Hope for the Giant Panda: Scientific Evidence and Conservation Practice 9811664773, 9789811664779

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Hope for the Giant Panda

Scientific Evidence and Conservation Practice WEI Fuwen

Science Press Beijing

Hope for the Giant Panda

Hope for the Giant Panda Scientific Evidence and Conservation Practice WEI Fuwen

Ecological Civilization–Building a Shared Future for All Life on Earth This book is dedicated to the 15th Meeting of the Conference of the Parties to the Convention on Biological Diversity Science Press Beijing

Fuwen Wei Key Lab of Animal Ecology and Conservation Biology Institute of Zoology, Chinese Academy of Sciences Beijing, China

ISBN 978-981-16-6477-9 ISBN 978-981-16-6478-6 (eBook) https://doi.org/10.1007/978-981-16-6478-6 © Science Press 2022 Jointly published with Science Press The print edition is not for sale in China (Mainland). Customers from China (Mainland) please order the print book from: Science Press. This work is subject to copyright. All rights are reserved by the Publishers, whether the whole or part of the material is concerned, specifically the rights of 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 publishers, 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 publishers nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publishers remain 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

Editorial Committee of the Series Consultants

HUANG Runqiu   Minister of the Ministry of Ecology and Environment of the People’s Republic of China HOU Jianguo   President of the Chinese Academy of Sciences

Directors

ZHAO Yingmin   Vice-Minister of the Ministry of Ecology and Environment of the People’s Republic of China ZHANG Yaping   Vice-President of the Chinese Academy of Sciences   Academician of the Chinese Academy of Sciences WEI Fuwen   Research Fellow of the Institute of Zoology, Chinese Academy of Sciences   Academician of the Chinese Academy of Sciences

Vice-Directors

CUI Shuhong   Director-General of Department of Nature and Ecology Conservation, Ministry of Ecology and Environment of the People’s Republic of China LIU Youbin   Director-General of Department of Communications and Education, Ministry of Ecology and Environment of the People’s Republic of China WEN Ya   Director-General of Bureau of Science and Technology for Development, Chinese Academy of Sciences LI Chunhong   Vice-President and Secretary-General of Chinese Society for Environmental Sciences

Members

SUN Ming   Deputy Director-General of Bureau of Science and Technology for Development, Chinese Academy of Sciences LIU Ning   Deputy Director-General of Department of Nature and Ecology Conservation, Ministry of Ecology and Environment of the People’s Republic of China   Negotiator for CBD COP15 Executive Committee LING Yue   Deputy Director-General of Department of Communications and Education, Ministry of Ecology and Environment of the People’s Republic of China HOU Xuesong   Deputy Secretary-General of Chinese Society for Environmental Sciences JING Xin   Director of Division of Biodiversity Conservation, Department of Nature and Ecology Conservation, Ministry of Ecology and Environment of the People’s Republic of China ZHOU Ju   Director of Division of Biotechnology, Bureau of Science and Technology for Development, Chinese Academy of Sciences ZENG Yan   Research Associate of Bureau of Science and Technology for Development, Chinese Academy of Sciences WANG Jing   Director of the Biology Branch of Science Press SHEN Jian   Vice General Manager of China Environment Publishing Group CHEN Yongmei   Director of Department of Science Popularization, Chinese Society for Environmental Sciences HU Feilong   Research Associate of Nanjing Institute of Environmental Sciences, Ministry of Ecology and Environment of the People’s Republic of China WAN Xialin   Deputy Section Chief of Foreign Environmental Cooperation Center, Ministry of Ecology and Environment of the People’s Republic of China

Foreword Ⅰ Giant panda, China’s “National Treasure”, has long been well-known to the public. As a “symbol of China’s openness to the world”, its innocent and endearing image has been disseminated all over the world. However, being a rare and endangered animal endemic to China, its survival and conservation have also garnered particular attention. The conservation of rare and endangered animals mainly focuses on habitat protection and captive breeding, but the origin of the species or their evolutionary histories are rarely explored. Every species has its own ancestors and descendants and is the product of adaptation to the Earth’s environment. Therefore, studying adaptation over evolutionary change is crucial for making appropriate conservation measures, and it is also one of the key components of conservation biology. The publication of Hope for the Giant Panda: Scientific Evidence and Conservation Practice provides us an opportunity to emphasize the importance of biodiversity conservation. The giant panda is a remarkable flagship and umbrella species and the research effort committed to its long-term survival unveiled it an ideal model to study unique adaptive evolutionary mechanisms. The giant panda has an 8-million-year evolutionary history, but humans have studied it scientifically for only 150 years. The author of this book is one of the most prominent scholars in the field of giant panda conservation research. He has been tracking the footprints of wild giant pandas in high mountains and dense forests for thirty years. Through applying the latest advanced technologies, he has shown us a completely different image of wild pandas from the languid state we have seen in the zoo. Besides being cute, giant pandas turn out to be very agile, intelligent and fierce. The establishment of giant panda molecular scatology techniques is one of the highlights of the author’s research achievements. Notably, he has integrated traditional ecological research with the rapidly developing fields of molecular bi-

ology and genomics, which elevated the scientific research of wild giant pandas to a new level. The significance of the book also lies in that it provides us a model for conservation biology research of endangered animals and corrects the false impression of giant pandas as weak species. Every species will inevitably experience birth and death, which only means the continuation of its pedigree rather than the end of evolution. This process is much longer than human history. The author’s research findings reveal that in the long history of evolution, giant pandas have made the greatest possible efforts to survive by continuously changing and adapting to new environments. It explains why giant pandas can survive while contemporary species were extinct due to the environment changes. At the same time, we also come to understand that the current endangerment of wild giant pandas is entirely caused by human activities. This may be thought-provoking: what should humans do? Why should we protect endangered species? This is because human beings as a biological species have also evolved in specific biotic and abiotic environments – protecting the environment is protecting ourselves. If we need to conserve a species, we must learn about its evolutionary history. Only by thoroughly understanding its ins and outs can we effectively put its conservation in practice. Given the rapidly evolving changes in the environment and consequences on endangered species, it is our mission to share this perspective with people outside the field of biology, and raise awareness on the status giant pandas are in. The author of this book has started his exploration. I would like to wish him successful and fruitful work!

Prof. CHEN Yiyu Evolutionary Biologist, Academician of the Chinese Academy of Sciences

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Foreword Ⅱ

Fuwen, is the youngest among my first postgraduates. At first sight, he has a small body frame like that of a teenager. However, I found him very active and refined in manners. Therefore, I call him Little WEI till today. In the 60 years of my research career, I have recruited more than 20 classes of postgraduates totaling over 100 students. Most of them worked on giant pandas in their postgraduate projects. However, only Little WEI continued carrying out comprehensive and in-depth research on giant pandas for over three decades after graduation. During these years, he has crossed mountains and rivers, braved wind and rain, and endured countless hardships. Finally, he has achieved significant innovative advancements in giant panda research. Giant pandas are rare and cryptic animals, and they ingest and defecate large amounts of bamboo. Their feces have a unique shape, with exfoliated intestinal epithelial cells found in the mucosa covering the surface, which acts as an exceptional material for ecological and evolutionary research. On this basis, Little WEI, together with his research team, pioneered the giant panda molecular scatology research using noninvasive sampling methods. He broke through the limitations of traditional techniques in ecological research and introduced advanced research methods including GPS collar tracking, infrared camera monitoring, molecular biology, genomics and metagenomics into his studies. Altogether, he has made a series of research breakthroughs in foraging strategies, spatial utilization, migration and dispersal, chemical communication, breeding and cub rearing, population history, adaptive evolu-

tion, genetic diversity, and evolutionary potential of the giant panda. These research achievements not only advance macroecology and conservation biology research of giant pandas to a new level but also provide an insight into conservation and management practice of this species, which represents an exemplary model for the research frontier of wildlife conservation biology in China. The book Hope for the Giant Panda: Scientific Evidence and Conservation Practice systematically introduces the history of giant pandas, their diet specialization and the underlying adaptative mechanisms, breeding strategies and dispersal patterns, population ecology, population genetics, and its evolutionary potential under environmental changes and human impacts. The content of this book stems from the first-hand scientific data and research findings obtained by years of hard work and continuous studies. In this book, readers will find well-structured informative content, expressed concisely and illustrated with vivid photographs. Time and time again you will find innovative ideas jumping “out of the box”. In my nineties, I am so glad to see Little WEI’s research outcomes assembled into a book and get published. Gratified by his success, I wrote this preface and hope Little WEI make further progress in his research!

Prof. HU Jinchu Zoologist, Professor of China West Normal University

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Foreword Ⅱ

Preface

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

  the campus of “Nanchong Teacher’s College”

In the scorching September of 1980, carrying a suitcase handmade by my elder brother, with my university admission notice and twenty Yuan RMB to support one month’s living expenses in my pocket, I stood sweating in front of the gate inscribed with the large red characters “Nanchong Teacher’s College” (now renamed as “China West Normal University”). I was sixteen years old that year, immature and thin, and my best hope for life was to become a teacher like my parents. The city Nanchong is located in the hilly area of eastern Sichuan, with the winding Jialing River passing through this famous “Silk City”. Inside the Yuping Park in the east of the city, there was a legendary “Ten Thousand Book Tower” where CHEN Shou, a Chinese historian of the Three Kingdoms period, wrote the book “Records of the Three Kingdoms”. My alma mater was on the northern edge of the city. Outside its narrow west gate were vast villages and farmlands. The campus enjoyed a tranquil environment with rows of tall and luxuriant camphor trees, while among the green vegetation were the small red buildings once served as offices for Mr. HU Yaobang, commissioner of North Sichuan Prefectural Administrative Office in the newly founded New China and later acted as the general secretary of the Chinese Communist Party during the 1980s. Next to Yaobangs former residence, clusters of blooming wintersweet sent forth fragrance under the cold moonlight.

Preface

  Prof. HU Jinchu

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 Prof. HU Jinchu (right) and WEI Fuwen (left) at the Denggouchi Church in Baoxing County

I majored in biology, and there is a well-known scientist in our department, Professor HU Jinchu. Prof. HU has lived in a humble abode all his life, but his footprints spread across the habitats of wild giant pandas. He is a pioneer of the wild giant panda research in China. It is my greatest fortune to be his student and to integrate giant panda research as an important part of my life. I still remember the first time I met Prof. HU. It was the early summer of 1984, with cicadas singing nonstop and loudly in the trees outside the window. On the table, the afternoon sun shone through the window and shone on books and a panda skull specimen. The room was stuffy; on the table, an old-fashioned fan was rotating while making cracking sounds. Prof. HU, with the spiky hair, looked a little thin yet hale and hearty. I was somewhat nervous and anxious about the upcoming graduate matriculation interview. However, Prof. HU’s eyes were full of kindness. He didn’t talk much but sent me several books before I left, telling me to study hard.

