kind. In fact, some pieces in the Otani collection in the NMK are precisely homologous to those of the Xiaohe cemetery recently reported by Chinese archaeologists in the 2002–2004 expeditions. Among the cultural artifacts, the felt hats, wooden stick figures, wooden carved facial masks, and woven baskets especially are similar to those described in these Chinese reports. As for the woven baskets found in the Xiaohe cemetery, previous analysis has revealed the identity of the materials found inside the baskets by a successful extraction of bovine casein and immunoglobulin. This shows that the Xiaohe cemetery users persistently employed dairy products in their funeral ceremonies (Liang et al. 2012: 81–85). Placing the woven baskets inside the grave, especially alongside the buried individual, is likely to be related to an Indo-Iranian tradition in which their staple dairy products were held sacred. The woven baskets might have been used as containers to hold the votive dairy products (Polos’mak 2001). At the Çatalhöyük site in Turkey, it is noteworthy that dairy products were found inside the woven baskets, without the use of any ceramic container. The absence of pottery in the Xiaohe cemetery might relate to a similar tradition. There is an excavated wooden, face-shaped sculpture in the NMK Lop Nur collection, whose function and context have never been identified. However, examples of the same wooden face type were excavated from Xiaohhe and Kongque cemeteries in situ, where the wooden face was located on the chest of the grave’s occupant. It is also noteworthy that part of the tooth is made from real tooth or ivory. This figure probably played a role as an amulet. This is an example of the ways in which the recent excavation of Xiaohe has helped scholars to figure out the previously unknown meaning of items in the NMK collection. It has generally been assumed that the Xiaohhe cemetery would have been used by residents of villages established next to the oasis. But no vestige of these villages has been discovered as yet. Thus we are unable to obtain specific information about the actual living conditions of the inhabitants contemporary with those interred in the cemetery. We know that these people inexplicably disappeared by 1500 BCE, and it is posited that they might have migrated due to climate change or for other reasons, or that they might simply have died out.

History of the Investigation of the Tarim Basin Graves and Cemeteries As described above, after the original investigation of the Xiaohe cemetery in the 1930s, the site was resurveyed by Chinese archaeologists in the early 2000s. However, recent investigations in Xinjiang have not been limited to the Xiaohe

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cemetery. Investigation updates revealed that similar early Bronze Age cemeteries were also discovered in the areas around the Kongque River of the east Tarim Basin area. The Keliyahe (Keriya river) northern cemetery located on the lower Keliya River is a site with similar archaeological significance (Zhang 2009). The people of the Xiaohe cemetery are therefore supposed to have been area inhabitants who lived at multiple oases within the Lop Nur region of 600 km radius. Due to the geographical traits of the Taklamakan desert, it is very difficult to access. Except for the above-mentioned investigations of the Xiaohe cemetery by Sven Hedin and Folke Bergman in 1930s (Bergman 1939, 1945) and Cemetery LF on a dune 4 km east of LE ancient castle from which Stein excavated eight graves in 1914, only brief surveys were carried out around Lop Nur in 1948 (Huang 1948). Only as late as 1979, when archaeological investigations resumed after the turmoil of the Cultural Revolution, the Gumugou (Qäwrighul) cemetery site at the Kongque River was investigated. However, full-fledged reputable investigation was not conducted on any other sites of this region for a long while. During and after the 2000s, along with the above-mentioned re-investigation of the Xiaohe cemetery, the Third National Cultural Relics Survey in China started to perform large-scale excavations in the Xinjiang region (NCHA 2012). With these achievements, the true pattern of the Tarim Basin Silk Road area of the early Bronze Age is becoming more and more clear. We summarize the recent updates of Tarim Basin archaeology as follows: (1) two mummies (a male and a female) were found at the LE northwest cemetery in 2003; (2) the Keliyahe northern cemetery, located at the Keliya riverside, 600 km west of the Xiaohe cemetery, was excavated in 2008. In the Keliyahe northern cemetery, approximately 60 graves had been robbed and destroyed; only 20 sets of remains were intact. The fundamental cultural heritages identified in this cemetery greatly resemble those of the Xiaohe cemetery and are assumed to have been built by settlers of the same culture (Zhang 2009). (3) In 2009, Grave 09LE50, located 9.4 km northeast of the LE ancient castle, was also investigated by the excavation team led by Idris Abdursal. A very well-preserved male mummy and four grass baskets were discovered, while no wooden coffins were found intact. Radiocarbon dating estimated the date of the site to be approximately 1700–1500 BCE or 2000–1800 BCE. (4) Grave 09LE4, located at 4.67 km north of the LE ancient castle, was investigated in 2009. A total of 13 graves were identified. (5) In 1979, 42 graves were discovered in the Gumugou cemetery. Of these, 36 graves contained wooden coffins. Timbers had been erected on the edges of the site. The grass basket and felt tigraxaudas discovered at this site look similar to those of the Xiaohe cemetery (Wang 1983). Other burials found in the Gumugou site are designated sun-shape burial, in which wooden pillars are erected radially around the grave. (6) Hanshuiquan, located 6 km north of the Kongque River, is unique because it was constructed in a different manner from the Xiaohe cemetery. Overall, seven burials were discovered at the site. Because such a site as Hanshuiquan showed a pattern distinct from that of the burials of the Xiaohe cemetery, there is a strong possibility that the culture of the Tarim Basin might henceforward be subdivided. Nevertheless, the overall cultural aspect of both is quite similar. The material culture here was represented by felt shoes

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and hats made out of wool and camel hair, the use of various types of grass baskets, bronze or copper ornaments, and wooden figures. Both populations are also thought to have shared living economies based on wheat agriculture and cattle breeding.

People from the West In recent archaeological reports, intense similarity could be observed between Altai stone statues in northern Xinjiang and those found in southern France, suggesting the expansion of Indo-European settlers in the early Bronze Age (Kovalev 2014, 2015). Such a broad expansion of European people from 3500 BCE throughout entire Eurasia has been proven by reports of language, domestication of horses, pastoral economy, bronze ware, etc. Based on the studies on well-preserved mummies and the various artifacts excavated, there is a strong consensus in academia that the early Bronze Age culture found in the Tarim Basin area might have been related to the rapid demic diffusion of Indo-Europeans who migrated from the Eurasian steppe to settle in the southern oasis area around the Taklamakan Desert by 2000 BCE. In the archaeological perspective, this European expansion had taken several stages. The first stage was incipient in ca. 3500 BCE with the expansion of nomadic culture. This diffusion is represented by the emergence of the Yamnaya Culture in Ukraine as well as the Afanasievo Culture in the Eurasian steppe. During the same period, such an expansion of the people spread eastward more to the northern Xinjiang area of China. The expansion can be corroborated by the archaeological identification of the Chemurchek culture (about 3000–2500 BCE, also known as Qiemuqieke or Ke’ermuqi in Chinese archaeology and Khemtseg in Mongolian archaeology). Next, the second stage is related to the diffusion of Andronovo culture with the invention and spread of chariots in the 2000 BCE. The people migrated, along with pastoral economy and highly developed bronze culture, into Central Asia. The Xiaohe cemetery might have been related to the expansion of Andronovo culture into the Xinjiang region. The reasons are as follows: First, the motifs drawn on woven baskets in Lop Nur and the Xiaohe cemetery resemble those of typical Andronovo culture. This is a very significant finding because the motif on woven baskets was one of the most distinguishing characteristics of Andronovo culture. Second, the dates of the Xiaohe cemetery, Gumugou, and Tiebanhe are all estimated to be 2000–1500 BCE, almost coinciding with the beginning of Andronovo culture expansion. Third, previous reports on the physical anthropology and ancient DNA analysis on the human remains of Xiaohe cemetery have concluded that these people might have been genetically very close to the people of Western Eurasia. In brief, the genetic analysis of the Xiaohe cemetery revealed that their maternal lineages were mainly of Eastern Europe origin, with some western European and Indian traits partly also included (Li and Zhou 2016). What is also noteworthy is that the people of the Xiaohe cemetery represent high genetic relatedness with the contemporary South Siberian settlers. This is clearly well matched with the expansion theory of Andronovo people in Eurasia during the Bronze Age.

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Meanwhile, Ancient DNA (aDNA) also indicated that the genetic traits of Indo-Europeans initially predominant among the Xiaohe cemetery tended to be changed into those of East or South Asian populations (Li and Zhou 2016). This result suggests that Indo-Europeans who originally migrated into this region actively interacted and merged with the aboriginal people by the process of marriage. Similar complex patterns could be also seen in the genetics of domesticated animals and ancient grains raised by the Xiaohe cemetery people. Actually, the cattle remains found at the Xiaohe cemetery site were also genetically close to the European and Middle Eastern domesticated ones. DNA analysis on wheat ingredients extracted from a bread remnant found at the Xiaohe cemetery also showed that its origin was highly likely to be of the Near East region. However, domesticated millet discovered at the Xiaohe cemetery was found genetically to resemble Chinese millet. This means that the origins of the crops and animals of the society were very complicated.

Research Updates on the Identity of Tarim Basin Mummies Arthur Aufderheide (2003), in his monumental book focused on examining the Tarim basin mummies, described the female mummy nicknamed the Beauty of Loulan and other mummies found in the Xinjiang province of China (Aufderheide 2003). In his visit to the Archaeological Research Institute at Urumqi (Wulumuqi in Chinese pinyin), Aufderheide saw six mummies, mostly from the Tarim River area. Of these, two were females, two males, and two infants. At Turpan (Tulufan in Chinese pinyin), he also found five mummies curated at the Turpan Museum (Aufderheide 2003). During his visit to the Shanghai Museum of Natural history, Aufderheide also noted that one female mummy found near Hami in the Tarim Basin was displayed and was in excellent condition. He and Dr. Victor Mair of the University of Pennsylvania, who visited later, were struck by the Tarim Basin mummies’ Caucasoid appearances. As seen in this chapter, the Caucasoid theory for the origin of these mummies has not been countered by any archaeological or anthropological evidence thus far. Regarding the so-called Cherchen Man, Aufderheidi described the male mummy as 2 m tall, having light-brown hair, and being spectacularly garbed in white deer skin boots, brightly colored trousers, and shirt, as well as felt leggings (Aufderheide 2003). Examining the weave pattern of some of Cherchen Man’s clothing, he thought that this might represent possible Celtic contact during the first millennium BCE. He also considered the Tocharians as a possible Caucasian affiliation (Aufderheide 2003). Recent archaeological reports further confirmed that the mummified people of the Tarim Basin graves are wholly of Indo-European origin. Indo-Europeans were distributed in the vast area from the western part of the Eurasian steppe to the Xinjiang area in China. Based on archaeological evidence to date, it is thought that European affiliation expanded to most of Eurasia in 3000 BCE with the usage of copper, kurgan building, and stockbreeding, having migrated into northern Xinjiang as well as western Mongolia in advance of that date. As for a second-stage migration of Indo-European people, the Andronovo Culture, represented by highly developed bronze tools and chariots,

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was diffused further into vast regions during 2000–1500 BCE. A sphere influenced by the culture was great in extent; even the chariots excavated from the late Chinese Shang dynasty (1600–1046 BCE) burials are related to that diffusion. The appearance of the Xiaohe cemetery is the earliest evidence supporting the diffusion of Andronovo culture in the second stage, expanding widely into southern areas on the Silk Road specifically. Recently, there is a new hypothesis correlating the prehistoric migration of Indo-Europeans into Xinjiang area with so-called proto-Tocharians. According to this hypothesis, Indo-Europeans who migrated into south-eastern Turkistan (Xinjiang, China) became the ancestors of the Tocharians who were the indigenous European settlers in central Asia. This concept is especially supported by Mallory and Mair (2000), Christopher Beckwith (2009), David Anthony (2010), and Sverchkov (2012). These researchers regarded Indo-Europeans of the Afanasievo culture who migrated into the northern route of Xinjiang Silk Road in 3000 BCE as the origin of Tocharians. The Yuezhi and Kushān kingdoms in later times are also considered as Tocharian affiliations. However, there is no vestige of influx of Afanasievo culture into the southern route of the Xinjiang Silk Road in 3000 BCE. Instead, the earliest archaeological evidence for an Indo-European influx into the southern route are the Xiaohe cemetery and the Lop Nur site in 2000 BCE as described above. It is generally accepted that the origins of the Tocharian language may lay with the proto-Tocharians who migrated from the West and then occupied the Silk Road routes. In brief, the influx of the ancient Indo-European nomads occurred along the Xinjiang Silk Road on two putative routes: the northern route occupied in 3000 BCE, and the southern route in 2000 BCE. Fortunately, the Tocharian language used in about 500 BCE is still preserved and is studied by researchers. According to these, Tocharian languages can be differentiated as A and B, although details of the languages are still obscure. Even so, Tocharian language A and B can be considered to be the final result of the cumulative migration and occupation of Xinjiang Silk Road by Indo-Europeans at different historical periods. Even after the extinction of the Xiaohe culture, the influx of such nomadic cultures constantly progressed. Notably, the Saka tigraxauda during 600–400 BCE flowed into various regions of northern China. Thereafter, until the Hunnic rise and invasion westward, for about 3000 years, Indo-Europeans with a pastoral economy migrated into regions on the Xinjiang Silk Road, introducing such new technologies as metallurgy, stockbreeding, dairy processing, and felt processing into East Asia. Thus, the Lop Nur artifacts of the initial Indo-European culture represented by the Xiaohe cemetery on the southern route of the Silk Road have deep significance in Eurasian archaeology in that they represent the commencement of cultural exchange between the east and the west, centering on the prehistoric Silk Road. The scientific analysis of the Lop Nor people reveals many novel findings that help us to understand their everyday life. With regard to their physical status, the percentages of tooth wear, dental calculus, and mandibular joint osteoarthritis were relatively higher in the Xiaohe culture people as compared to the other contemporary

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settlers (He et al. 2014). Some of the mummies found in this area were the inhabitants under such cultural and anthropological influences.

The Mummies in Xinjiang Museum Xinjiang Museum (or the Xinjiang Uyghur Autonomous Region Museum) is situated in the Shayibake district of Urumqi, Xinjiang Uygur autonomous region of China. The museum exhibits Xinjiang mummies from different temporal or spatial perspectives, explaining customs of various ethnic groups and the history of the Xinjiang region. Particularly attracting our attention is the exhibition room on the second floor, displaying “Death and immortality – the ancient mummy exhibition in Xinjiang.” Since September 2014, the exhibition has displayed the eight Xinjiang mummies that represent individuals from the prehistoric to the Tang dynasty period in Chinese history. Their estimated ages range from infants to adults with completely preserved cultural remains such as caftan, felt hat and shoes, and other garments. Of them, six are dated from the Bronze to early Iron Age, and two are known as Tang dynasty mummies. Of the mummies in Xinjiang Museum, the earliest is the female mummy excavated in 1980, at the Tiebanhe Cemetery site located on the tributary of the Kongque River. By the public, the female mummy is renowned as the Beauty of Loulan or the Loulan Beauty. In life she was 152 cm tall, with a high nose, deep eyes, long lashes, and a defined chin. Her nails, straight hair, and skin tissue are preserved very well. Her age at death is estimated to be 40–45 years. Her blood type was O. In 1995, her face was reconstructed by a Japanese researcher. Considering the associated artifacts and context in excavation, the site belongs to the Xiaohe Culture, dated ~1800– 1500 BCE (Qi et al. 2008). Another mummy, the so-called Princess of Xiaohe, is also displayed in the Xinjiang Museum. The female mummy was discovered at the Xioahe cemetery site. Her height was estimated to be 158 cm. The preservation status of the head and face is exceptionally good. The deep cavernous eyes and a sharp nose were a few of the descriptive characteristics. The mummy is dated 1800–1500 BCE (Bronze Age). Her related cultural heritage belonged to the Xiaohe culture, with its signature burial type and attributes of discovered artifacts such as felt hat, garments, and wooden facial masks. In the Xinjiang Museum, mummies presumed to be later in time are also on display. They are the mummies from the Ruoqiang cemetery site. The mummies of the infant, adult female, and male are displayed together. Considering the name of the site where the mummy was discovered (Cherchen or Qiemo County, Xinjiang province), the adult male mummy was nicknamed as Cherchen man. The adult female and male mummies were found together inside the grave, therefore, they are thought to have been wife and husband. In a DNA analysis, the female mummy showed the hybrid traits of ancient Europoid and Mongoloid peoples. Meanwhile,

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the infant mummy of the family, estimated to have been 8–12 months old at death, was also found in the grave #1. The infant mummy was dated to ~1000–800 BCE. Judging from associated artifacts, the mummy is assumed to be of the middle Bronze Age Yanbulake culture. In addition to these, several Loulan period mummies are displayed in the Loulan Museum in Ruoqiang County (Charkliq or Chaqiliq, Qakilik in Uyghur Language), China.

Conclusion Previous reports about the Xinjiang mummies by Aufderheide (2003) were extremely significant for mummy researchers because they provided fundamental data about one of the oldest Chinese mummies ever discovered, in the very arid desert area of the Chinese periphery. Since then, however, subsequent discussions among archaeologists in China and other countries indicate that they need more data about the mummies and their society. In this chapter, through descriptions of the Xiaohe cemetery, related Silk Road archaeological sites, and the mummies discovered therein, we have reviewed the present situation of research on the Bronze Age cultures left by ancient Indo-Europeans who drifted into northwestern China as early as 2000–1500 BCE. The mummies and the early Bronze Age artifacts associated with them found in the Xinjiang area have been investigated in the last century, but professional analyses based on substantive evidence has been limited. But because of archaeological information that has recently emerged from the Xinjiang Silk Road reinvestigation, the significance of the mummies has been validated anew. The Xinjiang mummies currently maintained in the local museums have significance in two aspects: their contribution to the history of Silk Road researches and to mummy studies worldwide. Especially for the latter, the mummies and their culture could provide invaluable information about the first demic diffusion of Indo-Europeans to the east in the prehistoric era. In general, research into the Silk Road has mostly been concentrated on the Han–Tang dynasty period. Nonetheless, actual cultural exchange between the east and the west pivoting on Central Asia initiated the expansion of nomadic culture in Eurasia as early as ca. 3000 BCE. What enabled nomadic people to pioneer in this desolate region was the newly invented pastoral economy. Through breeding herbivorous livestock, even uncultivable grassland can be made into a place habitable by humans. In addition, the climate at that time was more temperate then than at present. As oases and pastures developed in these areas, nomadic people could establish communities in different places along the Silk Road. In other words, the Silk Road did not unexpectedly emerge in the Han dynasty, but probably resulted as an outgrowth of existing footholds established as early as 3000 BCE, eventually connected to one another by the Silk Road as it consolidated the trade routes that persisted from antiquity. Such well-known cities as Loulan Kroraina and Turpan eventuated from the efforts of these nomads to set up their bases in the desert thousands of years ago.

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The Xinjiang mummies are also significant because of their superb preservation, even after the lapse of several millennia. As is well-known, naturally mummified examples provide the best research subjects for scientific analysis for revealing the life and health status of ancient peoples. Since the mummies are very rare cases that have been preserved in the remote periphery of China during ancient times, sensitive and detailed scientific studies on them in the future could provide wonderful data to archaeologists, anthropologists, and paleopathologists worldwide.

Cross-References ▶ Joseon Dynasty Mummies of Korea ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies in Japan ▶ Mummies in Siberia ▶ Mummies of Song-Ming Dynasty in China

References Anthony DW (2010) The horse, the wheel, and language: how bronze-age riders from the Eurasian steppes shaped the modern world. Princeton University Press, Princeton Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Beckwith CI (2009) Empires of the Silk Road: a history of Central Eurasia from the Bronze Age to the present. Princeton University Press, Princeton Bergman F (1939) Archaeological researches in Sinkiang. Publications of the Sino-Swedish Expedition 7, Stockholm Bergman F, Bexell G, Bohlin B et al (1945) History of the expedition in Asia 1927–1935. IV. General reports of travels and field-work. Elanders Boktryckeri Aktiebolag, Stockholm Cockburn A, Cockburn E, Reyman TA (1998) Mummies, disease and ancient cultures, 2nd edn. Cambridge University Press, Cambridge He L, Zhu H, Li WY et al (2014) Oral heath and diet of the Xiaohe cemetery populations in Lop Nur, Xinjiang Province. Acta Anthropol Sin 33(4):497–509 Huang WB (1948) Archaeological report of Lop Nor, vol 1. Science Press, Beijing. (Publication in Chinese) Kovalev AA (ed) (2014) Earliest Europeans in the heart of Asia: the Chemurchek (Qiemuerqieke) cultural phenomenon, vol 1. Book Antiqua, Saint Petersburg. (Publication in Russian) Kovalev AA (ed) (2015) Earliest Europeans in the heart of Asia: the Chemurchek (Qiemuerqieke) cultural phenomenon, vol 2. Book Antiqua, Saint Petersburg. (Publication in Russian) Li CX, Zhou H (2016) A study of the matrilineal genetic diversity of the human remains unearthed from Xiaohe cemetery. West Reg Stud 1:50–55. (Publication in Chinese) Liang YM, Liang YM, Yidilisi A (Idris A) et al (2012) Proteomic analysis of residues in grass basket excavated from Xiao-He graveyard. Sci Conserv Archaeol 24(4): 81–85. (Shanghai Museum; publication in Chinese) Mair VH (2006) The rediscovery and complete excavation of Ördek’s Necropolis. J Indo-Eur Stud 34(3–4):273–318 Mallory JP, Mair VH (2000) The Tarim mummies: ancient China and the mystery of the earliest peoples from the West. Thames & Hudson, London

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National Cultural Heritage Administration (NCHA) (2012) Atlas of Chinese cultural relics: Xinjiang Uyghur Autonomous Region, vol 1–2. Cultural Relics Publishing House, Beijing. (Publication in Chinese) Polos’mak NV (2001) Vsadniki Ukoka (Horsmen of Ukok). INFOLIO Press, Novosibirsk. (Publication in Russian) Qi XS, Wang B, Tang XM (2008) The ancient culture of Xinjiang along the Silk Road. Xinjiang People’s Publishing House, Xinjiang. (Publication in Chinese) Sverchkov LM (2012) Tokhary. Drevnie indoevropeitsy v Tsentral’noi Azii (Tocharians. Ancient Indo-Europeans in Central Asia). SMI-ASIA Publ., Tashkent. (Publication in Russian) Wang BH (1983) Preliminary report of ancient cemetery in Kongque River. Xinjiang Soc Econ 1:116–128. (Publication in Chinese) Xinjiang Institute of Cultural Relics and Archaeology (2004) The excavation report of Xiaohe cemetery. Res China’s Front Archaeol 3:338–398. (Publication in Chinese) Xinjiang Institute of Cultural Relics and Archaeology (2007) A brief excavation report on Xiaohe graveyard located in Luobubo, Xinjiang Autonomous Region. Cult Relics 10:4–42. (Publication in Chinese) Zhang YC (2009) The Northern Cemetery: a mystery lasting for thousands of years buried deep in the desert. Xinjiang Humanit Geogr 3:69–75. (Publication in Chinese)

Joseon Dynasty Mummies of Korea

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Dong Hoon Shin, Chang Seok Oh, Jong Ha Hong, Myung Ho Shin, Myeung Ju Kim, and Hye Jin Lee

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . How Could the Korean Mummies Be Formed? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Emergence of the Joseon Graves in History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Preservation Status and Histology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ancient DNA Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Successful Mummy Researches of Historians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea e-mail: [email protected] M. H. Shin Department of History, Pukyong National University, Busan, Republic of Korea e-mail: [email protected] M. J. Kim Department of Anatomy, Dankook University College of Medicine, Cheonan-si, Chungcheongnam-do, Republic of Korea e-mail: [email protected] H. J. Lee Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, Republic of Korea Ministry of National Defense Agency KIA Recovery and Identification, Seoul, Republic of Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_29

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Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1068 Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1069 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1069

Abstract

Joseon dynasty people’s life has been delineated by studies based on the extant historical literature in Korea. Notwithstanding this, over the recent several decades too, archaeologists have investigated the sixteenth- to eighteenth-century Joseon period mummies to glean corroborative or corrective data on that specific time period. In the studies, Joseon mummy’s almost-perfect preservation state has surprised anthropologists and attracted the attention of archaeologists, due to the high academic value of the artifacts interred with them. This chapter provides a general overview of the research work on Korean mummies that has been represented in academia. Keywords

Korea · Mummy · Joseon dynasty · History · Histology · Ancient DNA · Documents

Introduction Mummies unearthed at archaeological sites have been regarded invaluable to the disciplines of archaeology, anthropology, and paleopathology. Although the exact mechanism of mummification is as yet not fully understood, mummies are crucial repositories of scientific information. Today, studies on mummies have expanded by means of continuously renovated research techniques and archaeological methodologies. Worldwide mummy research continues to trace the sociocultural aspects of ancient peoples, by which multifaceted knowledge is acquired in much detailed spatiotemporal contexts (Cockburn et al. 1998; Aufderheide 2003; Lynnerup 2007; Shin et al. 2018a). In general, rich cultural endowments of East Asian countries have been largely inaccessible to the scholars of other continents, due mainly to language barriers. The same can be true for mummy researches. Mistaken assumption that very few mummies have been found or investigated in East Asian countries is based on the knowledge of environmental conditions not favoring a mummification process. Moreover, East Asia does not have any embalming tradition in history. These concepts caused the prevailing belief among the academia that mummification is uncommon in this region. All of this notwithstanding, the reality of mummy studies about East Asian countries is very different. Well-designed studies on mummies discovered in Korea, China, and Japan have been ongoing over the past several decades. Therefore, scientific news of mummy discovery and research are often reported from those countries. Nowadays, in the wake of the scholarly achievements thus far, academia agrees on the value of East Asian mummies in historical and scientific perspectives.

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Nevertheless, the brief reviews and infrequent reports aside (Peng and Wu 1998; Aufderheide 2003; Oh et al. 2017; Shin et al. 2018a), the details about the findings of East Asian mummies are still scarce. In this book, multiple chapters thus contribute to the achievements of mummy studies conducted in East Asia. Among the chapters dealing with cultural or biomedical characteristics of the Asian mummies, this chapter will describe the information about Korea mummies from the fifteenth- to nineteenth-century graves of the Joseon dynasty period (1392–1910 CE), which is rarely revealed by other historical or anthropological reviews.

How Could the Korean Mummies Be Formed? The Joseon dynasty mummies of Korea have been investigated more comprehensively than any other Asian mummies. Due to perfect preservation conditions, original scientific data concerning the health and disease status of the Korean people in history could be successfully obtained from the mummies (Lee et al. 2013). The basic information of Korean mummies reported so far is summarized in Table 1. Previous studies revealed that mummification in the world occurred at least in two different ways: natural or artificial mummifications (Shin et al. 2003a). Natural mummification generally occurs in harsh climatic conditions such as dry (e.g., desert) or permafrost environments. Such examples include Ötzi the Iceman (Hess et al. 1998), arctic mummies (Hart-Hansen et al. 1991), pre-Columbian South American mummies (Reinhard 1996), and mummies discovered in Taklimakan Desert (Spindler et al. 1996). At the earliest stage of researches, excellent preservation status of Joseon mummies certainly took the people by surprise (Lee et al. 2013; Song and Shin 2014). However, major inducing factors for natural mummification could not fully explain this phenomenon because the climate of Korea is not conductive to it at all. Then, have Korean mummies ever been formed by artificial embalming techniques? In fact, Joseon mummies were totally different from those embalmed in the other ancient civilizations. Korean people in history did not believe in the resurrection or immortality of the soul. Rather, the preservation of an ancestor’s corpse after death was likely an ominous sign to them. They must not have utilized any kind of embalming techniques for the funeral (Oh et al. 2017). Archaeologists thus presumed that the structure of the Joseon grave (called Hoegwakmyo in Korean) might have been closely related to the mummification. The Joseon graves were constructed throughout almost every corner of Korean peninsula during the fifteenth to nineteenth century (Shin et al. 2008). The graves were unique because they were surrounded by a lime-soil mixture barrier (Fig. 1). At archaeological sites in Korea, well-preserved mummies have rarely been found in the graves with the lime-soil barrier compromised. Instead, they were discovered when grave’s barrier remained intact at the time of discovery. The maintenance of barrier thus looks important for successful mummification inside Joseon graves (Lee et al. 2013).

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Table 1 Korean mummies investigated by 2020 Mummy Kunkook Yongin HD

Research institute concerned Dankook University Gyeonggi Cultural Foundation Jinju National Museum

Year 2001 2006 2006

Gangneung

Gangneung choi clan

2007

SN1-2 SN PK Gongju Seocheon Waegwan Dangjin HD2

Hangang Institute of Cultural Heritage Hangang Institute of Cultural Heritage Chungnam Institute of History and culture Gyeonggi Cultural Foundation Daedong Institute of Cultural Heritage Chungnam Institute of History and Culture Descendant of Onyang Jung clan

2007 2007 2007 2008 2008 2008 2009

Mungyeng

2010

Junggye Daegu_HS Cheongdo

Gyeongju National Research Institute of Cultural Heritage Dong-Seo Institute of Cultural Heritage Chungcheong Institute of Cultural Heritage Gyeong-Sang Cultural Heritage Research Center HanBeak Institute of Cultural Heritage Honam Institute of Cultural Heritage Honam Institute of Cultural Heritage DongGuk Institute of Cultural Properties Gyeong-Sang Cultural Heritage Research Center Han Ul Research Institute of Cultural Heritage Daedong Institute of Cultural Heritage Yeongnam Institute of Cultural Properties

Yeongweol Jangsung Gumi

Jungbu Institute for Archaeology Chungcheong Institute of Cultural Heritage Gumi City Hall, Korean Government

2015 2017 2019

Changwon

Center of History & Culture, Gyeongnam Development Institute Yeongnam Institute of Cultural Properties Hangang Institute of Cultural Heritage

Jinju Sapgyo Sacheon Hwasung YG2-4 YG2-6 Andong Dalsung

Gyeongsan GM-8

Sex Female Female Female

2010 2011 2011

Discovery Excavation Excavation Moving a grave Moving a grave Excavation Excavation Excavation Excavation Excavation Excavation Moving a grave Moving a grave Excavation Excavation Excavation

2012 2012 2012 2013 2014

Excavation Excavation Excavation Excavation Excavation

Male Female Female Male Female

2014 2014 2015

Male Female Male

2019

Excavation Excavation Moving a grave Excavation Excavation Moving a grave Excavation

2019 2020

Excavation Excavation

Male Male Male Male Female Male Female Female Female Male Male Female

Male Male Male Male Male Male

When Korean mummy studies began, researchers did not know why and how human remains were not decomposed but were mummified inside Joseon graves so perfectly. After a long period of research, however, they inferred that the perfect sealing property of lime-soil mixture might play an important role in the mummification (Oh and Shin 2014; Shin et al. 2018a). In addition, lime (calcium oxide) is

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Fig. 1 (a) The structure of the Joseon grave (called Hoegwakmyo). IC, inner coffin; OC, outer coffin; LSMB, lime-soil mixture barrier. (b–f) The Joseon graves were constructed in Korean peninsula during the fifteenth to nineteenth century (CREDIT: Dong Hoon Shin)

also known to have an affinity for water (Lee et al. 2013), augmenting an exothermic reaction. In experiments with miniature Joseon graves to simulate the mummification process in Korea, lime-soil mixture around the coffin, in the course of hardening, generated enough heat to eradicate bacteria inside the coffin (see the details in the chapter of this book: ▶ Chap. 48, “Mummies of Song-Ming Dynasty in China”). We presume that the same effect might have also occurred in actual graves constructed during the Joseon period (Oh and Shin 2014; Oh et al. 2018a). Nonetheless, it is unlikely that mummification in Korea was induced by a single cause. Rather, it is more likely that mummification could be achieved by the synergistic effect of multiple inducing factors. First, we note water infiltrating into the coffin through highly alkaline lime mixture layer. The pH of water inside the coffin also became alkaline, thus hindering bacterial growth required for decomposition of organic materials. Charcoal layer spread upon a bottom plate several inches thick might have been another cause for mummification due to its action in the removal of moisture (Fig. 2). The coffin wood might relate with the mummification too. Most coffins used for Joseon graves were made of a pine tree (Fig. 3). And the pine tree is known to delay or stop the decomposition due to its bacteriostatic or bactericidal effects (Spigelman

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Fig. 2 Mummy bundle and charcoal spread upon the bottom plate of coffin (CREDIT: Dong Hoon Shin)

and Donoghue 2003; Shin et al. 2008). Another potential factor is an anoxic condition inside the coffin. Korean mummies were often discovered in doublecoffined graves where many articles of clothing were discovered (Fig. 4). In those graves, oxygen was too scarce to be used for successful decomposition of organic materials (Shin et al. 2008; Shin et al. 2018a). We presume that complete sealing of a coffin, oxygen deficiency, extremely high pH, presence of charcoal, a coffin made of pine tree, and high temperature generated by the lime’s exothermic reaction might all be related to successful mummification in the Joseon graves (Shin et al. 2008; Oh et al. 2018a; Shin et al. 2018a).

Emergence of the Joseon Graves in History Mummification in Korea is more related to cultural aspects than to natural environments (Shin et al. 2018a). For whatever reason, the Joseon graves have surrendered mummified remains that were not identified in the other types of tombs investigated in Korea (Shin et al. 2008; Lee et al. 2013). In fact, the mummy-relating graves were not constructed before the Joseon dynasty period. Those unique structured tombs emerged abruptly by a political event: a rise of Confucians in Korean history (Shin et al. 2008). In the late fourteenth century, Korean Confucians criticized that the national crisis was caused by Buddhists. Among the Buddhists’ misdeeds, the worst

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Fig. 3 Coffin wood of Joseon grave. Most of them were made of pine tree (CREDIT: Dong Hoon Shin)

thing was the exemption from tax and military service. Since Korea at that time seriously suffered from repeated foreign invasions, such Buddhists’ evasions were vigorously blamed by the Confucians (Shin et al. 2008). Certainly, to fulfill their hopes of radical revolution in the country, the Confucians looked forward to the advent of a national hero (Shin et al. 2008). A general named Seong-gye Yi (1335–1408) was the man they wanted (Fig. 5). He was born in the northern frontier territory of the Korean kingdom. He was not of a noble birth. However, the political turmoil of the late fourteenth century gave him the opportunity for a great success. He won every battle against the foreign invaders, by which he became a symbol of national independence. With the general’s assumption of absolute authority over the entire kingdom, the Confucians toppled the extant hegemony of the Buddhists. They finally established their own Joseon dynasty (1392–1910 CE) as a necessary means to root out all the social evils caused by the Buddhists (Shin et al. 2008). After that time, Buddhist hegemony in Korean society began to be overturned. Among radical reforms undertaken, the rituals became one of the priority targets (Lee et al. 2013). Before the Joseon grave was generally accepted by the ruling elites, Korean noblemen used stone chambers for their burials (Fig. 6). The burial system was awfully expensive but easily plundered by tomb robbers. In this regard, the quick emergence of the Joseon graves in the fifteenth century appears to have been caused by the needs of the Korean people and society (Lee et al. 2013).

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Fig. 4 Many articles of clothing found in the Joseon grave (CREDIT: Dong Hoon Shin)

Fig. 5 A portrait of Seonggye Yi, the first King of Joseon dynasty. Courtesy of National Museum of Korea

In fact, the original structure of the Joseon graves was recommended by Zhu Xi (1130–1200 CE), the grand master of Confucianism (please see the details in this book’s chapter: ▶ Chap. 48, “Mummies of Song-Ming Dynasty in China”). Zhu Xi proposed the actual way on how the orthodox Confucians could build their graves (Lee et al. 2013): (1) a charcoal and lime-soil mixture was spread upon the pit

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Fig. 6 Stone chamber tomb. Courtesy of the National Museum of Korea

bottom, on which the coffin was laid; (2) the space around and topside of coffin were completely filled by the lime-soil mixture and charcoal; and (3) the lime-soil mixture was finally hardened to be a concrete-like block around the coffin. The grandmaster explained that the graves became proof against the grave robbers and insects once the lime-soil mixture would be hardened like a stone (Shin et al. 2008). The structural details of the early-stage Joseon graves were described in the provisions of the National Ritual Code 1474 (Gukjo-ore-ui) or other ritual guides published afterward (Lee et al. 2013). Nonetheless, the immediate adoption of Zhu Xi’s grave in the Joseon society was not easy. As is described above, prior to the Joseon period, the Korean elites favored the combined use of stone chamber tombs and duplicated (inner and outer) coffins. Joseon gentry still preferred double coffins in spite of Zhu Xi’s original recommendation (a single coffin). Even after the Joseon dynasty was founded, this age-old tradition (double coffins) was not abandoned with ease. However, this modification of Zhu Xi’s original idea was also intolerable to the orthodox Confucians at that time (Shin et al. 2008). They were not satisfied with the modification of Zhu Xi’s golden rule. They continuously argued that the Joseon graves must be changed in full accordance with Zhu Xi’s original design: a single coffin placed inside the grave. To the twenty-first-century people like us, this kind of debate may appear somewhat silly. However, no matter how ridiculous it may seem to us, the talks were very serious to religiously devout Joseon people (Shin et al. 2008). Eventually, the Joseon society continuously assimilated Zhu Xi’s ideology. The Joseon people respected Zhu Xi’s recommended grave as an ideal burial system until the fall of the dynasty (Lee et al. 2013; Oh et al. 2017; Shin et al. 2018a). In spite of their hopes to

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build ideal burials for Confucians, however, the Joseon graves unexpectedly served to mummify their parents, grandparents, and ancestors.

Preservation Status and Histology One of the most popular topics about the Joseon mummies was their preservation status (Shin et al. 2003b; Shin et al. 2018a). According to gross anatomical inspection, the Korean mummy’s overall state of preservation was very good (Figs. 7 and 8). The mummy skin showed elasticity, and the joints were freely movable. Creases and fingerprints were visible on the mummy skin. Nails on the hands and feet were well preserved. The mummies’ abdominal region was commonly depressed due to the contents of internal cavity being contracted and displaced. Radiological data also corroborated the Korean mummy’s excellent preservation status observed in anatomical inspection. Bone densities did not differ from living individuals. Muscles were well preserved in most parts of the mummified body. Internal organs were clearly seen on CT scan images. Mummified brains were found inside the skull, and other mummified organs such as the lung, liver, heart, etc. also remained intact in the body cavities (Shin et al. 2003a; Kim et al. 2006). Histological analysis revealed the preservation pattern of Korean mummies in more detail. In light and electron microscopy, the skin morphology of Korean mummy was somewhat different from those of living individuals (Shin et al. 2003a; Chang et al. 2006a). The mummy’s epidermis remarkably decreased in thickness, but the presence of melanin pigments was confirmed in the basal layer of the epidermis (Shin et al. 2003b; Chang et al. 2006a) (Fig. 9). Histology also showed many connective fibers filled in the skin dermis. The exact nature of connective fibers could not be defined by light microscopy. However, in electron microscopy, we observed banding patterns, the ultramicroscopic structure typically Fig. 7 A Korean child mummy found in Joseon grave (Yangju). (Shin et al. 2003b) (CREDIT: Dong Hoon Shin)

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Fig. 8 A Korean mummy found in Gangneung. (Lee et al. 2009) (CREDIT: Dong Hoon Shin)

Fig. 9 Histology of Korean mummy skin. (Shin et al. 2003a) (CREDIT: Dong Hoon Shin)

seen in collagen fibers (Shin et al. 2003a; Chang et al. 2006a) (Fig. 10). Microscopically, mummified skin was preserved better than expected. Histological research identified different patterns between mummy skins. It demonstrated differences according to the mummification process (natural and artificial) (Chang et al. 2006a). For instance, in case of the naturally mummified skin of ancient bog bodies (Germany), the epidermis almost disappeared; collagen

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Fig. 10 Electron microscopy revealed collagen fibers in mummified skin dermis. (Chang et al. 2006a) (CREDIT: Byung Soo Chang)

fibers were present in the dermis, but no cellular elements remained (Stucker et al. 2001). Similar histological pattern was also observed in naturally formed, 2500year-old Altai mummies (Romakov et al. 2002). By contrast, in artificially embalmed mummies, the skin exhibited different histological patterns. In Egyptian mummy skin, in addition to the connective fibers, subcellular structures such as the desmosomes, tonofilament bundles, and mitochondria could be identified (Hino et al. 1982; Montes et al. 1985; Perrin et al. 1994; Chang et al. 2006a). This means that artificially embalmed cases exhibited a better preservation status in skin histology than those of naturally formed mummies (Chang et al. 2006a). As for Korean mummies, the skin also exhibited loss of the epidermis and the presence of collagen fibers in the dermis. TEM images showed that the cells found in the skin dermis had compacted chromatin but very few cytoplasmic organelles. Adipocytes were visible in the hypodermis though their nuclei were not found in the cells (Shin et al. 2003a; Chang et al. 2006a). This means that subcellular structures were lacking in Korean mummy’s skin. The histology indicates that Korean mummy’s skin was more like naturally formed ones than artificially embalmed mummies (Chang et al. 2006a). Histological analysis was done on the Korean mummies’ hairs as well (Chang et al. 2006b). Despite archaeological hair samples looking intact, ultramicroscopic biodegradation actually occurred in them (Chang et al. 2006b). In general, weathering changes of hair proceed as follows: (1) a loss of hair scales; (2) a detachment of the cuticle layer that is protecting hair from biodegradation (Wilson et al. 2001); and finally, (3) the exposure of the hair cortex, the most important step in a hair weathering (Chang et al. 2006b). In this regard, the histology of Korean mummies’ hair was unique. The cuticle, cortex, and medulla of the Korean mummy’s hair were maintained well even after several 100 years of burial. Exo- and endocuticles were identified in the cuticle layer.

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No detachments or losses were found in the cuticle or cortical layers. No holes were observed in a hair medulla (Chang et al. 2006b). We also note that the scale edge of a Korean mummy’s hair exhibited a round border (Fig. 11). Considering a modern people’s hair, this was an impressive finding. The modern hair’s scale edge was generally sharp or angular due to repeated physical or chemical stimuli (Chang et al. 2006b). In the case of Korean mummy, calcium-rich crystalline coating might protect the hair surface against physicochemical damages (Chang et al. 2006b). Next, we will discuss the histology of the mummified internal organs. As for Korean mummy’s lung, histology showed collagen fibers in almost every part of pulmonary tissues examined. The remnants of alveolar cells were also visible in the mummified lung. The alveolar sac was delineated by degraded pneumocytes. Red blood cells (RBCs) and chondrocytes were found in the mummy bronchus (Fig. 12). The hepatocytes were preserved in the mummified liver (Shin et al. 2003a). In Korean mummy histology, the cells in superficially situated organs (e.g., skin) were much poorly preserved than those located deeper in body cavities (e.g., lung and liver, etc.). Therefore, when tissue sampling is needed for scientific research, specimens should be collected from the organs located deep in the mummies (Shin et al. 2003b). In many ancient civilizations, mummies’ intracranial contents were often removed by embalmers. However, the brains still remained in Korean mummy crania. The brains of Korean mummies retained original morphology in anatomical perspective. Histological analysis demonstrated that the cell bodies and nuclei disappeared in mummified brain. In histochemical and ultramicroscopic study of Korean mummy brains, the concentric structures, a typical TEM pattern of myelin sheath, were observed (Kim et al. 2008; Oh et al. 2013), as seen in mummified brain from the Tyrolean Iceman (Hess et al. 1998; Kim et al. 2008). Impressed by the excellent morphological, radiological, and histological data, scholars often presumed that Korean mummies’ preservation status is not so much

Fig. 11 Korean mummy hair. (Chang et al. 2006b) (CREDIT: Byung Soo Chang)

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Fig. 12 Red blood cells and chondrocytes in the lung and bronchus of Korean mummy (Shin et al. 2003a) (CREDIT: Dong Hoon Shin)

different from those modern individuals who just died. They even speculated that scientific experiments with the specimens of Korean mummies might be as productive as are the biomedical tests for living patients. However, no matter how impressive the mummy’s preservation was, delicate tests could not be easily applied to several 100-year-old Joseon mummy specimens.

Ancient DNA Analysis Over the past decades, the scope of ancient DNA (aDNA) analysis has expanded remarkably in the realm of archaeological science (Hofreiter et al. 2001). The technique is useful to obtain a knowledge about our ancestors’ medical traits in a more scientific way. The genetic background of specific diseases, the information useful for understanding worldwide disease history, could be also revealed by aDNA analysis. In case of Korean mummies, aDNA analyses were performed with mummified specimens obtained during autopsy or endoscopy. Such aDNA analyses were focused on ancient pathogens like ancient hepatitis B virus (aHBV) (Kahila Bar-Gal et al. 2012), Helicobacter pylori (Shin et al. 2018b), Ascaris lumbricoides (Hong et al. 2017), Clonorchis sinensis (Hong et al. 2019a), Paragonimus westermani (Hong et al. 2019b), and Trichuris trichiura (Hong et al. 2019c). The pathogen aDNAs were sequenced and analyzed to deduce phylogenetic data in spatiotemporal manner. The analysis of aHBV was done with a liver sample collected from a seventeenthcentury child mummy. This Korean mummy was discovered at the Joseon grave. Upon opening the coffin, the mummy bundle was moved to institute for more scientific research. The specimens of aDNA analysis were collected during an endoscopy. The analysis of aHBV genome revealed a sequence representing HBV

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genotype (HBV/C2) that was prevalent among the seventeenth-century Korean population. Comparing aHBV sequences from a Korean mummy with a modern counterpart, viral sequences showed genetic diversity between them (Kahila Bar-Gal et al. 2012). As for H. pylori, a bacterium causing gastric disease by proliferation in the mucosal epithelium of the stomach, modern genomes have been reported from all over the world. However, a historical picture of H. pylori is still lacking (Shin et al. 2018b). Since H. pylori aDNA could not be obtained from the skeletal series, there was a pressing need to acquire mummy stomach specimens for aDNA analysis. Especially as for H. pylori aDNA in East Asia, nothing is as good as stomach samples from well-preserved Korean mummies. The presence of H. pylori aDNA, especially of vacA alleles, was successfully identified in Korean mummy stomachs. This was the first report of H. pylori aDNA isolated from East Asian mummies (Shin et al. 2018b). The phylogenetic analysis of H. pylori DNA is summarized in Fig. 13. DNA analysis becomes an important tool, not only for the analysis of ancient pathogens but also for the genetic profiling of the people living in the past. There were several reports about successful DNA profiling of archaeologically obtained specimens. For instances, mitochondrial DNA (mtDNA) and short tandem repeats (STR) analyses showed genetic relationship between the individuals buried together in the third century Kazakhstan kurgan (Clisson et al. 2002). DNA analysis also replied to a great question concerning the fate of the Romanov royal family, revealing that none of the Russian Tsar’s wife, son, and daughters survived after 1918, thus closing the hitherto hot debate about the tragedy (Coble et al. 2009). Many researchers in Korea also performed genetic profiling to reconstruct the life history of mummified individuals (Oh et al. 2015). In brief, the genetic profiles of mitochondrial or Y-chromosomal DNA could be obtained from a child mummy discovered at Yangju. In aDNA analysis using internal organ specimens such as the lungs, liver, and muscles, a child mummy’s Y-STR profiles were successfully haplotyped (Kim et al. 2006). More recently, short tandem repeat (STR) and mitochondrial DNA (mtDNA) profiles could be also obtained from another Korean mummy (Oh et al. 2015). The identification of single-nucleotide polymorphism (SNP) pattern, the most common sequence variation in human genome, can provide researchers with a reference to uncover the individual’s genetic background for specific physical traits (Shastry 2002). Such traits include the ABCC11, ectodysplasin A receptor (EDAR), fibroblast growth factor receptor 2 (FGFR2), and ABO genotypes. The SNP located in the ABCC11 gene is genetically related to earwax type, also being associated with axillary osmidrosis, colostrum secretion, as well as the potential risk of mastopathy (Yoshiura et al. 2006; Ishikawa et al. 2012). Hair morphology is related with the SNPs in EDAR. The shovel shape of the upper incisors is also known to be EDAR gene dependent (Kimura et al. 2009). FGFR2 is the genetic determinants of hair thickness (Fujimoto et al. 2008, 2009), craniosynostosis (Ko 2016), and breast cancer (André and Cortés 2015). ABO blood type is another trait widely analyzed in human genetics. By combination of codominant A and B alleles and recessive O allele, ABO gene determines four different phenotypes: A, B, AB, and O

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Fig. 13 Phylogenetic tree of H. pylori vacA s-region. Red dots represent Korean Cheongdo and Dangjin mummies. Analysis was based on ML method. Numbers next to each node are bootstrap percentages (CREDIT: Jong Ha Hong)

(Lalueza-Fox et al. 2008). ABO genotyping is also useful for archaeological science because it can determine the blood type of ancient people without serological testing (Watanabe et al. 2013). Certainly, the reports on SNP research demonstrate its applicability to ancient human specimens. However, academic potential of this technique could not be evidently proven for the analysis of mummy specimens. With respect to the

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ABCC11, EDAR, FGFR2, and ABO genes, only a limited number of literatures were produced for mummified individuals. In this regard, the Joseon dynasty mummies could be an essential resource for the SNP analysis of ancient Korean people (Oh et al. 2018b). SNP analysis was thus focused on the ABCC11, EDAR, FGFR2, and ABO genes of Korean mummies (Oh et al. 2018b). In brief, SNP analysis indicates that Korean mummies’ ABCC11 genotypes were mostly for dry-type earwax and low risk for axillary osmidrosis. The variant alleles were also identified for EDAR and FGFR2 genes of Korean mummies. ABO genotypes (BO02, O01O02, AO01, BB, and O02O02) also were found in these cases. SNP genotyping on Korean mummies provide us with specific insights into the usefulness of this method in archaeological science (Oh et al. 2018b). Finally, the genetic analysis was also done for the diagnosis of a Korean mummy’s genetic predisposition to atherosclerosis. In a CT scan, multiple calcifications were identified in the aorta of the seventeenth-century Korean female mummy (Mungyeong) (Kim et al. 2015). The calcifications on radiography likely represent the atherosclerosis of the aortic wall; and the CT diagnosis was further confirmed by subsequent autopsy. The mummy must have had a coronary artery disease (CAD) as well because the left anterior descending (LAD) artery showed intimal thickening. This was a rare case of atherosclerotic cardiovascular disease (ASCVD) found in mummies (Kim et al. 2015; Shin et al. 2017). Despite its academic implications, the anatomical and histological research was not able to capture every aspect of ancient ASCVD occurring in the mummy. In aDNA of the Korean mummy, we identified the risk alleles of different SNPs for ASCVD among East Asian people (Shin et al. 2017). The reliability of the genetic analysis could be augmented by cross-validation of SNaPshot and Sanger techniques. The seventeenth-century Korean mummy was genetically predisposed to the risk of ASCVD. The paleogenetic study, the first of its kind outside Europe, confirms the validity of an adjunct SNP test in the definitive diagnosis of ancient ASCVD (Shin et al. 2017). In Korean mummy studies, a fluorescence microscopy equipped with a special filter set revealed autofluorescence emitted from the mummified structures. By laser capture microdissection, those fluorescence-emitting structures were proven to be cell residues with fragmented aDNA (Fig. 14). In mummy histology, due to the serious degradation of tissues over a long period, the morphology of cell residue is always hard to be differentiated, even for experienced histologists. Therefore, the detection of autofluorescence is a useful means to identify cell residues, thus enabling the selection of well-preserved mummy specimens for successful aDNA analyses (Lim et al. 2010). In spite of great successes in aDNA analysis among Korean mummies, we must note that there are still technical limitations to genetic analysis employed in the conventional studies. Actually, traditional aDNA analysis based on PCR and associated alignment of cloned sequences could not be completely free from modern DNA contamination (Hofreiter et al. 2001). Recently, next-generation sequencing (NGS) technique successfully challenges the claims about data authentication by its advantage on the analysis of ultrashort aDNA fragments (Orlando et al. 2015).

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Fig. 14 A fluorescence microscopy showing autofluorescence emitted from the mummified structures (Lim et al. 2010) (Credit: Dong Hoon Shin)

Therefore, in the forthcoming days, cutting-edge techniques such as NGS or DNA capturing will be used to fully exploit genetic information from Korean mummies.

Successful Mummy Researches of Historians Thanks to the growing body of literatures, the scientific community comes to recognize East Asian mummies in cultural perspective. In Korea, mummies and associated artifacts have been found inside the Joseon graves. Because they were found mixed up in the same coffin, experts from different research fields were called to do collaborative studies (Lee et al. 2013). Academic data gleaned therefrom can be accumulated as contextual information of the fifteenth- to nineteenth-century cultural remains and mummies. Today, interdisciplinary research on Joseon mummies has yielded invaluable data about the Korean people and society in history. One of such fascinating examples for multidisciplinary collaboration is the study of the 400-year-old Eung Tae’s grave (Lee et al. 2009, 2013). In April 1998, several Joseon period graves of the Kosung Yi clan were found during a rescue excavation. When one of the graves was investigated, a mummy bundle was discovered inside the coffin (Lee et al. 2009). The mummified person was a male. His name was Eung Tae. Respecting the descendants’ wishes, the mummified male was relocated to another cemetery on the same date. Therefore, no scientific research could be conducted on the mummy. During the dress historians’ work, many cultural artifacts were collected from Eung Tae’s grave, including different garments of the late sixteenth century (Lee et al. 2009). Inside the coffin, archaeologists also found many documents or mails. The preservation status of the documents was excellent. There were letters of a father to his son, expressing his grief of losing a son. The detailed profile of the buried individual could be revealed by a review of the documents. In the letter, Eung Tae

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was 31 years old when he died. Considering the letters, Eung Tae was the son of his father Yo Shin (1523–1611 CE) and the younger brother of Mong Tae (1551– 1642 CE) (Lee et al. 2009). The abovementioned information was not all that was available in the study of the letters or documents. A more sensational finding was the letter found spread upon Eung Tae’s face and chest. The letter was written in June 1, 1586. There was no overt identification of the letter’s writer (Lee et al. 2009). Judging from its contents, however, the letter must have been written by Eung Tae’s wife (Fig. 15). In the letter, she talked about an oath that they would be living together till death. She then deplored that her husband went to heaven alone, leaving their child behind. She wished to see her husband again in her dream; in the hope, she placed her letter within her husband’s coffin. Her letter ended with the following words: Please read my letter and come into my dream. Please show your face in my dream and say your words to me (Lee et al. 2009). In this letter, novel information about Eung Tae could be attained. In brief, he and his wife had children; and one of them might have been in her pregnancy. Considering the year of letter writing, Eung Tae must have died in 1586. Since Eung Tae was 31 years old when he died, his birth year was 1556 (Lee et al. 2009). Since this letter was written in the sixteenth-century Korean language, it must be academically important to concerned researchers. In another sense, this letter greatly impressed us because the Joseon period letter talked about the love story of a wife and her husband. Such explicit expression of love was likely difficult to be imagined for the sixteenth-century Joseon people who led an austere and ascetic life (Lee et al. 2009). However, a wife’s letter made us change our conception. Joseon people were very emotional and sensitive people like us.

Fig. 15 The wife’s letter spread upon Eung Tae’s face and chest. Courtesy of Andong National University Museum

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Fig. 16 The shoes made of Eung Tae’s wife’s hair. Courtesy of Andong National University Museum

Two more messages of Eung Tae’s wife were found inside the coffin. Among them, one was a brief memorandum on a wrapping paper for a lock of hair (Lee et al. 2009). Another message was also written on parcel paper to wrap the shoes that were found beside Eung Tae’s head. In the wrapping paper, scholars also noticed sentences: “Using my hair, I made these shoes. . .(illegible). . .You die before you can wear these shoes. . ..” (Lee et al. 2009). The shoes must be one of the most unique and culturally important remains discovered so far in the Joseon graves because they were likely made of human (the wife’s) hair, the first of such cases ever found (Lee et al. 2009; Kim 2010) (Fig. 16). Researchers wondered what may have happened to the surviving Eung Tae’s wife and children. About this, we do not have any idea or way to trace their destiny. There was no mention about her and her children in any historical literatures. This might have something to do with the political misfortune of Eung Tae’s clan in the seventeenth century. Due to a clan member’s rebellion against the King, a fierce revenge fell upon the clan. In the midst of political turmoil, the record about them might have almost disappeared (Lee et al. 2009; Kim 2010). The collaborative research on Eung Tae’s grave has made a big impact on Korean society and academia. Before the discovery, South Korean people did not regard the mummies and associated cultural remains as important cultural heritages worthy of academic research. However, Eung Tae family’s touching story changed many minds in South Korea. This report is one of the great successes ever achieved by mummy researchers in Korea, still making a profound influence on the academics (Lee et al. 2009; Kim 2010) (Fig. 17).

Conclusion As discussed above, Joseon mummy investigation in South Korea was problematic until quite recently, even in cases where permission for such study had been granted. However, the situation nowadays is in favor of researchers (Song and Shin 2014).

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Fig. 17 Another letter found in Joseon grave (Gwak Ju’s Letter; seventeenth century). Courtesy of Daegu National Museum

This chapter provided an overview and synthesis of the Korean mummy work, completed to date, along with some of the more fascinating cultural and biomedical information so acquired. We believe that future studies on Joseon mummies will add the final pieces of the puzzle in revealing the full extent of historical Koreans’ lives and lifestyles.

Cross-References ▶ Archaeological Findings of the Tarim Basin Graves and Mummies ▶ Joseon Dynasty Mummies of Korea ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies in Siberia ▶ Mummies of Song-Ming Dynasty in China Acknowledgment This research was supported by Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1D1A1B03030127).

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies of the Warring States and Western Han Periods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Excavation of Mawangdui Tomb No. 1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Biomedical Investigation of Mawangdui Mummy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Excavations on Mawangdui Tomb No. 2 and Tomb No. 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Excavation of Phoenix Hill Tomb No. 168 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Female Mummy of the Warring States Period . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Excavation of Double Dragons Tomb No. M1 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification in the Warring States and Han Period Tombs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

This chapter discusses four ancient Chinese mummies found in tombs of the Warring States and Western Han period from archaeological, cultural-historical, and medical perspectives. The most famous is the first, Western Han period female mummy excavated in 1972 from Tomb no. 1 at the Mawangdui site, Changsha city, Hunan province. Since the three Mawangdui Han tombs were found in extraordinary condition, many studies on the mummies themselves as well as various funerary artifacts have been carried out to understand still mostlyhidden, ancient Chinese culture. In 1975, another mummy of the Western Han period, a male, was excavated from Tomb no. 168 at the Phoenix Hill site, K. Joo (*) Department of Archaeology, Chungnam National University, Daejeon, Korea e-mail: [email protected] D. H. Shin Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_30

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Jiangling, Hubei province. The second female mummy was discovered in 1994 from Tomb no. 1 of the Guojiagang site, Jingmen region, Hubei province, which was thought to have been constructed in the Warring States period. This female mummy was the oldest mummy ever discovered in mainland China. In 2002, the third female mummy of Western Han period was excavated from Tomb no. M1 at the Double Dragon Tomb Site, Lianyungang city, Jiangsu province. This chapter summarizes the discoveries and academic research on these four ancient Chinese mummies as well as precious tomb artifacts such as lacquerware and fabrics. These four ancient Chinese tombs were found to have similar structures: they were covered with a thick kaolin clay layer, and contained lacquered wooden coffins in very deep ground soil. These characteristic structural features might be one of the most important factors affecting the near-perfect long-term preservation of those ancient mummies and organic artifacts. Keywords

Mawangdui Tomb · Mummy · China · Phoenix Hill Tomb · Double Dragons Tomb

Introduction Thanks to the rapidly growing body of archaeological reports, academic mummy research has come to recognize the distinctiveness of East Asian mummies in the cultural as well as biomedical perspectives. In the East Asian countries Korea, Japan, and China, there are many different kinds of mummies that have not been seen on the other continents. However, not many of these incredible findings and investigations in East Asia have been sufficiently represented or accounted for in the international academic world. The present chapter provides a general overview of the previous studies on ancient Chinese mummies found in the Warring States (475–221 BCE) and Western Han (202–9 BCE) period tombs. Among these mummies, the famous Mawangdui lady of the early Han dynasty is included. In this chapter, we delineate the mummies’ cultural and biomedical similarities and differences. Ancient mummies of the Tarim Basin region and those of the Chinese Song (960–1279 CE) and Ming (1368– 1644 CE) dynasties also have been discovered in Modern Chinese territory, but they are not included in this chapter. For those mummies, please see ▶ Chaps. 44, “Archaeological Findings of the Tarim Basin Graves and Mummies,” and ▶ 48, “Mummies of Song-Ming Dynasty in China” in this book.

Mummies of the Warring States and Western Han Periods In the spring and autumn period (770 to 475 BCE), many kingdoms in mainland China constantly waged warfare against each other, making for great political and social instability. However, the period also served as the background to remarkable

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progress in institutions, technologies, and philosophical and political ideas as well. By the fifth century BCE, the diverse culture of China was fully flourishing over the entire territory, despite its political and military complexity. Chinese civilization spread to areas of the Yangtze River Valley and parts of the southern territories of Manchuria at that time. During the next, Warring States period (475 to 221 BCE), small kingdoms merged into more powerful ones. Therefore, only seven major rival kingdoms (or states) contended for supremacy in China (Overy 2007, 88–89). After long political and military struggles, the Qin dynasty emerged as the final winner. The kingdom had continually attacked, destroyed, and annexed neighboring ones, finally ending rampant warfare that had been waged on a massive scale for several centuries. The first emperor of Qin, Shi Huangdi (re. 221 to 210 BCE), established a well-organized and powerful imperial system that had never been seen in East Asian history (Overy 2007, 88–89; The Metropolitan Museum of Art 2017, 1–11). Actually, the Han dynasty (206 BCE to 220 CE) of China was a beneficiary of the efficient politico-cultural system of its predecessor, the Qin dynasty. The elites of the Han dynasty enforced universal legal codes, centralized bureaucracy, and established strong military disciplines in the empire. With the explosive increase of the population, commerce and industry flourished, and big cities eventually emerged during the Han dynasty period (Overy 2007, 88–89). Mummies discovered in tombs dating to this period have fascinated worldwide researchers. Recently, Chinese archaeologists discovered four different mummies in very good condition from more than 2000-year-old Warring States and Western Han period tombs (Li 2016, 3). Three of them were discovered in the Hubei and Hunan provinces, and the other one was found in Jiangsu province. All of these mummies have been scientifically investigated, and they are currently displayed at local museums in China. Detailed information on the Warring States and Western Han period mummies is summarized in Table 1.

The Excavation of Mawangdui Tomb No. 1 The excavation of the Mawangdui (or Mawangtui) tombs was one of the greatest achievements of Chinese archaeology in the twentieth century. Besides its significance in academic aspects, the discovery and investigation of the Mawangdui tombs were so dramatic that the full story has come to be well known to the public throughout China. When the tombs were discovered, the world was still in the middle of the Cold War, and the communist China was not yet open to the outside world. The political situation of the country at that time thus inevitably affected the excavation of the Mawangdui tombs (Xiong and You 2006). The name “Mawangdui” comes from a name of a Changsha city hill that has hidden ancient tombs for a long time. Local tradition had wrongly identified the hill as the family tomb of the famous local king Ma Yin (852 to 930 CE) who ruled the Hunan region during the ninth to tenth centuries CE. Chinese archaeologists first surveyed this site in 1952, and evidently denied the authenticity of the local tradition. They concluded that the tombs must have been constructed during Han dynasty

1975

1994

2002

Guojiagang Tomb No. 1

Double Dragons Tomb No. M1

Year of discovery 1972

Phoenix Hill Tomb No. 168

Mummy Mawangdui Tomb No. 1

Lianyungang city, Jiangsu province

Jingmen city, Hubei province

Jiangling Region, Hubei province

Excavated sites Changsha city, Hunan province

Female

Female

25 BCE – 15 BCE

Male

Sex Female

The Warring States period

167 BCE

Estimated date of death After 168 BCE

Lianyungang Museum

Jingmen Municipal Museum

Jingzhou Municipal Museum

Collection Hunan Museum

Table 1 Mummies found in the Warring States and Western Han period Tombs in China Archaeological findings A wooden chamber Four nested wooden coffins Charcoal and kaolin clay on and around the wooden chamber A well-preserved mummy covered with silk and linens in acid water. Wooden figures, Lacquerwares, Pottery, Bamboo, and Wooden poles with inscriptions, Silk paintings A wooden chamber Two nested wooden coffins Kaolin clay on and around the wooden chamber A well-preserved naked mummy in reddish water Wooden figures, Lacquerwares, Pottery, Bamboo strips with inscription Destroyed wooden chamber and coffins A looted well-preserved mummy Lacquerwares, bronze artifacts, silk, and other fabrics Two wooden chambers Four parallel coffins with each corpse Kaolin clay on and around the wooden chamber One well-preserved mummy and water in the coffin no. 2 Lacquerwares, bronze seals, wooden poles with inscriptions

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period (206 BCE to 220 CE); thereafter, in 1956, the hill was designated a provincial historical heritage site. However, the tombs could not be properly investigated, due to the ensuing political turmoil of the Cultural Revolution (Xiong and You 2006). In December 1971, People’s Liberation Army soldiers at Changsha city accidentally came across the tomb when they were working at the hill to construct an air-raid shelter for an army hospital. In the process, they found uncertain structures emitting strange smelly gas from an underground cavity. Immediately, they reported the finding to Hunan Museum (previously known as the Hunan Provincial Museum). The museum staff reported the incident to central government officials in Beijing and got permission to excavate the site. This was in January 1972, when China was still in the mid of the Cultural Revolution (Xiong and You 2006). The archaeologists of Hunan Museum started to excavate the first tomb of the Mawangdui site on January 16, 1972. This excavation during high winter season was a very hard fieldwork. In addition, there was an insufficient budget and a lack of skilled labor for systematic archaeological excavation. Even high school students had to join the excavation as volunteer workers, as the tomb was too big and deep. More than 1500 workers, including the students, participated in the excavation (Yu and Wang 2016, 709). In April 1972, after a four-month excavation, the archaeologists could approach the core of the tomb, while not knowing whose tomb this was. They were very nervous when they found some evidence of tomb robbery, though fortunately, the tomb was still in an untouched state. The main burial of the tomb was discovered about 20 m below the surface of the mound. The shape of the burial pit was like an inverted step pyramid, and a wooden chamber was at the central bottom of the burial pit (a similar structure to that shown in Fig. 8 of this chapter). The archaeologists found a thick white kaolin clay layer under the soil. The lower part of the burial pit was full of the same kaolin clay, which covered and surrounded the wooden chamber. Beneath the kaolin clay layer, there was another layer, of charcoal, that wrapped the whole of the wooden chamber (Xiong and You 2006). These two unique materials, kaolin clay and charcoal, might have been used for the protection and preservation of the body (Fig. 1). The thickness of the kaolin clay layer covering the wooden chamber ranged from 1.0 to 1.3 m, and the thickness of the charcoal layer was between 40 and 50 cm. The total depth of the kaolin clay layer in the central pit of this tomb was around 3 m (Hunan Museum 1973a, 5). Kaolin clay is very sticky and plastic. In addition, its waterproof property is excellent. Charcoal is also a very good material to prevent humidity and putrefactive bacteria, as well as to preserve an overall good air condition. In fact, kaolin clay and charcoal were used for covering materials of noblemen’s burials in the Chu kingdom from the beginning of the Warring States period (Buck 1975; Xiong and You 2006). It is very interesting that mummy containing tombs of the Joseon dynasty in Korea have similar covering materials on and around their wooden coffins. As in the case of Joseon tombs with natural mummies, the wooden coffin was laid in the tomb pit and surrounded by a mixture of lime plaster and clay with a powdered charcoal layer. Therefore, an interment system using clay and charcoal materials might

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Fig. 1 Charcoal and white kaolin clay excavated from the Tomb no. 1 of Mawangdui site. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Kyeongmi Joo, Chungnam National University, Korea) Fig. 2 Wooden chamber of the Tomb no. 1 of Mawangdui site. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Kyeongmi Joo, Chungnam National University, Korea)

significantly affect the natural mummification process in tombs of ancient China and medieval Korea. For this theory, see the chapter on “Joseon Dynasty Mummies of Korea” in this book. The wooden chamber in Tomb no. 1 of the Mawangdui site was enormous (Fig. 2). As archaeologists continued the excavation, they found 26 sheets of rectangular bamboo strip mat made of weaving technique on the upper parts of the wooden chamber. The size of each bamboo mat is 2 m long and 1 m wide. Each bamboo mat has a handwritten Chinese character “Jia,” which means “home,” on each of the four corners (Hunan Museum 1973a, 121). The wooden chamber

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consisted of two layers made of many large timber planks, which were connected tightly to one another without any metal nails. The timber material was identified as “Cunninghamia lanceolata,” a kind of a Cypress tree native to China. The wooden chamber also consisted of double-stacked layers of timber planks, its overall shape resembling that of a giant rectangular box. The outer size of the upper part of the chamber was 6.73 m long and 4.81 m wide (Hunan Museum 1973a, 6–13). The total height of the wooden chamber was 2.8 m. Although many Chu and Han dynasty tombs had been discovered in the Changsha region before this excavation, the archaeologists had never before seen such a giant chamber. When the archaeologists in Changsha reported their findings of a giant wooden chamber in an undisturbed state to the Chinese government, the government immediately started to pay attention to this excavation. Thus, high-level governmental archaeologists in Beijing were dispatched to Changsha and joined the excavation. As the news attracted national attention, many media agents and the local public crowded into the excavation site, which numbers unfortunately hampered the excavation (Xiong and You 2006). The process of opening the wooden chamber started on April 15, 1972, and a few days later, the archaeologists discovered that the inner space of the wooden chamber was divided into five compartments (a similar shape to that shown in Fig. 9 of this chapter), each of which had not yet been opened. They were arranged centrically in the four directions of the central compartment. When the archaeologists removed the covers of the five compartments, they found that the main coffins had been buried in the central compartment (Hunan Museum 2017, 294–295). The four side compartments were full of many wooden figurines and burial items for the dead. The wooden figurines, numbering 163, were discovered in the north, east, and south compartments. They were divided into six groups according to their sex, style, and posture (Hunan Museum 1973a, 97–101; Hunan Museum 2019b, 237–249). The civil officials and most of the standard standing attendants were dispersed in the north, east, and south compartments. All of the female attendants, with costumes and including entertainers such as dancers and singers, were discovered only in the right section of the north compartment. A lacquer-painted wooden screen with a silk couch and a pillow, and a lacquer table on a reed mat were found in the left section of the north compartment (Fig. 3). In front of the screen and mat, there were arranged a set of lacquered trays and bowls with food for feasting (Fig. 4). Therefore, some scholars have speculated that the north compartment was a space for the ancestral rites of the dead. Many lacquerware, stoneware, bronze artifacts, vegetables, fruits, grains, meats, and spices were stored in the other three side compartments. Only the west compartment had no figurines, but there were many bamboo boxes with fully stuffed items, such as silk or other fabrics, animal bones, and plants or fruits. The four-directional compartments might have been used as a storage room or a treasure room for the daily life of the dead, who was the owner of this tomb. Most of the artifacts from the four side compartments were in an exceptionally fine condition and numbered over 1000 pieces. Among them, the most important ones are diverse kinds of lacquer artwork of the early Han dynasty. These days, much

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Fig. 3 Lacquered screen, lacquered table, and reed mat. Reconstructed display based on the findings of the left section in the north compartment of the Tomb no. 1 at Mawangdui site. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Kyeongmi Joo, Chungnam National University, Korea)

Fig. 4 Lacquered tableware set and lacquered ear cups excavated from the Tomb no. 1 at Mawangdui site. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Kyeomgmi Joo, Chungnam National University, Korea)

ancient Chinese lacquer artwork are being discovered from excavation sites representative of the Neolithic period. However, in the 1970s, when this excavation was being carried out, discoveries of ancient lacquer artwork of the Han and prior periods were very rare in China. Lacquer is a resinous sap product of natural lacquer trees, and it has been used as a coating, bonding, or decorating material for some core artifacts made of wood, bamboo, earthenware, metals, or fabrics (Frick and Kieser 2019). Lacquer artwork has been a very expensive product, as the production of lacquer requires much labors and exquisite technical skill, and maintenance of vast lacquer tree forests is labor-intensive as well. A total of 184 lacquer artworks were discovered in Tomb no. 1 of the Mawangdui site, and most of them had been

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preserved in a perfect condition. Some of them had lacquer paintings and inscriptions. The richness and splendor of the lacquer artworks in this tomb indicated that the tomb owner must have occupied a high status during her lifetime. The lacquer-painted wooden screen found in the north compartment is one of the rare and valuable lacquer furnitures of the Han dynasty (center of Fig. 3). It is adorned with double-sided lacquer paintings. The frontside painting is an abstract lozenge pattern painting with a round ritual jade “bi” in the center, and the backside painting is a flying dragon (Hunan Museum 2017, 95). The total height of the screen is 62 cm, and its width is 58 cm. However, the majority of the lacquer artworks of this tomb are colorful vessels and storage boxes. Most of the lacquer vessels of this tomb had been made of wood or bamboo cores, but some of them were made by the “dry lacquer” or “jiazhu” technique, which uses thin fabric cores with a lacquer bonding process (Hunan Museum 1973a, 76). Most of the lacquer vessels and containers were tableware for food and wine. Half of them were the “ear cup” type, which is an oval-shaped round bowl with two ears on each side. A total of 90 ear cups were discovered at this site and many of them have inscriptions in the center or at the bottom (front of Fig. 4). The most magnificent lacquer artwork is a cosmetic box set, which consisted of one big two-storied round box and nine small boxes inside (Hunan Museum 2019c, 156–163). It was discovered in the north compartment, and some small boxes of this set had been made by the dry lacquer technique. The main round box was decorated with colorful lacquer paintings representing flying clouds. The other small boxes were decorated with colorful lacquer paintings representing flying clouds or dragons. Another cosmetic box set in this tomb consisted of one big round box and five small boxes (Hunan Museum 2019c, 164–171). In those small inner boxes of the two cosmetic box sets, there were deposited several cosmetic ingredients and silk fabrics (Hunan Museum 2017, 186–187). These are among the oldest cosmetic examples for women in East Asia. The archaeologists started to open the central compartment with the main coffins at the excavation site on April 25, 1972. However, as the main coffins consisted of several layers of fragile materials, they were moved to the Hunan Museum a few days later, for it was desired that they be examined under conditions more conducive to their preservation. Although the museum might have been better than the excavation field site overall, it was not so suitable for scientific investigation. The temperature reached 30°C in the middle of the day, and the room lacked air conditioning. To reduce the temperature, the researchers had to surround the coffin with ice. The main coffin consisted of a total of four wooden coffins nesting together tightly. Many silk and cloth fabrics were discovered inside the coffins. The outermost coffin was a black lacquered wooden coffin, and there was no decoration on the surface (Hunan Museum 2019a, 6–9). The length of the first coffin was 2.95 m, the width 1.5 m, and the total height 1.44 m. The inside was all painted with red-colored lacquer, unlike the black outer surface. The second coffin and the third coffin were colorful lacquer-painted wooden coffins, but the base color and the themes of the paintings differed (Fig. 5). The second coffin had a black-lacquered base, and the

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Fig. 5 Lacquered wood coffin from the Tomb no. 1 of at Mawangdui site. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Kyeomgmi Joo, Chungnam National University, Korea)

four sides of the coffin showed free-floating or roaring clouds with many mythical beasts or gods. The size of the second coffin was 2.56 m long and 1.18 m wide, and the total height was 1.14 m (Hunan Museum 2019c, 3–8). The lacquer painting technique of this second coffin was very rare and unique, for many parts of the figures had been decorated in lacquer paste relief. This is one of the earliest examples of the later Chinese lacquered relief technique called “duiqi.” The third coffin had a red-lacquered base, and each painting of the three sides depicted several couples of confronted spiritual animals, such as dragons, tigers, deer, and birds. The only exception was one side that represented angled curvy white clouds. The size of the third coffin was 2.3 m long and 92 cm wide, and the total height was 89 cm (Hunan Museum 2019c, 9–13). The inner surfaces of both the second and third coffins were all colored with red lacquer. The fourth and innermost coffin was the smallest one, and was decorated with patterned silk fabrics attached by sticky lacquer as a glue. The size of this coffin was 2.02 m long and 69 cm wide, and the total height was 63 cm. The outside is painted with black lacquer and the inside is painted with red lacquer (Hunan Museum 1973a, 13–27; Hunan Museum 2019c, 14–15). All four coffins were in a very fine condition, for their surfaces had been fully coated by black and red lacquer. These coffins are perhaps the most magnificent lacquer artworks of the Han dynasty. The archaeologists found an extraordinary “T”-shaped silk painting on the last, innermost coffin cover in the afternoon of April 26, 1972, just before its opening. This is the first excavated silk painting and one of the best-preserved artistic representations of the Han dynasty. The represented images in this “T”-shaped silk painting are divided into three scenes in a vertical composition (Hunan Museum 2017, 286–287). On the upper part is represented the symbol of the Sun as a blackbird, the symbol of the Moon as a toad, a man on a flying dragon, five birds, two confronted dragons, one mythical tree with Sun discs, and spirits in Heaven. In the center of the painting, the portrait of the tomb owner as an old woman shows three standing female attendants and two seated male attendants (Silbergeld 1983).