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From then on, the giant panda has come into my life and never left untill this day. In the animal world, the panda is undoubtedly the most charming star, with black-and-white pelage, thick body, full of comedic appeal. People’s love for pandas has already surpassed the limitations of regions and races. However, as a scientist, what fascinated me most is its unique biological characteristics. In its evolutionary history, many members of the “Ailuropoda-Stegodon Fauna”, which was extremely prolific in the Pleistocene, have been long extinct. Why can the giant panda survive until today and become a “living fossil”? The giant panda is a member of Carnivora and retains the characteristics of the ancestral carnivorous digestive system. However, how does it live on bamboo, a low-energy food, to survive and breed through all these years? Sometimes I ponder, as the giant panda has evolved for 8 million years with all the ups and downs, how lucky I am to encounter it in my life!

Preface

  field work and the observation station

xv

In the summer of 1984, I went to the Wanglang National Nature Reserve and the Tangjiahe National Nature Reserve with Prof. HU to participate in a field survey. This was the first time I set foot in panda habitat. My heart was full of curiosity and excitement, but this feeling was quickly worn away by the tedious and arduous fieldwork. The Wanglang and the Tangjiahe reserves are located at the southern foothill of the Minshan Mountains. The mountains are high and steep, covered with dense vegetation, almost inaccessible to humans. Prof. HU was nearly sixty years old, but he still took us trekking through the mountains and rivers, set line transects and habitat plots, identified animal traces, and taught us to read the “wordless book” of nature. Years later, I realized that Prof. HU was explaining to us the need to be diligent, rigorous, and tireless on the avenue of science, as well as to undertake the role and responsibility of a teacher. After graduation, I stayed in the Department of Biology of Nanchong Teacher’s College for scientific research and teaching as an assistant to my mentor. I assisted in training his graduate students and participated in the panda ecological study conducted at the “Baixiongping Observation Station” in the Tangjiahe National Nature Reserve. In 1991, I went to the Mabian Nature Reserve in the Liangshan Mountains with Prof. HU and established the “Mabian Dafengding Observation Station”, where my nose was wounded by a fallen tree, leaving a permanent scar.

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

 Prof. WANG Zuwang (right) and WEI Fuwen (left)

  WEI Fuwen and three supervisors

In 1994, at thirty years of age, I came to the Institute of Zoology, Chinese Academy of Sciences for my Ph.D. study and still work here on the conservation biology of wild pandas to date. My supervisors were Prof. WANG Zuwang and Prof. FENG Zuojian. Prof. WANG is serious and rigorous while Prof. FENG is easy-going and witty, from whose guidance and teaching I benefited tremendously in my life and academic career. During my Ph.D. study, I went to the Yele Nature Reserve in the Xiaoxiangling Mountains and established the “Yele Ecological Observation Station” for comparative studies on the nutritional ecology and metabolism of giant pandas and red pandas. Entering the Institute of Zoology, Chinese Academy of Sciences to study and work was a pivotal turning point for me, since it provided an important platform for my later research. In the 1990s, the Institute of Zoology was in the tide of science and technology system reform of the Chinese Academy of Sciences. The older generation of ecologists encouraged students to solve macroscopic scientific questions with microscopic perspectives, and helped set up experimental and technical platforms to combine macro- and micro-scientific research. During my tenure as the Secretary General of the China Zoological Society, Academician CHEN Yiyu, the president of the Society encouraged me to explore the ins and outs of giant pandas from the adaptive evolutionary perspective, which then became the main storyline of my subsequent research. Through active communication with the older generation of scientists, my vision has been greatly expanded. This not only helped me to achieve important progress in the combination of macro-micro research but also enhanced my scientific thinking, providing benefits for a lifetime. From these experiences, I developed the general approach throughout my career: using state-of-the-art techniques and methods to tackle key scientific questions for better protecting and conserving giant pandas.

Preface

In the early 1980s, after the large-scale flowering and die-offs of Fargesia scabrida, F. denudate and Bashania faberi in the Minshan Mountains and the Qionglai Mountains, the entire society was filled with pessimistic despair about the future of giant pandas. A highly impact book entitled The Last Panda (Schaller, 1993) written by Dr. George Schaller, the famous western naturalist and panda expert, in the 1990s, also greatly worried about the future fate of wild giant pandas after several years’ study collaborated with my supervisor, Prof. HU Jinchu in Wolong and Tangjiahe reserves.

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With the increasing depth and breadth of research, our understanding of the giant panda, a species with a long evolutionary history, has become more comprehensive, yet unsolved mysteries always inspired me to continue the exploration. The giant panda once coexisted with animals such as the stegodon and formed the important “Ailuropoda-Stegodon Fauna” in history. The stegodon has long disappeared while the panda survives until today. Has the panda really come to the end of its evolutionary path? What has happened to the panda during its long evolutionary history? Was it fate that pandas made the transition from carnivory or omnivory diet to herbivory, and eventually to a specialized bamboo diet How does the panda digest and use bamboo, a highly fibrous food resource? What evolutionary adaptations characterize their morphology, ecology, behavior, genetics and even genomics? What are the impacts of bamboo flowering and subsequent die-off, habitat loss and fragmentation on giant pandas? These puzzles constantly attract me and inspire me.

Preface

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Giant pandas, as the “bamboo forest hermits” that inhabit dense forests in high mountains, are difficult to observe. Most of the time, we can only describe and infer their lives by observing their traces, leaving the mechanisms underlying many of my research outputs unknown. Fortunately, the rapid development of science and technology, especially advancements and wide applications of molecular biology and genomics, provided the means to unveil the biological mysteries of giant pandas. In 1999, I traveled to Cardiff University and the Institute of Zoology, Zoological Society of London to study the theory and methods of conservation genetics with Professor Michael Bruford. To carry out research on the conservation genetics of giant pandas, it was urgent to establish a unique molecular marker system for this species. At the same time, it was imperative to develop a sampling method that would not harm the animal and could meet the research needs. I set my sights on the panda feces, which is easily available in the wild. Speaking of the feces, people usually think it is stinky and disgusting. In fact, since pandas feed on bamboo, their feces are not smelly but scented with the fragrance of bamboo leaves. The feces of giant pandas are spindle-shaped in dark-green color, easily recognizable in the wild. The surface of the fresh feces is often covered with a thin layer of transparent mucus containing epithelial cells shed from the intestinal wall. These cells proved to be invaluable sources of DNA for application to the conservation genetics of wild pandas.

Preface

  fecal samples of the wild pandas

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  collecting samples in the field

  observing the feeding behavior of wild giant pandas

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

After returning to the Institute of Zoology, Chinese Academy of Sciences from the United Kingdom, I soon started to carry out research on the molecular scatology of giant pandas with my graduate students and the help of my friend, Professor Bruford. The process of exploration was full of hardships. From DNA extraction, primer screening, to microsatellite amplification, we tested the experimental conditions over and over again and repeated every experimental procedure step by step. After 6 years of trials, in 2006, our research articles on the evolutionary potential and population census of wild pandas were subsequently accepted for publication in the journals Molecular Biology and Evolution and Current Biology, respectively. These two articles are milestones for our research work representing the establishment of the molecular scatology technical system, which lays a methodological foundation for follow-up investigations. With these techniques in hand, we can accurately identify panda individuals using fecal samples widely found in the wild without disturbing the pandas. On this basis, we conducted an in-depth analysis of the genetic diversity, population genetic structure, and gene flow of extant wild panda populations in different mountain ranges and for the first time, discovering that wild populations of giant pandas still maintained high genetic diversity and evolutionary potential. The molecular scatology technical system has henceforth solved the practical problems long hindering conservation genetics research. In the following years, we made profound advances on how panda gut microbiota help digest bamboo cellulose, as well as the landscape genetics and dispersal patterns of giant pandas. The corresponding results were published in the journal Proceedings of the National Academy of Sciences of the United States of America, Ecology, Molecular Ecology, Conservation Biology and so forth. Since 2008, I turned my attention to emerging genomics technology and spent five years using population genomics methods to investigate the population history, process and cause of giant pandas’ endangerment over the eight-million-year evolutionary history. We found that the endangered status of the giant panda was caused by paleoclimate change, subsequently exacerbated by human-mediated environmental change and habitat loss. Our findings were published in the journal Nature Genetics.   equipments for molecular experiments

Preface

  s everal cover stories published on journals

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Preface

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While we continue to deepen our research on genetics and genomics, we still stick to macroecological studies and have never ceased looking for new scientific questions. In 2004, with the strong support of the State Forestry Administration and the Shaanxi Forestry Department, we established the “Qinling Giant Panda Field Research Base” at Sanguanmiao in collaboration with Foping National Nature Reserve and Dr. Ronald Swaisgood from San Diego Zoo Global of the United States, and my graduate students have conducted long-term field research ever since. We successfully applied new technical tools (such as the GPS collars that can automatically drop off, infrared cameras and video cameras, doubly labeled water method that can measure metabolism, etc.) and research methods (such as nutritional geometry, ecological stoichiometry, and stable isotope ecology) in field research. As a result, we gained a new understanding of energy metabolism, foraging behavior, communication behavior, reproductive behavior, mating system, mate choice and parental care of wild pandas. The related results were published in several journals such as Functional Ecology, Animal Behavior, and Biological Conservation. What excited me the most is that today, after 20 years, I finally carried out the measurement of the energy metabolism of giant pandas that I wished to do during my Ph.D. study, collaborated with Prof. John Speakman. We reported that the energy metabolism of giant pandas was very low and further revealed the mechanisms of maintaining such low energy metabolism, with the results published in the journal Science. Over thirty years of research, we have gradually unveiled the mysteries of the wild giant panda and illuminated our understanding of its future. Our research shows reasons for optimism of the panda’s future and it will likely coexist with us for a long time under the watchful eyes and care of human beings. This book tells the little-known stories of wild pandas through concise text and many photographs taken in our field research. As a starting point, I hope the publication of this book could open up new vistas in wildlife scientific research and science-based conservation practice in China.

  field work

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Our research was funded by the National Natural Science Foundation of China, the Chinese Academy of Sciences, the State Forestry Administration, San Diego Zoo Global, and Chengdu Giant Panda Breeding Research Foundation, without which none of this work would have been possible. Thanks to the State Forestry Administration, Sichuan Forestry Department, Shaanxi Forestry Department, Gansu Forestry Department, many nature reserves and panda captive breeding centers for their support of our research; special thanks to the Foping Reserve, Wanglang Reserve, Tangjiahe Reserve, Mabian Dafengding Reserve, Yele Reserve and Liziping Reserve for their support and cooperation in setting up field observation stations and conducting long-term field survey! Thanks to all my collaborators and students who have followed me in the research for years. I would like to dedicate this book to my supervisors who led me into science, Prof. HU Jinchu, Prof. WANG Zuwang, and Prof. FENG Zuojian! Due to the limitation of my knowledge, inadequacies in the book are inevitable. Suggestions for improvement will be gratefully appreciated.

Prof. WEI Fuwen Conservation Biologist, Academician of the Chinese Academy of Sciences

Abstract The giant panda is a well-known charismatic species endemic to China, but its fame as an iconic endangered species extends globally, attracting the attention of scientists and the public alike. A long-held prevailing view is that the giant panda is reaching the end of its 8-million-year evolutionary history and is doomed to extinction. Is this true? This book, Hope for the Giant Panda: Scientific Evidence and Conservation Practice, is a systematic summary of the current state of knowledge on all aspects of giant pandas’ evolutionary history and unique biological and ecological characteristics. It reviews the latest evidence and progress on panda’s population history, adaptive mechanisms of bamboo diet specialization, reproductive ecology, population ecology, and population genetics, unveiled by technological breakthroughs such as genetics, genomics and metagenomics. Using these emerging approaches, the book aims to evaluate the cause of the giant panda’s endangerment and inform and guide management and conservation practices based on the best available science. In general, the endangerment status of the giant panda should be attributed to extrinsic threats such as human disturbance rather than any intrinsic biological shortcomings. Not doomed to extinction by maladaptive traits, the giant panda has future evolutionary potential yet, and is deserving of our conservation efforts. Although much has been invested in the panda’s conservation, these investments return more value not just for the survival of this unique and important species itself, but also providing important ecosystem services that humanity relies on. Formidable efforts by the government, scientists and the public have had great effect, and today the giant panda is no longer “Endangered” on the IUCN Red List, and has been downlisted to “Vulnerable.” With continued support, the giant panda has a bright future yet. By detailing a plethora of conservation research amassed over the last three decades, this book provides a foundation for conservation practitioners and policymakers to enact effective management strategies, while also providing unique insights into panda biology and natural history for the broader public.

Contents Chapter 1 Population History of the Giant Panda

/1

1   Giant panda genome

/9 / 10 / 13 / 17 / 20

2   Population fluctuations of the giant panda 3  Population genetic structure of the giant panda 4  Population divergences of the giant panda 5   Uniqueness of Qinling giant panda

Chapter 2 Adaptive Mechanisms of Dietary Specialization of Giant Panda

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1   Adaptations in morphological function 2   Adaptations in foraging strategies 3   Adaptations in digestive strategies 4   Adaptations in space use and activity rhythm 5   Adaptations in habitat utilization 6   Adaptations in physiology and metabolism 7   Adaptations in genetics

/ 32 / 35 / 40 / 43 / 44 / 46 / 48

Chapter 4 Population Ecology and Population Genetics of the Giant Panda / 81 1   Age determination of the giant panda

/ 86 / 89

2  Life table and population viability analysis (PVA) 3  Individual identification and population survey of the

giant panda 4   Sex identification of the giant panda 5   Genetic diversity of the giant panda

/ 90 / 99 / 100

Chapter 5 Major Threats to Giant Pandas and Conservation Practices / 105 1  Impact of large-scale bamboo flowering and hunting on

giant pandas / 108 2  Population collapse mechanism of isolated small panda populations / 110 3  Rescue small populations of the giant panda: reintroduction and habitat corridors / 114

Chapter 3 Reproductive Strategy and Dispersal Pattern of the Giant Panda / 53

Epilogue  Hope for the Giant Panda Reference

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1   Communication behaviors of the giant panda

Afterword Ⅰ

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Afterword ⅠⅠ

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Afterword ⅠⅠⅠ

/ 135

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2  Reproductive patterns and sex hormone levels of the 3 4 5 6

giant panda / 65  Reproductive behavior and sexual selection of the giant panda / 66   Parturition and cub rearing of the giant panda / 71   Reproductive rate of the giant panda / 76   Natal dispersal pattern of the giant panda / 78

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Chapter 1

Population History of the Giant Panda

© Science Press 2022 F. Wei, Hope for the Giant Panda, https://doi.org/10.1007/978-981-16-6478-6_1

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

(a) historical distribution

(b) current distribution QIN

MS

ng

li Jia

Minjiang

iver XXL

Chengdu

g

jian

Dadu R

distribution of fossil records

QIO DXL LS

distribution in 18th ~ 19th centuries current distribution

  the historical (a) and current distribution (b) of wild giant panda QIN, the Qinling Mountains; MS, the Minshan Mountains; QIO, the Qionglai Mountains; LS, the Liangshan Mountains; DXL, the Daxiangling Mountains; XXL, the Xiaoxiangling Mountains

The giant panda (Ailuropoda melanoleuca) has an evolutionary history of approximately 8 million years, and its direct ancestors can be traced back to the primal panda (Ailurarctos lufengensis and Ailurarctos yuanmouensis) in the late Miocene (Qiu and Qi, 1989). In the late Pliocene to early Pleistocene, the pygmy panda (Ailuropoda microta) appeared and then evolved into the baconi panda (Ailuropoda melanoleuca baconi) during the middle-late Pleistocene (Wang, 1974). The extant species (or subspecies) of the giant panda (A. melanoleuca) is believed to have arisen in the Holocene. From pygmy panda to baconi panda and finally to extant panda, their body sizes have undergone changes from small to large and then to small again. A traditional view based on these changes holds that the giant panda is on the way to an “evolutionary dead end” (Pei, 1965, 1974).

r

ive

R tze ng a Y

Xi'an

Chapter 1  Population History of the Giant Panda

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

  fossilised skull of the pygmy panda

  skull of the extant panda

Chapter 1  Population History of the Giant Panda

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

However, the evolutionary history of giant pandas, especially their population fluctuations and divergences and the underlying mechanisms have long remained mysterious. Scientists try to infer their population fluctuations using the number and distribution of fossils excavated; however, it is difficult to reveal the true population dynamics due to the limited fossil numbers discovered. With the development of genomics, scientists now have the opportunity to accurately uncover patterns of population fluctuations and divergences with extensive genomic information passed down from their ancestors. Based upon the giant panda whole genome sequencing project in which we were involved (Li et al., 2010), we conducted the population genomics study of giant pandas for the first time. In this study, we unveiled their effective population size (Ne) changes and population divergence events from the primal, pygmy, baconi pandas to the extant panda. Furthermore, we clarified that the Quaternary climate fluctuations were primary drivers of their population fluctuations and divergences, while human activities since the Last Glacial Maximum and Holocene were responsible for severe population collapses and endangerment of the species. These results indicated that the current endangered status of giant pandas was mainly caused by the paleoclimatic changes and recent anthropogenic activities (Zhao et al., 2013; Wu et al., 2014).

Chapter 1  Population History of the Giant Panda

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Chapter 1  Population History of the Giant Panda

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  Giant panda genome

Sequencing the genome is vital to reveal the genetic mechanisms of an animal’s evolution. Using nextgeneration sequencing (NGS) technology, we successfully sequenced and assembled the giant panda genome. The genome comprises of 42 chromosomes (2n), totaling ~ 2.4 Gb, with a 23,371 annotated protein-coding genes. The genomic heterozygosity rate is 1.35 × 10−3, which is higher than most other endangered species. Loss of function of the umami receptor gene T1R1 implies that pandas may not sense the umami taste associated with high-protein foods. The panda genome is the first reported de novo genome assembly of a large eukaryote using next generation sequencing technology, and therefore demonstrates the feasibility of rapid de novo assembly (Li et al., 2010; Hu et al., 2017a; Wei et al., 2020a). By integrating approaches including flow sorting, 10X Genomics Chromium sequencing and reference-assisted assembly, we present a chromosome-level giant panda genome with a total size of 2.29 Gb, and demonstrate that the combined strategies employed can be used to generate efficient chromosome-level genome assemblies for other species (Fan et al., 2019). To understand the evolution of ancient giant pandas from the last glacial maximum, we used ancient DNA capture techniques and sequenced the complete mitochondrial genome of an ~ 22,000-year-old specimen from Cizhutuo Cave, Leye County, Guangxi Province, where no wild pandas currently survive. We found that the mtDNA lineage of the Cizhutuo panda coalesced with present-day pandas approximately 183 thousand years ago, much earlier than the time to the most recent common ancestor of mtDNA lineages shared by present-day pandas (~ 72 kya). Our results reveal a new panda mtDNA lineage from that of present-day populations in history, which unfortunately became extinct (Albert MinShan et al., 2018).

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  Population fluctuations of the giant panda

Using the panda genome sequence, we extracted information of genomic heterozygous loci. Subsequently, we reconstructed their demographic history with the pairwise sequentially Markovian coalescent (PSMC) model and identified two population expansions and two bottlenecks. The first population expansion occurred 2 ~ 3 million years ago, when pygmy pandas emerged and thrived, while the second expansion occurred 30 ~ 40 thousand years ago with the flourishing of baconi pandas. During these two periods, the mass accumulation rate (MAR) of Chinese loess was at low levels, which indicated it was warm and wet at that time in China. Suitable climate conditions might be the primary drivers of the panda expansions. The panda population reached its pinnacle in the second expansion, which coincided with the warm climate during the Greatest Lake Period (30 ~ 40 thousand years ago). Sufficient habitat and abundant food resources at this time contributed to the ability of baconi pandas to thrive. The first population bottleneck occurred ~ 0.2 million years ago, while the second occurred ~ 20 thousand years ago. The high MAR of Chinese loess during these two bottlenecks represented the relatively cold and dry climates, which might be the main cause for the rapid population decline.

primal panda

A. lufengensis and A.yuanmouensis

baconi panda

pygmy panda A. microta

A. melanoleuca baconi

effective population size (×104)

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years before the present   population fluctuations of the giant panda

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Chapter 1  Population History of the Giant Panda

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 Population genetic structure of the giant panda

We carried out whole-genome resequencing of 34 wild giant pandas (~ 2% of the estimated wild panda population size) and identified approximately 13 million single nucleotide polymorphisms (SNPs) across the panda populations. We applied several methods to analyze the population SNP data, including STRUCTURE method on population structure, principal component analysis (PCA) and phylogenetic tree inference, all of which yielded a consistent genetic structure. These results indicated that the Qinling (Shaanxi) population was significantly differentiated from the Sichuan population, while among the Sichuan population, the Minshan population formed a distinct genetic cluster, and the population from the Qionglai Mountains, the Daxiangling Mountains, the Xiaoiangling Mountains, and the Liangshan Mountains formed a single genetic cluster. Therefore, the extant giant panda populations can be divided into three distinct genetic clusters – Qinling (QIN), Minshan (MIN) and Qionglai-DaxianglingXiaoxiangling-Liangshan (QXL).

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The population genetic structure based on whole-genome SNPs is similar to the result of our previous study using 10 microsatellite loci. We analyzed 115 individuals from the six mountain ranges using ten microsatellite loci via molecular scatology (non-invasive genetics) techniques. The result from STRUCTURE supported a population structure of four genetic clusters — Qinling, Minshan, Qionglai, and Xiangling-Liangshan (Zhang et al., 2007a). The slight difference between this result and that based on the SNP data is that the Xiangling-Liangshan population split out from former Qionglai-Daxiangling-Xiaoxiangling-Liangshan (QXL), most likely due to the small sample sizes of Xiangling and Liangshan populations in the whole-genome SNPs study. In future research, we will collect more samples from the Liangshan, Xiaoxiangling and Daxiangling Mountains for genome sequencing and conduct an in-depth investigation of population divergences.