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The lower part is a ritual performance scene and several pairs of many mythical creatures. Along each side of the central and lower parts, two giant dragons intersected each other into a large round ritual jade “bi” and are flying upright, as if to protect the tomb owner and the funerary ritual represented in the central and lower scenes. Many scholars have agreed that these scenes represent Heaven at the top and the underworld in the central and lower parts (Wu 1992). Although there are many disputes on its usage, meaning, and interpretations, this “T”-shaped silk painting might be a special ritual “Name Banner” made for the funeral rite rather than a fey garment or flying garment (Wu 1992; Wang 2011; Gao 2019). When archaeologists carefully removed the silk painting banner and opened the last coffin, a large compound of silk and linen fabrics, which might have wrapped the corpse of the tomb owner, was laid in the water inside the coffin. It was covered and wrapped with many layers of silk and linen fabrics and garments (Hunan Museum 2017, 305; Hunan Museum 2019b, 202–214). These fabrics and silk garments were preserved in an extremely fine state, and many of them showed the high quality of silk weaving and embroidery technique typical of the Han dynasty. The remaining garments of the dead consisted of 20 layers tied nine times with silk bands. This is a traditional funeral rite that followed the Confucian literary traditions (Zhao and Gao 2002, 93–94). During the investigation of the innermost coffin, the archaeologists discovered a woman’s cadaver that was well preserved with no evidence of decomposition (Peng and Wu 1998; Xiong and You 2006). The mummified woman was wrapped with many layers of fabrics submerged in clear and transparent water 20 cm deep at the bottom of the coffin. The water assumed a brownish yellow color soon after excavation (Hunan Museum 1973a, 30). Although there were many disputes on the woman’s identification before the excavation of Tombs no. 2 and no. 3 of Mawangdui, most contemporary archaeologists and scholars have agreed that this lady is Xin Zhui, the wife of the first Marquis of Dai in the Western Han Empire, based on the inscriptions of a bronze seal and many lacquer artworks in the tomb. According to the Chinese historical records, there were four Marquis of Dai during the Western Han period. Her husband might have been the first marquis of Dai, Li Cang (or Li Tsang), who was the founder of the marquisate and the owner of the Tomb no. 2 at the Mawangdui site. There is no clear evidence for the construction date of Tomb no. 1 for Lady Dai, but it might have been built after 168 BCE, when the Tomb no. 3 was built (Hunan Museum 2004, 93– 94). The mummified Lady Dai and her belongings in the tomb were amazingly well preserved after 2000 years of burial (Aufderheide 2003). The structure of the two-layered wooden chamber and four nested wooden coffins comes from the traditional funerary rites of the Warring States period. Such a tomb structure was recorded in the Book of Rites, the Liji, compiled during the Zhou dynasty. Many excavated tombs in the Warring States and Han dynasties had such a structure. In addition, the tradition of wrapping the body in layers of fabric also was written in the Book of Rites. The ancient Chinese people also had a tradition of putting the personal belongings of the dead into the tomb, in order for the dead to be

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able to keep them forever. These ancient funeral rites and traditions wholly affected the construction of the Tomb for Lady Dai in the Han dynasty (Zhao and Gao 2002). However, in the mid of the Cultural Revolution, there were serious disputes on the treatment of the mummy. Some researchers thought that the artifacts and female mummy from the Mawangdui site would provide invaluable information on life and culture in ancient China. However, some politicians and researchers who supported the Cultural Revolution insisted that they need not preserve or even research findings from the Mawangdui site. Fortunately, it was finally decided that the Mawangdui mummy was to be preserved.

Biomedical Investigation of Mawangdui Mummy The Mawangdui mummy was subject to intensive biomedical investigations and subsequent conservation treatment (Correspondent 1973; Hunan Medical College 1980). To preserve the mummy, a formaldehyde-ethanol-glycerin mixture was infused into the blood vessels and body cavity (Xiong and You 2006). Then, the mummy was treated by preservatives filled inside a glass chamber. The Mawangdui mummy was once displayed to the public of Changsha city in such a state. However, the special exhibition at the museum failed to come to a successful conclusion, as a large disturbance occurred among the excited visitors during the museum display. To avoid further confusion, the exhibition came to an immediate halt, and the female mummy was moved from the museum to Hunan Medical College (Xiong and You 2006). After the treatment with preservatives, the female mummy was investigated by medical scientists. In an anthropological examination, the Mawangdui mummy’s stature was recorded as 154 cm, and her weight was 34.3 kg (Hunan Medical College 1980). The mummified lady’s preservation status was perfect. Her skin, hair, and eyelashes were preserved intact. The soft tissues’ elasticity had been maintained well. Fingerprints were clearly visible. Her joints were still movable. No evident signs of pathology were detected on her skin (Hunan Medical College 1980). Impressed by the excellent preservation status of the female mummy (Fig. 6), some scholars even presumed that the mummified body might have been embalmed in a new way that was not clearly proven until then. Other academics even argued that the water remaining inside the coffin might have had an effect on the mummy’s excellent preservation (Correspondent 1973). Next, the preservation of the skeleton was examined by X-ray (Hunan Medical College 1980; Peng and Wu 1998). In this study, due to the technical limitations in the 1970s, CT data was unavailable. Briefly, the preservation of the mummy skeleton was very good on X-ray images. Meanwhile, osteoporotic signs were found in the bones. In a comparison with the bone images of a modern female of similar age, the Mawangdui mummy’s osteoporosis on radiology did not seem to have come simply from the aging process; rather, the osteoporotic pattern could have been caused by post-mortem changes possibly due to the high acidic pH (5.18) of the coffin water (Hunan Medical College 1980). In the radius and ulna, there were signs of fracture

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Fig. 6 Excellent preservation status of Mawangdui mummy. Currently displayed in Hunan Museum, Changsha, China. (Photo courtesy of Taesik Kim, Yonhap News Agency, Korea)

and malunion. The mummified organs in the cranial, thoracic, and abdominal cavities were very well preserved, though seriously distorted or contracted. In the radiography of the cranial cavity, the mummified brain was reduced in size, and the dura mater was visible (Hunan Medical College 1980). The radiologists also found pulmonary calcifications possibly due to tuberculosis. A radiological sign of herniation of intervertebral disc (HIVD) was observed in the 4th lumbar vertebrae. Atherosclerotic changes were observed in the arteries of the lower extremity (Hunan Medical College 1980). Actually, the Chinese investigators considered and made preparations for the maintenance of the excellent condition of the Mawangdui mummy in perpetuity. To induce much easier penetration of fixative, they thus decided to open the mummy’s body cavity by autopsy and remove the mummified intestines, whereafter the body cavity would be filled by preservative and, finally, the incision would be closed by suturing (Xiong and You 2006). In December 1972, medical experts representing all of China finally gathered in Changsha city. Indeed, this work became a national event. After they first discussed the procedure of an autopsy, it was led by the staff of Hunan Medical College in Changsha city. Radiological information already obtained was helpful in deciding how an autopsy would proceed. They agreed that the autopsy should mainly focus on complete maintenance of the mummy organs and structures. Special care was also taken to maintain sterilization before and during the autopsy. The internal organs of the mummy removed during the autopsy would be subject to various scientific research (Xiong and You 2006). On December 14, 1972, the autopsy began. They started at the mummy’s head. When a part of the skull was removed, the operating surgeons discovered a wellpreserved brain and dura mater. The mummified brain was reduced to almost half of its original size. However, the general morphology of the cerebral hemisphere was still discernable (Xiong and You 2006). The preservation of the internal organs was much better than originally expected. The amount of fat in the subcutaneous layer

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led scholars to presume that the mummified female was overweight during her lifetime (Xiong and You 2006). In the autopsy, most of the internal organs were intact though collapsed and distorted (Peng and Wu 1998). Respiratory organs such as the trachea, bronchus, and lung were reduced in size too, but their appearance looked perfect. Neuroplexus of vagus nerve was identified in the mummy’s lung. The heart and diaphragm also were well preserved (Fig. 7). The digestive system was maintained well, though the intestinal walls had become very thin (Peng and Wu 1998). During the autopsy, a pathologist removed the internal organs for forthcoming scientific analyses. Then, the medical researchers’ autopsy was finally finished (Xiong and You 2006). Using the data obtained from the autopsied specimens, Chinese researchers replied to many questions about the mummy’s physical preservation status and cause of death, among others. According to their research, the Mawangdui female mummy’s estimated age was about 50 years. Her blood type was A, and her body’s mercury concentration was very high. They also found a total of 138 oriental melon seeds in her digestive tract. This means that the Mawangdui lady must have died at the time of the melon harvest. The diseases she allegedly suffered from during her lifetime include atherosclerosis, coronary artery disease, multiple cholelithiasis (gallstones), HIVD, fracture and malunion, pulmonary calcification, anthracosis, and gallbladder anomaly (Hunan Medical College 1980). An obstetrician also found evidence of perineal tearing possibly due to delivery of a child. A parasitological examination also indicated Schistosoma japonicum, Trichuris trichiura, and Enterobius vermicularis infections (Li 1984; Hunan Medical College 1980; Yeh and Mitchell 2016). Actually, the discovery of S. japonicum eggs in the Mawangdui mummy allowed the origin of this parasitic disease to be traced back to about 2100 years ago (Hunan Medical College 1980). As for her cause of death, they ruled out the possibility of murder, as no signs of acute trauma were observed. During the autopsy, the cerebral vascular accident was

Fig. 7 Mawangdui mummy’s heart and diaphragm once displayed in Hunan Museum, Changsha, China. (Photo courtesy of Taesik Kim, Yonhap News Agency, Korea)

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excluded as a cause of death as well, given the lack of any relevant signs in the mummified brain. It is also unlikely that she had suffered from chronic wasting disease, considering her rich subcutaneous fat and no signs of bedsores on her back (Hunan Medical College 1980; Xiong and You 2006). Considering the discovery of oriental melon seeds in the stomach, food was likely ingested shortly before her death (Hunan Medical College 1980). All of the data indicated that the Mawangdui lady had not died of chronic illness, but had been subject to a sudden death of unknown cause. The most likely candidate for her cause of death is myocardial infarction or heart attack, possibly due to biliary colic. In fact, in the autopsy, the blood supply of the mummy’s coronary artery was found almost blocked by atherosclerosis (Hunan Medical College 1980; Lee et al. 1986; Peng and Wu 1998).

The Excavations on Mawangdui Tomb No. 2 and Tomb No. 3 The archaeologists discovered two more tombs at the Mawangdui site, just beside Tomb no. 1 of Lady Dai, during its excavation. Tomb no. 2 was at the west side of Tomb no. 1, and Tomb no. 3 was at the south side of Tomb no. 1. These two tombs were excavated from November 1973 to January 1974 (Hunan Museum 1974). Since many artifacts and a well-preserved mummy were found in the first tomb, the Chinese government highly anticipated that the excavations of the two other tombs could also be great archaeological achievements. Impressed by the worldwide attention to its spectacular artifacts from Tomb no. 1, the excavations of Tomb nos. 2 and 3 could proceed with more funding and professional participation. The excavation of Tomb no. 3 started in November 1973, just after the excavation of Tomb no. 1. The archaeologists found that this tomb had similar burial structures to Tomb no. 1 (Fig. 8). The main burial was laid at the bottom of the deep burial pit in the shape of an inverted step pyramid, and the main wooden chamber was wholly Fig. 8 Mawangdui Tomb no. 3 Site. Inverted step pyramid shape deep in the original mound surface. Mawangdui Tomb Site Museum, Changsha, China. (Photo courtesy of Joo Kyeongmi, Chungnam National University, Korea)

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covered with kaolin clay and charcoal layers. The thickness of the charcoal layer was 15–30 cm, and the general thickness of the kaolin clay was around 60–70 cm. However, the kaolin clay layer in Tomb no. 3 was not regularly spread on and around the chamber. Some parts of the kaolin clay layer were only 10 cm thick, and several parts of the chamber had no kaolin clay layer (Hunan Museum 2004, 28). The tomb structures of these two tombs were very similar, and both were discovered in the undisturbed state. Tomb no. 3 had a two-layered wooden chamber and three nested wooden coffins. The inner space of the wooden chamber was divided into five compartments, and the main coffins were buried in the central compartment (Fig. 9). The division of the wooden chamber was in the centralized format, similar to the case of Tomb no. 1, but the north compartment was much wider than the other side compartments, and was divided into two sub-divisions. All four side compartments were full of burial artifacts, such as many wooden figurines and lacquer artworks (Hunan Museum 2004, 28–47). The main coffins of Tomb no. 3 consisted of lacquered wood, but they had no lacquer paintings. All of them were painted in brown-colored lacquer on the outside, and in red-colored lacquer on the inside. The size of the first outmost coffin was 2.57 m long, 1.16 m wide, and 1.13 m high. The size of the second, middle coffin was 2.34 m long, 0.92 m wide, and 0.88 m high. The size of the last, third innermost coffin was 2.14 m long, 0.72 m wide, and 0.67 m high (Hunan Museum 1974, 42). Dark brownish colored water at a depth of 30 cm filled the last coffin. The water had no odor and contained many minerals such as Si, Mg, Na, K, Al, Hg, Fe, Mn, Cu, Ti, and Ba (Hunan Museum 2004, 41). A name banner in the form of a “T”-shaped silk painting was discovered in the upper part of the last coffin, and the portrait of the tomb owner was represented as a man. The representational style and composition were very similar to the name banner from Tomb no. 1 (Wang 2011). In Tomb no. 3, there were three more silk paintings and many silk manuscripts preserved in an excellent state. In addition, Fig. 9 The wooden chamber model of the Tomb no. 3 at Mawangdui site. Currently displayed in Mawangdui Tomb Site Museum, Changsha, China. (Photo courtesy of Kyeongmi Joo, Chungnam National University, Korea)

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many bamboo and wooden poles with inscriptions had been deposited in this tomb (Hunan Museum 2017, 192–194). These manuscripts and inscriptions were identified as old books on Confucianism, Daoism, and ancient Chinese medical studies, as well as other literary traditions in ancient China. Most of these old and rare books were stored in the east compartment. Among them, the most important ones are the traditional Chinese medical books, including “Purls Law (or Maifa),” “Prescriptions of fifty-two diseases (or Wushier bingfang),” and “Drawings of Guide and Pulling (or Daouintu).” “The Drawings of Guide and Pulling” is a silk painting consisting of 44 drawings of human figures performing exercises (Hunan Museum 2017, 266–267), which were the earliest exemplary book related to the meditational and hygienic practices for gaining spirit and energy in the Chinese tradition. These old and rare book studies have flourished all over the world after being published in Chinese and English (Harper 1998; Liu 2016; Lo 2018). In addition, many traditional natural medicines were discovered in the tomb (Hunan Museum 2004, 274–280). It is important that the “Inventory of Burial Object,” written on 411 pieces of wooden and bamboo poles, was discovered in the east and west compartments of Tomb no. 3. The exact date of the tomb construction, written in the inventory poles, was identified as the year 168 BCE, the twelfth year of Emperor Wen of the Han dynasty (Hunan Museum 1974, 43). The lacquer artworks from Tomb no. 3 totaled 319 works with colorful lacquer decorations. Almost half of them were 148 pieces of ear-cup-type vessels (Hunan Museum 2004, 117; 126). In addition, there were five musical instruments, many silk pieces and garments, and many weapons made of wood, horns, and bronze in Tomb no. 3. Despite the finding of many precious artifacts in Tomb no. 3, the body of the tomb owner had not been mummified. The skeletonized body wrapped in 18 layers of silk and other textiles and garments was laid in the brownish water inside the innermost coffin (Hunan Museum 2004, 41). The incomplete mummification of this man might have been due to some imperfect condition of the kaolin clay layer outside of the wooden chamber. The excavation of Tomb no. 3 of Mawangdui confirms that a perfect wrapping condition of the kaolin clay layer outside of the wooden chamber or coffin is very crucial for the intactness of the organic materials in the burials. By anthropological analysis, the skeletonized body turned out to be a man in his thirties (Hunan Museum 1974, 42). The archaeologists were disappointed by the lack of any mummy in Tomb no. 3. Thereafter, they immediately began to excavate Tomb no. 2 to the west. Tomb no. 2 was the earliest constructed tomb at the Mawangdui site, but it was heavily damaged by ancient and modern tomb raiders. The archaeologists identified that the tomb had been built in the same wooden chamber structure with kaolin clay and charcoal layers, but the ground plan of the tomb was in the oval shape, unlike the quadrangular plan of Tombs no. 1 and no. 3. It might have been made of a two-layered wooden chamber and two nested wooden coffins, but most of the timbers were decayed and had merged to the extent that the original structure could not be identified (Hunan Museum 2004, 9–10). The only identifiable parts of the coffins were the bottoms, and unfortunately, there was no trace of the body or its garments. The outer

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coffin was 3 m long and 0.95 m wide. The inner coffin was 2.65 m long and 0.7 m wide. Both coffins were painted with black-colored lacquer on their outside, and with red-colored lacquer on their inside (Hunan Museum 2004, 11). However, especially during the excavation of the Tomb no. 2 site, the archaeologists found important historical artifacts by which the three tombs of the Mawangdui site could be identified. In the north compartment of its wooden chamber, the archaeologists discovered three inscribed small seals made of bronze and jade. The most important one was a light-gray colored jade seal with the name of “Li Cang,” the founder of the Dai marquisate (Hunan Museum 2017, 48). This seal might be the private seal of the tomb owner. The other two seals were official seals with turtle handles. One gilt-bronze seal has the inscription of “Changsha chengxing,” which means, “the Prime Minister of Changsha.” The other, bronze seal has the inscription of “Dai hou zhi yin,” which means, “Seal for the Marquis of Dai” (Hunan Museum 2004, 24–25; Hunan Museum 2017, 49). According to these seal inscriptions, the archaeologists were able to determine that the three tombs of the Mawangdui site had been built for the first Marquis of Dai, Li Cang, and his family. According to the Chinese historical records, Li Cang became the first Marquise of Dai in 193 BCE and died in 186 BCE (Hunan Museum 1974, 46). Thus, Tomb no. 2 probably was built at that time. However, there have been many disputes on the identification of the owner in Tomb no. 3. Although the owner was identified as a young man in his thirties, and a possible son of Li Cang and his wife Xin Zhui, there was no concrete evidence of his name. Considering the artifacts found inside the tomb, he might have been a high-level soldier. In recent research, many scholars have agreed that he might be the second Marquis of Dai named Li Xi, who actually died in 168 BCE. Although the ancient Chinese Historical record tells us that the second Marquis of Dai died in 165 BCE, it might have been a textual error or mistake on the exact date of his death (Fu 2016). Chronologically, we can confirm that Li Cang, the first Marquise of Dai, died in 186 BCE and was buried in Tomb no. 2; then, his son Li Xi, the second Marquise of Dai, died next in 168 BCE and was buried in Tomb no. 3; finally, Xin Zhui, the Lady Dai of that time, who was the wife of Li Cang and the mother of Li Xi, died a few years after 168 BCE and was buried in Tomb no. 1. On this basis, the Mawangdui site was a family cemetery for the Marquis of Dai during the second century BCE. Thousands of years after being buried, they finally reappeared, showing the economic and political power they had enjoyed while alive. The excavation of the Mawangdui site occurred during the Cultural Revolution period. At the time of the tombs’ discovery, political strife between opposing factions was rife. Advocates of the Cultural Revolution never supported this research. The Gang of Four even questioned whether the Mawangdui lady was a worthwhile subject of scientific investigation. They also insisted that the female mummy be cremated immediately without proper research. Meanwhile, the reformers in China consistently supported an appropriate investigation of the Mawangdui mummy (Xiong and You 2006). Despite the difficulties and limitations, the research level for the Mawangdui mummy was very high, even compared with developed countries at that time.

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Fig. 10 Permanent display of Mawangdui mummy in Hunan Museum, Changsha, China. (Photo courtesy of Taesik Kim, Yonhap News Agency, Korea)

At that time, preservation of the mummy was of great concern to the Chinese leadership. Chinese scholars developed their own technique for preservation of the female mummy. The Mawangdui lady is now displayed permanently at Hunan Museum in Changsha city, China (Fig. 10). Many articles on the mummy and the cultural artifacts from the Mawangdui tombs have been published to date (Hunan Museum 2017, 2018, 2019a, b, c).

The Excavation of Phoenix Hill Tomb No. 168 Actually, tombs similar to the Mawangdui cases had often been reported from Changsha and nearby regions, but the excavation reports were published without any comments on the mummies and organic materials like silk or lacquered works. The subsequent discovery and research on the Mawangdui mummy have had a tremendous impact on Chinese society and academia. After the excavation of the Mawangdui site, Chinese archaeologists have paid more attention to well-preserved mummies and cultural remains from the Warring States and Western Han period tombs. In 1975, archaeologists in Hubei province excavated Tomb no. 168 at the Phoenix Hill site in Jiangling Region. Phoenix Hill was a part of the ancient Jinan cityfortress site, one of the ex-capital remains of the Chu Kingdom (1030 BCE – 223 BCE). There are many ancient tombs at this Phoenix Hill site, and archaeologists have excavated more than 180 tombs from the Qin and Han dynasty

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periods since September 1973. Those tombs were made for Chu people of varying social status and periods (Ozawa 2006). One of them was Tomb no. 168, which was constructed during the Western Han period. Its excavation began on 30th March and finished on June 20th, 1975 (Jinancheng 1975; Hubei Institute 1993). To the north of Tomb no. 168 was Tomb no. 167, and to the south of it was Tomb no. 169. Tombs no. 168 and no. 169 might have been built for a couple. Tomb no. 168 was situated in a deep burial pit 7.9 m below the ground (Jinancheng 1975, 1). The tomb structure was similar but much simpler than the Tombs of Mawangdui. The burial pit was full of blue-grey colored clay 5.26 m deep. Under the clay layer, the wooden chamber was laid in the center of the burial pit, wholly and regularly covered with a white greenish kaolin clay layer 14 cm thick (Hubei Institute 1993, 457–458). This kaolin clay layer seems to have sealed the coffin off from oxygen and maintained an aseptic condition inside of it. The wooden chamber was made of one layer of tightly bound wooden timbers. The length of the chamber as 4.62 m and the width was 3.17 m. The total height of the wooden chamber was 2.19 m. The chamber was divided into four compartments by thick timbers at the top, but the lower parts actually consisted of only three compartments. This tomb had its head to the east, unlike the tombs of Mawangdui, which had their head to the north. The main coffins were deposited in the southwestern side compartment, which was the biggest compartment of the chamber (Hubei Institute 1993, 459–464). The east compartment, which was the upper part of the owner’s head, or a head section, was full of wooden figurines and lacquer artworks. The north compartment seems to have been divided into two sections, but actually, they were connected as one long side section (Hubei Institute 1993, 464–465). The main coffins consisted of two nested wooden coffins. The outer coffin was wholly painted with black colored lacquer on both the inner and outer surfaces, and every joint line of the timbers was meticulously coated with many layers of hemp cloth band and lacquer. The size of the outer coffin was 2.56 m long, 0.97 m wide, and 1.02 m high. The outside of the inner coffin was painted with black colored lacquer. The size of the inner coffin was 2.23 m long, 0.76 m wide, and 0.71 m high. The inner coffin was also tightly coated with many layers of hemp cloth band and lacquer. A bamboo screen and two bundles of dry straw were laid on the upper part of the outer coffin (Hubei Institute 1993, 462–463). During the excavation, a well-preserved male mummy was discovered inside the tomb (Wu 1982; Peng 1995). When the tomb was opened, archaeologists found dark red fluid or water in the innermost coffin. A mummified old man was discovered submerged in this water (Hubei Institute 1993, 462; Peng and Wu 1998). During the inspection, a white jade seal was discovered inside the mummy’s oral cavity. His name was revealed to be “Sui,” as inscribed in the jade seal (Hubei Institute 1993, 507). Although his full name was uncertain, according to the inscriptions of the bamboo slips, his social status was that of a local governor of Shiyang village in the west part of Jiangling County. He died in 167 BCE, the thirteenth year of Emperor Wen of the Han dynasty (Hubei Institute 1993, 509–510). He was almost

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contemporaneous with the female mummy of the Mawangdui site, but his social status was much lower than hers. The burial goods from Tomb no. 168 of Phoenix Hill were much less numerous than those from Tombs no. 1 and no. 3 at the Mawangdui Site. In this tomb, a total of 165 pieces of fine lacquer vessels were discovered in the north compartment (Hubei Institute 1993, 466). The north compartment was filled with water, and so many of those artifacts were slightly moved from their original positions by the water. All of the lacquer vessels were made of wood and decorated with lacquer paintings. Among them, a total of 100 ear cups were found (Hubei Institute 1993, 474–476). The most interesting artifacts from this tomb were those in a set of stationery for the nobleman. The set, one of the earliest examples of ancient Chinese stationery, was perfectly stored in a bamboo box at the side section (Zhong 1975, 20–22). It consisted of an ink stone, two ink sticks, a brush, six empty bamboo slips, a bronze knife, and other things (Zhong 1975; Hubei Institute 1993, 491). In addition, an “Inventory of Burial Object” written on many bamboo poles was discovered in the middle of the north section (Hubei Institute 1993, 499–503). Anatomical and histological analyses showed a perfect preservation status for the male mummy from the Phoenix Hill site. He looked to have been aged in his sixties. His skin was soft and elastic, but no hair remained on his skin. The joints were highly flexible. All of his teeth were in fine condition. The mummy’s body measured 167.8 cm in height and 52.5 kg in weight (Jinancheng 1975, 3; Peng and Wu 1998). His blood type was AB. Radiography and autopsy also revealed well-preserved internal organs. Even the tympanic membrane was found in the left ear. The brain (weight: about 970 g) occupied approximately 4/5 of the cranial cavity (Jinancheng 1975; Peng and Wu 1998). The dura mater was intact. Vessels and cranial nerves remained. The thyroid gland, very rarely found in mummies, was also discovered. The mummy’s intestine, biliary tract, gall bladder, liver, heart, muscle, lung, and others were all examined (Jinancheng 1975; Wei et al. 1981). In general, the histology of this male mummy showed abundant collagen fibers. The main component remaining in the mummified brain was myelin remnants showing a typical lamellar structure on electron microscopy (Zhou 1981; Peng and Wu 1998). In the paleopathology, there were no osteoporotic changes. This mummy of the Phoenix Hill site might have had liver cirrhosis, gallstones, cholecystitis, and/or atherosclerosis. Sui’s cause of death was likely acute peritonitis caused by a chronic gastric ulcer and a complicated perforation at the lesser curvature of the stomach (Zhou 1981; Cheng 1984; Aufderheide 2003). Signs of pleuritis and pericarditis were also found during the autopsy (Jinancheng 1975). A parasitological examination showed ancient eggs of S. japonicum, Clonorchis sinensis, Taenia sp., and T. trichiura (Wei et al. 1981; Zhou 1981; Li 1984; Aufderheide 2003; Yeh and Mitchell 2016). The result confirmed that the above-mentioned parasite infections were prevalent in the ancient Jiangling region of China (Wei et al. 1981). The mummified body of Sui and his burial properties are currently curated at Jingzhou Museum in Hubei province.

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The Female Mummy of the Warring States Period Han dynasty tombs of the Changsha region were culturally influenced by the tombs of Chu, the ancient Chinese kingdom that once ruled the Yangtze River basin during the Warring States period (402 BCE–221 BCE). Even prior to the discovery of the Mawandgui mummy, partial human remains were found in Chu period tombs. However, no complete Chu-period mummy was investigated until 1994. The only Warring States or Chu period mummy was accidentally discovered in a tomb located in Guojiagang village in Jingmen city, Hubei province in March 1994. Since Jingmen city was one of the ex-capitals of the ancient Chu kingdom, many Warring States period tombs have been discovered in and around the city. Unfortunately, this mummy from the Guojiagang site was not discovered by archaeologists, but was accidentally found by a group of tomb raiders. When they plundered several ancient tombs there, a mummified woman was discovered. Those tomb raiders removed the female mummy and reburied her in another location. They stole silk fabrics, wooden figurines, lacquer artworks, and bronze artifacts from the tomb (Tang et al. 1994; Wu et al. 1996). The thieves were arrested the following month, but many cultural artifacts were already lost. When archaeologists reinvestigated the tomb site, there were very few artifacts left inside. Fortunately, the archaeologist of Jingmen Municipal Museum rediscovered the female mummy during an excavation near the site in May 1994 (Liu and Zhao 2013, 37). According to the archaeologists, the tomb was constructed in the Warring States period or Chu kingdom period during the third or fourth century BCE, much earlier than the Mawangdui and Phoenix Hill tombs (Peng and Wu 1998; Liu and Zhao 2013). This estimation was confirmed by carbon dating analysis (Wu et al. 1996), which means that this case represents the oldest ancient mummy discovered in China (Wu et al. 1996). Archaeologists confirmed that the tomb had doublenested coffins. When the female mummy was rediscovered, she was scientifically investigated by experts (Wu et al. 1996; Peng and Wu 1998). According to their reports, her height was 160 cm. She had dark-colored skin and long hair. Her teeth were preserved, but dental attrition was serious. According to her dental age estimation, she might have been 65–70 years old. The soft tissues were elastic but subcutaneous fat was absent. Her joints were still freely movable (Wu et al. 1996). Like the female mummy of Mawangdui, the Guogiagang mummy was very finely preserved as well. An autopsy subsequently was performed on the mummy. The intestines could be identified but had severely shrunk and collapsed. Specimens for a parasitology study were collected from the intestine, and C. sinensis and T. trichiura infections were evident (Wu et al. 1996). According to a chemical analysis, the mummified woman’s blood type was AB (Wu et al. 1996; Peng and Wu 1998). In a histological study, Wu et al. (1996) confirmed that the mummified tissue was mainly composed of connective tissue (collagen) fibers. Jingmen Municipal Museum of Hubei province is currently curating this female mummy.