Chapter 1  Population History of the Giant Panda

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Chapter 1  Population History of the Giant Panda

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  P opulation divergences of the giant panda

Through summary statistics of the SNP data, we obtained a Joint Site Frequency Spectrum (JSFS) of three panda populations, then analyzed the population divergence histories by population analysis software ∂a∂i. This software simulates SNPs frequencies in populations via diffusion approximation and generates a demographic history model most likely to satisfy the existing JSFS distribution under the given population mutation rate. The population divergence history model showed that the QIN and non-QIN (Sichuan) populations diverged ~ 0.3 million years ago, corresponding with the onset of the Penultimate Glaciation (0.13 ~ 0.30 million years ago), whereas the non-QIN cluster diverged into the MIN and QXL populations ~ 2,700 years ago, likely resulting from anthropogenic activities that fragmented habitat and reduced connectivity. The genetic uniqueness of the QIN population is presumably due to the population divergence event of approximately 0.3 million years ago and later habitat and genetic isolation caused by anthropogenic activities in this region.

Ne QIN

MIN

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38 879

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QXLXXL-

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years before the present   population divergences of the giant panda

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

Revealing the population history of the giant panda helps understand the causes and processes of its endangerment, and provides scientific evidence for effective conservation and management. Firstly, the QIN population is unique mainly due to its distinct population history, hence, policy makers should consider it as a separate management unit and inhibit anthropogenic gene flow between QIN and other populations. Secondly, anthropogenic gene flow among the three genetic populations should be avoided. Lastly, when developing strategies of captive breeding and reintroduction, decision makers should also take the genetic populations into account and choose individuals with similar genetic backgrounds for reintroduction into the target wild populations.

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 U niqueness of Qinling giant panda

The uniqueness of Qinling giant pandas also lies in a rare pelage phenotype — brown and white (known as the brown giant panda), which is clearly distinguished from the typical black and white pelage of giant pandas. Up to now, there are only three cases of brown giant pandas with physical or photographic evidence; together with all other claimed observations there are at least eight cases. The first brown panda, Dandan (female), was found in the Foping National Nature Reserve in 1985 and rescued to Xi’an Zoo where it lived for 15 years. She gave birth to a male cub (QinQin) with normal pelage color. Dandan died in 2000, and specimen is kept in the Museum of Foping Reserve. The second brown panda was captured by a research team led by Professor Pan Wenshi of Peking University in the Changqing National Nature Reserve adjacent to the Foping Reserve in 1992. They put a radio-telemetry collar on it and released it back to the wild for daily scientific tracking (Pan et al., 2001). The third brown panda, Qizai (male), was found in our research area in the Foping Reserve in 2009, and recognized as the descendant of a collared panda as part of our long-term tracking program. Qizai was later rescued to the Shaanxi (Louguantai) Rescue and Breeding Center for Rare Wildlife and now lives in captivity. Considering the rarity of this special pelage phenotype, we are using genomics methods to reveal the mystery of its genetic mechanisms.

 Qizai

Chapter 1  Population History of the Giant Panda

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  Qizai (left)

  baby Qizai

 Dandan

Chapter 2

Adaptive Mechanisms of Dietary Specialization of Giant Panda

© Science Press 2022 F. Wei, Hope for the Giant Panda, https://doi.org/10.1007/978-981-16-6478-6_2

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The giant panda belongs to the Order Carnivora and has a digestive system typical of carnivores. Compared with typical herbivores, it has neither a large stomach nor cecum for microbial fermentation, nor cellulase enzyme to decompose celluloses. However, the giant panda has evolved to live on a specialized bamboo diet over its long evolutionary history, which provides an ideal model to explore the adaptive evolutionary mechanisms of dietary specialization.

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  Bashania fargesii

  Fargesia qinlingensis

  Fargesia rufa

Chapter 2  Adaptive Mechanisms of Dietary Specialization of Giant Panda

  Fargesia denudate

  Chimonobambusa szechuanensis

  Yushania mabianensis

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Bamboo belongs to the subfamily Bambusoideae of the family Poaceae with wide distribution and high species diversity. More than 50 bamboo species are known to be edible for giant pandas, among which about 20 species are preferred. Giant pandas are distributed across six mountain ranges from southern to northern China where the number of bamboo species varies greatly, resulting in significant regional differences in the composition of bamboo species that giant pandas live on. In the Qinling Mountains, giant pandas mostly feed on Bashania fargesii and Fargesia qinlingensis, but their conspecifics in the Minshan Mountains mainly use F. scabrida, F. denudate and F. nitida. In the Qionglai Mountains, pandas’ primary food sources are B. faberi and F. robusta and in the Xiangling Mountains, they are B. spanostachya and Yushania lineolata. In the Liangshan Mountains, their preferred bamboos are Qiongzhuea macrophylla, Y. glauca and Y. mabianensis (Hu and Wei, 2004; Wei et al., 2011).

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As the exclusive food source for the giant panda, bamboo comprises 70% ~ 80% cellulose, hemicellulose and lignin, while protein, fat and soluble carbohydrate only account for 20% ~ 30%. Bamboo is a low nutrition and low energy food. Some authors have argued that this dietary specialization was a key driver of population decline leading to endangerment of giant pandas. Our research on this topic sheds light on this viewpoint. When do giant pandas shift their diet to bamboo? How do they live on the so-called low-quality bamboo? Have they adapted to bamboo diet during the process of dietary specialization? Why has the panda still retained a digestive system with gut morphology and digestive enzymes similar to carnivores rather than herbivores? Does dietary specialization indeed cause their population decline and endangerment? Our long-term research revealed that the giant pandas have evolved multilevel adaptations to bamboo diet during their evolutionary history, including morphological adaptations, ecological and behavioral adaptations in foraging strategies, digestive strategies, nutritional utilization, activity rhythm and habitat use, physiological adaptations in energy metabolism and hormone regulation, and genetic adaptations such as the changes in umami taste receptor genes. In addition, bamboo resources are abundant in their natural habitat. Therefore, despite the minimal evolutionary changes in gut morphology, pandas are exquisitely adapted to “making a living” on bamboo, and their specialized bamboo diet was not a crucial factor causing decline and endangerment.

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Stable isotope analysis is widely used to understand dietary changes in animals. Carbon and nitrogen isotopic values in bone collagen can reflect the dietary composition in ancient animals. Isotopic signals can also reveal dietary changes between modern and historical populations (Boecklen et al., 2011). Previous evidence suggested that pandas switched to bamboo feeding about two million years ago, however, the first detailed description of their bamboo diet is only a few hundred years old. In order to address their evolution of dietary preferences and habitat change, we measured and compared stable carbon (13C/12C) and nitrogen (15N/14N) isotope ratios in bone collagen, and carbon (13C/12C) and oxygen (18O/16O) isotopes in tooth enamel from extinct and extant pandas and contemporary sympatric carnivorous and herbivorous mammals. We show that giant pandas have had a diet dominated by C3 resources over time and space. Carbon and nitrogen isotope ratios in bone collagen from ancient and modern pandas and sympatric species, indicate that the trophic niches of ancient and modern pandas are distinctly different. We find that the isotopic trophic and ecological niche widths of ancient pandas are approximately three times larger than those of modern pandas and infer that ancient pandas hada more complicated food structure and habitat type than their modern counterparts. During diet specialization, we propose that this genus underwent two major dietary shifts, the first was completed before the Pleistocene from carnivory or omnivory to herbivory, and the second from herbivory to a 99% bamboo diet, unfinished as at the mid-Holocene (Han et al., 2016, 2019). We are much surprised and curious about this interesting finding because it is difficult to be understood why it looks like this. In the future, we need to garther more panda fossil samples to further confirm this finding.

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  Adaptations in morphological function

Bamboo is a lignified plant with high concentrations of lignin and cellulose. How can the giant panda easily grip, bite off and chew the tough stems of bamboos? Previous research found that the forepaw of giant pandas evolved a pseudothumb, an enlarged radial sesamoid that facilitates pandas to grasp bamboo between the pseudothumb and the opposing palm. Besides, pandas have an enlarged zygomatic arch to accommodate strong masseter muscle attachment, and a complex cuspidate molar masticating surface with many dental facets that help to crush refractory fibers in bamboo. By comparing skulls of hypercarnivores (tiger and leopard), omnivorous carnivore (black bear), and herbivorous carnivores (giant and red pandas), we find that the skull of giant pandas had evolved functional adaptations for bamboo eating (Wei et al., 1990; Zhang et al., 2007b).

pseudothumb

1 cm

  the pseudothumb of panda's forepaw

  panda's hindpaw

Chapter 2  Adaptive Mechanisms of Dietary Specialization of Giant Panda

  the skull and teeth of the giant panda

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  the feeding site of the giant panda

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  Adaptations in foraging strategies

What foraging strategies do pandas employ to meet their nutritional and energy needs from the “poor-quality” bamboo food? Since the 1980s, our studies conducted in different mountain ranges have documented that giant pandas adopt an optimal foraging strategy to maximize nutrition and energy from the bamboo diet. Bamboo resource in panda habitats is so abundant that it is unlikely to be a limiting factor for population growth of giant pandas (He et al., 2000a, 2000b; Wei et al., 1996a, 1997a, 1999a, 2011). The main foraging strategies of pandas are summarized as follows: ingest large amounts of bamboos, the daily consumption of fresh bamboo can reach 10 ~ 20 kg of bamboo leaves and stems, as much as over 30 kg of shoots, therefore, pandas can maximize the intake of nutrition and energy from bamboo ingested; choose the most nutritious bamboo species, for example, in the Mabian Dafengding National Nature Reserve, giant pandas prefer nutritious Q. macrophylla and Y. glauca rather than Chimonobambusa pachystachys; select the most nutritious bamboo tissues – shoots and leaves, among different bamboo tissues, giant pandas favor the most nutritious and fresh bamboo shoots and leaves; select moderate size and young bamboo stems, for the bamboo stems of the same species, pandas usually prefer young stems of a medium size with higher nutritional value than older stems; switch bamboo diets seasonally to achieve nutritional balance, pandas often switch food seasonally between different bamboo species and different bamboo tissues (such as bamboo shoots, bamboo leaves, and bamboo stems) to meet nutritional and energy needs, and hence achieve nutritional balance. Our research shows that in the Qinling Mountains, giant pandas undertake a seasonal elevational migration to access B. fargesii at low altitude and F. qinlingensis at high altitude at times of year when these species are most nutritious. Using nutritional geometry and ecological stoichiometry approaches (Simpson and Raubenheimer, 2012; Sterner and Elser, 2002), we find that this foraging pattern guarantees a balanced intake of important nutrients such as nitrogen, phosphorus, and calcium which have different contents in different bamboo species and tissues. Combined with studies on panda reproductive ecology, we show that pandas utilize two bamboo shoot periods, with differing characteristics to meet their nutritional needs. During pregnancy pandas utilize bamboo with high nitrogen and phosphorus contents and concentrate foraging on new leaves during the early period of lactation after cubs are born. This seasonal food switch is thus of great significance to panda reproduction (Nie et al., 2015a). Food resources are patchily distributed in the environment and carnivores and

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herbivores have adopted different foraging strategies to maximize feeding efficiency. We used global positioning system collars to track wild giant pandas at high resolution (50%) and lower carbohydrate content than any other herbivore. We also show that the composition of panda milk is quite similar to that of carnivores, indicating nutritional similarity of panda diets with carnivores. These findings suggest that the evolutionary transition from carnivory to a bamboo diet was not nearly as harsh as it might be, because two diets are very similar in major nutritional components (macronutrients). It also sheds new light on why giant pandas have retained a digestive system resembling that of carnivores. Our study provides the first empirical illustration of how multilevel diet niche theory can help to solve fundamental challenges in nutritional ecology. It demonstrates that a species that is an extreme specialist herbivore at the level of the foods it eats, can at the level of macronutrition be adapted to a diet typical of higher trophic levels (Nie et al., 2019).