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The Excavation of Double Dragons Tomb No. M1 The Double Dragons site is in a small village named “Shuanglong” in the Haizhou district of Lianyungang city, Jiangsu province, China. The site was excavated in July 2002 by archaeologists from Lianyungang Museum. According to the excavation report, there were two Han dynasty tombs at the site. Between them, a wellpreserved female mummy was discovered in Double Dragons Tomb no. M1 (Lianyungang 2012, 4). Tomb no. M1 of the Double Dragons site consisted of two wooden chambers in a great burial pit (Fig. 11). In the wider and large north chamber, there were three coffins in juxtaposition. The size of the north chamber was 3.46 m long and 2.34 m wide. All three coffins in the north chamber had traces of the dead. The female mummy was discovered in the third coffin of the north chamber, which was laid at the northernmost side. The smaller south chamber had only one coffin inside. The size of the south chamber was 2.7 m long and 1.26 m wide. The total length of the two chambers was 4.2 m, and the width was 3.6 m. The height of the remaining wooden chamber was about 1.5 m. The upper part of the tomb was severely damaged, but the kaolin clay layers were preserved on and around the wooden chamber. The thickness of the kaolin clay layer was about 10 cm (Lianyungang 2012). All of the four coffins in this tomb were made of wood and painted with black colored lacquer on the outside. Among them, the second coffin in the middle of the north chamber was for a man, and the other three coffins were for women. There remained some fragments of bones from the man in the second coffin, and a female mummy was discovered in the third coffin. The other two coffins had no body fragments of the tomb owner, but the remaining artifacts in the coffins indicated that the owners might be women. Therefore, it was concluded that this tomb had been made for a man and three women of the Western Han dynasty (Lianyungang 2012). The burial goods of this tomb were less abundant than those of other Han dynasty tombs. In the first coffin, there was only a bronze mirror, a few wooden combs, and Fig. 11 The wooden chambers and coffins excavated at the Tomb no. M1 of the Double Dragons site, Lianyungan, Jiangsu. Currently displayed in Lianyungang Museum, Lianyungang, China. (Photo courtesy of Minku Kim, Chinese University of Hong Kong, Hong Kong)

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others. The size of the first coffin was 2.15 m long, 0.72 m wide, and 0.64 m high. The second coffin had many burial goods such as two iron swords, a stationery set with a brush and an inkstone, many wooden poles with inscriptions, a bronze mirror, and two bronze seals. The bronze seals might have been the property of governmental officials, but their inscriptions were in an illegible state. However, there was an inscription of “Donggong” on the cover of the coffin, which means “Lord of the East.” The second coffin was 2.28 m long, 0.76 m wide, and 0.76 m high. The fourth coffin in the south chamber was the same size as the first coffin, and the burial goods were scanty, including a bronze mirror, a lacquered cosmetic box, and a few wooden figurines (Lianyungang 2012, 4–5). Among these four coffins, the most important one is the third coffin from the north chamber, where the female mummy with the most luxurious burial goods was discovered. The size of the third coffin was similar to that of the second coffin. When the archaeologists opened the cover, water was found. The burial goods of the coffin were a silver ring, a bronze mirror, a few wooden hairpins, a few lacquer containers, and a bronze seal. The seal discovered inside a lacquer cosmetic box had the inscription, in four characters, “Ling shi Hui ping,” which indicated the name of the owner to be “Ling Huiping” (Lianyungang 2012, 5–6). In June 2003, scientists from Nanjing Medical University investigated the female mummy of the Double Dragons site. According to their reports, the mummy was generally similar to the other Han dynasty mummies mentioned in this chapter. She was 1.58 m tall and 25.5 kg in weight. She had 29 teeth in a fine state. Her hair was very thick (Fig. 12). She seems to have passed away at the age of about 55 (Chen et al. 2008; Lianyungang 2012). According to the burial goods and inscriptions in the tomb, Double Dragons Tomb no. M1 was built between 25 BCE and 15 BCE during the Western Han dynasty. The main tomb owner was the man in the second coffin of the north chamber, who must have been the father of the Marquise of Quping of that time. He might have been the prior Marquise of Quping, but his name remains uncertain. The three females in the same tomb might have been his wives, but they were not of the same social status according to the burial goods and coffin size. Ling Huiping, Fig. 12 Female mummy named as Ling Huiping excavated at the Tomb no. M1 of the Double Dragons site, Lianyungan, Jiangsu. Currently displayed in Lianyungang Museum, Lianyungang, China. (Photo courtesy of Minku Kim, Chinese University of Hong Kong, Hong Kong)

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the female mummy of the third coffin, might have been the formal wife, given her highest social status. The other two women might have been the concubines of the Marquise (Zhao 2014). Neither the full excavation report on the tombs of the Double Dragons site in Lianyungang city nor the scientific analysis report on this new female mummy of the first century BCE has been published. The construction and burial style of this tomb are very different from those of the Hunan and Hubei provinces tombs. Here, at the Double Dragons site, a total of four persons were buried together in the same wooden construction. In addition, the burial goods were much simpler than in the second century BCE Hunan and Hubei provinces tombs. Such differences may have come from the later date and lower social status of the people at the Double Dragon sites relative to those at the Mawangdui and Phoenix Hill sites. However, the well-preserved state of the female mummy was very similar to those earlier mummies. It might be important that those tombs with well-preserved mummies were constructed in fine kaolin clay layers and contained fully lacquered coffins. There is no mention on the usage of a charcoal layer at the Double Dragons site in the brief excavation report, and so this issue will have to await future studies.

Mummification in the Warring States and Han Period Tombs Actually, several historical records include accidental discoveries of ancient wellpreserved human bodies in ancient Chinese tombs. People in past times believed that such an antidecaying phenomenon might have been caused by a certain magical power of some natural ores such as jade. In the twentieth century, scholars have considered that bodies later found mummified might have been subject to artificial embalming techniques since lost to history (Aufderheide 2003). Peng and Wu (1998) presumed that mummification might have been induced by the interaction of multiple external conditions such as humidity, temperature, and pH, among others, inside Western Han period tombs. When the tomb of Mawangdui was opened, the water inside the coffin attracted the researchers’ attention. Some claimed that cinnabar (HgS) included in coffin water might be the major factor inducing mummification. In fact, ancient Chinese people believed that mercury might improve one’s longevity and health, and might have had some role in keeping bodies from decomposing. However, it is clear that the bactericidal effect of cinnabar is too weak to form mummies; alternatively, rather than having been infused by embalmers, coffin water might have originated from body liquefaction or condensation of humid air (Hunan Medical College 1980). The most noteworthy factor causing mummification is the air-tight sealing effect of kaolin clay around the coffin. Lee et al. (1986) noted fine kaolin clay used for sealing Western Han dynasty tombs. Wherever the kaolin clay layer has been absent from ancient tombs, well-preserved human or cultural remains could not be retrieved therefrom. Kaolin clay sealing might have prevented percolation of air and water into coffins, thereby causing anoxic conditions, and, finally, halting the decomposition process therein. Such a completely sealed coffin

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would have formed an unfavorable condition for proliferation of putrefying bacteria (Kuo 1972; Hunan Medical College 1980; Lee et al. 1986). The charcoal layer might also have contributed to mummification in the Mawangdui tomb, especially by absorbing moisture that otherwise would have seeped into the coffin (Lee et al. 1986). Mummies were always wrapped in many layers of fabrics, which might also have caused the anaerobic condition inside coffins. Regardless, subsequent proliferation of anaerobes finally turned coffin pH into a highly acidic, very unfavorable condition for growth of putrefactive bacteria (Hunan Medical College 1980).

Conclusion Chinese scholars have emphasized the similarity of various Chinese mummies of different historical periods. For instance, the Western Han period mummies and those from the Song and Ming period tombs were not considered as separate entities in China (Shin et al. 2018). The mummification in the Warring States and Western Han period tombs was likely due to the kaolin clay layer around the coffin (Chung 1972; Editorial Department 1972; Lee et al. 1986). Similarly, the presence of a limemixture coating in Song and Ming period tombs might be related to a mummification-related sealing effect (Wang and Zhang 2012). In the biomedical perspective, those mummies have indeed shown considerable similarities. For example, they did not undergo embalming or evisceration. Their anatomical and radiological appearances also were very similar. The histological patterns looked alike as well. The mummies discovered in Chinese tombs can therefore be included in the same category: “wet corpses,” “ancient corpses,” or “Mawangdui-type cadavers” (Hunan Medical College 1980; Peng and Wu 1998; Wang and Zhang 2012). In fact, Chinese archaeologists have considered that well-preserved corpses from the Warring States and Western Han period tombs are different from conventional mummies (e.g., Egyptian mummies), and that therefore, they must be distinguished by the nomenclature, “Mawangdui-type cadaver” (Hunan Medical College 1980; Peng and Wu 1998). However, it is not true that only artificially embalmed bodies can be called “mummies.” Rather, there are different kinds of naturally mummified bodies as well, such as Bog bodies or Joseon mummies that were not artificially embalmed but nonetheless are classified as mummies. In the same regard, the introductory books of Cockburn et al. (1998) and Aufderheide (2003) identified the Mawangdui lady as a natural mummy. In summary, the present chapter provides an overview of the research work completed on four famous ancient Chinese mummies from the Warring States and Western Han period tombs. Excavated in the Hubei, Hunan, and Jiangsu provinces, they are well-known symbols of Chinese culture of the past. These mummies and the related artifacts rediscovered during the past decades can provide much invaluable understanding of the uniqueness of Chinese history and culture.

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Cross-References ▶ Archaeological Findings of the Tarim Basin Graves and Mummies ▶ Joseon Dynasty Mummies of Korea ▶ Mummies of Song-Ming Dynasty in China Acknowledgments This research was supported by Basic Science Research Program through the National Research Foundation of Korea by the Ministry of Education (2020R1A2C1010708).

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Mummies in Japan

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Hisashi Fujita, Jun Koike, Hiroki Sugimori, and Dong Hoon Shin

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies of Buddhist Monks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Fujiwara Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Japanese Mummies in East Asian History . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Although Japan’s natural environment is not suitable for mummification process, at least two types of Japanese mummies have been reported so far. The detailed historical facts about the Japanese mummies are available in the literatures or temple traditions of the verse. The mummies of Japan are mostly those of Buddhist monks, the Sokushinbutsu. They were estimated to have been mummified in 1362–1903, and then, having been revered as the religious saints at their respective Buddhist temples that were mainly distributed in the prefectures of East Japan. The Fujiwara family mummies are another type of mummies found in Japan. They were the medieval political leaders ruling the northeast territory of

H. Fujita (*) Doshisha University, Kyotanabe, Japan e-mail: [email protected] J. Koike · H. Sugimori Department of Nursing, Daito Bunka University, Higashimatsuyama, Japan e-mail: [email protected]; [email protected] D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_31

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Japan during the twelfth century. The mummies of this family over four generations are housed in the Chuson-ji, the Buddhist temple built by Fujiwara no Kiyohira (1056–1128 CE). Thanks to previous academic research, scholars have come to admit the differences between Japanese mummies and those of other East Asian countries. Actually, the mummies of Korea and China have little in common with the mummies of Japan; and the similar findings could be often represented in Japanese history that has taken a separate path in East Asia. Keywords

Mummy · Japan · Fujiwara family · Sokushinbutsu · Buddhism · Chuson-ji

Introduction Although the discovered number is not so many and the exact mechanism of phenomena is as yet not fully comprehended, mummies unearthed in East Asia provided academically invaluable information to the disciplines of anthropology, archaeology, and history of the countries (Hunan Medical College 1980; Li 1984; Kim et al. 2008, 2015; Kahila Bar-Gal et al. 2012; Oh et al. 2013; Shin et al. 2013; Seo et al. 2014; Yeh and Mitchell 2016; Hong et al. 2017; Seo et al. 2017; Shin et al. 2017; Oh et al. 2018a; Shin et al. 2018). In general, East Asia has a long history. Many ancient peoples lived in complicated and delicate societies in a highly organized manner. Just as ancient Egyptian mummies were influenced by the idea of afterlife, East Asian mummies were also the definite outcome of a historical belief prevailing the region (Shin et al. 2008; Lee et al. 2013; Oh and Shin 2014; Song and Shin 2014; Oh et al. 2017, 2018a). Therefore, to understand the mummies discovered in East Asia, we need the general information of the people and society at the time and place, which could be revealed by the collaborative research of archaeologists, historians, and anthropologists. East Asian countries are known to have very few traditions of artificial embalming in history. However, this does not mean that there were not possibilities regarding the artificial mummification of dead bodies. For instance, archaeologists in China have suspected that the Han Dynasty mummies (e.g., Mawangdui) found in Yangtze River Valley might have been embalmed or treated with unknown chemicals for mummification after death. Of course, this might have actually happened in history. However, we must note that there are no clear historical or scientific evidences revealed so far, especially in case of mummies found in Korea and China. The scholars’ opinions on this matter are therefore somewhat skeptical. In this regard, Japanese mummies are unique. The mummies of Japan exhibit a very distinct pattern of preservation and had different historical background that could not be traced as seen among the mummies of the other East Asian countries. Although not completely proven, artificial treatment might have been involved in mummification in Japan though not everyone agrees. Considering the exact process of mummification that could be revealed by the researches of the historical records, at least for

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some cases found in Japan, archaeological or anthropological evidence suggestive of artificial embalming or mummification could not be completely denied. Indeed, at present, mummy research is sparse in Japan. However, well-designed studies on the Japanese mummies were already being conducted several decades ago. At that time, by means of variable scientific methodologies and techniques, Japanese investigators traced the general preservation pattern of mummies as well as the evidences of their physical or pathological traits. Due to the pioneering researches, even before the mummies in other East Asian countries began to have been researched in the 1970s, the academic significance of Japanese mummies was already revealed in multi-faceted perspectives (Ogata 1998; Sakurai 1998; Aufderheide 2003). Regardless of these perspectives, to date the reality of Japanese mummies is not well known to the outside world. The brief reviews and infrequent reports (Ogata 1998; Sakurai 1998; Aufderheide 2003) aside, there have been very few introductions to or summaries about the studies on Japanese mummies and related cultural heritages. Knowledge of Japanese mummies therefore remains limited for the scholars outside of Japan. Therefore, this chapter deals with the Japanese mummies of the Fujiwara family and Buddhist monks discovered so far in Japan. It first summarizes the past achievements of mummy studies conducted in Japan. It will then visit certain Buddhist temples to comment on how the Japanese mummies have been preserved and currently displayed there. The information has not been properly presented in any books or articles of mummy research so far. In this chapter, it is also worthwhile to note the cultural as well as bioarchaeological similarities and differences between the mummies found in Japan and the other East Asian countries.

Mummies of Buddhist Monks Like Korea and China, Japan’s natural environment or climate is never suitable for proper natural mummification processes. Nonetheless, at least two types of Japanese mummies are known to archaeologists and anthropologists so far. Although many stories about the mummies around the world have been lost in the course of history, the detailed historical facts about the Japanese mummies are available to the researchers through literature or temple traditions of the verse that are still passed down to us. This means that the cultural background of Japanese mummies could have been successfully recovered with the help of historical or archaeological studies. The Japanese mummies reported so far are mostly those of Buddhist monks. They are called Sokushinbutsu. As well known, Buddhism is one of the largest and influential religions in the world. Although it encompasses variable beliefs, traditions, and spiritual practices, the fundamental teaching of Gautama Buddha, the founder of the religion, is to overcome worldly suffering by breaking the cycle of death and rebirth, and therefore, finally achieving the Nirvana, the ultimate goal of every Buddhist. Various practices have been recommended to the Buddhists as a correct path to the truth: observance of moral precepts, monasticism, meditation, etc. Sometime before fourth century BCE, two major teaching branches came out from

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the Buddha’s original school: Theravada and Mahayana. Theravada Buddhism has been widely worshipped in South and Southeast Asian countries such as Sri Lanka, Cambodia, Laos, Myanmar, and Thailand. Meanwhile, the Mahayana has been popular in East Asian countries including Japan, Korea, and China. In history, the first introduction of Buddhism to Japan occurred in 552, by a Korean missionary sent to Emperor Kimmei’s court. After serious strife between powerful clans concerning the acceptance of the newly introduced religion, Buddhism could finally be embraced by the Japanese people and society. From the sixth century on, Japanese conceptions of death and funeral customs changed remarkably under the influence of Buddhism. Many Buddhist temples were constructed throughout Japan. Buddhist priests and nuns remarkably contributed to the evolution of spiritual life and culture. Even after the Meiji Restoration in 1868, Buddhism could be successfully modernized and therefore maintained its significant presence in the country as one of the major religions. Actually, there are many different belief traditions of Buddhism in Japan. Among the traditions, in some Buddhist temples, we note the presence of Buddhist monk mummies. For Buddhists, the monk mummies are not simply dead individuals who died hundreds of years ago. Rather, they have been revered as the Buddhist saints at their respective temples (Ogata 1998). According to historical traditions, the estimated dates of these mummies range from 1362 to 1903 (Ogata 1998). All of them are mostly distributed in the Prefectures of Niigata, Yamagata, Nagano, Fukushima, Gifu, Ibaraki, and Kyoto, the east part of Japanese archipelago. Detailed information is summarized in Table 1 and Fig. 1. As is well known today, these Buddhist monk mummies, Sokushinbutsu, are not naturally formed ones. Nevertheless, this does not mean that the monk’s bodies have been artificially embalmed after his death. Actually, the monks mummified themselves by their own intention and efforts with little help of the others. According to the tradition, the monks prepared for self-mummification for a long time. Very well laid-out plans were prepared in order to be mummified successfully after death. Other than very unique cases such as Tetsuryukai (Fig. 2), in which evisceration was performed and the body cavity filled with lime, Sokushibutsu mummies were mostly mummified by a process called self-immolation (Ogata 1998; Aufderheide 2003). The process was initiated by deliberate dietary control. For example, the cessation of cereal consumption and eating pine needles, resins, and seeds were recommended. Such a diet control, extending over the long course, reduced the monk’s fat content and muscle mass, thus keeping his weight down. At the final stage of selfmummification, the monk would die from severe dehydration. This process greatly promoted the monk’s mummification by keeping moisture, fat, and muscle in the dead body to a minimum (Morimoto 1993; Aufderheide 2003). After death, the body was placed in a double coffin that was interred in an underground stone chamber. Inside the coffin, charcoal was filled around the monk’s corpse. The stone chamber was installed with drainage to keep it dry. A few years later, the mummified body was removed from the stone chamber. Additional drying was induced thereafter by the exposure to wind, sunlight, or smoking. The drying process reminds us of similar mummification processes adopted in other

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Table 1 List of Buddhist monk mummies in Japan Mummy Kochi Tansei Junkaia Hommyokai Yuteia Shugi Zenkai Gyojun Chieia Chukai Shukaia Shinnyokai Myoshin Enmyokai Tetsumonkai Komyokaia Myokai Tetsuryukai Kankaia Bukkaia a

Age 82 63 78 61 92 78 85 45 ca. 30 58 62 96 36 55 62 ? 44 62 ? 76

Year of death 1363 1613 1636 1683 1683 1686 1687 1687 1736 1755 1780 1783 1817 1822 1829 1854 1863 1868 1878 1903

Location of mummy preserved Saishoji, Teradomari, Niigata Amidaji, Sakyo-ku, Kyoto Gyokusenji, Tsugawa, Niigata Honmyoji, Asahi, Yamagata Kanshuji, Asakawa, Fukushima Myohoji, Iwase, Ibaraki Kannonji, Kanose, Niigata Zuikoin, Anan, Nagano Buppoji, Suwa, Nagano Kaikoji, Sakata, Yamagata Shinjuin, Kashiwazaki, Niigata Dainichibo, Asahi, Yamagata Yokokuraji, Tanigumi, Gifu Kaikoji, Sakata, Yamagata Churenji, Asahi, Yamagata Zokoin, Shirataka, Yamagata Myojuin, Yonezawa, Yamagata Nangakuji, Tsuruoka, Yamagata Daienji, Niigata, Niigata Kanzeonji, Murakami, Niigata

The mummies in poor preservation status

Asian countries (Piombino-Mascali et al. 2016). Finally, the mummies were dressed as Buddhist monks and exhibited as religious objects of worship in Buddhist temples (Morimoto 1993; Aufderheide 2003). The idea of Buddhist monks’ self-mummification in Japan seems to have been imported from the Asian continent, along with the Buddhist faith itself (Aufderheide 2003). Sokushinbutsu, the title for mummified monks, means that they became enlightened while leaving their bodies as they were. Actually, the followers and believers of this practice do not regard this as a suicide at all. Rather, the Buddhists in Japan stressed the importance of ascetic practices to reach the enlightenment. In Buddhism, the Sokushibutsu, the mummification of the enlightened monk, is a symbol of great self-sacrifice and dedication to the religious truth (Fig. 3). To date, a series of scientific research has been conducted on such mummies. A full or part of the details on them was summarized in the previous reviews in English (Ogata 1998; Sakurai 1998; Aufderheide 2003). Among Buddhist monk mummies, nine individuals have been scientifically examined so far by the collaborative team of different research fields (Ogata 1998). In brief, those monk mummies have a lot in common with each other. They were all in sitting positions. Their skin was well preserved. There were few organs remaining inside the body cavities. However, it is not evident whether the internal organs have been artificially removed or not. Radiological studies also revealed pathological signs such as kyphosis, luxation, compression fracture, periostitis, ankylosing spondylitis, osteoporosis, spondylosis,

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Fig. 1 The map of Japan showing the prefectures where Sokushinbutsu mummies are located. Note many Sokushinbutsu Mummies mostly found in eastern Japan but very few in western Japan (Credit: Hisashi Fujita)

Fig. 2 Tetsuryukai died and mummified in 1868 (Credit: Yasuhiko Okuno)

osteophytes, and osteosclerosis caused by osteoarthritis (Ogata 1998). Thanks to the scientific researches, Buddhist monk mummies’ preservation status could be known to academia as well as the public. In fact, those Buddhist monk mummies could be the outcome of artificial mummification, in that it was planned and effected before the monks’ death (Morimoto 1998). However, Buddhist monk mummies in Japan could be also distinguished from the other embalmed mummies around the world because they were not mummified by full-pledged artificial embalming techniques. We presume that neither evisceration nor

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Fig. 3 Enmyokai died and mummified in 1822 (Credit: Yasuhiko Okuno)

embalming was a part of that mummification process of the Sokushibutsu. Despite the entire mummification process for Sokushibutsu mummies not being completely understood, the obvious fact about the mummies is that they are very unique mummies that have been rarely found in the other countries.

Fujiwara Mummies In Japan, another type of mummy also has been reported (Sakurai 1998). The Fujiwara family, one of the political leaders in the late Heian period (794– 1185 CE), ruled the northeast territory of medieval Japan, until the final conquest of the Kamakura Shogunate in 1189 CE (Fig. 4). The mummified bodies of this family noblemen over four generations are currently housed in the Konjiki-do of Chuson-ji, a Buddhist temple located in Iwate Prefecture. In the temple legend and tradition, Chuson-ji was constructed by Ennin, the famous Buddhist priest of the eighth to ninth century. However, according to more reliable historical literatures, the temple was founded as late as the twelfth century. It was Fujiwara no Kiyohira (1056–1128 CE) who built this temple. Kiyohira was the founder of the Northern Fujiwara clan in Mutsu Province. His family was one of the noble descendants and his father was a mid-level bureaucrat in the territory. The early life of Kiyohira was a series of miseries and misfortunes. His

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Fig. 4 Territory of the Oshu Fujiwara family. They ruled almost a quarter of Japan, excluding Hokkaido. The headquarter was located at Hiraizumi, Iwate Prefecture. Their political stronghold was famous for excellent horses and gold mines. The Fujiwara family’s economic wealth became enormous from these (Credit: Hisashi Fujita)

Table 2 List of Fujiwara clan mummies Mummy Kiyohira Fujiwara Motohira Fujiwara Hidehira Fujiwara Tadahira Fujiwara

Age 73 54? 66 23

Year of death 1128 1157 1187 1189

Location of mummy preserved Chusonji, Hiraizumi, Iwate Chusonji, Hiraizumi, Iwate Chusonji, Hiraizumi, Iwate Chusonji, Hiraizumi, Iwate

clan was in trouble with other clans so that his mother became the concubine of his enemy, and Kiyohira himself was also raised by the enemy clan. Nevertheless, Kiyohira finally won the final battle in 1087 CE, thus returning to his old town in what is now Hiraizumi of Iwate Prefecture as a victor. After this point, Kiyohira’s clan prospered like an independent kingdom, for about 100 years. When he grabbed the power in the northeast territory of Japan, to comfort the miserable souls who died in many battles and to wish for his own easy passage into Buddhist eternity, he made a huge contribution to construct the temple of Chuson-ji (1105 CE). Konjiki-do is a subsidiary building of the Chuson-ji temple. In 1124, the building was built as the mausoleum of Kiyohira himself and his family members. Inside the building, the mummified bodies of Fujiwara no Kiyohira, Motohira (1105–1157), Hidehira (?1122–1187), and the head of Yasuhira (1155–1189) are present (Table 2). Among them, Motohira, the second ruler and the son of Fujiwara no Kiyohira, extended Northern Fujiwara clan’s political influence to a vast area nearby. Next, Hidehira, the third ruler of Northern Fujiwara, was the grandson of Kiyohira and the son of Motohira.

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Around the Genpei War (1180–1185 CE) between the warlords of medieval Japan, he was successful in defense of his territory, thus being appointed as the imperial governor of Mutsu Province (1181 CE). However, in the reign of Yasuhira, the son of Morihira, the Northern Fujiwara clan was finally defeated by the Kamakura Shogunate. The rule of Kiyohira’s clan finally ended in 1189. In the building Konjiki-do, there are three altars (central, left and right). And Fujiwara’s mummies and mummified head were placed inside each altar. According to the temple tradition, the mummies of central, left, and right altars are those of Kiyohira, Motohira, and Hidehira, respectively. The proper scientific investigation of the Fujiwara mummies and related cultural remains that were placed inside the Konjiki-do was not performed until the 1950s. The academic investigation in 1950, supported by The Asahi Shimbun Foundation, was therefore an epoch-making event. The investigators were Japanese scientists of different research fields: Tei-ichi Asahina (chemistry), Kotondo Hasebe (anthropology), Torao Ohtsuki (microbiology), etc. The investigation of 1950 revealed many interesting academic findings. First, radiological analysis showed that the mummy placed in the central altar looked to be the oldest one, possibly over 70 years old at the time of his death. Researchers presumed that the old man might have experienced hemiparesis due to the possible sign of cerebrovascular accident in his brain. Next, the mummy of the right altar was estimated to be between 60 and 70 years old. He might have been suffered from spondylitis. Finally, the mummy of left altar was likely to have been around 60 years old. He might have died of acute illness due to idiopathic etiology because any signs of chronic diseases were not found during investigation. Different from the temple tradition, the investigation of 1950 revealed that the mummy in the left altar might be Hidehira, instead of Motohira who was instead placed in the right altar. Yasuhira’s mummified head was also proven to be placed inside the right altar along with Motohira’s mummy. In the report of 1950, there were few signs of evisceration in the bodies of the Fujiwara family mummies; accordingly, no artificial embalmment was evidenced for the mummification. Hanihara (1996) claimed that Fujiwaras might have been mummified under natural conditions after simple treatment for desiccation. Regardless, others also argued that the mummies might have been treated by a certain kind of artificial embalming technique. From the perspective of forensic medicine, Tanemoto Furuhata thought that the general condition of the Fujiwara mummies could not rule out the possibility that they were subjected to evisceration or other artificial treatment after death. Some people even speculated that the mummification of Fujiwara noblemen might have been significantly influenced by the embalming tradition of the Ainu, an ancient native people in the northern territory of Japan. Actually, it is still unclear how the Fujiwara clan members were mummified (Sakurai 1998).

Japanese Mummies in East Asian History As shown in the other chapters of this book, Chinese mummies date mainly to two historical periods. The first corresponds to the Warring States and Western Han periods (circa 2,000 years ago) (Fig. 5). The mummies representative of these

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Fig. 5 Timetable of Korean, Chinese, and Japanese mummies. Green in Korea, the mummies of Joseon Dynasty; Blue in China, Warring States and Western Han dynasty mummies; Brown, mummies of Song-Ming dynasty period; Red in Japan, Fujiwara mummies; Yellow, Buddhist monk mummies in Japan (Credit: Dong Hoon Shin)

periods have been mostly uncovered in the graves of Hubei and Hunan Provinces (Editorial Department 1972; Zheng et al. 1979; Hunan Medical College 1980; Wei et al. 1981; Zhou 1981; Wu 1982; Wu et al. 1996; Cockburn et al. 1998; Yeh and Mitchell 2016). Next, during the second millennium (especially of the Song and Ming dynasty periods), another type of mummy was found in unique types of graves around the downstream basin of Yangtze River (Suzhou Museum and Culture Center of Jiangyin 1982; Taizhou Municipal Museum 1986, 2013; Zhou et al. 1999) (Fig. 5). The details of these Chinese mummies are dealt in the other chapters of this book. Curiously, Chinese scholars do not consider the mummies of two distinct historical phases (the Warring States and Western Han periods versus the Song/Ming dynasties) as separate entities. Rather, they classified all the ancient or medieval Chinese corpses into a single group (except for Tarim basin mummies), calling them wet corpses (Wang and Zhang 2012), ancient corpses (Peng and Wu 1998), or Mawangdui-type cadavers (Hunan Medical College 1980; Lee et al. 1986; Peng and Wu 1998). Rather than emphasizing each other’s differences, more attention was given to the similarities found among the Chinese mummies. Meanwhile, Korean mummies unearthed from Joseon period graves (Fig. 5) are also closely related to the Chinese mummies in terms of historical and cultural

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origin. Actually, their graves of mummies were constructed in a very similar way. The inside of coffin was completely isolated from the outside environment by means of the sealants such as kaolin clay or lime-soil mixture (Lee et al. 2009a; Oh et al. 2018b; Shin et al. 2012). Related funeral customs were also similar. The coffin was tightly packed with lot of clothes, causing an oxygen deficiency during mummification. The mummified individuals did not undergo either embalming or evisceration after death (Lee et al. 2009b). Therefore, the visual or radiological appearances of the Korean and Chinese mummies also look very similar: their mummified organs being seriously shrunken, distorted, and dorsally displaced inside the body cavity (Hunan Medical College 1980; Wu 1982; Shin et al. 2003b; Lim et al. 2008; Lee et al. 2009b; Kim et al. 2014; Kim et al. 2016). The histological findings of them were very similar too (Hunan Medical College 1980; Shin et al. 2003a). Since the mummified corpses exhumed from Korean and Chinese graves share a number of cultural and biological characteristics, they could be classified into the same category whatever they will be called. From an academic point of view, the mummies in both countries are identical beings with almost similar characteristics. As seen in this chapter, interdisciplinary research has so far yielded invaluable data about Japanese mummies’ preservation pattern, and other cultural or biological backgrounds relating to them. Comparing with other East Asian mummies, there are remarkable aspects to be considered exclusively for Japanese mummies. The mummies found in Japan so far apparently differ, to some extents, from those of Korea and China, both in terms of the ideological background, mummification process, and preservation pattern, etc. (Fig. 5). Different from the other East Asian mummies, the burial pretexts of the mummified body in Japan mostly followed Buddhist traditions or ideas. Preservation status is also different from Chinese and Korean mummies. Unlike Korean or Chinese ones, Japanese mummies were dried mummies. They did not have soft, elastic, and moist skin; and their joints did not move freely as seen in Chinese and Korean mummies. After mummification, they were mostly placed at Buddhist temples, being worshipped by the religious believers. This is also a different point considering Chinese or Korean mummies were always discovered at the graves of archaeological sites. The photos of Enmyokai and Tetsuryukai used in this chapter were reproduced based on the plate published in Masashi Hijikata (2018) Shinpen Nihon no Miirabutsu wo tazunete (Publisher: Tenmujin, Tokyo). We are grateful to the generous permit of the author and photographer (Mr. Yasuhiko Okuno) for using those photos in this chapter. This research was supported by Basic Science Research Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2017R1D1A1B03030127).

Conclusion Thanks to the academic achievement, the scholars and the general public in Japan have come to recognize the similarities and differences between their mummies and those of other East Asian countries. The mummies of Japan have little in common

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with Korean and Chinese mummies. Therefore, it can be said that the history of Japan, which has taken a unique and separate path in East Asia, could be also represented in the mummies discovered in the country. All of this notwithstanding, the scientific investigations of the mummies in Japan are not sufficiently complete. Many years have passed since the last study was conducted on Japanese mummies, and while new scientific techniques have continued to evolve, to date very little additional investigation has been carried out on the Japanese mummies. This, of course, is largely because of religious taboos in Japanese society. However, at some point in time, we would like to have more opportunity to investigate the Japanese mummies once again, with much highly renovated research tools. The present chapter thus intends to synthesize the previous reports to date in order to provide the general overview for future related researches on Japanese mummies.