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  a gaint panda is eating bamboo leaves

  a gaint panda is eating bamboo shoot

  a gaint panda is eating bamboo pole

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 Adaptations in digestive strategies

Although giant pandas ingest 10 ~ 20 kg of fresh bamboo each day, they can only digest a small part (~ 17% of dry matter). The gut passage time of the bamboo is relatively short: approximately 8 ~ 9 hours (Schaller et al., 1985; Hu, 2001). Our study found that giant pandas could maximize nutrition and energy intake from bamboo through an optimal digestive strategy by ingesting as much bamboo as possible, with rapid passage through the digestive tract, and frequent defecation (Wei et al., 1999a). Previous research revealed that giant pandas digested 80% ~ 90% of crude protein and crude fat in bamboo, as well as some cellulose (8%) and hemicellulose (27%) (Dierenfeld et al., 1982). However, we did not identify any gene encoding celluloseor hemicellulose-digesting enzymes in the giant panda genome (Li et al., 2010). Thus, how pandas digest and utilize cellulose and hemicellulose of bamboo has long remained a mystery.To address this question, we conducted the 16S rRNA gene sequencing of panda gut microbiomes, and found cellulose- and hemicellulose-digesting Clostridium bacteria. Further metagenome analysis identified the genes encoding cellulase, beta-glucosidase, xylan 1,4-beta-xylosidase and 1,4-β-xylanase in panda gut microbiota, whose protein products could decompose cellulose and hemicellulose into glucose. These results reveal the mystery of how symbiotic gut microbes of giant pandas help to digest cellulose and hemicellulose in bamboo diet (Zhu et al., 2011a; Wei et al., 2015a, 2015b).In addition, we investigated the effects of seasonal nutrient variation on the composition and function of panda gut microbiomes by nutritional and metagenomic analysis. Our results show significant differences in gut microbiome abundance between shoot-feeding and leaf-feeding periods. Metagenomic functional analysis also indicated that the gut microbiomes of pandas helped to enhance crude fiber utilization efficiency during the nutrient-deficient leaf period, whereas they expanded their functional capacity to reinforce the utilization of crude protein in the protein-rich shoot period (Wu et al., 2017). The gut microbiome plays a critical role in wildlife health, nutrition and physiology, and metagenomic analysis provides a valuable tool to identify communities. Based on our research, for the first time, we propose a new concept of conservation metagenomics, a subdiscipline of conservation biology (Wei et al., 2019a). It aims to understand the roles of the microbiota in evolution and conservation of endangered animals, and address main scientific issues related to host evolution, physiology, nutrition, ecology and conservation to improve management of wildlife.

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 A daptations in space use and activity rhythm

We used global positioning system collars to automatically record location data of free-ranging giant pandas to study their activity patterns and space use. The data we obtained show their home ranges were generally smaller than those of other carnivores, and activities within home ranges were also relatively stable. Giant pandas rarely make long-distance movement, instead they usually spend days in the same feeding patch, typically sleeping at the feeding sites or under a large nearby tree. The time spent on resting can reach 12 hours each day, and a single rest is approximately 2 ~ 3 hours. We concluded that the small home range and large proportion of time spent on resting may effectively contribute to energy saving, which helps pandas adapt to low-energy bamboo diet (Zhang et al., 2014).

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Hope for the Giant Panda: Scientific Evidence and Conservation Practice

 Adaptations in habitat utilization

Suitable habitats provide pandas with a stable food supply and living environment. We applied landscape ecology methods to study habitat selection and utilization of giant pandas from smallscale microhabitats to large-scale landscape patterns, and found that giant pandas preferred open primary forests with gentle slope and moderate bamboo density. The preference for gentle slopes likely contributes to energy conservation for adapting to the low-nutrition/energy bamboo diet. The identification of the panda’s preference for old-growth forest highlighted its importance for habitat protection and helped guide management decisions and forestry policy (Wei et al., 1996b, 1999b, 2000; Feng et al., 2009; Zhang et al., 2004a, 2006a, 2009, 2011a; Qi et al., 2009, 2011, 2012a, 2012b). To further reveal the gender differences of giant pandas in habitat selection, we used a combination of field surveys, sex identification through fecal DNA and ecological niche factor analysis modelling. We found that both males and females preferred areas at high altitudes with high forest cover. However, significant sex differences in habitat selection were also observed. Habitat preferences of females are more restrictive than those of males, and females have a stronger association with high altitude conifer forest and mixed forest. The more restricted habitat preference of females could be explained by their needs for dens for giving birth to their cubs. Therefore, effective conservation and management strategies should consider these differences in habitat selection of females and males (Qi et al., 2011).

  bamboo forest with low vegetation density

Chapter 2  Adaptive Mechanisms of Dietary Specialization of Giant Panda

  bamboo forest with moderate vegetation density

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  bamboo forest with high vegetation density

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 Adaptations in physiology and metabolism

For a long time, we speculated that the giant panda maintains a low metabolic rate to achieve a daily energy balance as a bamboo specialist. However, no one ever measured the metabolic rates of wild pandas. To address this knowledge gap, we adopted a method using doubly labeled water (Speakman, 1997) to measure the daily energy expenditure of captive and free-ranging pandas and found an exceptionally low daily energy expenditure (Nie et al., 2015b). Measurements of daily energy expenditure across five captive and three wild pandas averaged 5.2 megajoules (MJ)/day, only 37.7% of the predicted value (13.8 MJ/day) for mammals of the same weight. It is generally believed that the metabolic rate of humans is low. But the metabolic rate of a panda with a body weight of 90 kg is even less than 50% that of a human with the same weight. More importantly, we revealed the mechanisms that enable giant pandas to maintain such a low metabolism. A combination of

morphological, behavioral, physiological and genetic adaptations, leads to low energy expenditure, and enables them to survive on a bamboo diet. Morphologically, sizes of energy-consuming organs related to resting metabolic rate, such as brains, livers, and kidneys, are relatively reduced by 20% ~ 30% compared with other mammals of the same weight. If you have ever been to a zoo to see pandas, you must have noticed that they are usually lazy and do not move much. The GPS collar tracking data indicated that wild pandas spent about half of the day resting and moved only about 27 meters per hour on average, both of which helped save a large amount of energy. Further, the secret of low metabolism also lies in the low levels of thyroid hormones, thyroxine (T4) and triiodothyronine (T3), which regulate the resting metabolic rate. Our measurements of T4 and T3 were lower than those of hibernating black bears. A unique mutation found in the panda dual oxidase 2 (DUOX2) gene critical for thyroid hormone synthesis might

  thermal images of giant pandas

Chapter 2  Adaptive Mechanisms of Dietary Specialization of Giant Panda

explain the low thyroid hormone level in giant pandas. DUOX2 encodes a transmembrane protein that catalyzes the conversion of water to hydrogen peroxide, which is used in the final step of T4 and T3 synthesis. DUOX2 gene of giant pandas contains a single substitution of C to T in the 16th exon, which results in a premature stop codon (TGA). This mutation results in loss of its gene function. Loss-of-function mutations in DUOX2 could lead to hypothyroidism in humans and mice. A challenge posed by low metabolism is how to sustain a

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stable body temperature. Our research showed that pandas had a deep pelage able to trap their body heat. Thermal imaging measurements suggested the body surface temperatures of giant pandas were significantly lower than those of other black-and-white animals such as zebras and spotty dogs exposed to the same environmental temperature. Therefore, maintaining the low energy expenditure enables pandas to survive on bamboo diet with a high fiber, representing profound physiological adaptation (Nie et al., 2015b).

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 Adaptations in genetics

The giant panda genome sequencing project reported the pseudogenization of umami taste receptor gene Tas1r1 in panda genome. This might hinder pandas from sensing umami-rich chemicals such as proteins, demonstrating a case of genetic adaptation to the bamboo diet (Li et al., 2010). Furthermore, by comparing the taste receptor genes in different panda populations via population genomics methods, we found that two nonsynonymous sites, A52V and Q296H of the bitter taste receptor genes Tas2r49 in the Qinling population were under positive selection compared with the Sichuan population. This could be explained by the fact that, compared to the Sichuan population, pandas in the Qinling population consume more bamboo leaves, which contain more bitter components than bamboo stems. These results revealed the evolutionary local adaptation to the differences in bamboo nutrients among different mountains (Zhao et al., 2013).

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We further used functional expression in engineered cells to detect agonists of pTAS2R20 and checked the perception differences in the in vitro responses of pTAS2R20 variants to agonists. We find that pTAS2R20 is specifically activated by quercitrin and the receptor variant with A52V and Q296H found in Qinling pandas confers a significantly decreased sensitivity to quercitrin. We quantified contents of quercitrin in bamboo leaves from the Qinling Mountains and found that they were significantly higher than those of bamboo leaves from other mountain ranges. The decreased sensitivity to quercitrin in Qinling pandas results in higher-quercitrin-containing bamboo leaves tasting less bitter to them and thus makes them more palatable. Our findings confirm the local genetic adaptation of Qinling pandas to their habitats based on the functional experiment analysis. (Hu et al., 2020a). We also investigated the genetic mechanism of adaptive convergent evolution of the giant panda and the red panda (we recently identified two red panda species, the Himalayan red panda and Chinese red panda, Hu et al., 2020b) by comparative genomics methods. The giant and red pandas belong to families Ursidae and Ailuridae within the superfamily Musteloidea, respectively, with an estimated divergence time of approximately 43 million years ago. Although the giant and red pandas are phylogenetically distant, they have evolved specialized bamboo diets and adaptive pseudothumbs for handling bamboo, representing a classic model of convergent evolution. Through de novo sequencing the red panda genome and improving the giant panda genome assembly, we carried out comparative genomics analyses and identified genes under adaptive convergent evolution potentially involved with their bamboo diet and pseudothumb development. Amongst these genes, limb development genes DYNC2H1 and PCNT are important candidates for pseudothumb development, and genes involved in the digestion and utilization of bamboo nutrients such as essential amino acids, fatty acids, and vitamins are under adaptive convergent evolution. Meanwhile, we also found that the umami taste receptor gene TAS1R1 was convergently pseudogenized in both pandas, which resulted in the failure to recognize meat. These findings present interesting genetic convergence events driven by the specialized bamboo diet (Hu et al., 2017a).