Cross-References ▶ Archaeological Findings of the Tarim Basin Graves and Mummies ▶ Joseon Dynasty Mummies of Korea ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies in Siberia ▶ Mummies of Song-Ming Dynasty in China

References Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Cockburn TA, Cockburn E, Reyman TA (1998) Mummies, disease, & ancient cultures, 2nd edn. Cambridge University Press, Cambridge Editorial Department (1972) Summary of discussions on the Mawangtui Tomb number 1 of the Han Dynasty. Archaeology 5:37 Hanihara K (1996) An additional study on the mummies of the Fujiwaras in the medieval age. Bull Int Res Cent Jpn Stud 13:11–33 Hong JH, Oh CS, Seo M, Chai JY, Shin DH (2017) Ancient ascaris DNA sequences of cytochrome B, cytochrome C oxidase subunit 1, NADH dehydrogenase subunit 1, and internal transcribed spacer 1 genes from Korean Joseon mummy feces. J Parasitol 103(6):795–800 Hunan Medical College (1980) Study of an ancient cadaver in Mawangtui Tomb no. 1 of the Han Dynasty. Cultural Relics publishing House, Beijing Kahila Bar-Gal G, Kim MJ, Klein A, Shin DH, Oh CS, Kim JW, Kim TH, Kim SB, Grant PR, Pappo O, Spigelman M, Shouval D (2012) Tracing hepatitis B virus to the 16th century in a Korean mummy. Hepatology 56(5):1671–1680 Kim MJ, Oh CS, Lee IS, Lee BH, Choi JH, Lim D-S, Yi YS, Han W-J, Kim Y-S, Bok GD, Lee SD, Shin DH (2008) Human mummified brain from a medieval tomb with lime-soil mixture barrier of the Joseon Dynasty, Korea. Int J Osteoarchaeol 18(6):614–623 Kim Y-S, Lee IS, Jung G-U, Kim MJ, Oh CS, Yoo DS, Lee W-J, Lee EJ, Cha SC, Shin DH (2014) Radiological diagnosis of congenital diaphragmatic Hernia in 17th century Korean mummy. PLoS One 9(7):e99779

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Kim MJ, Kim Y-S, Oh CS, Go J-H, Lee IS, Park W-K, Cho S-M, Kim S-K, Shin DH (2015) Anatomical confirmation of computed tomography-based diagnosis of the atherosclerosis discovered in 17th century Korean mummy. PLoS One 10(3):e0119474 Kim Y-S, Lee IS, Oh CS, Kim MJ, Cha SC, Shin DH (2016) Calcified pulmonary nodules identified in 350-year-old-Joseon mummy: the first report on ancient pulmonary tuberculosis from archaeologically obtained pre-modern Korean samples. J Korean Med Sci 31(1):147–151 Lee CF, Oscarson DW, Cheung SCH (1986) The preservation of a cadaver by a clay sealant: implications for the disposal of nuclear fuel waste. Nucl Chem Waste Manag 6:65–69 Lee EJ, Shin DH, Yang HY, Spigelman M, Yim SG (2009a) Eung Tae’s tomb: a Joseon ancestor and the letters of those that loved him. Antiquity 83(319):145–156 Lee IS, Lee EJ, Park JB, Baek SH, Oh CS, Lee SD, Kim YS, Bok GD, Hong JW, Lim DS, Shin MH, Seo M, Shin DH (2009b) Acute traumatic death of a 17th century general based on examination of mummified remains found in Korea. Ann Anat 191(3):309–320 Lee E-J, Oh CS, Yim SG, Park JB, Kim Y-S, Shin MH, Lee SD, Shin DH (2013) Collaboration of archaeologists, historians and bioarchaeologists during removal of clothing from Korean mummy of Joseon dynasty. Int J Hist Archaeol 17(1):94–118 Li Y (1984) Summary of parasitic research on corpses in China. Acta Anthro Sin 3:407–411 Lim DS, Lee IS, Choi KJ, Lee SD, Oh CS, Kim YS, Bok GD, Kim MJ, Yi YS, Lee EJ, Shin DH (2008) The potential for non-invasive study of mummies: validation of the use of computerized tomography by post factum dissection and histological examination of a 17th century female Korean mummy. J Anat 213(4):482–495 Morimoto I (1993) Buddhist mummies in Japan. Acta Anat Nippon 68:381–398 Morimoto I (1998) Japan: recent developments. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, disease & ancient cultures, 2nd edn. Cambridge University Press, Cambridge Ogata T (1998) Japan: scientific research. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, disease & ancient cultures, 2nd edn. Cambridge University Press, Cambridge Oh CS, Shin DH (2014) Making animal model for Korean mummy studies. Anthropol Anz 71(4):469–488 Oh CS, Lee SJ, Lee SD, Kim MJ, Kim YS, Lim DS, Shin DH (2013) Amplification of DNA remnants in mummified human brains from medieval Joseon tombs of Korea. Anthropol Anz 70(1):57–81 Oh CS, Kang IU, Hong JH, Slepchenko S, Park JB, Shin DH (2017) Tracing the historical origin of Joseon mummies considering the structural similarities between the burial systems of Korean and Chinese dynasties. Pap Anthropol XXVI(2):68–81 Oh CS, Hong JH, Park JB, Lee W-J, Bianucci R, Piombino-Mascali D, Shin DH (2018a) From excavation site to reburial ground: a standard protocol and related ethics of mummy studies in South Korea. Asian J Paleopathol 2:1–8 Oh CS, Kang IU, Hong JH, Park JB, Shin DH (2018b) An experimental assessment of the cause of mummification in the Joseon period Burials, Republic of Korea. Archaeol Ethnol Anthropol Eurasia 46(1):117–122 Peng LX, Wu ZB (1998) China: the Mawangtui-type cadavers in China. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, disease & ancient cultures, 2nd edn. Cambridge University Press, Cambridge Piombino-Mascali D, Beckett RG, Abinion OV, Shin DH (2016) The mummification process among the Ibaloy of Kabayan: a paleohistological note. In: Krutak L, Deter-Wolf A (eds) Marked for life: the archaeology of tattooing. University of Washington Press, Seattle Sakurai K (1998) Japan: a research and cultural history. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, disease & ancient cultures, 2nd edn. Cambridge University Press, Cambridge Seo M, Araujo A, Reinhard K, Chai JY, Shin DH (2014) Paleoparasitological studies on mummies of the Joseon dynasty, Korea. Korean J Parasitol 52(3):235–242 Seo M, Oh CS, Hong JH, Chai J-Y, Cha CS, Bang Y, Cha IK, Wi YG, Park JM, Shin DH (2017) Estimation of parasite infection prevalence of Joseon people by paleoparasitological data updates from the coprolites of pre-modern Korean mummies. Anthropol Sci 125(1):9–14

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Dong Hoon Shin, Chang Seok Oh, and Jong Ha Hong

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . History of Song and Ming Dynasties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chinese and Korean Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Origin of Korean Graves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . The Structure of Song and Ming Dynasty Graves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientific Studies of Song-Ming Period Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummification Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Comparison with Mawangdui Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Abstract

Over the past several decades, mummies have been found along with very wellpreserved cultural artifacts (e.g., clothing) in tombs representative of the Song and Ming dynasties from Chinese history. Despite their academic significance, SongMing period mummies have not been reported in detail outside China. Although

D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_32

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the structure of Song-Ming tombs was very diverse, most that held mummified remains had been sealed by a lime-soil-rice paste mixture likely implicated in the mummification that occurred therein. We presume that Song-Ming period tombs of China and those of the Joseon period in Korea have the same cultural origins. Both peoples must have accepted this tomb style, particularly as it effectively defended against grave robbers and was cost-efficient to construct. Keywords

Mummy · China · Song dynasty · Ming dynasty · Joseon dynasty · Grave

Introduction The discovery of mummies is not a phenomenon specific to certain regions, countries, or continents. Mummies have been reported from research in many Asian countries as well. For example, there are mummies of Buddhist monks of China, Southeast Asia, and Japan (Aufderheide 2003; Oh et al. 2017). Monks voluntarily attempted self-mummification from religious motives, as they were desirous of reaching the spiritual realm by escaping their worldly existence. Ascetic self-preparation of monks often caused them to have been mummified gradually, a process that began while they were still alive. The monks who were successful at selfmummification were kept in temples and worshiped thereafter by Buddhist followers (Pringle 2002). Information regarding Japanese monk mummies is addressed in another chapter in this book (▶ Chap. 47, “Mummies in Japan” by Fujita and Shin). The study of Siberian mummies also has contributed to our knowledge of native peoples of the Eurasian arctic or subarctic areas (Oh et al. 2017). Through scientific research of such mummies, many hard-to-understand facts in the historical literature have been made abundantly clearer (Sato et al. 2011; Bagashev et al. 2017; Slepchenko et al. 2019). In addition, pre-modern mummies of the Joseon dynasty (1392–1910 CE) in South Korea have been discovered, scientifically studied, and reported to the international academia community. The excellent preservation status of Korean mummies has allowed for detailed and authentic studies that could not be conducted on any other mummies (Seo et al. 2014; Oh et al. 2018a; Shin et al. 2018a; Hong et al. 2019). In China, different kinds of mummies have been formed under a variety of circumstances. The actual pattern of mummification in China is somewhat unique when compared with mummies reported from Europe or the Americas. One example are the mummies found in tombs dating to the Warring States (403 BCE-221 BCE) or Han dynasty (206 BCE-220 CE) periods (Editorial Department 1972; Zheng et al. 1979; Hunan Medical College 1980; Zhou 1981; Wu et al. 1996; Peng and Wu 1998; Aufderheide 2003; Yeh and Mitchell 2016; Shin et al. 2018a). In those studies, the cultural artifacts and the mummies from these tombs were found in a perfectly preserved state and amazed the Chinese people. From the academic information gleaned from these mummies and the associated cultural remains, scholars have revealed many facts that were not possible to obtain from conventional historical

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research (please see ▶ Chap. 46, “Mummies in China: Mawangdui and Other Related Mummies” by Joo and Shin). Other examples of mummies found in China are the naturally formed mummies discovered in the dry region of the Tarim Basin, Xinjiang province. The arid climate and salty soil conditions are likely contributors to mummification (Aufderheide 2003; Oh et al. 2017). Because of their excellent preservation and archaeological significance, the Tarim Basin mummies have attracted worldwide attention throughout academia as well as the public in general (Pringle 2002). An in-depth discussion on the Tarim Basin mummies can be found in another chapter in this volume (▶ Chap. 44, “Archaeological Findings of the Tarim Basin Graves and Mummies” by Kang et al.). Although the Chinese mummies described above are well-known to the general public, there are still others that have not been well recognized by the public at large, especially outside of China. For example, it is important to note that while the mummies of the Song (960–1279 CE) and Ming (1368–1644 CE) dynasties are less well-known outside of China, over several decades, news media in China have covered the accidental discoveries of mummies and cultural artifacts that were found to be perfectly preserved inside their tombs. This news has sometimes been leaked to Western countries but not to any great degree. Aufderheide (2003) described these Chinese mummies dating to the second millennium in his book, The Scientific Study of Mummies. With the exception of Aufderheide’s brief, pioneering introduction and occasional, scarce media coverage, the scientific and cultural properties of Chinese mummies formed during the Song and Ming dynasties have not been well-known to academia (Oh et al. 2017).

History of Song and Ming Dynasties China has a long and well-documented history. A number of dynasties rose and fell throughout Chinese history. Various peoples lived together and often fiercely competed with each other to attain hegemony in the Middle Kingdom. Great empires like the Qin (221–206 BCE) and Han (202 BCE to 220 CE) dynasties once unified China and prospered with a centralized bureaucracy and highly developed social systems. However, long after such a consolidation period, the collapse of the Han Empire brought disunity (220 CE). The Tang dynasty (618–907 CE) was the final outcome of the resultant long-term political turmoil. It became a highly centralized kingdom, facilitating Chinese cultural hegemony in all of East Asia (Overy 2015). In 755 CE, the rebellion of Lu-shan An, a famous frontier general, harshly impacted the political stability of China. Although this revolt was finally suppressed, the imperial authority of the Tang emperors was remarkably reduced by a series of uprisings on the part of magistrate-generals. Political power subsequently was slipping through the fingers of the central government, and each province in China became wealthy and prospered. In fact, each became independent kingdoms with large and bustling cities. Successive short-lived dynasties rose and fell repeatedly, thus ushering in the years of territorial fragmentation in Chinese history (Overy 2015). At the end of the ninth century, the political situation of China changed once again. By the coup d’etat of General Zhao Kuangyin (927–976 CE), the politically unstable

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five dynasties’ period ended. General Zhao then founded the Song dynasty (960– 1279 CE), which finally reunified all of China except for a small portion of the northeastern area lost to the Khitans (Liao dynasty). From a military point of view, the Song empire was not as powerful as its predecessor the Tang dynasty. However, they developed a national examination system to recruit well-educated imperial officials, thus reaching the apex of bureaucratic evolution in Chinese history. During the Song dynasty, economic growth continued, the population increased, trade prospered, and cities became medieval metropolitan centers. Scientific advances reached new heights as well. Education became popular due to highly developed printing techniques and economic growth in both urban and rural societies (Overy 2015). Eventually, new scholar-bureaucrats emerged to rule the Song empire. These men were gentry-class intellectuals who exerted a great influence on the course of Chinese history. They were politicians, philosophers, Confucians, and even military strategists. They served the central government as imperial officials. They were fully convinced of their political and cultural roles regarding their country’s prosperity (Overy 2015). Fan Zhongyan (989–1052 CE), one of the most famous grandmasters of the gentry class at that time, claimed that he would be “the first one” to worry about the world’s hardship and “the last one” to enjoy its happiness after all others. Life during the Song dynasty was not always peaceful and happy but was exposed to a constant threat from the north. At first, the danger was from the Khitans and, then, from the Jurchen Jin dynasty (1115–1234 CE), which occupied the northern territory of China. The latter even expelled the Song dynasty from most territory north of the Huai River. Because of the Jin dynasty’s occupation of the northern territory, the Song Dynasty was subdivided into Northern Song (960–1127 CE) and Southern Song (1127–1279 CE) periods. Even after being pushed south by the Jin dynasty, the Southern Song still prospered economically and culturally. In the thirteenth century, however, an even more serious threat came from the north. The Mongols, pastoral nomads, conquered almost the entire Eurasian continent. They built a vast empire never before seen in history. From Korea to the eastern border of Europe, all political and economic power became theirs. The Mongols even destroyed the Southern Song dynasty, replacing it with their own Yuan (1271– 1368 CE) dynasty. After about a century of rule by the Mongols, Zhu Yuanzhang (1328–1398 CE), one of the rebel leaders, established a new Chinese dynasty, the Ming. When the Mongols were pushed out of China in 1388, the Ming controlled the Chinese mainland. This new kingdom prospered until 1644, when the Manchus conquered China once again, this time founding the Manchurian dynasty of Qing (Overy 2015). As we have described, the Chinese dynasties have changed ceaselessly, but the gentryclass intellectuals continued to be the leading figures in Chinese society. Most mummified individuals found inside Song-Ming period graves were such men.

Chinese and Korean Mummies The reports on the Song and Ming dynasty mummies in China are of special interest to scholars in South Korea. In Chinese archaeology, not all of these mummies were found in a singular type of tomb but have mostly come from graves that were

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uniquely structured and sealed by a lime mixture (Taizhou Municipal Museum 2013). Archaeologists in South Korea have speculated that the lime mixture was more than a simple adhesive or filling material. They noted strong exothermic reactions when lime met water and a resultant hard, sealing barrier resistant to outside elements (Fig. 1). This property of the lime-soil mixture barrier served to cut off the free exchange of water or air between the inside and the outside of the coffin. They presumed, therefore, that there might be a significant causal relationship between the lime mixture and the mummification that occurred inside the graves (Oh et al. 2017; Shin et al. 2018a). Over several decades, Korean mummies have been reported for graves dating to the Joseon dynasty (1392–1910 CE). The most remarkable feature of Joseon mummies is their perfect preservation status after hundreds of years of burial (Oh et al. 2017) (Fig. 2). Various aspects of the Korean mummies, either in cultural or biological perspectives, have been investigated by specialists representing different research fields (Shin et al. 2003; Lee et al. 2013; Song and Shin 2014; Seo et al. 2017; Oh et al. 2018a, b; Shin et al. 2018a). With anatomical, histological, radiological, and molecular techniques, scientific information primarily focused on the physical and pathological traits of the sixteenth- to eighteenth-century Korean people has been successfully gleaned from the Joseon period mummies (Oh et al. 2013; Kim Fig. 1 Structure of Joseon dynasty grave with lime-soil mixture barrier in East Asia (Korea). Note the presence of the barrier around the coffin (Credit: Dong Hoon Shin)

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Fig. 2 Heavily packed clothing in Joseon dynasty grave of East Asia (Korea). The preservation status was very good (Credit: Dong Hoon Shin)

et al. 2014; Seo et al. 2014; Kim et al. 2015, 2017; Seo et al. 2017; Kim et al. 2018; Shin et al. 2018b; Hong et al. 2019). This multidisciplinary research approach is invaluable to anthropologists who have endeavored to reveal the interaction between men, environments, and pathogens in East Asian history. The research techniques employed in mummy research in Korea have improved our understanding of Joseon people and society to an unexpectedly high degree (Oh et al. 2017). The details of the Korean mummy research are discussed in another chapter in this book (▶ Chap. 45, “Joseon Dynasty Mummies of Korea” by Shin et al.). Despite Korean mummies’ significance, their cultural and historical origins are not yet completely known. The only subject that has been agreed on by researchers is that the distinctive mummy tomb type started to be used from the early Joseon period, possibly under the strong influence of the Confucian ideology imported from China (Shin et al. 2008; Oh et al. 2017; Shin et al. 2018a). Indeed, historically, the Korean mummies have been discovered in the graves that were used by the Confucians of the country.

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Armed with intense intellectual convictions and religious beliefs, Confucians blamed the Buddhists for the evils prevalent in late fourteenth-century Korea (Shin et al. 2008; Oh et al. 2017). From 1392 CE, when the Confucians finally toppled the Goryeo dynasty (918–1392 CE) and founded their own Joseon dynasty (1392– 1910 CE), every Buddhist ritual was thoroughly purged under the new leadership (Shin et al. 2008; Oh et al. 2017). Indeed, the Confucians in Korea were so fundamental that they strictly adhered to even the most trivial Confucian rituals. Therefore, the adoption of Confucian rituals deeply influenced Korean people, society, and culture altogether (Oh et al. 2015, 2017) (Fig. 3). In this context, the concept of a specific tomb style was eventually introduced into Korean society. In the twelfth century, Zhu Xi (1130–1200 CE) of China (Fig. 4), one of the great innovators of Confucian ideology, argued that a grave sealed by a mixture of lime, soil, and sticky rice water would be the best option for Confucianists (Oh et al. 2017). His writing in this vein (Jujagare in Korean; Zhouzijiali in Chinese) subsequently was imported to Korea. Perhaps he might have chosen this kind of grave because it was commonly used in South China when Zhu Xi was still alive. As Joseon Confucians respected him as their great mentor (Fig. 5), Zhu Xi’s recommended grave style was soon accepted enthusiastically among the Confucians of the Joseon dynasty (Shin et al. 2008).

Fig. 3 The importation of Confucian rituals influenced Korean society and people very deeply. Every aspect of state administration and rituals had to abide by the Confucian axioms. The photo here is of the Temple of Confucius at Seoul City (Credit: Dong Hoon Shin)

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Fig. 4 Portrait of Zhu Xi (courtesy of National Museum of Korea)

Numerous tombs that faithfully followed Zhu Xi’s principles were built throughout Korea. This approach formed a major axiom of Confucian funeral rituals in Joseon society (Oh et al. 2017; Shin et al. 2018a) (Fig. 6). Several 100 years later, Korean archaeologists discovered thousands of such graves, sometimes with mummies inside of them (Fig. 7). These tombs had been sealed with a lime-soil mixture (Lee et al. 2009). Such Joseon graves are known as Hoegwakmyo in Korean.

The Origin of Korean Graves The emergence of Joseon period graves in Korean history seems to have been deeply influenced by Chinese cultural heritage. Therefore, it is not surprising that similar graves and mummies have also been reported from Chinese archaeology (Oh et al. 2017; Shin et al. 2018a). In fact, we show in this chapter that there have been discoveries of far-earlier Song-Ming period burials on the Chinese mainland

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Fig. 5 Portrait of Shi-Yeol Song (1607–1689 CE), champion of seventeenthcentury Confucian fundamentalists in Korea. He supported Zhu Xi’s recommendations enthusiastically (courtesy of National Museum of Korea)

Fig. 6 Sangryebiyo (published 1648), a summary of Zhu Xi’s ritual book Jujagare (courtesy of Daegu National Museum)

in graves of generally the same type as used later in Korea (Oh et al. 2017; Shin et al. 2018a). The Song-Ming period mummies archaeologically discovered thus far have been reported from this type of burials.

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Fig. 7 Joseon grave (Hoegwakmyo in Korean) at archaeological site. The structure is similar to the graves of the Song-Ming period in China (Credit: Dong Hoon Shin)

It is not clear when this type of tomb (sealed by the lime-soil mixture) began to be constructed in China. As discussed above, Zhu Xi’s book (Jujagare) described the prototype grave that would be constructed thereafter in Korea. The graves must have been actively constructed in China while he was still alive (1130–1200 CE). Therefore, scholars mostly have traced the cultural origin of the Korean mummy and related graves only to the Southern Song period (1127–1279 CE) (Oh et al. 2017; Shin et al. 2018a). Nevertheless, in Chinese archaeology, one of the earliest tombs sealed by the lime-soil mixture was the Sun Siniangzi mu (tomb) reported from Jiangyin city of Jiangsu province (Suzhou Museum and Culture Center of Jiangyin 1982; Oh et al. 2017; Shin et al. 2018a). According to the archaeological report, it was built in 1055 CE as the grave for a Northern Song bureaucrat’s wife (Oh et al. 2017). This discovery is important to the attempts to trace the cultural origin of the Joseon dynasty tombs in which the Korean mummies have been found. Since the Sun Siniangzi mu tomb sealed by the lime-soil mixture appeared far before Zhu Xi was born (1130 CE), the possible prototype of Korean graves must have been used by the gentry of the Northern Song period (960–1127 CE) as well (Oh et al. 2017; Shin et al. 2018a). In this respect, at least the Northern Song people must have built similarly structured tombs for their funerals. Thus, due to the recent archaeological findings in China, the cultural origins of Korean tombs sealed by the lime-soil mixture can be traced farther back in time. We can therefore readjust the historical and cultural origins of Korean mummy-related graves based on the estimated date of the Sun Siniangzi mu tomb (Oh et al. 2017; Shin et al. 2018a).

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The Structure of Song and Ming Dynasty Graves For many years after its introduction, Zhu Xi’s recommended tomb was not mainstreamed in Korean society. Over time, the burial system soon overcame the strong resistance of Korean burial traditions and became favored by most upperclass people of the period. During the earliest period of its introduction, however, the grave was constructed in a somewhat different way from Zhu Xi’s original recommendation. Briefly, the coffin was first placed inside the burial pit, and then the periphery was sealed by the lime-soil mixture (Fig. 8a) (Kim 2016; Oh et al. 2017). This procedure was never included in Zhu Xi’s original recommendations. In this regard, Joseon Confucians who wished to strictly follow the principles of Zhu Xi were opposed to such a modification. Finally, their fundamentalist claims were widely accepted by the Korean society in the seventeenth century. In brief, following the original procedures of Zhu Xi, they first built a lime-soil mixture wall inside the burial pit, after which only one coffin was placed inside the space between the walls (Kim 2016; Oh et al. 2017) (Fig. 8b). As discussed above, the Confucians in Korea established Zhu Xi’s recommended grave as the principal burial system of Joseon society (Oh et al. 2017). There were some variations in structure among graves in the earliest period of construction. However, they soon became standardized, regardless of the construction date or the place of discovery. Such archaeological uniformity was likely due to the fact that the Confucians in Korea built their graves not by oral or technical traditions but by strictly respecting Zhu Xi’s original ritual codes as summarized in his book (Oh et al. 2017; Shin et al. 2018a). Nonetheless, the situation regarding grave style homogeneity in China was different. According to Chinese archaeology, the Song-Ming period tombs sealed by the lime-soil-rice paste mixture were very diverse in structure. Unlike Korean graves, the coffins placed inside the Chinese graves were mostly doubled. Some coffins were even surrounded by bricks, which practice is not seen in Zhu Xi’s original ritual book. Certainly, Zhu Xi’s recommendation was not followed universally in China; rather, the Chinese must have constructed graves following the popular trends at the given time (Oh et al. 2017; Shin et al. 2018a). Despite this structural variety, we found that some of the Song and Ming period graves in China do look structurally very similar to Zhu Xi’s recommended grave style as seen during the Joseon dynasty of Korea. The Song and Ming dynasty graves did have one thing in common: the coffin perimeter was sealed by the prescribed lime-soil-rice paste mixture. Further, the structure must have influenced the mummification process inside the graves, just like the cases observed in South Korea (Oh et al. 2017; Shin et al. 2018a). Let’s see the Xu Fan couple tomb of the Ming dynasty (1533 CE) (Fig. 8c). Inside it, the couple’s coffins were laid side by side. When Chinese archaeologists excavated the tomb, the outer coffin was sealed by a lime-soil mixture (Taizhou Municipal Museum 1986). The structure of this tomb is obviously similar to Zhu Xi’s recommended grave style found at Korean archaeological sites (Oh et al. 2017). Other Ming dynasty tombs of similar structure are those of the Li Xian Fang couple

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Fig. 8 Comparison of Joseon period Korean (a and b) and Song-Ming period Chinese (c to f) graves. The area marked in gray color is part of the lime-soil mixture. The green color indicates the coffin. In (a), the coffins were placed first in the burial pit; and then the lime-soil mixture walled the coffins. Two coffins were present inside the grave. In (b), the lime-soil wall was made first; and then the coffin was put down inside the secured space. Only one coffin could be found. The figures are redrawn based on those in Kim (Kim 2016) and Oh et al. (2017). The Song-Ming period tombs of (c) to (f) are similar to the Joseon Korean tombs. (c) Xu Fan couple tomb (Taizhou, Jiangsu); (D) Li Xian Fang couple tomb (Shanghai); (e) Liu Xiang couple tomb (Taizhou, Jiangsu); (f) Sensenzhuang tomb (Taizhou, Jiangsu). These figures are redrawn based on the plates in Shanghai Shi Wen Wu Guan Li Wei Yuan Hui 2009; Taizhou Municipal Museum 1986, 1992, 2013 (Credit: Jong Ha Hong)

(Shanghai) (Fig. 8d), Liu Xiang (Taizhou) (Fig. 8e), and Sensenzhuang village (Taizhou) (Fig. 8f). Like the Xu Fan couple tomb, the coffins in those cases had been completely sealed with a lime-soil mixture, further demonstrating that the prototype of Zhu Xi’s recommended Korean graves was present in Song-Ming period China (Shanghai Shi Wen Wu Guan Li Wei Yuan Hui 2009; Taizhou Municipal Museum 2013; Oh et al. 2017; Shin et al. 2018a).

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Scientific Studies of Song-Ming Period Mummies Archaeologists have found very well-preserved mummies in Song-Ming period graves sealed by lime mixture. In the case of the Xu Fan couple described above, the mummified bodies were found in a perfectly preserved condition (Oh et al. 2017; Shin et al. 2018a). The mummy’s soft tissue was still elastic, and the skin color was light yellow. On histological examination, adipocytes, tracheal cartilage, and cell nuclei, etc. were identified in the mummified tissue (Taizhou Municipal Museum 1986; Oh et al. 2017). There was also another case of a Song-Ming period mummy investigated by Chinese archaeologists. In 1988, a Southern Song dynasty grave of the thriteenth century was discovered at De’an County in Jiangxi province. The grave had been constructed in 1274, just a few years before the final fall of the Southern Song Empire to the Mongols’ invasion. Inside the coffin, archaeologists found a mummified female who had been the wife of Wu Chou, an official of the Southern Song dynasty (Zhou et al. 1999; Oh et al. 2017; Shin et al. 2018a). As in the cases of the Xu Fan mummies, the mummified wife’s organs and tissues were perfectly preserved (Oh et al. 2017). Chinese scholars also have reported impressive results from paleopathological research conducted on Song-Ming period mummies. Using scientific techniques, they uncovered a pathological sign of atherosclerosis in the Xu Fan mummy (Oh et al. 2017). Also, in a paleoparasitological study on mummy coprolite samples, they found that the Song-Ming period individuals must have suffered from various parasitic infections (Taizhou Municipal Museum 1986; Oh et al. 2017). Paleoparasitological research thus far conducted on Song-Ming period mummies has detected ancient Clonorchis sinensis, Fasciolopsis buski, Ascaris lumbricoides, and Trichuris trichiura eggs (Li 1984; Yeh and Mitchell 2016) (Table 1). These archaeoparasitological results are very similar to those for Korean mummies of the Joseon period. Most scientific research methods that have been applied to Korean mummies also have been successfully performed in research on Song-Ming period mummies.

Mummification Process What then was the actual mechanism of mummification occurring inside the SongMing period graves? Like South Korea, China also lacks the climatic conditions conducive to natural mummification (Oh et al. 2017). Aufderheide (2003) suggested that the good preservation status of these mummies might have been caused by an antiseptic technique once popular in China (Oh et al. 2017). However, we note that no embalming technique of the Song-Ming period has been discovered so far by any historical or archaeological investigations. In the Confucian tradition of East Asia, it was never desirable for the dead body to remain intact even after death. To explain the phenomenon of near-perfect preservation of mummified remains therefore, we must find another hypothesis and evidence supporting it.

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Table 1 Song-Ming period mummies and archaeoparasitological results (Li 1984; Yeh and Mitchell 2016) Location (province) Jintan (Jiangsu) Hengyang (Hunan) Guangzhou (Guangdong) Guangzhou (Guangdong)

Yangzhou (Jiangsu) Yangzhou (Jiangsu) Fuqing (Fujian)

Shaowu (Fujian) Longyan (Fujian) Fuzhou (Fujian)

Parasite A. lumbricoides T. trichiura C. sinensis A. lumbricoides T. trichiura C. sinensis Fasciolopsis buski A. lumbricoides T. trichiura A. lumbricoides T. trichiura C. sinensis F. buski T. trichiura A. lumbricoides A. lumbricoides T. trichiura A. lumbricoides C. sinensis T. trichiura

Time period Song dynasty Song dynasty Ming dynasty Ming dynasty

Ming dynasty Ming dynasty Ming dynasty

Ming dynasty Ming dynasty Ming dynasty

Various hypotheses and speculations have been proposed to explain the cause of mummification inside Song-Ming period graves. Above all, Chinese archaeologists have argued that mummified bodies might have been perfectly preserved due to the complete sealing of the grave from the outside environment (Taizhou Municipal Museum 1986; Oh et al. 2017). Mummification might also have been in part caused by materials such as lime or mercury found inside graves during archaeological investigations (Zhou et al. 1999). The maintenance of constant temperature and humidity inside the coffin also has been convincingly posited as a cause of SongMing period mummification (Taizhou Municipal Museum 1986). Other investigators have pointed to the densely packed clothes found inside Song-Ming graves as a key component of mummification (Wang and Zhang 2012; Taizhou Municipal Museum 2013). In general, Song-Ming period coffins were filled with clothing so tightly that there was no empty space left inside, thus inducing an oxygen-deficient environment. This evidently inhibited the active proliferation of bacteria therein (Wang and Zhang 2012; Taizhou Municipal Museum 2013; Oh et al. 2017). We presume that Song-Ming period mummification likely was induced by the structure of the graves and/or the unique funeral traditions. It is likely that mummification occurring inside Song-Ming graves was not so simple a process but rather was caused by a very complicated action of multiple factors such as the complete sealing of the graves, maintenance of constant temperature and humidity, and others (Taizhou Municipal Museum 1986; Oh et al. 2017). The above hypotheses are not new to Korean scholars. Similar arguments regarding the causes of mummification also have been raised in Korea. For example, no

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embalming tradition was known in Korean history (Kim et al. 2006). The country is not an ideal place for natural mummification, because it is neither a dry land nor a permafrost region. As in the case of the Song-Ming period mummies, scholars often think that mummification in Korea might have been influenced by heavy packing of clothing inside the coffin (Fig. 2). Animal experiments with miniature graves might also have implications for the possible causes of Song-Ming period mummification (Oh and Shin 2014; Oh et al. 2018c). The researchers built miniature graves mimicking Joseon period tombs that were, like the Song-Ming period ones, surrounded by a lime-soil mixture barrier (Fig. 9). The experimental results were astonishing. Very strong exothermic reactions occurred when the lime contacted with moisture. The average temperature inside the coffin rose to 130.8  23.5 °C for 141.0  64.7 min. This means that such a high temperature maintained for a long time could be enough to kill nearly all microbes that might have been initially present inside graves (Oh et al. 2018c). Considering that microorganisms remaining in a dead body proliferate rapidly after death and ultimately induce decomposition of organic materials (Gill-King 2000), a high and long-sustained temperature must have contributed to mummification inside Joseon period graves (Oh et al. 2018c). This hypothesis was further corroborated by another experiment. In an animal model experiment, samples from rat intestines were cultured on MacConkey (MAC) or blood agar plate (BAP) media (Oh et al. 2018c). After the bacteria had been incubated, neither aerobic nor anaerobic bacteria were cultured on the MAC or BAP plates. This means that the strong heat generated by the lime-soil mixture had sterilized the microbes inside the intestines, thus hindering the decomposition of organic materials inside of the miniature graves (Oh et al. 2018c). In this regard, we note that, like Korean graves, Chinese Song-Ming period graves were surrounded by similar kinds of materials (a lime-soil-rice paste mixture). Therefore, high temperature probably was generated by that lime mixture type as well, thus slowing down the process of decomposition caused by bacterial proliferation and finally inducing mummification.

Fig. 9 Miniature grave mimicking Joseon period tomb. A lime-soil-mixture wall surrounded the coffin. An experimental animal was placed inside the coffin. The experiment revealed that the contact of lime with water generated intense exothermic reactions (Credit: Dong Hoon Shin)

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Comparison with Mawangdui Mummies In general, scholars have noted certain features shared by ancient and medieval mummies discovered in China. In ancient Chinese nobles’ graves of the Warring States (402 BCE to 221 BCE) or Western Han (202 BCE to 8 CE) period, very wellpreserved mummies were discovered. Like the Song-Ming period mummies, their skins were soft and wet. Their joints could still move freely even after an exceptionally long time of burial. Histological analysis conducted on the mummies also showed an excellent preservation status like that of the Song-Ming period mummies. Indeed, the preservation pattern of the Xu Fan mummies of the Ming dynasty was similar to that of the ancient Chinese mummies of the Warring States or Western Han period. The gross anatomical and histological morphologies reported for the Xu Fan mummies were not unique at all relative to those of the Mawangdui mummy of the Han dynasty. In comparing those ancient (Warring States or Western Han period) and Song-Ming period mummies, scholars speculated that, due to their similarities, they do not properly belong in separate classifications. Since Chinese mummies of different historical periods share many patterns that are not identified in any known mummy category throughout the rest of the world, they have all been assigned to a single classification: wet ancient corpses or Mawangdui-type cadavers (Hunan Medical College 1980; Lee et al. 1986; Peng and Wu 1998; Wang and Zhang 2012). Thus, if the Song-Ming and Joseon period mummies belong to the same type due to their shared characteristics, the Mawangdui-type classification also could be applied to the Korean mummies (Shin et al. 2003). Whatever we call these types of mummies, we need to study them more comprehensively across borders. Collaborative efforts among scholars from both China and Korea would greatly enhance research outcomes. To date, reports on the possible relationship between mummies of ancient China and Korea are insufficient in number.

Conclusion Through the studies conducted on the Song-Ming period graves and mummies, scholars have acquired invaluable information on the health and disease status of Chinese people living very long ago (Oh et al. 2017). Important cultural remains such as clothing from centuries ago have become important subjects of historical research through archaeological investigations of Song-Ming period graves. Based on those achievements, academics can re-evaluate the importance of Song-Ming period graves and mummies in order to better uncover and understand the lives of people living hundreds of years ago. Scholars have noted similarities between mummies of the Song-Ming period in China and those of the Joseon period in Korea. The cultural origin of the Joseon graves and mummies could be traced back to the Northern Song period of China. The bodies in both cases became mummified inside very uniquely structured graves surrounded by a lime mixture barrier. We surmise that mummification in both the

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Song-Ming and Joseon period tombs might have been induced by similar mechanisms sharing features of preservation and pathological status. As in the cases of Joseon graves and mummies, considerable amounts of clothing and other cultural artifacts have been discovered during the excavations of the SongMing period graves (Taizhou Municipal Museum 1986, 2013; Oh et al. 2017). Some such artifacts and a mummy are displayed in Taizhou Museum, Jiangsu province. The Song-Ming period mummies were once displayed in the Shanghai Natural History Museum as well, but the exhibition is no longer available. What we have discussed in this chapter mostly amounts to a presumption rather than a definitive conclusion. To corroborate our hypotheses, more discoveries and discussions should be made on Chinese mummies of Song-Ming period graves. For example, researchers must answer questions regarding whether the Song-Ming mummies discovered thus far were actually formed by the same process as seen in the cases of Joseon period mummies. While much is known, many details on SongMing graves and mummies are as yet unavailable. Pending the obtainment of such information and a final consensus on its significance in respect of East Asian mummies, this chapter might prove meaningful to the concerned scholars.

Cross-References ▶ Archaeological Findings of the Tarim Basin Graves and Mummies ▶ Joseon Dynasty Mummies of Korea ▶ Mummies in China: Mawangdui and Other Related Mummies ▶ Mummies in Japan ▶ Mummies in Siberia Acknowledgment This research was supported by Basic Science Reseach Program through the National Research Foundation of Korea by the Ministry of Education (2020R1A2C1010708).