Chapter 3

Reproductive Strategy and Dispersal Pattern of the Giant Panda

© Science Press 2022 F. Wei, Hope for the Giant Panda, https://doi.org/10.1007/978-981-16-6478-6_3

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Reproduction, an important part of the animal life cycle, is fundamental for population growth and species persistence. In the 1980s, due to the difficulty of captive breeding of giant pandas, it was generally believed that its endangerment was closely related to their poor reproductive capacity. However, is it the case for wild giant pandas? Wild giant pandas live in dense bamboo forests in high mountains and remain solitary outside the mating season. During the mating season, how do male and female pandas communicate with each other and how do they choose their mates? Do males win mating opportunities through fighting and how is the winner determined? Is their reproductive rate really low? How do female pandas rear their cubs? How long will the cubs live with the mother before they disperse? Which sex disperses, males or females? Answering these questions on reproductive strategies and dispersal patterns is of great significance for captive breeding and reintroduction of giant pandas.

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With the development and application of new technologies and methods such as high-frequency GPS/VHF radio collars, infrared camera trapping, and non-invasive genetics by fecal DNA analysis, we can now reveal the reproductive strategies and dispersal patterns of the giant panda from behavioral, physiological and genetic aspects at both individual and population levels.

  infrared surveillance systems

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  infrared surveillance systems

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  Communication behaviors of the giant panda

Giant pandas are typical solitary animals and rarely come in direct contact with each other. As they are adapted to living in dense, dark bamboo forests, their vision is poorly developed. Instead, they communicate with conspecifics mainly via olfactory and auditory cues, particularly via scent marks.

  marking behavior of giant pandas

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(1) Chemical communication Chemical communication is a dominant mode of communication which has several advantages. Chemosignals are long-lasting and can be broadcast over a wide area. Chemosignals allow communication among individuals without direct visual contact, and can take place over large distances and time spans. Many chemical signals originated from urine and specialized scent gland secretions play an important role in intraspecific communication of many mammals (Swaisgood et al., 1999; Smith and Harper, 2003). Giant pandas can exchange chemical information through two types of scent markings: anogenital gland secretions and urine marking. Pandas do not deploy scent marks randomly in the environment, they mainly use tree trunks with specific characteristics but occasionally use rocks or the ground. Our research found a significant selectivity for characteristics of marked trees when pandas deposit different scent marks. For anogenital gland secretions, pandas favor trees with rough bark and moderate diameter at breast height, but for urine marking, trees with larger diameter at breast height, rougher surface or more moss-covered are preferred. This selectivity helps increase chemical signal persistence and/or the size of the odor field in the environment, enhancing the likelihood of detection by conspecifics (Nie et al., 2012a). We also found that several chemical constituents (especially volatile compounds) of anogenital gland secretions (AGS) were only pre-

  scent mark of giant pandas

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  sent marking behaviors

sented in the mating season, whereas nonvolatile compounds were lower in the mating season. The seasonal variations in chemical composition of AGS would play an important role in governing the reproduction of giant pandas, such as mate location, attraction, and male-male competition (Zhou et al., 2019). Further research also found significant seasonal and sex differences in the scent marking frequency. The scent marking peak occurs in the mating season, indicating its important role in reproductive activities. Female pandas mark scent trees the most in the mating season to advertise their reproductive status and attract more males to participate in the mating competition (Nie et al., 2012a).

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(2) Vocal communication Giant pandas are solitary for most of the year, and only come together for mating purposes during the mating season, at which time most vocal communications occur. We have collected data on the vocal behaviors of giant pandas during the mating season for several years. The results of this long-term investigation revealed that the vocal communications between male-male or female-male play important roles in advertising individuals’ competitiveness, exchanging reproduction-related information and synchronizing reproductive status (unpublished data).

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  the GPS radio collar does not affect the moving, mating, littering and parental behaviors of pandas

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 Reproductive patterns and sex hormone levels of the giant panda

In the Foping Reserve in the Qinling Mountains, we found an obvious seasonal reproductive pattern of the giant panda. These findings were the result of multiyear observations using GPS/VHF radio-collared individuals and noninvasive endocrine assays of sex hormones in fresh feces. Mating happens from March to April each year when females experience a single estrus period. They advertise their reproductive status via scent marks and vocal communications, which generally attract 3 to 5 males that congregate around the female to engage in reproductive competition (Nie et al., 2012b). In contrast, males participate in reproductive activities several times during one mating season. Males encountered multiple potential mates and competed for reproductive access to females. The male testosterone levels showed significant seasonal fluctuations, with the highest taking place during the mating season. However, males could not maintain elevated levels of testosterone throughout the mating season; instead, it peaked during encounters with potential mates. Maintaining a high level of testosterone is metabolically expensive, while male pandas enter the mating season during a period of low food availability in spring. Taking these factors into account, we speculate that reproductive activities of pandas may be energy-limited, and the pattern of testosterone fluctuations may be a reproductive strategy to adapt to the energy constraints (Nie et al., 2012b, 2012c).

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 Reproductive behavior and sexual selection of the giant panda

We have been continuously tracking and observing the reproductive activities of wild pandas for many years and found that the estrus period of female pandas can last from several days to over ten days. During the estrus period, females spent most of the time squatting or lying on a tree with several males (from 2 to 7) aggregating under the tree. Males competed for access to estrous females through fighting, which was the main feature of their reproductive activities (Yong et al., 2004; Nie et al., 2012b).

 the female chooses the mating site and waits for the male winner

 the male follows the female’s scent

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 males compete for the mating opportunity

The number of males involved in mating competition may increase during female’s estrus period and their competitive rank will change through time. Males that temporarily win the competition often show typical mate guarding behaviors. Positioned under the tree containing the estrous female, the male monitors the female and prevents her escape and blocks access by other males. Only the male who wins the final competition has the opportunity to mate with the female. By analyzing the relationship between male body size, male hormone levels, and the results of competition, we concluded that the competitive outcome was generally determined by male body size. There was no significant correlation between competition results and level of androgens. Larger males were often more aggressive in competition and ultimately earned the opportunity to mate with the females (Nie et al., 2012c).

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Sexual selection is a hot topic in ecology and evolutionary biology. Two types of sexual selection, male-male competition and female mate choice, are the most common in nature. With the rapid development of population genetics and molecular biology, evolutionary biologists interested in sexual selection not only study the phenotype of animals but also go deep into the underlying genetic basis. More and more studies reveal that genetic factors play a key role in sexual selection. In an 11-year study on the reproductive strategies of wild pandas in the Qinling Mountains, we combined reproductive behavior observation and molecular scatology methods to analyze the relatedness for 19 mating-pairs from mating sites and 11 parent-pairs based on paternity identification. The results showed that pandas’ mate choice was not consistent with inbreeding avoidance, indicating that the mating-pair formation of wild pandas was not influenced by relatedness. Our studies on the effect of genetic heterozygosity (i.e. genetic diversity) in male-male competition indicated that genetic heterozygosity did not explain the establishment of male dominance (Hu et al., 2017b), which further confirmed that the dominance status of male was closely related to body size. A stronger individual could0 more easily establish dominance and obtain mating opportunities (Nie et al., 2012c).

 mate guard

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 Parturition and cub rearing of the giant panda

Giant pandas mainly give birth in August and September every year. Due to the differences in the timing of the estrus and mating of pandas in different mountain ranges, the corresponding delivery time is not synchronous. Through direct tracking and infrared camera monitoring in the Foping Reserve in the Qinling Mountains, we found that the birth period peaked in mid-late August. Before parturition, female pandas need to find a suitable birth-den, which can provide a relatively protected and safe environment for the mother and the young during parturition and cub rearing. Depending on the environment, pandas mainly use tree dens or stone caves as birth-dens. In the Minshan, Qionglai, Liangshan and Xiangling Mountains, most pandas live in oldgrowth forests in high mountains where large trees are available, thus, female pandas often den in tree cavities. In the Qinling Mountains, most of the huge trees have been logged, so female pandas mainly use stone caves as dens.

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The selection of a suitable birth-den will directly affect pandas’ reproductive success. To better understand the selection and availability of birth-dens by giant pandas, we investigated each den site used and unused in our study area. The results indicated that wild pandas have high requirements for the quality of birth-dens, characterized by a narrow entrance with deep roomy interior chambers, steep slope, and close to water. However, stone cave dens meeting the above conditions are limited in a region, so it is speculated that the quantity and quality of birth-dens may be an important limiting factor for the population growth of giant pandas. We suggest that nature reserve managers should construct artificial dens to provide more suitable birth-dens for giant pandas (Zhang et al., 2007c; Wei et al., 2019b). In addition, we also observed several times giant panda rearing young cubs outside of dens in a bamboo thicket, which may further confirm the lack of suitable birth-dens in the region.

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  t h e b i r t h den of the gaint panda

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Female pandas generally stay in the birth-dens for about two weeks after parturition, during which time they mainly feed their cubs and maintain fasting. Later on, the mother begins foraging and frequently moves the cub to shallow, well-lit caves on the edge of the cliff — what we call “rearing dens”, or leaves them in a safe place in the forest; once the old leaves the cub typically climbs a tree to await the mother’s return. When female pandas leave their cubs alone for foraging, they will stay in close proximity to their cubs when the cub is still young. As the cubs grow up, the mothers will increase foraging distances to hundreds of meters from the cub. Cubs usually live with their mothers until they are about one and half a year old (some up to two and half a year old), and then separate from their mothers and begin to live on their own (unpublished data). Because the cub is often left alone for long periods, people developed the misconception that the mother abandons her cub, inappropriately rescuing the cub and taking it to a breeding center. Such actions should be prohibited.

 cub rearing of the giant panda

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  Reproductive rate of the giant panda

The reproductive rate of wild giant pandas has an important impact on their population growth and has attracted public attention. The panda cubs usually live with their mothers until the cubs are 1.5 years old, so wild giant pandas normally breed once every two years. However, if the cubs fail to survive that year, the mothers can breed again in the next year. Breeding data collected on wild giant pandas in Wolong from 1978 to 1992 showed that females came into estrus at 6.5 years old but usually mated and bred at 7.5 years old. Males were able to successfully compete to gain mating opportunities at 8.5 years old. The maximum reproductive age for both sexes was about 20 years old. Theoretically, a female giant panda could rear up to seven cubs in her lifetime. The annual reproductive rate of wild giant pandas in our study was 62.5% (58.33% producing a single cub and 4.17% producing twins (Wei and Hu, 1994). Since 2007, we have monitored nearly 20 mating sites in the Sanguanmiao region of the Foping Nature Reserve in Shaanxi Province and recorded eight cubs by direct observation, including one pair of twins (Hu et al., 2017b). These data suggest that the reproductive rate of wild giant pandas is not as low as previously thought.

 cub rearing of the giant panda

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 cub rearing of the giant panda

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  Natal dispersal pattern of the giant panda

Migration and dispersal are important life-history characteristics of animals that influence individual spatial distribution and population genetic structure. How do cubs establish their home ranges after they grow up and leave their mothers? Due to the difficulty of individual identification in the wild, the natal dispersal pattern of giant pandas has remained enigmatic for a long time. However, by applying molecular scatology techniques and spatial genetics analysis, for the first time, we reveal that the giant panda, unlike most of the mammals, has a surprising female-biased natal dispersal pattern. Our population-genetic studies indicate that female subadults leave their mothers’ home ranges and disperse long distances to establish their own home ranges (Zhan et al., 2007; Hu et al., 2010a). Field observations further confirmed this dispersal pattern at individual level. For instance, Chunchun, a GPS-collared female subadult, dispersed nearly 20 km within a short time from her birthplace in the Sanguanmiao region to the Xihe region to establish her own home range (Zhang et al., 2014). Combined with our studies of pandas’ birth-dens, we put forward the hypothesis of “birth den resource competition”, suggesting that female-biased natal dispersal is closely related to the limited birth-den resources in the environment rather than bamboo resources (Zhang et al., 2007c). Inbreeding (mating between relatives) has negative consequences on animal fitness, which may be counteracted by inbreeding avoidance mechanisms. Inbreeding results in the increase in genetic homozygosity and inbreeding depression, reducing evolutionary potential for animals to adapt to a changing environment. Our research revealed that female-biased natal dispersal drives inbreeding avoidance of giant pandas. We analyzed the relatedness for 19 mating pairs from mating sites and for 11 parent pairs based on paternity identification in Foping and Changqing reserves and found that although there was moderate-level inbreeding (21.1% of mating pairs, 9.1% of parent pairs and 7.7% of panda cubs), no high-level inbreeding was observed. More significant levels of inbreeding could be passively avoided by female-biased natal dispersal rather than by breeding dispersal or active relatedness-based or MHC gene-based mate choice mechanisms. It is worth noting that even in the Foping and Changqing reserves with a large number of giant pandas, there is still some moderate inbreeding. This highlights the risk of inbreeding faced by small panda populations and emphasizes the importance and urgency of improving habitat connectivity and promoting gene flow among isolated populations (Hu et al., 2017b; Ma et al. 2018; Yu et al., 2018).

  an example of female-biased natal dispersal pattern—Chunchun

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Chapter 4

Population Ecology and Population Genetics of the Giant Panda

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Population size, sex ratio, and age structure are basic parameters of population ecology and are fundamental for studying population dynamics and viability analysis. The population dynamics (increase or decline) of giant pandas has long been a concern. The traditional view suggested that population collapse and low genetic diversity were important drivers that led to pandas’ endangerment. Is this really the case? To estimate the population size of wild pandas, the Chinese government conducted four national population censuses in 1974 ~ 1977, 1985 ~ 1988, 2000 ~ 2004 and 2012 ~ 2014. The corresponding census results were 2459, 1114, 1596 and 1864 individuals, respectively, suggesting a population trend of decreasing first and then increasing numbers.

 

 

  field investigation

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The conventional panda survey relies on fecal characteristics to distinguish individuals, such as the bamboo bite size in feces (the size of bamboo stem or leaf fragments undigested in feces) or fecal size (diameter of feces). These methods remain controversial due to large intraindividual variation in fecal characteristics, which makes it difficult to accurately identify individuals. However, molecular genetic techniques provide new opportunities for panda population ecology and population genetics. Our research demonstrated that the traditional census methods underestimated panda population size, that the panda population is increasing, and that the genetic diversity remains high, indicating that the giant panda population is recovering and the genetic diversity is not the cause resulting in its endangerment.

  field investigation

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 Age determination of the giant panda

Accurate age determination is critical for studying population dynamics. By analyzing the panda tooth slice, we can accurately determine the age based on tooth growth lines (cementum annuli), just like tree rings. Combining the number of growth lines and the wear of molars, the age group can be determined (Wei et al., 1988). We used this method to determine panda age and identified the maximum age as 26 years old in the wild. However, this method is only be used where we have access to teeth, that is, with captive or dead pandas. For living wild pandas, we developed another method using telomere length of chromosomes as an alternative. Determining the age of wild individuals will be a focus of our future research.

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  Life table and population viability analysis (PVA)

The population trend can be predicted from the life table compiled with the mortality rate of oneyear-old cubs and the natural mortality rate of different age groups identified by tooth wear. Combined with the reproductive lifespan (usually at age 20) and the birthrate per female panda, we can calculate the net reproductive rate and intrinsic rate of natural increase. Our research showed that the net reproductive rate of giant pandas (R0) was larger than 1, and the intrinsic rate of natural increase r > 0, both indicating a potential population growth trend (Wei et al., 1989). To understand the population dynamics of isolated small populations, we used these population parameters to perform population viability analysis (PVA) of isolated small populations via VORTEX software. We found that even without inbreeding or severe catastrophes such as bamboo flowering and die-off, these small populations would still suffer from high extinction rates. Therefore, we suggested the government should pay more attentions to the conservation of isolated small populations (Wei et al., 1997b).

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 Individual identification and population survey of the giant panda

(1) Molecular scatology method (non-invasive fecal DNA method) Because giant pandas are elusive, wary and very hard to be observed in their complex habitat, it is difficult to carry out accurate individual identification and population surveys in the wild. The traditional individual identification method based on fecal bamboo bite sizes may provide results with low accuracy. Therefore, it has long been a challenge to establish an accurate individual identification and population censusing method. Since 1999, we have attempted to establish molecular scatology methods based on fresh fecal DNA. Using field-collected fresh feces as materials, we explored techniques of fecal DNA extraction, microsatellite primer optimization, PCR amplification system and genotyping error rate estimation, and successfully set up a molecular scatology technique system to accurately estimate wild panda population sizes (Zhang et al., 2004b, 2006b, 2007a; Wu et al., 2009; Zhan et al., 2006, 2010; Wei et al., 2001, 2012).

  giant pandas’ fecal samples

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From 2003 to 2004, we searched for fresh panda feces valley by valley and slope by slope in the Wanglang National Nature Reserve and its surrounding areas in the Minshan Mountains, and collected more than 300 fresh fecal samples and one blood sample left in the snow. Applying 9 panda-specific microsatellite loci for genotyping, we successfully identified 66 individuals in the Wanglang Reserve, doubling the previous estimate (27 individuals) in the Third National Survey of Giant Panda using traditional methods (Zhan et al., 2006, 2009). This implies a significant underestimation for the actual population size by traditional survey methods. Due to the low-quantity and easily degradable features of fecal DNA, estimation of error rates of microsatellite genotyping is a routine and important task. Thus, we developed a mathematical algorithm-based method to estimate the genotyping error rate and found the average error rate per locus was as low as 1.57 × 10−6 ~ 9.01 × 10−4 in the above study. Based on these estimates, we expect that there were fewer than four erroneous genotypes in our dataset (Zhan et al., 2010).

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  handle fecal samples

The giant panda molecular scatology technique system has been recognized by the State Forestry Administration and was introduced as an auxiliary method into the Fourth National Survey of Giant Panda and routine population monitoring. Although this method is very effective for population genetics research, it has some limitations in the national survey of panda populations. First, its successful application depends on the quantity and freshness of field-collected feces. Usually, only feces defecated within a week

will meet demands of genetic analysis and individual identification, with feces defecated within 1 ~ 3 days being the best. Second, this method is very time-consuming and costly. Therefore, it is necessary to further improve the traditional bamboo bitesize method in the national survey, while the molecular scatology method could play an important role in areas with denser giant panda populations, such as the Qinling Mountains.

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（a） bridge of the nose ear

ear

face

shoulder

face nose posterior limb

mouse forelimb

front view （b） shoulder

neck

forelimb testis hindlimb

hindlimb

back view

  front and back “blank” pattern of the giant panda

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(2) Camera trap method Infrared camera trapping, as a non-invasive sampling method in behavioral ecology, can be used to obtain data on activity rhythm, distribution, breeding and cub rearing behavior of wild pandas without disturbing them. In recent years, with the popularization of infrared cameras in panda reserves, it is no longer difficult to obtain a large number of clear photographs of wild pandas. Nevertheless, except for applications in species diversity surveys or public conservation education, these photographs can rarely be used as an effective tool for estimating populations. Therefore, it is of great scientific and practical significance to mining the infrared camera data for individual identification, population census and monitoring. Unlike some other wild mammals such as tigers and leopards, pandas do not have a pelage color pattern suitable for individual identification. So it is necessary to establish a species-specific identification system with camera trap photos. Based on the observation of morphological characteristics of several captive pandas, we summarized the body areas with some potential identification features that can be used for individual identification. Then, we constructed the panda “blank” pattern to fill in different identification features for standardization comparison. We also prioritized these features according to differences in their feature stability (i.e. characters which may change over time, or be obscured), therefore precluding the use of unstable features in individual identification.

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We placed life-size (1∶1) conspecific panda decoys in the wild to attract pandas to remain for longer periods of time in view of camera trap arrays, so that the same individual can be photographed by multiple cameras from several angles for individual identification. Using CameraBase software to convert and combine image data, we then selected photos with good visibility of the individually identifiable information. We examined each photo, documenting unique features at each anatomical location of each individual, using multiple angles, which together were compiled to create a unique suit of individually recognizable features for each panda.

We obtained 12,871 photos from 192 encounters over five months. After excluding photos with poor quality or unfavorable angles, 133 encounter events were successfully used for individual identification, and 11 adult pandas (3 females, 7 males and 1 sex-unknown) were identified. We examined these results using a double-blind test with 12 volunteers and achieved an average repeatability of 89%. Although this individual identification method is not as accurate as the molecular scatology method, it is easily implementable for reserve patrols and has important application value for panda population survey and routine monitoring (Zheng et al., 2016).

trail A panda decoy

infrared camera

~90°

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detection field   c  amera trap set-up at decoy sites

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  monitoring photos from different angles(face, left, right, buttock)

  integrated feature information for individual identification

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  Sex identification of the giant panda

Directly determining the sexes of wild pandas is almost impossible. The advent of molecular scatology makes it less challenging to accurately identify panda sexes and investigate the sex ratio. We used self-developed Y chromosome-specific SRY gene primers to conduct sex identification of 66 wild pandas in the Wanglang Reserve based on fecal DNA, and identified 35 males and 31 females, with no significant deviation from the 1:1 sex ratio (Zhan et al., 2006). The sex determination will promote further studies on panda population ecology. For example, it can help with the understanding of the patterns of sex-biased dispersal, sex-specific spatial genetic structure and habitat utilization by male and female pandas (Wei et al., 2012).