References Aufderheide AC (2003) The scientific study of mummies. Cambridge University Press, Cambridge Bagashev AN, Razhev DI, Poshekhonova OE, Slepchenko SM, Alekseyeva EA (2017) An anthropological study of mummified remains from the Zeleny Yar cemetery on the lower Ob, Western Siberia. Archaeol Ethnol Anthropol Eurasia 45(1):135–145 Editorial Department (1972) Summary of discussions on the Mawangtui Tomb number 1 of the Han dynasty. Archaeology 5:37 Gill-King H (2000) Chemical and ultrastructural aspects of decomposition. In: Haglund WD, Sorg MH (eds) Forensic Taphonomy. CRC Press, Boca Raton Hong JH, Oh CS, Chai J-Y, Seo M, Shin DH (2019) Cytochrome c oxidase subunit 1, internal transcribed spacer 1, Nicotinamide adenine dinucleotide hydrogen dehydrogenase subunits 2 and 5 of Clonorchis sinensis ancient DNA retrieved from Joseon dynasty mummy specimens. J Korean Med Sci 34(20):e149 Hunan Medical College (1980) Study of an ancient cadaver in Mawangtui Tomb no. 1 of the Han dynasty. Cultural Relics Publishing House, Beijing

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Kim WL (2016) Research on the tomb culture of the gentry during the Joseon dynasty. Publishing Company Minsokwon, Seoul Kim SB, Shin JE, Park SS, Bok GD, Chang YP, Kim J, Chung YH, Yi YS, Shin MH, Chang BS, Shin DH, Kim MJ (2006) Endoscopic investigation of the internal organs of a 15th-century child mummy from Yangju, Korea. J Anat 209(5):681–688 Kim Y-S, Lee IS, Jung G-U, Oh CS, Yoo DS, Lee WJ, Lee E-J, Cha SC, Shin DH (2014) Radiological diagnosis of congenital diaphragmatic Hernia in 17th century Korean mummy. PLoS One 9(7):e99779 Kim MJ, Kim Y-S, Oh CS, Go J-H, Lee IS, Park W-K, Cho S-M, Kim S-K, Shin DH (2015) Anatomical confirmation of computed tomography-based diagnosis of the atherosclerosis discovered in 17th century Korean mummy. PLoS One 10(3):e0119474 Kim YS, Kim MJ, Hong JH, Oh CS, Bianucci R, Shin DH (2017) The scientific and ethical background of the invasive studies on the Korean mummies of the Joseon dynasty. Asian J Paleopathol 1:5–11 Kim Y-S, Oh CS, Kim MJ, Hong JH, Shin DH (2018) Post-factum autopsy to confirm cardiac structures visible on computed tomography images of Korean mummies: the radiological basis of paleo-cardiology. Anthropol Anz 75(4):339–350 Lee CF, Oscarson DW, Cheung SCH (1986) The preservation of a cadaver by a clay sealant: implications for the disposal of nuclear fuel waste. Nucl Chem Waste Manage 6:65–69 Lee IS, Lee EJ, Park JB, Baek SH, Oh CS, Lee SD, Kim YS, Bok GD, Hong JW, Lim DS, Shin MH, Seo M, Shin DH (2009) Acute traumatic death of a 17th century general based on examination of mummified remains found in Korea. Ann Anat 191(3):309–320 Lee E-J, Oh CS, Yim SG, Park JB, Kim Y-S, Shin MH, Lee SD, Shin DH (2013) Collaboration of archaeologists, historians and bioarchaeologists during removal of clothing from Korean mummy of Joseon dynasty. Int J Hist Archaeol 17(1):94–118 Li Y (1984) Zhong guo gu shi ji sheng chong xue yan jiu zhi zong shu. Acta Anthropol Sinica 3(4):407–411 Oh CS, Shin DH (2014) Making animal model for Korean mummy studies. Anthropol Anz 71(4):469–488 Oh CS, Lee SJ, Lee SD, Kim MJ, Kim Y-S, Lim D-S, Shin DH (2013) Amplification of DNA remnants in mummified human brains from medieval Joseon tombs of Korea. Anthropol Anz 70(1):57–81 Oh CS, Koh B-J, Yoo DS, Park JB, Min SR, Kim Y-S, Lee SS, Ge J, Seo SB, Shin DH (2015) Joseon funerary texts tested using ancient DNA analysis of a Korean mummy. Anat Rec 298:1191–1207 Oh CS, Kang IU, Hong JH, Slepchenko S, Park JB, Shin DH (2017) Tracing the historical origin of Joseon mummies considering the structural similarities between the burial systems of Korean and Chinese dynasties. Pap Anthropol XXVI(2):68–81 Oh CS, Hong JH, Park JB, Lee WJ, Bianucci R, Piombino-Mascali D, Shin DH (2018a) From excavation site to reburial ground: a standard protocol of mummy studies in South Korea. Asian J Paleopathol 2:1–8 Oh CS, Shin DH, Hong JH, Lee E (2018b) Single nucleotide polymorphism analyses on ABCC11, EDAR, FGFR2 and ABO genotypes of mummified people of Joseon dynasty, South Korea. Anthropol Sci 126(2):67–73 Oh CS, Kang IU, Hong JH, Park JB, Shin DH (2018c) An experiment assessment of the cause of mummification in Joseon period burials, Republic of Korea. Archaeol Ethnol Anthropol Eurasia 46(1):117–122 Overy R (2015) The times complete history of the world. Collins Bartholomew, Edinburgh Peng LX, Wu ZB (1998) China: the Mawangtui-type cadavers in China. In: Cockburn A, Cockburn E, Reyman TA (eds) Mummies, Disease & Ancient Cultures, 2nd edn. Cambridge University Press, Cambridge Pringle HA (2002) The mummy congress: science, obsession, and the everlasting dead. Hyperion, New York

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Sato T, Razhev D, Amano T, Masuda R (2011) Genetic features of ancient West Siberian people of the middle ages, revealed by mitochondrial DNA haplogroup analysis. J Hum Genet 56(8):602– 608. https://doi.org/10.1038/jhg.2011.68. PMID: 21753768 Seo M, Araujo A, Reinhard K, Chai JY, Shin DH (2014) Paleoparasitological studies on mummies of the Joseon dynasty, Korea. Korean J Parasitol 52(3):235–242 Seo M, Oh CS, Hong JH, Chai J-Y, Cha SC, Bang Y, Cha IG, Wi YG, Park JM, Shin DH (2017) Estimation of parasite infection prevalence of Joseon people by paleoparasitological data updates from the coprolites of pre-modern Korean mummies. Anthropol Sci 125(1):9–14 Shanghai Shi Wen Wu Guan Li Wei Yuan Hui (2009) Shanghai Ming Mu. Wen wu chu ban she, Beijing Shin DH, Choi YH, Shin KJ, Han GR, Youn M, Kim CY, Han SH, Seo JC, Park SS, Cho YJ, Chang BS (2003) Radiological analysis on a mummy from a medieval tomb in Korea. Ann Anat 185(4):377–382 Shin MH, Yi YS, Bok GD, Lee E-J, Spigelman M, Park JB, Min S-R, Shin DH (2008) How did mummification occur in bodies buried in tombs with a lime soil mixture barrier during the Joseon dynasty in Korea? In: Pena PA, Martin RM, Rodriguez AR (eds) Mummies and science world mummies research. Academia Canaria de la Historia, Santa Cruz de Tenerife, pp 105–113 Shin DH, Bianucci R, Fujita H, Hong JH (2018a) Mummification in Korea and China: Mawangdui, Song, Ming and Joseon dynasty mummies. Biomed Res Int:6215025. https://doi.org/10.1155/ 2018/6215025 Shin DH, Oh CS, Hong JH, Lee H, Lee SD, Lee E (2018b) Helicobacter pylori DNA obtained from the stomach specimens of two 17th century Korean mummies. Anthropol Anz 75(1):75–87 Slepchenko SM, Ivanov SN, Gusev AV, Svyatova EO, Fedorova NV (2019) Archaeoparasitological and palynological analysis of samples from intestinal contents of a child mummy from Zelyeny Yar burial ground (XII-XIII centuries AD). Archaeol Res Asia 17:133–136 Song MK, Shin DH (2014) Joseon mummies identified before bioanthropological studies began in Korea. Pap Anthropol XXIII(1):117–134 Suzhou Museum and Culture Center of Jiangyin (1982) The tomb of sun Siniangzi of the Norther Song dynasty at Jiangyin. Wenwu 12:28–35 Taizhou Municipal Museum (1986) Excavation of the Ming dynasty tomb of Xu fan and his wife at Taizhou, Jiangsu. Wenwu 9:1–15 Taizhou Municipal Museum (1992) Excavation of the Ming joint burial tomb of Liu Xiang and his wife at Taizhou City, Jiangsu. Wenwu 9:66–77 Taizhou Municipal Museum (2013) The excavation of a tomb of the Ming dynasty at Sensenzhuang in Taizhou City, Jiangsu. Wenwu 11:36–49 Wang WG, Zhang W (2012) Analysis of antisepsis techniques of the Ming-period corpse excavated in Taizhou. Zhonghua Yi Shi Za Zhi 42(1):12–14 Wu Z, Guan Y, Zhou Z (1996) Study of an ancient corpse of the warring states period unearthed from Tomb no. 1 at Guo-Jia Gang in Jingmen City (a comprehensive study). J Tongji Med Univ 16(1):1–5 Yeh HY, Mitchell PD (2016) Ancient human parasites in ethnic Chinese populations. Korean J Parasitol 54(5):565–572 Zheng GZ, Feng WH, Boa YH, Xue JN, Ying YS (1979) Microscopic and submicroscopic studies on the peripheral nerve and the skeletal muscle of the female cadaver found in the Han tomb no. 1. Sci Sinica 22(9):1095–1098 Zhou D (1981) Study of oral diseases and other aspects of the ancient corpse of the Western Han dynasty unearthed from tomb no. 168 on Phoenix Hill at Jiangling County. Nihonshikaishigakkaishi 8(4):38–41 Zhou D, Zhou Y, Yang M (1999) De’an Nan Song Zhou Shi Mu. In: Nanchang: Jiangxi ren min chu ban she. Jiangxi People Publishing House, Jiangxi

Part IX Cultural Aspects of Mummy Studies

Fake and Alien Mummies

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Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Historical Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Extraterrestrial Life and Forged Remains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Ata, the Atacama Alien . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Nazca Alien Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Scientists and the Alien Mummies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusions and Recommendations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Mummies’ popularity and attractiveness is a well-established fact for all museums that have placed them on display. Public fascination is rooted in the apparently counter natural avoidance of decay these bodies have achieved in some unfathomable ways. Nevertheless, this morbid attraction has also fueled the creativity of some people in wrong ways, probably from time immemorial. In recent academic memory, though, it is known that purported mermaids and makeshift Egyptian mummies, for instance, toured Europe and North America from the mid-1800s to the early 1900s, as part of circuses’ side and freakshows. Although most of visitors saw them tongue-in-cheek, their continuity was guaranteed by their display being profitable. It was education and respect of

G. Lombardi (*) Cátedra Pedro Weiss, Universidad Peruana Cayetano Heredia, Lima, Perú C. Rodríguez Martín Instituto Canario de Bioantropología and Museo Arqueológico de Tenerife, OAMC-Cabildo de Tenerife, Santa Cruz de Tenerife, Spain e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_36

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fellow humans’ demise, which slowly phased out these once prevalent elements of popular culture. The academic world has not been spared by the manipulation of human remains, in order to escalate positions in an ill manner, or as an easy way to obtain quick fame, popularity, and even income. Though the Piltdown skull case stands as the foremost from a historical perspective, sadly, it is far from standing alone. Recent on-line and television appearance of purportedly extraterrestrial remains excavated in unknown circumstances in the Southern Peruvian coast has revealed itself as the end product of a long process which joins grave looting, yellow press, and some disoriented professionals. Organizers and attendees to the IX World Congress on Mummy Studies held in Lima, Peru, faced a disinformation campaign by presenting a declaration preventing the public and the media of this hoax. Since then, the producers of this scam program have kept, unfortunately, trying to validate their claims by using fallacious strategies, fueled by the revenue of selling the show. Forgeries are obscure in their own nature. Scammers purposefully hide the true origin of their products. This has been the case of these false alien mummies supposedly found near Nazca by grave looters. The location remains in secrecy, the discoverer only accessible to the production ring; and any attempt to oppose their plot faces attacks and more deception build up on, namely, conspiracy theories. In the time of artificial intelligence development to unprecedented levels, and of deep fake creations quickly distributed online, we could only foresee newer attempts to not only fool the media and the public, but also academia. The main aim of this chapter is to prevent this from happening, through exposure and education. Keywords

Mummies · History · Hoaxes · Alien Mummies

Introduction Although the title of this chapter may suggest the contrary, most fake mummies are indeed composed of mummified bodies or body parts rather than solely handicraft or other inert materials. Therefore, a common element these mummies share is that they have been processed and assembled, in modern times, to mislead the public. In sharp contrast, museum mummy replicas are not real either, but are both made to protect delicate original corpses, as well as to educate the public. Therefore, the definition of this category focuses more on a deceptive intentionality behind their production rather than on the materials used to compose each particular item. It is also important to take into account that there are ancient assemblages that, at first sight, could seem to fit into this category, but for very different reasons, such as symbolic burials or hasted reconstructions made by grave robbers.

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Bioarchaeological studies are intrinsically complex and multidisciplinary. Nevertheless, the dealing with forgeries adds up other elements not unfamiliar to forensics, such as criminal psychology, police research, as well as press coverage and science ethics. It is considering this palindromic complexity that this chapter unfolds, attempting to provide a panoramic view of a problem that seems to be growing.

Historical Background Without a doubt, hoaxes and deceit are probably as old as civilization. Ancient oracles in different cultures had special middle people who translated the wills and decisions purportedly communicated in hidden ways by supernatural deities, unreachable to commoners. No wonder among successive ancient religions, new ones have usually downgraded the preceding as mythology, and any revelation of out-of-the-limb facts that contradicted the official set of beliefs faced scorn, punishment, even death; as happened in the well-known cases of Galileo and Hypatia. Though the chasm between current religions and science is still there; fortunately, nothing that serious happens nowadays due to discrepancies. A respectful truce prevails. Both the Enlightenment and the Reform fell mental and physical walls, paving the ways for the development of individual freedoms, among which, learning about the others drove travelers and explorers to discover new lands. Consequently, naturalistic knowledge grew and fueled literature and other arts. Public interest in seeing and even collecting antiquities, including human remains, stemmed out of this trend, epitomized by the Egyptomania that seized Europe in the early nineteenth century (Brier 2013). Genuine interest focused on traveling to the land of the pharaohs or visiting the growing collections of the soon-to-become greatest museums in the world. But, alas, a black market quickly formed, and also an underground craft for falsifying valuable exotic objects did, too (Riggs 2016). After some time, true trade networks were formed, skillfully linking greedy grave looters, callous middlemen, and an uneducated fraction of the public avid for an easy way to materially based prestige. Over time, these parties have remained, and though their deeds keep unseen, the cases and variety of their crimes have just gotten more complex (Sotiriou 2018). Apparently, soon after the appearance of this illegal activity, a really shocking twist occurred when black-market sellers faced scarcity of goods at the source: good quality mummies were just running out in Egypt. Looting, of course, intensified, but also mimicking the real products made its debut. Fake Egyptian mummies entered the market, with little to do to detect or deter their presence. Body snatching had occurred in the western world before, fueled by an intensified need for medical schools’ dissection cadavers; this newer form was just an extension of that independently developed craft (Ross and Ross 1979). Counterfeit mummies and forged extraordinary discoveries have not spared anthropology, being the purported Piltdown man’s skull the most famous example

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of such deceptions, again fueled by a desire to festinate prestige and academic progression (Fig. 1). On a more lucrative end, touring nineteenth-century circus sideshows, such as P. T. Barnum’s, prompted the display of falsified objects such as, for instance, mermaid mummies and petrified people (Williams 2017; Fig. 2). Fig. 1 “Piltdown man” skull cast. Use of photography available without restriction from https://commons.m. wikimedia.org/wiki/File: Sterkfontein_Caves_1. jpg#mw-jump-to-license

Fig. 2 Fake mermaid from the Moses Kimball collection. Use of photography permitted without restriction from https://commons.m.wikimedia.org/wiki/File:Feejee_Mermaid,_shown_in_P.T._ Barnum%27s_American_Museum,_1842,_as_leased_from_Moses_Kimball_of_the_Boston_ Museum,_papier-mache_-_Peabody_Museum,_Harvard_University_-_DSC06154.jpg

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In the year 2000, the alleged discovery of a Persian mummy in Pakistan attracted public attention once again. Though researchers reacted very cautiously, as Egyptian-style mummification is absent from local archaeological records, intrigue was sowed and fueled by early claims for an authentic Persian royal, which, in the end, proved wrong. Nevertheless, this case was unique in its degree of sophistication: its makers most probably had professional training or support to perform autopsies and even to assemble a complete but fake archaeological context to lever validity to the find. Fortunately, the scaffolding crumbled down and the hoax became fully exposed as a complex criminal case involving a lively black market for antiquities. Moreover, the case involved a contemporary person whose body, irregularly obtained for the purpose of this forgery, could only be properly buried about 12 years after her demise (Romey and Rose 2001). Although the exact size of the antiquities black market is difficult to obtain, estimates consider it second only to the international illegal drugs’ traffic. Therefore, a mostly invisible but important fake mummies’ trade must be active, and the aforementioned case could just be the tip of the iceberg. It is disheartening to consider missing people might have actually ended up entangled in such criminal networks. From a historical perspective, two periods in counterfeit mummy production could be devised. The first one focuses on Egyptian-looking forgeries which were at its prime in the nineteenth and early twentieth centuries. The second is still a relatively new, ongoing phenomenon, which aims at creating ancient extraterrestriallooking remains. The fake Persian mummy represents a rare contemporary example belonging to the first period. The remaining part of this chapter focuses more on the second period, mostly a South American issue that has not been fully addressed by academia and which represents a threat to science as a whole. Another contemporary and important case of mummy forgery comes from Germany. In 2013, an ancient Egyptian looking assemblage – the Diepholz Mummy – was discovered; though authenticity was promptly dispelled for accompanying objects, the body required further analyses. Understandably, the mysterious mummy captured mediatic attention, but there was a novel twist: an early claim for the body to be an animal-human hybrid created by aliens (Von Schade et al. 2013). Later research confirmed the mummy’s skeleton was made up of plastic bones, being the skull the only authentic, though of unknown origin, human part.

Extraterrestrial Life and Forged Remains Very few things in the history of science would probably draw more attention than the irrefutable demonstration of the existence of extraterrestrial life, or even better, the demonstration of the presence of such organisms in earth’s geological or archaeological record. Very disappointingly, not a single serious report supports possible candidates for such claims so far. Experimentation has revealed that the molecular basis of life, as we know it, most probably developed in warm fresh water ponds on a primeval earth. Moreover, the discovery of earth-like exoplanets suggests

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this process could have succeeded many times in the Universe. Despite these optimistic data, all serious efforts directed to detect ET civilizations’ radio signals from outer space, remain futile. Even much publicized UFO footage released by the Pentagon seems very inconclusive (West 2020; Yuhas 2020). In addition, the evolutionary model of life on our planet seems sound and logical, with species competing and adapting to local, variable constraints and niches, in an almost seamless perfection for millions of years. Therefore, it would make it very easy to spot any outsider to the system. Cladistic distribution of life is consistent up to molecular level, not only genomic but also proteomic and even at the level of the coevolution of bacteria and parasitic biomes. Apparent exceptions to the cell theory, such as viruses and prions, represent nevertheless mere shortcuts to the genetic paradigm. Ever since the primordial soup, life on Earth reveals itself as a welltuned orchestra playing the most beautiful symphony to perfection. Yet, over the last few years, and perhaps connecting antiquities’ black-market networks and media moguls eager to sellout the discovery of alien life, a notorious claim has appeared involving South America: the alleged discovery of a set of ancient extraterrestrial remains. The sudden rise of a corpus of such bodies, produced through real but manipulated archaeological remains, came as no surprise. The region has witnessed, on foreign yellow-press TV, a chronic denial for native cultures’ most distinct features and achievements. Some productions still present Inca megaliths as impossible to be assembled by human means; also, they have proposed an extraterrestrial or hybrid origin for the people who practiced skull remodeling, despite all archaeologic and ethnographic evidence supporting the development of this trait as a cultural product. Forgeries involving pseudo archaeological remains are not new in this part of the world either, being the best-known cases the Ica engraved stones, the fake gold tumi knives, and uncountable false pieces of pottery (Polidoro 2002). In most of these cases, the hoaxers’ ultimate goal was to sell their products to inexpert tourists and collectors. In the case of the engraved stones, promotion of a private museum and festinated prestige were also evident objectives. Although the cases that have been more publicized involve Ata, the Atacama alien and the Nazca alien mummies, there is no doubt many more pseudo alien bodies have been produced and traded. South America appears ideal to take forgeries of this kind to another, almost industrial level for two reasons. First, the continent holds, as mentioned in this book (South American Mummies), the largest collection of naturally mummified bodies, and second, grave looting and corruption are more prevalent and even more socially accepted there than in other continents.

Ata, the Atacama Alien Over 20 years ago, European tourists were offered a very small body in Northern Chile, resembling an alien-looking creature, stretched in a pen set box (Fig. 3). Following the purchase, the new owners, decided to verify their supposedly groundbreaking discovery. Unfortunately for them, but understandably, very few academics

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Fig. 3 Fetus-turned-‘alien’ mummy (Credit: Marco Rodas)

took them seriously. It was in 2007, over the VI World Congress on Mummy Studies held in Lanzarote, that, upon peer pressure, one of us (G.L.) had the chance of looking closely to the creature. Its holder had been looking for wider attention, perhaps dreaming of using the conference’s support as a springboard to global attention; despite his wandering, no one had accepted. Once carefully observed through a magnifying glass, a verbal report was offered to the owner: it was a dehydrated human fetus, stretched and modified, so to look like what their sellers intended it to represent. No doubt its realization required great artisan’s skill, but at the same time, that kind of business, with a human cadaver, was and is a real crime. Moreover, the owner was assured that if DNA research would be done on the fetus, its parents could probably be located, and perhaps prosecuted. Despite the owner disliking the assessment, academic honesty is simply non-negotiable. Later, on-line references to the mummy were found, which by then had been given the moniker of Ata. There were many publications in ufology pages and numerous but suspicious comments celebrating and supporting the revolutionary find. The ill-fated Chilean fetus had become an Internet character. Later on, it was verified that the creature was indeed human, but carried genetic anomalies, as many miscarriages do (Bhattacharya et al. 2018; Nolan and Butte 2018; Hayden 2018).

Nazca Alien Mummies Since 2017, mystery-themed websites and local tabloids started to cover the apparent discovery of a sizable set of alien-looking remains in Peru, with the participation of North American media producers. In contrast to full exposure of the alleged finds,

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the origin of the set remained elusive. Released information emphasized images, even radiographs, but avoided providing data on archaeological project, site, or paraphernalia associated. When enquired, the producers conceded the discovery was made by grave looters in the Nazca area and justified not exposing site details, in order to protect it from those against “the truth being revealed.” Available imagery shows remains that could be divided into two groups: the first one is composed by about a dozen fully stretched, slim plaster figurines, ranging on children’s sizes, with big heads and large eye sockets, all of them sporting three equally stretched fingers and toes. X-ray plates revealed crude anatomical inconsistencies, such as the use of carved animal skulls or the addition of upside-down phalanges, and even child’s long bones to elongate reassembled fingers and toes. The second group, to the trained eye, was unmistakable: they were pre-Columbian Andean mummies, naturally dehydrated and flexed in their typical sitting position, but totally divested from their funerary bundles, elaborate multilayered fabric cocoons that both helped to protect the bodies and, typically, left imprints in their skin. This second group shared with the first two characteristics: First, their white coloration, as if they had been caulked with a plasterlooking substance – released information by the producers later conceded the substance was diatomite, a fossil white sand readily available in the Nazca area. Second, these mummies also presented three fingers and toes in their hands and feet. They also shared large eye sockets and lacked ears. Observing these preColumbian Nazca dwellers could not be sadder. There was no doubt: these people’s remains had been mutilated. The set of remains presented above, therefore, comes indeed from Nazca, a region famous for its namesake pre-Columbian culture, celebrate desert lines, and Maria Reiche’s studies on them. But the area is, unfortunately, also known for insidious grave looting carried out for generations and, more recently, as background setting for ancient alien theories. Therefore, discovering alien corpses in Nazca would fill the gap, as ultimate proof to dispel all incredulity, particularly by academia. A set of elements was just there in Southern Peru to be profited by. The latest twist merely represents, therefore, grave looting with an added value (Fig. 4).

Scientists and the Alien Mummies Right after the 9th World Congress on Mummy Studies, held in Lima, Peru, in 2017, a press conference was called by associates of a North American TV production company to release the news of the discovery of a set of extraterrestrial mummies in Nazca. Several researchers, including the organizers of the 9WCMS, after evaluating all evidence available (mostly on-line photographs, including X-ray plates), decided to prevent the public from the hoax by publishing on-line documents, such as the Momia Nazca series at cientificos.pe and the following:

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Fig. 4 A desecrated Nazca mummy (Credit: Steve Mera)

DECLARATION OF THE SCIENTIFIC COMMUNITY REGARDING THE FRAUD OF EXTRATERRESTRIAL MUMMIES The subscribers of this document, members of the national and international scientific community, experts in the study and conservation of human remains (mummies and skeletons), communicate the following: 1.- In the last few months, the alleged discovery of “extraterrestrial mummies” in our country has been publicized through an irresponsible, organized disinformation campaign. 2.- Testimonies and published images on this case, allow to assert that these findings correspond undoubtedly to pre-Columbian human remains – Cultural Patrimony of the Nation –, maliciously manipulated and even mutilated to obtain an “ad hoc” appearance for commercial exploitation. Moreover, the exclusion of the entire related archaeological contexts is absolutely contrary to the scientific investigation of this kind of cultural finds. 3.- It is upon our authorities to make the corresponding accusations, since this “production” has violated numerous national and international norms that watch for the defense of Cultural Heritage. We hope that these same authorities will seek the protection and correct investigation of these remains and their place of origin, in order to stop looting and trafficking of human remains. We also expect exemplary sanctions for those responsible of this depredation of heritage that is of all Peruvians and Humanity as a whole. 4.- Finally, the criminal abuse of corpses for petty ends violates human dignity in a profound way. Thus, exploitation of pre-Columbian mummies carried out by this organization, attacks and particularly offends the Andean Culture, implying that its achievements were due to an alleged “alien aid”. We offer our best offices to collaborate with the authorities to demonstrate our statements in the corresponding instances. We also offer to participate in activities that would defend our patrimony and help educate the public about our ancestors and their legacy.

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Lima, July 10, 2017. Sonia Guillén, Guido Lombardi, Elsa Tomasto-Cagigao, María del Carmen Vega, Mellisa Lund, Patricia Maita, Martha Palma, Carlos Herz, Alejandra Valverde, Marcela Urizar; more signatures and accessions follow.

The result of this declaration was very positive, so the fraud became limited to the group of subscribers of the production (Heaney 2017). Nevertheless, the last few years have revealed new strategies, encompassing the sad involvement of trained researchers supporting the forgers’ claim. Unfortunately, biased reports, based on financial sources’ orientation, are not foreign to scientific publications.

Discussion Some South American landmarks, such as Machu Picchu, are often presented as being built by aliens or with their help. Despite serious research demonstrate otherwise, some public seem to prefer fantastic, otherworldly options. A subset of popular media producers has profited on this public’s preference, following on the pioneering steps of bestseller science-fiction writer Erich von Däniken. Perhaps entrusted by the level of perfection reached by moviemaking 3D imaging, special effects, and the application of AI technology, these producers, apparently, are daring to go for the ultimate deception: to fool science itself. Obtaining official backing, or merely getting partial support by accepting a reasonable doubt, would give them access to a long-cherished mankind goal, proving contact with alien civilizations, even from prehistoric times. Excellent TV programs and movies have always honestly profited of the fictional extraterrestrial theme, and some of them have become part of the fondest films ever made, such as Steven Spielberg’s Close Encounters or E.T., The Extraterrestrial. On the other end of the fiction continuum, mockumentaries, which could affect the sensitivity of some unaware viewers, are less damaging than productions thoroughly aimed at fooling the public for a revenue (Wallace 2019). Though freedom of choice protects scammers, in the aforementioned cases, the situation of this well-organized and planned scam is more complex and goes beyond a marketing matter. The use and abuse of both animal and human remains – including well-preserved Nazca human mummies – have been proven by our careful observation of public images of the Nazca Alien Mummies and through research done on a few samples that reached, through a voluntary donation by a ring’s regretful member to the local police and studied by forensic archaeology expert Flavio Estrada. As noticed, the scammers’ circle has kept access of the remains to themselves, disregarding the experts’ committee offer to study them if surrendered to the authorities. Complete assessment of the manipulated anatomy of the constructs or dummies reveals the use of dog and probably llama skulls turned around so that pseudo-orbits have been carved in their occipital bones, use of a mixture of hand and feet phalanges to lengthen the purported tridactyl fingers. In some cases, these phalanges were glued

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together – with an instant synthetic product and a mixture similar to papier maché – in wrong anatomical positions (Salas–Gismondi 2017) (Fig. 5). These cases fit to to a pattern: (a) Mostly human and/or animal remains, manipulated in order to resemble popular extraterrestrial icons. (b) An initial set of claims about extraordinary discoveries made by ufologists and allied researchers on non-scientific online media. (c) On-line addicted ufology sites inundated by – most probably fake – reviews that provide bogus validation of the claims and boldly rejects any counterargument. (d) Avoidance of independent assessment. Beside economic support provided by illegal trade networks, press allies grant news coverage, offer positive feedback to the find claims, and cast doubt on science in general and rebuts any criticism as part of a conspiration. Science is open to scrutinize any claims, always. The scientific method guarantees absolute transparency, yet its pace could be too slow or its ways not as picturesque as some media would prefer. Nevertheless, some minimum criteria should be met. In the case of the aforementioned manipulated fetus, a serious police investigation should determine the origin of this human cadaver. Trading or diluting

Fig. 5 Incoherent anatomy of purported alien “tridactyl mummy” (Momia Nazca on cientificos.pe) (Credit: Rodolfo Salas-Gismondi)

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the original crime behind ET arguments is plainly wrong. Science should never accept any wrongdoings. Besides the daring anatomical inconsistencies, there are several missing elements that the producers of this hoax just decided to bypass: the study of the archaeological context and paraphernalia of the bodies found. The adamant neglect to follow the archaeological method, applicable even in the case of fortuitous finds by lay people, is very revealing. Most of the assembly appears covered by a coat of dusty white diatomite powder which is otherwise inexistent in the Peruvian archaeological record. Nevertheless, despite its supposedly ancient age, the coat is perfectly clean, and as seen on images posted online by the producers, it is detaching very easily, revealing the true dark color beneath, characteristic of Andean mummies. Moreover, over some protruding parts of the bodies, such as the knees, imprints from the original textiles wrapping the sitting cadavers, are visible. Where are the textiles? The inconsistencies and fabrications of this assembly are just grotesque. Besides mutilations and manipulations, another novel level of abuse should not be disregarded, as the whole development of the Andean culture is placed under a question mark and belittled; its achievements all of the sudden denied by a hypothetical participation of a somehow superior alien race (Heaney 2017). Finally, to dare to remove the condition of human beings to perfect human remains by altering them, even by mutilation – cutting their ears, fingers, and toes, reshaping the orbits – to produce tridactyl creatures, constitutes a crime of the vilest nature. The confabulated production ring has apparently even dared to request from the artisans to implant stones resembling eggs into the pelvic area of a female construct: probably true fossil eggs smuggled out of China and readily available online, so to assert the creature was a pregnant reptilian alien! Unfortunately, such possibilities are not impossible to consider since there is a well-established international black market for cultural and particularly archaeological objects, real or not, with links to wider and more dangerous organized crime networks (Sotiriou 2018). With these forgeries, pseudoscience has, no doubt, reached new lows.

Conclusions and Recommendations Other long-held forgeries, not involving human remains, have also been very common over time, and some have appropriately been debunked after decades of publicized deception. Among them, it is worth remembering the Loch Ness monster, the crop circles, and the ancient crystal skulls (Everts 2013). Again, science-fiction is acceptable and inspiring; many accomplishments in science have been preceded by artistic dreams in different branches of the arts. Nonetheless, con-artists just deter legality, missing the chance of orienting their skills in a fruitful, honest manner. Their lack of ethics, expressed by their systematic disrespect of scientific ways – not following the scientific method and local laws – favoring the use and abuse of looted archaeological items, merely supports their true nature and their ultimate publicityseeking objective.

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Nevertheless, there is also responsibility on the academic side of this problem. Researchers focus on their work and follow, in many cases, tenure-oriented careers which make them look down at popularizing their findings. This system has created a gap between the holders of advanced scientific knowledge and the general public. Sadly, sometimes daring to reach out to the media has prompted, in fact, punishment from university officials (Martinez–Conde 2016). In the same line of the aforementioned declaration by mummy studies’ experts in Lima, it would be advisable to incorporate the problem of forgeries and deception to the academic discussion, so as to perhaps define steps to orderly follow in cases such as those mentioned in this chapter. Our failure to face this issue appropriately could reinforce ill-oriented activities, and even inadvertently, keeping silent could be erroneously interpreted as tacit permissiveness. It is clear that the scientific method should always be followed, along with compliance with local laws and respect for the deceased. In this regard, it is important to consider the Vermillion Accord as a reference to foster respect for any handling of human remains, even in the case of a dubious human affiliation. As mentioned before, for different reasons, some disoriented professionals end up serving hoaxers’ activities. In these delicate cases, an appeal should be made for each individual to stick to the codes of ethics of each profession, as well as to maintain moral integrity. Breaches of codes of ethics should be handled internally by each profession’s guild according to its own parameters and guidelines. The certain existence of life in other planets or elsewhere in the Universe is not at stake here; honesty and respect for all humans, our ancestors, and their legacy is. Justice is slow everywhere and Peru is no exception; we hope that, in the end, sensibility and decency will prevail, along with setting jurisprudence over cases like these. These altered Nazca mummies and manipulated skeletons will certainly end up enriching Peru’s cultural patrimony, at the Nacional Police Museum as the most extravagant and shameful scam involving human remains in history.

Cross-References ▶ Showing Respect to the Dead: The Ethics of Studying, Displaying, and Repatriating Mummified Human Remains ▶ South American Mummies

References Bhattacharya S, Li J, Sockell A, Kan M, Bava F, Chen SC, Ávila-Arcos M, Ji X, Smith E, Asadi NB, Lachman R, Lam H, Bustamante C, Butte A, Nolan G (2018) Whole-genome sequencing of Atacama skeleton shows novel mutations linked with dysplasia. Genome Res 28:423–431 Brier B (2013) Egyptomania: our three-thousand-year obsession with the land of the pharaohs. St. Martin’s Publishing Group, New York, NY Dias N (2021) Human remains in the history of archaelogy. In: Oxford handbook of the history of archaeology. Oxford University Press, Oxford, UK

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Everts S (2013) Crystal Skulls Deemed Fake. Chem Eng News 91(9):48–49 Hayden E (2018) Tiny Mummy’s ‘alien’ appearance finally explained. National Geographic, March 22 Heaney C (2017) The racism behind alien mummy hoaxes: pre-Columbian bodies are once again being used as evidence for extraterrestrial life. The Atlantic, August 1st Martinez–Conde S (2016) Has contemporary academia outgrown the Carl Sagan effect? J Neurosci 36(7):2077–2082 Nolan G, Butte A (2018) The Atacama skeleton. Genome Res 28:607–608 Polidoro M (2002) Ica stones: Yabba-Dabba-Do! Skeptical Enquirer 26(5):24–25 Riggs C (2016) Beautiful burials, beautiful skulls: the aesthetics of the Egyptian mummy. Br J Aesthet 56(3):247–263 Romey KM, Rose M (2001) Special report: Saga of the Persian princess. Archaeology 54(1):24–25 Ross I, Ross CU (1979) Body snatching in nineteenth century Britain: from exhumation to murder. Br J Law Soc 6(1):108–118 Salas–Gismondi R (2017) Momia de Nasca, parte 2: Análisis de las manos y pies. http://www. cientificos.pe/index.php/2017/06/28/momia-de-nasca-parte-2-analisis-manos-y-pies/ Sotiriou KO (2018) The F words: frauds, forgeries, and fakes in antiquities smuggling and the role of organized crime. Int J Cult Prop 25(2):223–236 Von Schade M, Wolf K, Schäfer C, Winterstein T (2013) Die Kinder-Mumie vom Dachboden. Erich von Däniken sicher: “Es ist ein Mischwesen”. Bild. 5 August Wallace P (2019) Phony facts and eco-media: fake nature and the call for widespread media literacy. J Environ Commun 13(6):790–803 West M (2020) Explained: new navy UFO videos. https://www.youtube.com/watch? v¼Q7jcBGLIpus&feature¼youtu.be Williams J (2017) A new American freak show. In: Media, performative identity, and the new American freak show. Palgrave Macmillan, London, UK Yuhas A (2020) The Pentagon released U.F.O. videos. Don’t hold your breath for a breakthrough. New York Times, April 28. https://www.nytimes.com/2020/04/28/us/pentagon-ufo-videos.html

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Dong Hoon Shin, Chang Seok Oh, Jong Ha Hong, and Mi Kyung Song

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummies and Clothing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A Mummy Bundle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Interdisciplinary Collaboration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mummy’s Teeth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . An Example of Mummy Clothing Research in South Korea . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Cross-References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

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Very well-preserved clothing and textiles often have been discovered in East Asian mummy studies. Despite their significant academic value, however, fullfledged scientific studies on mummy clothing began just recently in South Korea. D. H. Shin (*) Laboratory of Bioanthropology, Paleopathology and History of Diseases, Department of Anatomy and Cell Biology, Institute of Forensic and Anthropological Science, Seoul National University College of Medicine, Seoul, South Korea C. S. Oh Department of Mortuary Science, College of Health Industry, Eulji University, Seongnam, Gyeonggi-do, South Korea e-mail: [email protected] J. H. Hong Department of History (Archaeology), Institute of Korean Archaeology and Ancient History, Kyung Hee University, Seoul, South Korea e-mail: [email protected] M. K. Song (*) Department of Fashion Design and Marketing, Seoul Women’s University, Seoul, South Korea e-mail: [email protected] © Springer Nature Singapore Pte Ltd. 2021 D. H. Shin, R. Bianucci (eds.), The Handbook of Mummy Studies, https://doi.org/10.1007/978-981-15-3354-9_37

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Thanks to the pioneering work of dress historians, clothing collected from Joseon period graves has grown into a large historical repository at institutions such as museums. Utilizing the mummy clothing collected so far, scholars have traced historical changes in clothing fashions on the Korean peninsula. Every finding obtained by mummy clothing research becomes an invaluable resource for scholarly reconstruction of pre-modern dress and textiles. Given the recent technological development in the field, long-asked questions about actual clothing history could finally be answered, thus bringing about a great deal of progress in knowledge. In this light, the academic value of mummy clothing has come to be highly re-appreciated among academics and the public at large in South Korea. Keywords

Korea · East Asia · Clothing · Textile · Mummy · Joseon period

Introduction Archaeologists have discovered a number of mummy cases that surrendered many cultural artifacts. They are an essential resource of academic research into various aspects of premodern societies and peoples worldwide. As in other places where the mummies have been discovered, East Asian scholars also have carried out collaborative work in cases where well-preserved mummies and other cultural remains were discovered together in the archaeological field. By their studies on artifacts and mummified remains inside graves, research groups in East Asia could catch vivid glimpses of ancient or medieval peoples’ lives (Lee et al. 2009a, 2013, 2014; Seo et al. 2014, 2017; Song and Shin 2014; Arguelles et al. 2015; Oh et al. 2015, 2017, 2018; Shin et al. 2018). Among cultural remains, well-preserved textiles or clothing is one of the most important subjects of East Asian mummy research. Actually, many well-preserved articles of clothing have been collected and examined by interdisciplinary working groups. This holds particularly true for archaeological investigations all over the world. For example, mummies of ancient Peru, which had been subjected to a thorough embalming process entailing the removal of internal organs and delicate treatment of the skin (Ostolaza 2009), were wrapped in a shroud or simple clothing to form a funerary bundle. Those cultural monuments could be invaluable subjects of research for dress historians. Actually, during the last century, data gleaned from PreColumbian textiles and mummies has been among the most remarkable achievements of South American archaeology (Ostolaza 2009; Doig 2013). By studies on textiles or clothing, researchers were able to identify detailed aspects of changes in ancient Peruvian society (Doig 2013). Pioneering studies on mummy textiles or clothing are achievements not only of New World countries. In Old-World regions too, such as East Asia, dress historians have conducted serial research on mummy clothing discovered in the archaeological field. The most notable examples of such countries are South Korea and China. Over a wide swath of the Korean peninsula, most of the mummies examined have been

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found in several-hundred-year-old Joseon dynasty (1392–1910 CE) graves. Actually, such discoveries initially surprised academics and the public alike, because no embalming techniques as practiced at that time were known (Song and Shin 2014). In South Korea alone, premodern clothing has been collected in approximately 100 or more archaeological cases (Lee et al. 2013; Song and Shin 2014).

Mummies and Clothing Many articles of clothing and other textiles were buried in Joseon period tombs. Indeed, in the course of archaeological investigations, it has not been rare to find such articles in very well-preserved states of preservation. Despite the obvious academic value of mummy clothing, research of the history of dress did not begin in South Korea until 1968, and scientific studies on mummies did not start in earnest until the 2000s. Therefore, we do not know how many Korean mummies and clothing were lost to decomposition without proper academic investigations of Joseon graves. Fortunately though, reports of the discovery of mummies and clothing rapidly increased in recent years because a law requiring archaeological surveying prior to construction projects was enacted (Lee et al. 2013). Pioneering dress historians, utilizing clothing collected from Joseon graves, traced changes occurring in clothing fashions during the past several centuries (Song and Shin 2014). Their work did not proceed wholly without difficulty, however. In the earliest days of mummy clothing research, in cases where perfectly preserved graves and mummies were discovered, descendants seriously curtailed or outright denied access to their ancestors or their clothing (Oh et al. 2018). Even in cases where researchers received permission, it was often granted only after considerable delay, and their investigation in many cases had to be wrapped up in a hurry so that mummified remains could be reburied as soon as possible. Nonetheless, as time passed and social attitudes changed in South Korean society, dress historians were able to collect and gain access to their research materials more readily than before. Also, whereas earlier research had been conducted in the open excavation field, concern about contamination of archaeological samples therefrom (Hermann and Hummel 1994; Melchior et al. 2008; Roberts and Ingham 2008) led to the recognition that sampling should be done under contamination-free conditions in well-equipped lab facilities (Hofreiter et al. 2001; Willerslev and Cooper 2005; Lee et al. 2013). It was acknowledged that every researcher in the lab should wear sterilized gowns, head caps, gloves, and masks (Fig. 1), and that the examination tools employed must always be sterilized before use (or that disposable ones be used instead) to prevent modern contamination. To date, in case of South Korea, the articles of clothing collected by dress historians have grown into large collections. Articles numbering 10,000 or more are currently maintained in institutions and museums throughout the country (Fig. 2). Since every finding obtained came to be documented and classified in detail, the collection has become an invaluable resource for a scholarly reconstruction of dress history in South Korea that otherwise might never have been possible (Song and Shin 2014). By their research, dress historians have isolated changes in

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Fig. 1 Dress historians and mummy clothing (Credit: Dong Hoon Shin)

Fig. 2 Mummy clothing maintained in museum (Courtesy of Korea University Museum)

clothing patterns throughout the historical timeline. They also have gained access to reliable information on the funeral rites of the Joseon period. As long-asked questions about actual clothing history began finally to be answered, the academic value of mummy clothing rose steeply in the estimations of both academia and society in East Asia (Lee et al. 2009a, 2013; Song and Shin 2014).

A Mummy Bundle When mummies have been discovered in East Asia, whether South Korea or China, they often are swaddled in a mummy bundle. The mummy bundle generally was composed of a mummified individual, his (her) clothing, and other cultural remains. To dress historians, this is very important because many pieces of clothing, such as jackets, undershirts, trousers, and sashes, were packed together in the large bundle. Unraveling this bundled object is an important moment not only for dress historians

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but also for other relevant scholars, because it includes so much academic information representative of the era of the mummified person and his (her) society (Lee et al. 2009a). In the earliest days of mummy research, specifically in the 1840s for Egyptian mummies, bandage removal was performed even in front of a large audience. At the time, clothing removal was not necessarily done for academic purposes, but rather as a kind of entertainment for the audience (Pringle 2001). Well-designed academic research on mummy clothing has been carried out only recently. As mentioned above, there have been many outstanding studies, especially on the famous South American mummies. For instance, careful and deliberate, scientific removal of mummy clothing from a mummy bundle was performed on a 1600-year-old, elitestatus Peruvian female mummy reposed in full regalia (Williams 2006; Lee et al. 2013). The information obtained during the process proved invaluable to dress historians around the world (Williams 2006). Over the past decade, in South Korea, unraveling of mummy bundles has been reported much more frequently and in more detail than in other mummy cases (Fig. 3). In general, the amount of clothing collected from a mummy bundle is enormous. As many as 200 pieces of clothing were discovered in the case of one well-preserved upper-class Korean grave (Koh 2006). Table 1 summarizes the mummy bundles academically investigated by dress historians in South Korea thus far. Based on the knowledge accumulated so far, the general arrangement of clothing inside of mummy bundle can be revealed. Briefly, at the initial stage of unraveling, various articles of space-filling clothing are found. These garments were intended to keep the mummy bundle from moving around inside the coffin. The outermost part of the mummy bundle was wrapped in broad textiles and tied with hemp ropes. When the wrapping textiles are removed, dress historians typically find that many pieces of clothing were stacked inside the mummy bundle (Fig. 4). A shrouded body (mostly mummified) finally is exposed after the removal of stacked clothing and fabrics. Many coats, jackets, undershirts, drawers, and skirts were used as shrouds. Shoes, socks, and other garments also are found inside a mummy bundle (Fig. 5).

Fig. 3 Mummy bundle of South Korea (Credit: Dong Hoon Shin)

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Table 1 Dismantling cases of mummy bundles in South Korea Mummy ID Yangju (Child) Papyung Yoon Yongin Hadong 1 SN1-2 SN3-7-1 SN2-19-1 SN2-19-2 GJ1-2 Gangneung SN PK Dangjin Seocheon Waegwan PJ SM Hadong 2 (HD-2) Mungyeong Jinju Hongsung Sacheon Sapgyo Hwasung YG2-4 YG2-6 Andong Dalsung Junggye Daegu-HS Cheongdo Yeongweol Jangseong Gumi Changwon Gyeongsan Konkuk Seoul

Date of excavation 2001 2002 2005 2006 2007 2007 2007 2007 2007 2007 2007 2008 2008 2008 2008 2009 2010 2010 2011 2011 2011 2012 2012 2012 2013 2014 2014 2014 2014 2014 2017 2019 2019 2019 ? ?

Mummy bundle dismantling X O X X O X X X O O O O O O X O X O O O O O O O O O O O O O O  O O X X

 means dismantling stopped unfinished

A veil covered the mummy’s face. A detailed example of clothing removal from Korean mummies is summarized at the end of this chapter. According to dress historians, the clothing found inside the mummy bundle was not randomly inserted, but instead, had been orderly arranged according to a certain principle. Dress historians have argued that such uniformity might have been due to

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Fig. 4 Inside of mummy bundle (Credit: Dong Hoon Shin)

Fig. 5 Garments found inside a mummy bundle (Credit: Dong Hoon Shin)

the very strong control of Joseon society over funerals. Actually, ritual guidebooks (Garye books) that were highly respected and popular among Joseon people must have been related to bundling uniformity. The Garye books have a special meaning to dress historians, because the funeral ceremony of the Joseon period, including the arrangement of clothing in the mummy bundle, was performed in a very deliberate and regulated way according to their recommendations (Lee et al. 2013). This means that whenever dress historians investigate a mummy bundle, they often discover clothing and textiles that were packed and layered in a very formalized way that was generally accepted by the people at that time.

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Interdisciplinary Collaboration When researchers in each specialty independently conduct their own investigations on mummies, they often face the difficult situation of cultural or human remains being co-mingled inside the coffin (Lee et al. 2013). In those cases, archaeologists need dress historians’ help so as not to incur damage to cultural remains during their investigation. In brief, anthropologists’ analyses could not proceed smoothly without dress historians’ involvement, and vice versa. We note that in the case of Joseon tombs, perfect sealing of the coffin could be maintained over the long period until final excavation. And since the mummified individual found inside the mummy bundle normally is heavily clad, indeed sealed, in clothing and textiles, their careful removal by dress historians under sterilized conditions can ensure that contamination continues to be kept to the absolute minimum (Lee et al. 2007, 2009a, b, 2013; Kim et al. 2008; Lim et al. 2008; Seo et al. 2008). Another advantage that archaeological scientists can gain from collaboration with dress historians is the latter’s data obtained during the unraveling of the mummy bundle, which helps the scientists to make better, more contextual interpretations of their data (Lee et al. 2013). For instance, the mummy’s sex, as estimated by dress historians’ reference to clothing or ornaments, could be helpful to bioarchaeologists, whose own sex estimation is based mostly on simple presumption from regression or discriminant function analysis. Conversely, interdisciplinary collaboration can benefit dress historians’ work as well. In general, the personal profile of a mummy is investigated by archaeologists. Most mummified individuals of the Joseon period belonged to rich and well-educated clans. They had a detailed record about the history of their clans, called Jokbo (the genealogy book). By referencing the Jokbo, the name, age, sex, dates of birth and death, government career, etc., could be revealed. The information inscribed on the tombstone and the other historical documents also provided archaeologists with valuable context on the given individual. All of this data, certainly, could be very enlightening with respect to dress historian’s research. Various theories and presumptions of dress historians are continually corrected by such information. In the case of a seventeenth-century mummy (HD-2) unearthed in Hadong County, South Korea, we can see how greatly the information of archaeological scientists affected dress historians’ interpretation of their data (Lee et al. 2013). Archaeologists had revealed that the mummy was a female who had been the second wife of a member of the county gentry. Many articles of clothing, textiles, and others were found in the grave (Fig. 6). By the historical literatures, archaeologists estimated that she had lived in the late sixteenth to the early seventeenth century, because her husband was alive in 1610–1650, and her third son was born in 1627 (Lee et al. 2013). Indeed, acquiring a personal profile on a mummified individual is a starting point for the faithfulness and reliability of dress historians’ research. As for this case (HD-2), additional information on the mummified female was acquired by anthropologists as well. During the examination, fetal bones were found inside her trousers, which indicated that she was pregnant at the time of death (Lee et al. 2013). When anthropologists examined the mummy’s body, the skin at the abdomen was somewhat distended, which also would suggest pregnancy (Lee et al.

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Fig. 6 Shoes found in seventeenth century grave (Lee et al. 2013) (Credit: Dong Hoon Shin)

Fig. 7 Loosely fitting skirt of female mummy (Lee et al. 2013) (Credit: Dong Hoon Shin)

2013). Since the estimated age of the fetal bones was about 32 weeks, the mother probably did not die of labor dystocia during parturition but rather, she almost certainly experienced a stillbirth. Actually, no extant historical references mentioned that the woman was pregnant at the time of her death (Lee et al. 2013). Only anthropological work could reveal this. This fact was significant to dress historians, because it made them reconsider why the woman’s skirts had been worn so loosely around her waist (Fig. 7), a fact that had previously been ignored by dress historians (Lee et al. 2013).

Mummy’s Teeth In East Asian countries like Korea, the funeral ceremony was very thoroughly and strictly stipulated by the ritual codes prevailing at that time. Actually, the types and amounts of clothing collected from graves closely correlates with the social status of the individuals found therein. At the hierarchical apex of Joseon society were the landowning gentry called Sadaebu. Below them, middling people took various jobs

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such as medical doctor, interpreter, or government clerk. At the bottom of the social pyramid were freemen or cottagers, specifically peasants, merchants, and craftsmen, among others (Shin et al. 2008). Actually, most Korean mummies discovered have been Sadaebu, the people of the highest rank in Joseon society, which means that the mummy clothing collected from those graves had been worn by the ruling class elites at the time (Shin et al. 2008). In general, the types of clothing found have been stereotypic of those codified during the Joseon dynastic period (Lee et al. 2013). Nonetheless, in actual excavation cases, there are sometimes also exceptional findings that do not match the instructions mentioned in the ritual book. Such exceptions, even if they are rare, often spark serious disputes among scholars about the proper interpretation of findings (Kim et al. 2011). For instance, recently, a male mummy was discovered in Seoul city, South Korea. A tree ring test showed that the tomb likely had been constructed between 1605 and 1733 CE. A mummy bundle was discovered inside the coffin (Oh et al. 2015). CT radiography of the mummy bundle revealed small radiopaque materials present beside the mummy’s head. They looked similar to teeth, but the radiologist was not sure of the substance. Dress historians, meanwhile, found many articles of clothing stacked on or wrapped around the mummy. They also discovered a pouch that was tethered to the side of a mummy bundle (Fig. 8). Inside the pouch, they identified many teeth: 32 permanent ones and one deciduous (Oh et al. 2015). The teeth corresponded to the radiopaque materials shown on CT images. Since the alveolar sockets of the mummy were healed, the individual might have lost his teeth long before his death. A histomorphometry of the mummy’s estimated age using a

Fig. 8 Pouch tethered to mummy bundle (Credit: Dong Hoon Shin)

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bone biopsy indicated 67.5  7.0 years. By tooth attrition, the male mummy’s estimated age was 49.53  7.0 years old. Considering this discrepancy of the age estimations, the authors presumed that the mummified man must have kept the teeth until his death, possibly for about 20 years after he lost them in his late 40s (Oh et al. 2015). Dress historians in Korea knew of historical literature describing pouches in which a person’s lifetime’s worth of lost teeth, hair, and clipped nails were collected (Oh et al. 2015). Such pouches were kept throughout his/her lifetime; and finally, when the individual died, they were interred post-mortem inside the coffin. Nonetheless, when dress historians discovered the pouch in a mummy bundle during excavation, they doubted whether the teeth actually belonged to the mummified person (Oh et al. 2015). The researchers applied ancient DNA techniques in order to solve this problem. The results for the teeth in question (Q) and confirmed brain and bone (K) samples were compared. Such a Q–K comparison entailed in STR analysis did not exhibit full concordance of loci profiles due to accidental allelic drop-outs in some loci. However, the authors proved, by appropriate statistical analysis, that the teeth in the pouch likely corresponded to those of the mummified individual. A comparison of the mitochondrial (mt) DNA sequences also exhibited 100% matches between the Q–K samples. This means that the original presumption (the teeth were the mummified individual’s) could not be denied by statistical reasoning (Oh et al. 2015).

An Example of Mummy Clothing Research in South Korea Based on the typical and detailed collaboration pattern between dress historians and other specialists, we summarize another example of mummy clothing research performed in South Korea. Historians, in reviewing the available literature, traced a seventeenth-century male mummy (Gangneung)’s chronology as follows: The mummified male, named Gyeongsun, was born in 1561 (as the first son of Unwoo) and died in 1622 at the age of 61. Unwoo was a very famous scholar, belonging to one of the most distinguished families in the county. Unwoo hoped that his son (Gyeongsun) could likewise become a scholar or official of the kingdom. Gyeongsun tried to pass the National Examination for the classics licentiate degree, but was not successful. Historians found a record of 1586 in which Unwoo seriously worried about Gyeongsun’s failure in the National Examination. Instead, Gyeongsun could have got a special appointment for an honorary official post available for prominent men’s sons. According to the historical record, he almost drowned at sea in 1591. Except for this, he seems to have spent his life peacefully as a prominent gentleman in the countryside, not involving himself in any kind of political controversy. In 1602–1605, great famine and flood struck the city in which he lived. In 1605, his father died. He had three sons and three daughters. Upon his death, the King’s Court conferred posthumous honours of Commander on him. (Lee et al. 2009b)

Subsequently, the radiologists took CT axial images of the mummy bundle (Fig. 9). Since the mummy was wrapped in many articles of clothing, the preservation status inside had to be estimated by CT imaging prior to unwrapping. As seen on

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Fig. 9 CT scan of Gangneung mummy (Credit: Dong Hoon Shin)

Fig. 10 Mummy clothing seen on CT scan (Credit: Dong Hoon Shin)

the radiological images, the mummy’s internal organs were preserved well. The images also confirmed that many pieces of clothing had in fact been wrapped around the body (Fig. 10). In the next step, the dress historians removed all of the clothing wrapped around the mummy. This unraveling process reflected the CT findings. All artifacts were numbered at the time of their recovery. The textiles and clothing collected from the mummy bundle are summarized in Table 2. The clothing in the mummy bundle had been arranged in the same way as mentioned in Joseon dynasty’s ritual ethics codes. Briefly, we found some space-filling clothing stacked around the mummy bundle, around which, in turn, a broad textile was wrapped. Inside the wrapping textile, a man’s coat with straight collar, a man’s coat with round collar, a man’s coat with waist pleats, a man’s coat with back slit, a man’s coat with side slit, and a man’s coat with waist pleats were found. Another wrapping textile was found in a deep layer of the mummy bundle.

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Table 2 List of clothing collected from mummy bundle (Lee et al. 2009b)

Number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37

1165 Name of clothes Open drawers Socks Open drawers Drawers Open Drawers Hemp ropes (Horizontal) Hemp ropes (Vertical) Outer comforter Man’s coat with round collar Man’s coat with straight collar Man’s coat with side slit Man’s coat with waist pleats Man’s coat with back slit Man’s coat with waist pleats Hemp ropes (Horizontal) Hemp ropes (Vertical) Inner comforter Man’s coat with wide sleeves Man’s coat with round collar Man’s coat with waist pleats Nobleman’s coat Man’s coat with straight collar Pillow Waist cord Nobleman’s coat Man’s coat with side slit Man’s coat with side slit Jacket Long Sleeved Undershirt Hand Wrapper Open drawers Drawers Cap Veil for face Man’s headband Ear plugs Beads for headband

Material Cotton Cotton Spun Silk Tabby Hemp Hemp Hemp Hemp Satin Damask Blended Tabby Ramie Ramie Ramie Spun Silk Tabby Spun Silk Tabby Hemp Hemp Cotton Spun Silk Tabby Cotton Cotton Spun Silk Tabby Ramie Spun Silk Tabby Silk Threads Ramie Satin Damask Ramie Spun Silk Tabby Ramie Satin Cotton Cotton Spun Silk Tabby Satin Horse Tails Silk Cotton Glass

Inside of the bundle, a man’s coat with straight collar (#22), a noblemen’s coat (#21), a man’s coat with round collar, a man’s coat with waist pleats, and a man’s coat with wide sleeves were discovered. The enshrouded individual was finally exposed when the dress historians removed the clothing covering the trunk. The shrouds included a jacket, a nobleman’s coat (#25), a man’s coat with side slit (#26), another man’s coat with side slit, a long-sleeved undershirt, a waist cord, a hand

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Fig. 11 Removal of clothing from mummy bundle. The numbers correspond to the numbers in Table 2. NC, Noblemen’s Coat; MCSC, Man’s Coat with Straight Collar; WC, Waist Cord; MCSS, Man’s Coat with Side Slit (Credit: Dong Hoon Shin)

wrapper, open drawers, and drawers (Fig. 11). On the mummy’s head, a cap and a face veil were discovered. The mummy wore the headband with beads (#37) (Fig. 12). Ear plugs filled both ears. The collected clothing and other remains were washed, repaired, or reconstructed in the dress historian’s lab. Depending on the clothing, either wet or dry cleaning was employed. Wet cleaning could be done in a large water tank with a detergent, using mesh bags to prevent damage caused by contact between articles. Meanwhile, in cases where damage to clothing was serious, or either metal or paper had been used for the fabric, dry cleaning was preferred. Next, the researchers recorded the shapes of the objects using a line-drawing system with an Adobe Illustrator program (Adobe Inc., San Jose, California). After all of these processes were completed, the repaired articles were kept in a large neutral paper box in which they could, to the extent possible, be spread flat without folding. Figures 13 and 14 are examples of repaired original mummy clothing and a replica of the man’s coat, respectively.

Conclusion The research on mummy clothing plays a leading role not only in mummy research but also in the dress history of South Korea. Clothing excavated from Joseon graves has been investigated in collaboration among researchers representative of many academic fields; and indeed, revealing the changes in Joseon dynasty clothing is the greatest achievement of South Korean dress historians thus far. A variety of information necessary to an understanding of the history of clothing in East Asia can be obtained, including the actual uses and designs of specific articles. Moreover, through joint South Korean/Japanese/Chinese research, international trade routes in East Asia and exchanges of textile fabrication techniques among them could be revealed. Such successes and overall progress in the field have been made possible by recent technological developments of archaeological science. Thanks for example to technological advances in cleaning techniques, the preservation of excavated clothing has become

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Fig. 12 Mummy’s headband. (Credit: Department of Anatomy and Cell Biology, Seoul National University)

Fig. 13 Repaired original mummy clothing (Nobleman’s coat; #21) (Credit: Seoul Women’s University Museum)

more successful and consistent. Scientific-analytical processes also can be applied to the estimation of original colors and the natural dyes used therein. Using such techniques and the information yielded thereby, replica making has become easier. Repaired or reproduced mummy clothing is currently on display in many museums and other institutions in Korea (Fig. 15). During the past decades, appreciation of the value of mummy clothing has grown among both academics and the general public in East Asia.

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Fig. 14 Replica of man’s coat with side slit (#26) (Credit: Seoul Women’s University Museum)

Fig. 15 A replica of Joseon dynasty female’s clothing (Credit: Heeseon Kim)

As long as mummy research continues in East Asia, the study of clothing excavated from graves will continue to inspire the researchers involved.

Cross-References ▶ Joseon Dynasty Mummies of Korea

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Acknowledgments This research was supported by Basic Science Research Program of the National Research Foundation of Korea, the Ministry of Science, ICT & Future Planning (2020R1A2C1010708).

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Shin MH, Yi YS, Bok GD et al (2008) How did mummification occur in bodies buried in tombs with a lime soil mixture barrier during the Joseon Dynasty in Korea? In: Pena PA, Martin RM, Rodriguez AR (eds) Mummies and science. World Mummies Research, Santa Cruz de Tenerife, pp 105–113 Shin DH, Bianucci R, Fujita H et al (2018) Mummifications in Korea and China: Mawangdui, Song, Ming and Joseon Dynasty mummies. Biomed Res Int 2018:6215025 Song MK, Shin DH (2014) Joseon mummies before bioanthropological studies began in Korea. Pap Anthropol XXIII(1):117–134 Willerslev E, Cooper A (2005) Ancient DNA. Proc Biol Sci 272:3–16 Williams AR (2006) Mystery of the tattooed mummy. Nat Geo 215:70–83

Index

A 1984 Aboriginal and Torres Strait Islander Heritage Protection Act, 98 Abydos, 592, 594, 639 Acarology, 497–500, 510 Acetolysis, 536 Acquiring mummies buying, selling and trading human mummies, 68–69 museum acquisition, 67–68 Adapter sequences, 304 Adapter trimming process, 306 Adenocephalus pacificus, 416 Adenomatous polyposis coli (APC), 278, 789 Adipocytes, 1060 ADMIXTURE, 376, 377, 379, 380 aDNA analysis, 1027 Afanasievo culture, 1042, 1044 Agarose gel electrophoresis, 847 Age, 397 Age-at-death determination, 583–585 Ainu, 104 Akhmim, 603, 605–608, 610 Alexandria, 571, 611, 613 Alien bodies, 939 Alien mummies, Nazca, 1148 Alluvial soils, 950 Alpacas, 646 Amalgam, 634 Amenhotep II, 631 Ameniryirt, 571–580, 582, 585–589, 606 American Association of Physical Anthropology (AAPA), 90 American trypanosomiasis, 333–334 Amulet, 582, 584, 598, 599, 601–604, 606– 608, 610, 615, 616 Amun, 572, 607–609 Ancestry, 398, 399, 404

Ancient DNA (aDNA), 97, 102, 318, 319, 344, 352, 403, 405, 779, 790, 1043 analysis, 449–455, 1062–1066 archaeogenetics in Africa, 289–290 assessment of contamination level, 308–310 authentication, 320 authenticity criteria, 328 biases and errors, 302 cloning-based studies, 327 demic diffusion, 286 DNA degradation, 353–356 domesticated animals, 293–294 Eastern Eurasians and coastal migration hypothesis, 290–291 fragmentation, 326 genomic diversity in wildlife, 294 genotype calling, 310–312 geographic locations, 288 LCN DNA samples, 356–357 mapping reference genome sequence and read filtering, 305–306 migration and admixture, 286 molecular degradation, 326 mummified soft tissues, 286, 302 next generation sequencing (NGS) methods, 302 Paleogenetic Lab, 325 PCR, 319 population genomics of archaic hominins, 287–289 “population Y” hypothesis, 291–293 post-mortem chemical damages, 306–308 quantification and assessment, 326 quantitative analysis, 326 recovery, 319, 322 sequencing adapter and barcode sequences, 302–305 TB infection, 329

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1172 Ancient DNA (aDNA) analysis, mummy research, 278–280 HLA-DRB1 gene, 278 mitogenome, 273–274 smallpox, 277 Ancient Greeks, 881 and Egyptian embalming practice, 878–879 and Egyptian mummies, perception of, 879–880 Ancient Native Americans, 403 Ancylostoma duodenale, 417 Ancylostomiasis, 417 Andean mummy(ies), 21, 22, 204 analyses, 206 Andes, 5, 12, 15, 22, 25, 27, 32 Anga culture, 986 ANGSD program, 310 Animal domestication, 293, 294 Animal mummies, 4, 18, 30 artifact types, 638 baboon and hunting dog, 631 carbon 14 dating, 642 chemical test, 640 desiccation, 642, 644 DNA, 641 experimental mummification, 642 imaging technology, 640 macroscopic examination, 640 museum contexts, 636 pet animal, 630 poultry, 632 sacred animal, 632 socio-economic role, 645 vervet monkey, 631 veterinary and animal management practices, 639 votive mummies, 643 Ankhhap, 608, 609 Anthropogenic mummies, 61, 81 Anthropogenic mummification, 50–53, 215, 744–746 Anthropological research on human skeletons, 90, 91 Anthropologists, 1160 Anthropology, 99, 213 Antipathy, 181–182 Antisuyu, 13 Anubis, 569, 580 Apis Bull, 632, 634 Archaeoacarology, 497, 498, 510 Archaeological Resources Protection Act of 1979 (ARPA), 95, 97 Archaeological Survey of Nubia Projects, 31

Index Archaeologists, 1160 Archaeology, 94, 96, 99, 497 ancient Greece (see Ancient Greeks) Archaeoparasitology, 276, 548, 557, 559, 560 of Korean mummies (see Korean mummies) Archaic hominin genomes, 287 Archaic Period, 6 Arctic mummies, 1013 Argentina, 937 Armant, 632 Artefacts, 71 Artemidoros, 617 Arteriosclerosis, 730, 737 Arteriosclerotic cardiovascular diseases disease evolution, 149 histological evidence, 152 identification, 151 infra-red spectroscopy, 159 limitations and pitfalls, 159 literary evidence, 150 molecular evidence for, 159 non-biological sources, 149 palaeoradiological evidence, 155 pathogenesis and clinical manifestations, 149 Arthropods, 496–499, 503, 504, 506, 510 Artificial embalming, 656 Artificial Intelligence (AI), 265 Artificial mummies, 23 Artificial mummification, 42, 45, 55, 885 Ascaris, 321, 337–339, 341 Ascaris lumbricoides, 418, 450–451 Ascetic self-preparation, 1118 Aseki, 984, 987, 991, 992, 1002, 1007 Astigmatid mites, 500 Asyut, 639 Atacama Desert, 646, 932 Atherosclerosis, 120–121, 935 Atherosclerotic cardiovascular disease (ASCVD), 278 Attel mummies, 764 at Tuna al-Gebel, 638 Autolysis, 43 Autopsy, 498, 501, 852, 856–858, 863, 866, 874 atherosclerosis, 120–121 calcified pulmonary nodules and tuberculosis, 121–122 cardiac structure, 123–125 checklist and ethics for mummy, 125–126 cross-check, 113 intestines and coprolites, 116–118 paleopathology, 118–119

Index B Baboon(s), 631, 634 Bacteriophage, 280 Bastet, 633 Beer, A., 786 Beeswax, 640, 656, 685 Berlin, 764 Bioanthropology, 895 Bioarchaeology, 92, 214, 217, 218, 225, 245, 246 Bioarchaeology of care (BOC), 247–248, 934 Bioarchaeology of mummified human age-at-death determination, 583–585 biological sex determination, 585 life and death of Ameniryirt, 585–589 Bioinformatics, 366 Biological analyses, 885 Biological anthropology, 90, 96 Biomedical methods, 29 Bitumen, 576, 616, 689 biomarkers, 655 Blood vessel, 149–152 Body conservation/cultivation, 866, 873 Bog bodies, 72, 808, 834 challenges in research, 818–821 characteristics of, 817–818 geographic distribution, 809–810 historical and cultural contexts, 811–812 potential intents, 812–814 preservation, 828–829 Bog mummies, 845–848 Bollaert, William, 22 Bone Biopsy needle, 260 Book of Abraham, 27 Botanical analysis, 535 Brain histology, 852, 855, 856, 859 Brandenburg, 764 Brazil, 946 archaeological sites, 947–953 Caverna da Babilônia, 949–953 Church of Santo Antonio Aparecido, 948–949 climate and soil, 947 Gruta do Gentio II, 949 intestinal parasites, 334–343 Lapa do Boquete, 947–948 mummification, 946–947 treponematoses, 331–333 tuberculosis, 328–330 Brazil’s National Museum (BNM), 937 Bremer Bleikeller, 763 British Museum, 571, 574, 580, 581, 586, 588, 589, 591, 600–602, 604, 616, 618, 621, 891

1173 Bronze Age, 1042 Buddha-shaped statue, 75 Buddhism, 1105, 1107 Buddhist monk mummies, 1105–1109 1857 Burial Act, 98 1981 Burial Act, 98 Burial laws, in the United States, 95–97 Burial laws and regulations in Australia, 98 in Canada, 97, 98 in United Kingdom, 98 Burrows-Wheeler Aligner (BWA) backtrack program, 305

C Cabildo de Tenerife, 890 Calcifications, 157, 158 Canadian Association for Physical Anthropology, 98 Canales, Pedro P., 23 Canary Islands, 11–16, 19, 25, 32, 33, 887 Cancer, 587 carcinogens in history, 143 incidence of, 132 molecular evidence, 142 paleo-epidemiology, 141–142 pathobiology of, 133–137 radiological techniques, 133 Canopic, 576, 578–579, 610 Capillariid, 324, 337, 342, 343 Capture, 365 Carbon 14 dating, 642 Carcinogenesis, 133–135, 142, 143 Cardiovascular disease, 588, 601 Castor oil, 657 Catacomb, 632 Catacombe dei Cappuccini, 756 Caverna da Babilônia, 949–953 Cedar wood tar oil, 656 Central Yakutia mummies, 1018 Chagas disease, 276–277, 420 and Chinchorro, 422–423 oral transmission, 421 pathoecology of Chiribaya, 428 in Texas-Coahuila border, 430–431 Chancay mummy, 938 Charlotte de Savoie, 916–918 Chemical damage, 306–308 Chemistry, 926 Cherchen Man, 1043 Chickens, 632 Chile, 630

1174 China, parasitic findings in, 518–519 Chinchaysuyu, 13 Chinchorro mummies, 421, 464, 934 and Chagas disease, 422–423 Chiribaya Alta, 425 Chiribaya Baja, 424–425 Chiribaya of Sothern Peru, 423–424 El Yaral, 425 San Gerónimo, 424 Chinchorro mummies dietary analysis, 421 Chinese liver fluke (Clonorchis sinensis), 524 Chinese mummies, 558–559 Chinese troops, 534 Chiribaya culture, 423 Christian Era, 26 Christianity, 7 Chumash descendants, 103, 104 Chuson-ji, 1109, 1110 Claude Gaillard, 637 Cleopatra, 614, 615 Climate change, 818 Clonorchis sinensis, 441–444, 449, 450, 453–455 Clothing, mummy dress historians, 1155, 1156 dry cleaning, 1166 mummy bundle, 1156–1159 in museum, 1156 South Korea, mummy clothing research in, 1163–1166 wet cleaning, 1166 Coca, 935 Coffin, 569, 571, 573, 578–580, 589, 592–597, 599, 601, 607, 608, 615, 620 wood, 1055 Colatoi, 756 Collagen, 818 Collasuyu, 13 Combination mummies, 61 Commission of Arts and Sciences of Egypt, 18 Communist Chinese forces, 534 Computed tomography (CT) analysis, 113–115 Computed tomography (CT) scanning, 71 Computer-assisted post-processing, 183 Computer-generated three-dimensional (3D) modeling approach, 168 Computerised Axial Tomography, 256, 266 Confucian rituals, 1123 Congenital diaphragmatic hernia, 118 Coniferous resin, 655, 656 Consent, 64–67 Conservation, 908, 915 Contamination estimation, 310

Index contamMix program, 309 Coprolite(s), 324, 327, 535, 537, 538, 540, 541, 543 Coprolite and mummy intestine analysis coprolite residues, 465–473 Dan Canyon burial, 482–485 diet and environment, 464 gut level data, 463 microscopic and molecular analyses, 463 recovering coprolites and sectioning intestines, 465 reporting result, 473–474 Skiles Mummy, 485–488 Ventana Cave Mummy, 475–482 Copyright law, 80 Corpernicus, 401 Corpocenosis, 496 Co-sequenced common adapter sequences, 304 Cossack mummy archaeological information, 836, 837 CT radiography, 840–841 dental prosthesis in 3D model, 841 histology, 841–844 historical considerations, 837–838 mitochondrial DNA analysis, 845 preservation status of, 848–849 radiocarbon dating, 838 stable isotope analysis, 843 Cranial sutures, 585 Craniofacial reconstruction (CFR) accuracy of, 171–172 computer-generated three-dimensional (3D) modeling approach, 168 CT and anthropological data, 167 facial appearance of mummified individuals, 167 Homo floresiensis skull, 167 of mummy, 168–170 portraits, 173–175 Richard III, 167 skull analysis, 168 Crypt, 780 Crypt and catacomb mummies, 743, 751–756 anthropogenic mummification, 744 Austria, 766–768 Czech Republic, 769 France, 771 Germany, 763–766 Hungary, 769–770 Italy, 756–763 Lithuania, 770–771 Spain, 771 Switzerland, 771

Index CT scan/scanning/scanner, 573, 575, 577, 580–584, 586–588, 598, 600–602, 605, 606, 613, 615, 640 Cultural and social anthropology, 872 Cultural artifacts, 887 Culture, 65 Cuntisuyu, 13 Cytosine deamination, 359

D 3D, 581–583 Dakhla, 618, 620 Dannenfeldt, Karl H., 10 Deaccessioning mummies, 68 Death, 42–44, 48, 52–55 Decalcified bones, 817 Decomposition, 42–45, 47–49, 53 Dedifferentiation, 133 De-gloving, 43 Degradation parameters, 359 Degraded DNA, 361, 367 Degraded grains, 537 Dehydration, 932 Deir el-Bahari, 595–597, 616 Deir el-Bersheh, 594, 596 Deir el-Medina, 598 Demography, 289, 292 Denisovan, 402, 403 Dental attrition, 588, 900 Dental prosthesis in 3D model, 841 Dermestid beetles, 644 3D geometric surface analysis, 171 Diane de Poitiers, 918 Dicom Reader software, 264, 265 Diet, 198–201, 463, 464, 470–472, 474, 475, 482, 487, 535, 541, 543 Digital imaging and communications in medicine (DICOM), 581, 602, 605, 609 Digital volume tomography (DVT), 229 Dilaceratio corporis, 909 Diodorus Siculus, 570 Diptah, 608 Diterpenoids, 655 Djehutynakht, 594 Djer, 592 3D multi-detector computed tomography (MDCT), 182, 226 DNA, 578, 641 analysis, 64, 102, 103 crosslinking, 355 damage patterns, 359 extraction, 795

1175 extraction methods, 363 fingerprinting, 274–275 fragmentation, 262 quantification, 357 recovery, 363 DNA-based prediction, of pigmentation traits, 388–389 eye color prediction, 389–395 hair color prediction, 395–397 skin color prediction, 397–399 DNA degradation, 353–356 DNA recovery, 363 massive parallel sequencing, 363–366 mitochondrial DNA analysis, 360–361 small-sized PCR products, 361 DNA quality and quantity gel electrophoresis, 357–358 MPS data, 359–360 quantitative PCR (qPCR), 358–359 Documentation, 81 Documents, 1066 Dog, 631 Double Dragons Tomb, excavation of, 1095–1097 Double-stranded library preparation, 364 3-D prints, 640 3D reconstruction, 263 Drents Museum, 823 Dry cleaning, 1166 Dry embalming, 745 3D Skull model, 169 Dual energy computed tomography (DECT), 226 Dual energy CT scanning, 585 Dual-purpose machines, 264 Ducks, 632

E East Asian mummies, 455, 1050 Echinostoma, 419 Ecological fitting, 433 Eggs, 634 Egypt, 630 Egyptian animal mummies, see Animal mummies Egyptian embalming practice, 878, 879 Egyptian medical profession, 9 Egyptian mummy(ies), 5–7, 67, 91, 181, 204, 205, 360, 879–880 scientific observations in South America and artificial mummies, 21–24 skin, 1060 19th century scientific study of, 20–21

1176 Egyptian Museum, 637 Egyptology, 30, 31 Egyptomania, 19 Embalmers, 938 Embalming, 50–53 King Henri IV of France, 919–923 King Louis XI and Charlotte de Savoie, 916–918 Louis XIII and Louis XIV of France, 923–925 Louis XVIII of France, 925–926 modus operandi, 910–913 origin in France, 908–913 procedures, 878–879 Sorel’s, A., 915–916 techniques, 913–915 Endogenous nucleases, 353 Endoparasites, 520, 522 Endoscopy, 825 history, 184 methodologies, 223–224 Enterobius vermicularis, 321, 327, 334, 338, 340, 341, 448 Enzymatic inhibition, 44–45 Era of Exploration, 12, 32 Ethics, 61, 91, 92, 103, 106 ethical concerns, 100, 101, 106 ethical considerations, 91, 93, 99–101, 103 ethical dilemmas, 91, 94 ethical issues, 91, 93, 100, 103, 105 Étienne Geoffroy St. Hilaire, 637 Euphemia, 621 Europe, 809 Experimental mummification, 642 Experimental paleoparasitology, 321 Eye color prediction, 389–395

F False, 634 eye, 600, 601 Fan Zhongyan, 1120 Fayum, 600, 613, 616, 618 Female Mummy, 1096 Field imaging, 224–225 Final foods, 475 Fire mummies, in Kabayan region cave burials and coffins, 967–970 cultural aspects, 964–967 death ritual practices, 976 death rituals, 970 headhunting, 963 mummification procedure, 971

Index mummification science, 974–976 symbols of status, 960 village and burial rituals, 978 First Intermediate Period, 6, 568, 592–594, 596 Fleas, 420 Flies, 644 Fluorescence microscopy, 1066 Food, 632, 640, 645, 646 Forensic acarology, 497–500, 510 Forensic anthropology, 92 Forensic entomology, 499 Forensic science, 497 Fragmentation of skeletal elements, 913 Frankincense, 658 Freeze drying, 829 Frézier, Amédée François, 21 Frozen mummies, 1012–1014, 1017 Fujiwara mummies, 1109–1111 Functional analysis, 402, 405

G Gangneung mummy, 1164 Gas chromatography (GC)/mass spectrometry (MS), 655–658 Gebelein, 573, 575, 585, 590, 593–595 Geese, 632 Genetic relatedness, 373–376 Genome-wide association studies (GWASs), 388, 395, 398, 399, 405 Giza, 593 Gliddon, George R., 29 Glycyphagidae, 507, 508 Grave, 1120–1133 Greek medical practice, 9 Greenland mummies, 181 Gruta do Gentio II, 949 Guanche culture, 33 Guanche mummies, 11 anthropogenic mummification, 898 Athanatos, 896 CRONOS project, 895 demographic data, 898 dental pathology, 900 diet analysis, 899 history of, 888 markers of physical activity, 900 metabolic stress markers and pathology, 900 natural/natural intentional mummification, 897 paleoparasitological studies, 902 repatriation, 893–894 skeletal pathology, 900

Index soft tissue pathology, 902 spoliation, 889–891 types of burial, 896 Guinea pigs, 646 Gumugou cemetery, 1041 Gut content analysis, 463, 535 “Gut-level” data, 463 Gut microbiomes, 280 Gymnophalloides seoi, 441, 444

H Hair color prediction, 395–397 Han Dynasty, 514, 515, 517, 525–528 achievements, 515 agricultural development, 515 anatomical studies on mummies, 517 ancient physicians in, 526 animal husbandry, 516 flukes in, 524–525 isotopic analysis, 516 pinworm (Enterobius vermicularis), 522–523 roundworm (Ascaris lumbricoides), 517–520 tapeworm (Taenia sp.), 520–522 whipworm (Trichuris trichiura), 523–524 Hardy-Weinberg equilibrium, 379 Hawaiʻi Department of Land and Natural Resources, 97 Hawaiʻi Revised Statutes (HRS), 97 Hawara, 611, 616 Helicobacter pylori, 277–278 Heliocentric theory, 31 Heliopolis, 632 Helminthiasis, 340 Henutmehyt, 578 Hepatitis B virus (HBV), 277 HERC2, 393, 394, 396 Herodotus, 4, 5, 7, 8, 18, 32, 570, 573, 575, 577, 595 Hetepheres I, 593 Hierakonpolis, 573, 592 HIrisPlex, 396, 398, 400–402 HIrisPlex-S, 398, 400, 402–404 Histology, 1058–1062 analysis, 886 of West Siberian mummy, 1026 Histomorphometry, 1162 History of mummy studies Canary Islands, colonization of, 11–16 Egyptian mummies, 5–7 Egyptomania and unrolling mummies, 19–20 influence of Egyptian mummification, Egyptian and Greek medical practice, 8–10

1177 Medieval Period and Renaissance, 10–11 Napoleon and savants, 17–19 North America, collecting and viewing mummies in, 24–28 17th and 18th centuries, 16–17 19th century American scientific contributions, 29 19th century scientific study of Egyptian mummies, in UK, 20–21 Hoaxes and mummies, 1144 Hoegwakmyo, 1053 Homer, 8 Homo floresiensis skull, 167 Hookworms, 417 Horemheb, 631 Hornedjitef, 607, 608 Horse, 631 Hospice, 475–488 Hottentot venus, 83 Huaycan, 941 Human genetic history, 286 Human mummies, 4, 18 Human remains, 90–95, 98–102 analysis of, 103 burial laws (see Burial laws and regulations) destructive methodologies, 100–103 ethical issues in scientific research on, 91, 93 ethical issues of studying, 90 ethical research on, 106 ethical treatment of, 94 during excavations, 93 (see also Egyptian mummies) human cadavers, 90 human skeletal remains, 97 human skeletons, 94 non-destructive methodologies, 99–100 skeletons, 90 study of, 94, 98, 99 Human sacrifice, 813 Hungarian mummies, 181 Hybridization enrichment procedure, 365 Hydrolytic damage, 354 Hypervariable segment I (HVS-I), 329

I Ibaloy, 959–961, 963, 964, 967, 970, 971, 975, 978, 979 Ice Man, 66, 204 ICOM Code of Ethics, 70 Identity-by-descent (IBD) segments, 374 Illumina adapter sequence, 303 Inca, 934

1178 Inca mummies, 206, 207 Incision, 574, 575, 593, 595–597, 599, 601, 613, 614 Incomplete Lineage sorting, 372 Indigenous groups, 65 Institutional Review Board (IRB), 450 Intermediate eye color, 393, 394, 404 Intermediate mummification, 215 Internal barcode sequences, 303 Intestinal helminths, 427–428 Intestinal infection, 729 Intestinal parasites, 321, 323, 334–343 IrisPlex, 393, 394, 396, 398, 400, 403 Irthorru, 603, 606, 607, 610 Isotopes analysis of mummies, 204–206 carbon isotope ratio, 199–201 definition, 198 nitrogen isotope ratio, 199–201 stable isotopes and radioisotopes, 198 standard values (STD), 199 strontium, sulfur, hydrogen, and oxygen isotope ratios, 201–203 type of body tissue, 203 Isotopic analysis, 515, 516, 642

J Japanese monk mummies, 1118 Japanese mummies, 1104 Buddhist monks, 1105–1109 in East Asian history, 1111–1113 Fujiwara mummies, 1109–1111 Jeneval system, 537 Johann-Friedrich-Blumembach Institut für Zoologie und Anthropologie, 890 Joint POW/MIA Accounting Command, 535 Jomon people, 200 Joseon Dynasty, 173, 174, 557, 1118, 1121, 1155 mummies, 105 Joseon dynasty mummies of Korea, 1068, 1069 ancient DNA (aDNA) analysis, 1062–1066 coffin wood, 1053 emergence of, 1054–1058 exothermic reaction, 1053 natural or artificial mummifications, 1051 preservation conditions, 1051 preservation status and histology, 1058–1062 researches of, 1066–1068 Joseon grave, 1126 Joseon-Korean mummies, 557–558

Index Joseon mummies, 441, 445, 453, 455 Joseon period, 1155, 1156, 1159, 1160 Korean, 1128 Joseon period tomb, 1131 Julia Pastrana, 83

K ka, 569 Kahun papyrus, 9 Keliyahe northern cemetery, 1039, 1041 Kennewick Man, 96, 97 Kharga, 611, 618 Khnum, 633 Killed in Action (KIA) soldiers, 534 King Louis XI, 916–918 King Richard III, 362 Koenig, 254 Korea, 168 Korean child mummy, 1058 Korean mummy(ies), 170, 181, 535, 1120–1124 ancient DNA analyses, 449–455 ancient parasite species, 448–449 archaeoparasitological research on, 441–442 cardiac structures, 123 congenital diaphragmatic hernia, 118 hair, 1061 location of, 442 parasitological infection, Joseon period to 21st century, 444–445 scientific research, 125 skin, 1059 trematode infection in, 442–444 Korean War, 534 Kukukuku, 986, 988, 994 Kuntskammera Museum, 891 Kushite, 572, 603

L Lapa do Boquete, 947–948 Lardoglyphus robustisetosus, 502 Late Period, 568, 577, 579, 599–607, 610, 622 Late Pleistocene megafauna, 262 LCN DNA samples, 356–357, 367 Legitimacy, 75 Lenin, Vladimir Il’ič artificial embalming, 863, 865 autopsy, 852 embodiment of, 872–874 fate of Lenin’s brain, 860–863

Index illnesses, 857 living sculpture, 870–872 medical history, 857 official autopsy report, 853–857 official version of cause of death, 859–860 regular treatment and re-embalming, 868 re-sculpting” mix, 868 tissue preservation, 867 Lenin’s illness, 856, 873 Lice, 419–420 Life histories Beer, A., 786 Borsodi, T., 792 Hausmann family, 788–789 Kustár, A., 796 Simon, A., 793 Stefanovits, G., 790–791 Tauber, A., 786 Waitzenbach, T., 790 Weiskopf, J., 791 Ilamas, 646 Llullaillaco mummies, 206, 937 Local communities, 938 Locus-by-locus analysis, 311 Loretum, 781 Louis Lortet, 637 Low copy number (LCN), 352

M Maat, 569 Maatkare, 631 Macrocheles muscaedomesticae, 502 Magnetic resonance imaging (MRI), 117, 230, 258 Major facial muscles, 169 Mammoth Cave, 26 Manchester Mummy Project (MMP), 30 Manchester museum mummy project, 654 Manipulation, 55 Maori culture, 84–85 MapDamage pattern, 307 Marker mutations, 137 Markulis, Juozas, 505 Massive parallel sequencing (MPS), 353, 363– 366, 388, 399, 401–403, 405 Mawangdui mummy(ies), 1085, 1091, 1132 biomedical investigation of, 1084 excavations on, 1087–1091 Mawangdui site, 1078, 1080, 1088 Mawangdui tomb, excavation of, 1075–1084 Mawangdui Tomb Site Museum, 1087 MC1R, 393, 395, 396, 402, 405

1179 Media, 79 Medical imaging, 213–218, 220, 225, 226, 235, 248 for archaeology, 260 data capture and storage media, 254 Digital Xray, 257–258 Dual energy machines, 257 historic X-ray films, 254 modalities, 182, 217 MRI, 259 mummies, 63 phase contrast imaging, 265 radiation damage to ancient DNA (aDNA), 261–262 terahertz imaging, 259 ultrasound imaging, 259 x-rays, 255–257 Medical mummies, 216 Medici Project, 761 Medieval, 589, 620–621 Mediterranean cultures, 888 Mégnin, Jean Pierre, 499 Melanesian blond, 404 Melanogenesis, 389 Merit, 598 Metagenomics, 279 Metastatic carcinoma, 586, 587 Methanolic extract, 655 Michael, 621 Michaeler crypt, 766 Microbiome, 279, 280 Micro CT scanning, 265 Microfossils, 536 Micro-haemorrhages, 153 Microthoracius spp., 427 Middle Kingdom, 568, 594–596 Migration, 198, 201–203 Mineralization, 538 Ming Dynasty, 517 See also Song and Ming dynasties Minimally destructive analyses, 63–64 Mite affiliations, with mummies in archaeological contexts, 502–504 in forensic contexts, 498–502 Mitochondrial DNA (mtDNA), 273, 274, 319 analysis, 360–361, 845 sequences, 361 Modified SHEP 1, 2 & 4, 398 Molecular abnormality, 135 Molecular biology strategies, 327 Molecular paleoparasitological methodology, 325–328 Molecular Paleoparasitology Lab, 324–325

1180 Mongols, 1120 Monkey, 631 Morality, 61 Multimodal imaging, 825 Mummy(ies), 60, 168–170, 286, 293, 302 acquiring, 67–69 Ata, 1144 Atacama alien, 1144 bundle, 1054 caring for, 66 consent, 64–67 counterfeit, 1141, 1143 definition, 60–61 Diepholz, 1143 display of, 69–78 disposal of, 80 educational purpose for displaying, 71 Egyptian-style, 1143 ethics, 61–62 excavation, analysis and conservation of, 62–63 fake, 1140 fake Persian, 1143 forgery, 1143 fraud of extraterrestrial, 1147–1148 good quality, 1141 medical imaging, 63 mermaid, 1142 minimally destructive analyses, 63–64 Nazca alien, 1144–1146 Persian, 1143 photography, 78–80 repatriation, 81–83 science, 30, 214 scientists and alien, 1146 settings, 60 study research, 61 teeth, 1161–1163 Mummies of the World, 79 Mummification, 42, 54, 356, 946–947, 958, 960, 964–967, 971, 973, 974, 979, 1119 anthropogenic, 50–53 enzymatic inhibition, 44–45 post-mortem decay, 42–44 spontaneous, 45–49 Mummification in the Nile valley Ameniryirt, 572–578 bioarchaeology (see Bioarchaeology of mummified human) CT scanning, 581–583 Late Antique and medieval period, 620–621 New Kingdom, 596–599 Old Kingdom and First Intermediate Period, 592–594

Index Predynastic period and Early Dynastic Period, 589–592 Ptolemaic Period, 607–611 radiology, 581 Roman Period, 611–620 Second Intermediate Period, 597 significance, 569 texts, 570 Third Intermediate Period and Late Period, 599–607 Mummification processes and preservation acidic soil conditions, 216 classification of, 214 decomposition, 216 smoking method, 216 spontaneous mummification, 215 temperature, 216 Mummified arterial vessels, 160 Mummified soft tissues, 286 Mummy clothing, in East Asia dress historians, 1155, 1156 mummy bundle, 1156–1159 in museum, 1156 South Korea, mummy clothing research in, 1163–1166 Mummy studies, forensic significance of, 53–54 Municipality of Itacambira, 948 Musée de l’Homme, 890 Museo Nacional de Antropología, 890 Museum, 60 visitors, 74 Museum of Archaeology and Anthropology, 891 Mycenaean mummy, 880 Mycobacterium tuberculosis, 275, 276, 328, 797 endemic strains of, 785 infection, 121 molecular analysis, 786 paleomicrobiological analysis, 791 PCR, 796 tuberculosis infection, 797–800 Mycobacterium tuberculosis complex (MTC), 327 Myianoetus spp., 500

N Nationalism, 645 National Memorial Cemetery of the Pacific (NMCP), 534 National Museum of Korea (NMK), 1039–1040 National Museum of the American Indian Act of 1989 (NMAIA), 95–97 Native American artifacts, 93

Index Native American Graves Protection and Repatriation Act of 1990 (NAGPRA), 95 NAGPRA, 95–98, 103 Native American human remains, see Human remains Natron, 216, 575, 592, 595, 596, 613, 620, 640, 643–645 Natural History Museum in Lyon, 637 Natural intentional mummification, 885 Naturally mummified bodies, 946 Natural mummification, 42, 44, 46–48, 55, 181, 884 Natural mummy, 575, 590 Nazca alien mummies, 1145, 1146 Neanderthal, 402, 403, 405 Necator americanus, 417 Neolithic glacier mummy abnormalities of hair, 731 age at death, 723 anthropological examination, body size and stature, 723 dating of mummy, 722 detection of body, 721 diseases of teeth and oral cavity, 730–731 evidence for physical activities, 726–727 examination of, 721 intestines, 727–729 life and death, 736–737 non-traumatic skeletal pathology, 730 nutrition of, 725 potential therapeutic treatment, 732–733 provenance and genetic origin, 724–725 pulmonary pathology, 729 sequelae of traumatic injuries, 733–736 systemic infectious disease, 731 type of mummification, 723–724 vascular pathology, 729–730 Nesmin, 610 Nespamedu, 610 Nestawedjat, 601, 603 New Kingdom, 568, 577–579, 596–599 Next-generation sequencing (NGS), 273, 279, 280, 287 methods, 302 Nidality theory, 415 Nigrovits, A., 793 Nile Delta, 566 Nile valley, 4, 6, 7, 566, 572, 585, 588, 611, 618, 620 Non-destructive methods, 63 Non-invasive mummy research, 182–183 Non-invasive techniques, 184 Northern Song, 1120 North Korean Army, 534

1181 χ2 test, 702 Nubia, 572, 621 Nut, 578

O Oasis/Oases, 566, 611, 613, 618, 620 Oculocutaneous albinism (OCA), 392 Odyssey, 8 Offering, 632–634, 640, 645, 646 Oils, 640 Old Kingdom, 568, 592–596 Old Kingdom Period, 6, 9 Opisthorchis felineus eggs, 1028 Organic balms in Egyptian mummification chemical investigation, 659–661 fat, beeswax, bitumen, coniferous resins and vegetable tannins, 655–658 Fourier transform infrared spectroscopy, 656 Fourier transform Raman spectroscopy, 656 gas chromatography/mass spectrometry, 655–658 lipid extraction and analysis, 685 material type, variation of chemical composition with, 706–708 mummy sample description, 661–684 percentage of balm compositions, 685–700 preparation of balms, 699–704 solvent extraction, 656 thermal desorption and pyrolysis GC/MS, 657 variation in balm composition over time, 693–698 variation on balm composition with age of individual, 703–705 variation on balm composition with body location, 706 variation on balm composition with gender, 705 Oryza pollen, 541 Osiris, 569, 572, 577–580, 598, 605 Osteoarthrosis, 726, 727 Otani collection, in Seoul, 1039 Oxidative damage, 355 Oxisols, 949 Oxyurid, 322, 341, 342

P Packing, 573–575, 585, 593, 597, 599, 601, 603, 606, 615 Padiamenet, 603 Padiamun, 603 Paired-end (PE) sequencing design, 304 Palaeoradiology, 149

1182 Paleogenetic(s), 318, 320–323, 325, 326, 328– 334, 337, 338, 340, 341, 344 collection approach, 322–324 Paleoimagers, 219 Paleoimaging, 213, 819 data collection, 214 Paleo-oncology, 133 Paleoparasitological analysis, 820 Paleoparasitological research, 1129 Paleoparasitology, 515, 521, 522, 528 experimental, 321 and paleogenetics, 321–322 Paleopathology, 132, 136, 137, 143, 934 change, 785 examination, 910, 915, 916 Paleoradiology, 213, 214 bioarchaeological context, 217 case and population imaging studies, 238–239 data archiving, 239 data collection, 242 data collection methods, 218 diagnosis by consensus, 244–245 digital volume tomography (DVT), 229 dual energy computed tomography (DECT), 226 endoscopic methodologies, 223–224 field imaging, 224–225 in field settings, 224–225 funerary and burial practices, 218 instrumentation accessibility, 241 interpretational challenges, 242–243 magnetic resonance imaging (MR), 230 MDCT, 226 medical versus bioarchaeological approach, 243 micro-CT technology, 228–229 multiplanar reformatting and 3D postprocessing, 227 opportunities, 245–248 teleradiology, 236 training challenges, 245 triage and ethical decision making, 236–238 value added data, 231–233 workflow design, 234–236 Palermo catacombs, 756 Palynological procedures, 537 Palynology Laboratory, 535 PaMinSA, 941 Papua New Guinea, 984, 986, 989, 1000, 1002, 1005–1007

Index Paracas peninsula, 24 Paragonimus westermani, 441, 443, 444, 449, 450, 452–453, 558 Parasites, in Han Dynasty flukes in, 524–525 pinworm (Enterobius vermicularis), 522–523 roundworm (Ascaris lumbricoides), 517–520 tapeworm (Taenia sp.), 520–522 whipworm (Trichuris trichiura), 523–524 Parasitism, 548, 549, 556–560 Parasitology Adenocephalus pacificus, 416 Ascaris lumbricoides, 418 destructive mummy autopsy, 413–414 Echinostoma, 419 fleas, 420 hookworms, 417 laboratory analysis, 414 lice, 419–420 parasite life cycle and nidality, 414–416 pinworms, 418–419 Trypanosoma cruzi, 420–421 whipworms, 417 Parrots, 646 Pathoecology, 412 Chinchorro, 421–423 Chiribaya, 426–428 of infectious diseases, 415 isolated mummy, 429–430 prehistoric human migrations, 431 Pazyryk mummies, 1015–1016 Peat bog characteristics of, 815 definition, 814 mummies, 834, 835, 845, 847, 848 Pennsylvania Museum, 5 Perimortem, 820 Permafrost mummies, 1013 Personal protective equipment (PPE), 323 Peru, 630, 938 Pet, 630 Petamenophis, 614 Pharonic Period, 6 Phase Contrast Imaging, 265 Phenotype, 388, 389, 392, 394, 395, 402–405 Philippine culture, 958 Phoenix Hill Tomb, excavation of, 1091–1093 Phoretic mites, 500 Photography, 213

Index Phytoliths, 536 Pigeons, 632 Pigmentation traits, 388 ancient samples, 402–404 DNA-based prediction, 388–401 historical figures and forensic identification cases, 399–402 Pilgrims, 633 Pinworm (Enterobius vermicularis), 522–523 Pinworms, 418–419 Pistacia resin, 656, 657, 705, 710 Plaques, 149 Plato, 879, 880 Pleistocene sub-fossil bones, 183 Polymerase chain reaction (PCR), 183, 262, 272, 273, 276, 280, 306, 319, 325–327, 352, 366, 794, 796 Population genetics of ancient genomes genetic relatedness and consanguinity, 373–376 population structure, 376–379 F4 statistics, 380–383 Post factum autopsy, see Autopsy Post-mortem chemical damages, 306–308 Postmortem interval (PMI), 499 Post-trimming read length filtering, 304 Potentilla, 539–541, 543 Poultry, 632 Pre-Columbian textiles, 1154 Predynastic, 568, 573, 589, 590, 592, 595 Prehistoric human migrations, 431 Preservation, 42, 45–49, 52–55 Principal component analysis (PCA), 376 Private collectors, 64, 67, 68 1986 Protection of Moveable Cultural Heritage Act, 98 Proto-Tocharians, 1044 Protozoa, 318, 319, 324, 339 Provenance, 68 Proxy, 64 Pseudo, 634 Pseudo-haploidization, 376 Pseudopathology, 886–887 Psychotropic plants, 935 Ptah, 632 Pthirus pubis, 419–420 Ptolemaic mummy, 656 Ptolemaic period, 568, 597, 606–611, 622 Pubic symphysis, 584, 585, 590, 591, 600, 621 Pulex irritans, 420 Putrefaction, 43

1183 Q Qilakitsoq mummies, 1013 Qin, 1119 qpAdm, 382, 383 Quality related issues, 361 Quantitative analysis, 537 Quantitative PCR (qPCR), 358–359 Quaternary period coprolites, 465 Queen Isitemkheb D, 631 R Ra, 569, 579 Radiocarbon, 590, 621 analysis, 96, 102 Radio-frequency (RF), 230 Radiography, 214, 634 Radiology, 113, 115, 116, 118, 120, 581 Rams, 633 Ramses I, 597 Random allele sampling approach, 312 Random sampling, 311 Ranefer, 593 Reconstructive Polymerization (RP), 327 Rehydration, 535 Reliquary, 909, 922, 924 Renaissance, 10 Repatriation, 65, 68, 81–83, 94–98, 103, 104 Julia Pastrana, 83 Saartjie Baartman, 83–84 Toi moko, 84–85 Resin(s), 574–576, 582, 583, 592, 593, 596– 604, 606–611, 613–616, 618, 620, 640 Rheumatoid arthritis (RA), 278 Rhind Magical Papyrus, 571 Rhizomes, 543 Richard III, 402 Rigor mortis, 916 Ritualistic sacrifice, 813 Ritual of Embalming, 571, 577 Roman period, 7, 568, 571, 579, 607, 611 Root hairs, 540 Rosaceae pollen grains, 541 Rosalia Lombardo mummy, 757 Roundworm (Ascaris lumbricoides), 517–520 Russia, 549 Russian immigrants, 556, 558 S Saartjie Baartman, 83–84 Sacred animal, 630, 632, 633, 636 mummies, 632

1184 Sacred Animal Necropolis (SAN), 638 Sadaebu, 1161 Safety measures, 937 Saltpeter Cave, 26 Saqqara, 592, 593, 610, 632 Scanning electron microscopy (SEM), 342 Scarab beetles, 634 Schistosoma japonicum, 524 Schistosomiasis, 31, 524 Schliemann, Heinrich, 880 Schmorl’s Nodes, 589 Schwerin, Wolgast, 764 Scientific techniques, 1129 Scythian mummies, 181 Second Intermediate Period, 6, 596–599 Segmentation, 581–583 Self-mummification, 1118 Senenmut, 631 Sensaos, 614 Seong-gye Yi, 1056 Seqenenre, 596 Sequential analyses, 198 Serra da Capivara National Park (PNSC), 951 Seth, 569 SHEP 1, 2 & 4, 396 SHEP 1 & 2, 394 Shi-Yeol Song, 1125 Short tandem repeat (STR) analysis, 1163 genotyping, 274 Shrunken heads, 69 Siberian mummies, 1012, 1118 anthropological study, 1014 frozen mummies, 1012–1014 geographic locations, 1013 mummies of Oglakhta, 1016–1017 preservation of, 1023–1027 scientific analyses, 1027–1028 from Ukok Plateau, 1014–1016 West Siberian Arctic Graves, 1018–1023 Yakut frozen bodies, 1017–1018 Siberian peoples, 549–557 Sicily Mummy Project, 757 Siculus, Diodorus, 8 Single base extension (SBE) assays, 394, 405 Single nucleotide polymorphism (SNP), 372, 388 DNA-based prediction, of pigmentation traits, 389–390 SNP-based phenotype prediction, in mummy studies, 404 Single-stranded library preparation, 364 Skin color prediction, 397–399

Index Skull analysis, 168 Small fragment, 363 Smallpox, 277 Small-sized PCR products, 361–362 Smith, Grafton Elliott, 8 Smoked Body mummification method body fluids, 1000 burial and funerary practices, 992 context and culture, 984 cultural aspects, 987–991 ethnographic aspects, 1005–1007 fire and smoke impact, 1001 miroclimates in burial cliff galleries, 1003 physical barrier, 1002 smoking hut, 1000 taphonomic change/impact, 1002 warrior culture of Anga, 994–996 Smudging, 73 Snipper, 394, 396, 403, 404 Social media, 78 Soft tissues, 280 Soil surfaces, 202 Soil-transmitted helminths (STHs), 334 Sokushinbutsu mummies, 1105, 1106, 1108 Solidification, 538 Sommersdorf mummies, 764 Song and Ming dynasties Chinese and Korean Mummies, 1120–1124 history of, 1119–1120 Korean Graves, origin of, 1124 and Mawangdui mummies, 1132 mummification process, 1129–1131 scientific studies of, 1127–1128 structure of, 1127–1128 Sons of Horus, 578–579 Sorel, A., 915–916 South American mummies, 24, 69, 645, 1157 anthropological collections, 932 bioarchaeological review, 933–934 chronic and metabolic diseases, 935 environmental studies, 934–935 infectious diseases, 933 pharmacological studies, 935 Southern Song, 1120 South Korea, 549, 557 Spatial relationship, 817 Spectroscopic methods, 657 Sphagnan, 816 Sphagnum, moss, 815 Spontaneous-enhanced mummification, 215 Spontaneous mummies, 214 Spontaneous mummification, 45–49, 215, 947 Spontaneous (natural) mummification, 808

Index St. Michan’s Church, 77 Stable isotope analysis, 102, 843 Stakeholder, 62 Standard pollen concentration, 537 Standard values (STD), 199 State Historic Preservation Division (SHPD), 97 F4 statistics, 380–383 Steneotarsonemus bancrofti, 508 Sternal end approach, 585 Stochastic effects, 357 Stockholm paradigm, 433–434 Stone chamber tomb, 1057 Storage, 941 Strangulation, 634 Strongyloides, 334 Strontium, 201, 202 STRUCTURE program, 378 Sulfur isotope ratio, 202 Surprising Chinchorro mummies, 938 Symbolic Space for Ethnic Harmony complex, 104 Synchrotron medical imaging (SMI), 231 Syphilis, 331, 332

T Taenia, 339 Taklamakan Desert, 1034, 1041, 1042 Tamut, 601, 610 Tang dynasty, 1119 Tapeworm, 520–522 Tarepit, 607 Tarim Basin graves and mummies identity of, 1043–1045 investigation of, 1040–1042 Tarim Basin mummies, 1119 Tarsonemidae, 507–509 Tashtyk culture, 1016–1017 Tattoo, 590, 592, 595, 598, 621 Tauber, A., 786 Taxonomic parameters, 342 Teleradiology, 236 Tell el-Amarna, 599 Temples, 645 Tenerife island, 887 Terahertz imaging, 259 Terebinth, 644 Tetisheri, 597 Textiles, 1154, 1155, 1157, 1159, 1160, 1164, 1166 Thebes/Theban, 572, 597, 600, 603, 606–608, 613–616, 618

1185 The Boston Journal of Philosophy and the Arts, 25 The Crusader, 77 Theodosius, 7 The Scientific Study of Mummies, 5, 1034 The Weerdinge Couple, 821–827 Third Intermediate Period, 7, 568, 573, 579, 599–606, 610, 618, 620, 622 Thoth, 633 Tjayasetimu, 600, 618 Transnasal method, 608 Trauma sequels, 733–736 Treaty of Alexandria, 19 Trematode infection, in Korean mummies, 442–444 Treponema pallidum, 331 Treponematoses, 327, 331–333 Trichomes, 538, 540 Trichostrongylus, 337, 341 Trichuris, 322, 337, 339, 340 T. trichiura, 324, 334, 338, 339, 417, 451 Trimethylsilylated TLE, 690, 692, 694, 696, 700 Triterpenoids, 657 Trypanosoma cruzi, 276, 321, 333, 420–421 Tsantsa, 69 Tsimane, 936 Tuberculosis (TB), 275–276, 318, 327–330 Tumour diagnosis, 134, 136, 139 in mummies/skeletonised mummies, 137–141 in paleopathology, 137 Tutankhamun, 578, 597, 632 Tutankhamun’s mummy, 91 Tuya, 598 Tyrophagus longior, 498

U Uhle, Max, 23 Ukok Plateau, 1014–1016 Ultrasound, 258 Ultrasound imaging, 259 UN troops, 534 “Unwrapping” mummies, 254 USSR, 852, 857, 858, 874 Utrecht, 891

V Valley of the Kings, 597, 598, 631 Valley of the Queens, 616

1186 Vác church and civic archives, 786–796 crypt, 780 crypt entrance, 782 discovery of crypt, 782–783 under dominant church, 780–782 Dominican church of, 780 history of, 779–780 Loretum, 781 ossuary, 782 vent holes, 782 Vác mummy project life histories (see Life histories) molecular analysis for tuberculosis, 794–797 mummification mechanism, 783–786 Vegetable tannin, 655 Venzone mummies, 762 VG studio max volume rendering, 582 Victual mummies, 644 Victual or food mummies, 631 Violence, 813 Virtual autopsies, 33 Visual inspection, 232 Vitamin D–folate hypothesis, 397 Votive mummies, 636

W Wackerstein mummies, 765 Wah, 595 Warring States; mummification in, 1074, 1097 Warring States Period, 515 Waty, 593 Wedjat, 598, 601, 606, 608, 610

Index Western Han Periods female mummy of, 1094 mummies of, 1074 mummification in, 1097 Western Siberian peat bogs, 835 See also Cossack mummy Wet cleaning, 1166 Wet embalming, 745 Whipworm (Trichuris trichiura), 417, 523–524 Whole genome sequencing (WGS) data, 403 Wildlife genomics, 294, 295 Wolfstein mummies, 764 Wrapping, 577, 581, 582, 592–595, 597, 598, 603, 608, 610, 613, 618, 620, 622

X Xiaohe cemetery, 1036–1038, 1040–1044 Xinjiang Archaeological Institute, 1038 Xinjiang Museum, 1045–1046 X-ray(s), 581–589, 617, 640

Y Yamalo-Nenets Autonomous Okrug (YaNAO), 1019 Yamnaya Culture, 1042 Yaral, 646 Yuya, 598

Z Zeleny Yar, 1027 Zhu Xi, 1124