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 G enetic diversity of the giant panda

Genetic diversity is one of the most important components in biodiversity conservation. It is generally recognized that species with higher genetic diversity have greater abilities to adapt to environmental changes, and thus have higher evolutionary potential. For a long time, the giant panda has been considered to have very low genetic diversity, contributing to its endangerment. However, by reviewing previous studies on panda genetic diversity, we found that the small sample sizes or limited genetic markers used in those studies are possibly responsible for the low genetic diversity estimated (Wei et al., 2012). We re-evaluated the genetic diversity of wild pandas through molecular scatology and large-scale genetic sampling. Our results indicated that regardless of wild or captive pandas, mitochondrial markers, microsatellite markers or whole-genome single nucleotide polymorphisms (SNPs), the giant panda still retains relatively high level of genetic diversity (Zhang et al., 2007a; Hu et al., 2010a, 2010b; Zhao et al., 2013; Zhu et al., 2010a, 2010b, 2011b, 2013a; Shan et al., 2014). We have extensively collected the fecal, hair, tissue, blood and skin samples from wild pandas in the six mountain ranges for genetic diversity analysis. Using molecular methods, we obtained the DNA sequences of mitochondrial control region of 159 pandas and the genotypes of 10 microsatellite loci of 115 individuals. The results did not show low genetic diversity, instead, indicating that pandas still have a high level of genetic diversity and evolutionary po-

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tential compared with other bears (Zhang et al., 2007a). We also conducted population genomic research on different geographic populations by large-scale genome resequencing, confirming our previous conclusion. Among six extant populations, the Qionglai population had the highest genetic diversity, while the Qinling population had the lowest (Hu et al., 2010a, 2010b; Zhu et al., 2010a, 2010b, 2011b, 2013b; Zhao et al., 2013). The captive genetic management of giant pandas strives to preserve genetic diversity and avoid inbreeding to ensure that populations remain large, healthy, and viable for future reintroduction. We also evaluated the genetic diversity of captive populations by genotyping 19 microsatellite loci, and found high levels of genetic diversity and low levels of inbreeding, similar to wild panda populations. Through effective population genetic management, the overall inbreeding level is relatively low for populations in Chengdu Research Base of Giant Panda Breeding (Chengdu), China Research and Conservation Center for the Giant Panda (Wolong) and Beijing Zoo (Beijing). However, the population in Shaanxi Rescue and Breeding Center for Rare Wildlife (Louguantai) is facing a high risk of inbreeding, thus, measures should be taken into account to avoid inbreeding depression. Based on the genetic structure analysis, we detected a significant genetic structure among these four captive populations, wherein Louguantai and Chengdu populations clustered together, while Wolong and Beijing populations clustered into another cluster, which reflected the gene flow among these captive populations. Taking the genetic structure of wild pandas into account, we suggested that the Louguantai population should be managed as an independent captive population to avoid gene flow with other captive populations to represent the genetic uniqueness of their Qinling origin. Considering that the population founders of Chengdu, Wolong and Beijing populations were all from Sichuan, the gene exchange among them should be promoted (Shan et al., 2014). In addition, the selection of captive individuals for reintroduction should consider their geographic origin, genetic background, and genetic contribution to wild populations in the future.

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Rapid human population growth and increasing human activities have severely affected the survival of giant pandas. On the one hand, large-scale deforestation, road construction, and economic development along these roads have greatly altered panda habitat, causing serious habitat loss and fragmentation and hindering free dispersal and gene flow. On the other hand, poaching, hunting, and zoo collection of wild pandas have directly caused the population decline. In addition, the periodic natural phenomenon of bamboo flowering and die-off (with a 40-to-100-year cycle) has further aggravated the adverse situation. However, the ecological and genetic mechanisms for these negative factors influencing giant pandas are still unclear.

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 I mpact of large-scale bamboo flowering and hunting on giant pandas

During a period of nearly 50 years before the enactment of Wildlife Protection Law in 1988, the population size of wild pandas dropped rapidly due to a series of ecological and anthropogenic events such as large-scale bamboo flowering and dieoff, habitat loss, hunting/poaching and zoo collection. According to incomplete statistics, at least 1,000 wild pandas died or were removed from wild populations, especially during large-scale bamboo flowering and die-off in the Minshan Mountains (MS) in the 1970s and the Qionglai Mountains (QL) in the 1980s. These events aroused worldwide attention to the giant panda’s fate, and it was believed that the rapid population declines would seriously impact its population viability. We compared genetic variation in mitochondrial DNA and microsatellite markers of historical and extant panda populations before and after 1988 when the Wildlife Protection Law was implemented and the last large-scale bamboo flowering event occurred. The results showed that the rapid population decline did not significantly reduce the overall effective population size and genetic variation of this species. Giant pandas could feed on nonflowering bamboo species or disperse long distances to cope with the adverse effects of bamboo flowering when the habitat is connected. However, a simulation of genetic diversity changes in the next 200 years shows that if the present habitat fragmentation and restricted migration of isolated populations continue, genetic diversity loss will inevitably occur and detrimentally affect the population viability. Therefore, we propose to further enhance habitat protection and increase habitat connectivity (such as constructing habitat corridors) to promote gene exchange among isolated populations, preserve extant genetic variations and maintain long-term evolutionary potential (Zhu et al., 2013b).

  bamboo flowering

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  highway within the giant panda habitat

  logging

 Population collapse mechanism of isolated small panda populations

Habitat loss and fragmentation are one of the main causes of species vulnerability and extinction, and understanding these threats is a key step for devising effective conservation strategies. The Fourth National Survey showed that the panda population was subdivided into approximately 33 local subpopulations of various sizes, of which 22 were at risk of extinction, especially 18 populations with less than 10 individuals at high risk. However, how habitat fragmentation will drive the extinction of isolated small populations is not yet clear. Using molecular scatology and landscape genetic approaches, we studied panda populations in the Daxiangling (DXL), Xiaoxiangling (XXL) and Liangshan Mountains (LS) where habitat loss and fragmentation were most severe. We found that the XXL population experienced a strong, recent demographic reduction (60-fold), starting approximately 250 years ago. Human population expansion and use of non-native crop species (e.g. potatoes and maize) at the peak of the Qing Dynasty resulted in land-use change, deforestation, and habitat fragmentation, which likely have led to the drastic reduction of the most-isolated XXL population (Zhu et al., 2010a). Further analysis indicated that the natural barrier of the Dadu River might have induced the significant genetic divergence between the XXL and DXL populations. The National Road 108 crossed the XXL and DXL habitats further divided each of these populations into two subpopulations. Our results implied that large rivers and highways can severely impede gene flow, and demographic, genetic, and environmental factors would lead to a local extinction if the population remains isolated. Therefore, we recommend constructing habitat corridors and propose panda reintroduction projects as important ways to rescue these isolated small populations (Zhu et al., 2011b). Additionally, the LS panda population has suffered a serious recent population decline of 95% starting 1000 years ago when agricultural activities began to flourish, and consequent habitat heterogeneity affected the dispersal and gene flow of the LS population (Hu et al., 2010a, 2010b).

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  felling bamboo

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  the released panda “Taotao”

Habitat fragmentation restricts the migration, dispersal, and gene flow among panda populations, therefore accelerating genetic divergence among isolated small populations, increasing inbreeding probability, and leading to population collapse (Hu et al., 2010a; Zhu et al., 2010a, 2010b). To tackle this issue, we suggested to the government that panda reintroduction and habitat corridor construction should be implemented as soon as possible, which has received significant attention from the State Forestry Administration. Several pandas have been released to the XXL population, and simultaneously the Nibashan Habitat Corridor in the DXL is under construction by the Sichuan Forestry Department.

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3

 Rescue small populations of the giant panda:    reintroduction and habitat corridors

Reintroduction is an important measure for conserving endangered species which can help maintain their long-term persistence in the wild. In recent years, several reintroduction practices targeting endangered species have been proposed and successfully implemented, among which the most representative cases include black-footed ferrets (Mustela nigripes), American black bears (Ursus americanus), and Pere David’s deer (Elaphurus davidianus). Considering the high degree of panda habitat fragmentation and the fact that small isolated populations are affected by random factors within and outside the populations, in situ conservation measures alone cannot guarantee their long-term survival in the wild. Instead, releasing captive individuals to the targeted populations could be an effective way to increase the population size and genetic diversity. With advancements in captive breeding techniques, captive panda populations have increased rapidly, reaching 600 individuals by the end of 2019, providing a foundation for reintroduction. After making sure criteria were met for reintroduction (Zhang et al., 2006c), we submitted a national-level panda reintroduction plan to the State Forestry Administration. We suggested a two-step strategy: first, to translocate rescued wild pandas into target populations to determine if they can survive and breed in a new habitat. This approach will enhance our experience to better inform the subsequent release of captive pandas. Then, to release captive-bred pandas into targeted populations after prerelease training in different-sized wild enclosures, and conduct long-term post-release monitoring. On this basis, the State Forestry Administration officially implemented the panda reintroduction project, and successively translocated two rescued wild pandas, “Shenlin No. 1” and “Luxin”, to the Longxi-Hongkou Reserve in the Minshan Mountains and the Liziping Reserve in the Xiaoxiangling Mountains, respectively. We participated in the post-release monitoring of these reintroduced pandas and confirmed that they both survived in the wild and have established their own home ranges, one of which, “Luxin”, even successfully reproduced and produced offspring (Yang et al., 2018). Later, several captive individuals, such as “Xiangxiang”, “Taotao”, “Zhangxiang”, and “Xuexue”, have been released into the wild. Although “Xiangxiang” and “Xuexue” died in the wild, “Taotao” and “Zhangxiang” have gradually adapted to the wild environment and survived well.

Chapter 5  Major Threats to Giant Pandas and Conservation Practices

Scientific planning is a prerequisite and vital for successful reintroduction. This includes choice of release sites and individuals (gender and genetic background), disease control, and post-release monitoring. According to our findings on wild pandas, we proposed the most important criterion to guide the scientific implementation of reintroduction. First, the priority should be given to small and isolated populations (such as XXL and DXL populations) because new released individuals are urgently needed to improve long-term population persistence (Zhu et al., 2010a, 2010b). Second, female pandas are preferred as candidates for reintroduction because their female-biased natal dispersal pattern found in the wild indicates they are likely preadapted to reintroduction, which is similar to a dispersal event. Thus, releasing female individuals could improve the survival rate (Zhan et al., 2007). Last but not least, selecting individual with a genetic background compatible with target populations should be taken into account (Zhao et al., 2013). The panda reintroduction is a long-term process which faces many challenges. Fortunately, current reintroduction programs are progressing well, and more captive individuals will be released into the wild in the future to save isolated small populations or to reestablish new panda populations. This project has been noted by the prestigious journal Science as Hope for Wild Pandas (Schenkman, 2010). Another approach to rescue isolated small populations is to construct habitat corridors. The establishment of habitat corridors can connect isolated habitat patches to facilitate migration and dispersal of individuals, thus increasing interpopulation gene flow. In addition to improving their population viability through reintroduction, habitat corridor construction is also of great significance in connecting isolated habitat patches and increasing interpopulation gene flow for small populations threatened by habitat fragmentation (Hu et al., 2010a; Zhu et al., 2010a, 2010b). At present, several corridors are being planned or under construction, such as the Nibashan Corridor in the Daxiangling Mountains, and Huangtuliang Corridor and Tudiling Corridor in the Minshan Mountains. We believe that these corridors will play pivotal roles in the conservation of these isolated small populations.

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© Science Press 2022 F. Wei, Hope for the Giant Panda, https://doi.org/10.1007/978-981-16-6478-6

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Considering multiple factors, such as body size changes from fossil to extant species, diet specialization, breeding difficulties, low genetic diversity, population decline, habitat loss, and bamboo flowering and die-off, a prevailing view emerged that the giant panda has gone down an evolutionary dead-end and is doomed to extinction. In 2009, Mr. Chris Packham, a BBC journalist, even concluded that because pandas have gone down an “evolutionary cul-de-sac”, we should give up on conservation efforts and let them go (http://www.radiotimes.com/ news/2009-09-22/chris-packham-let-pandas-die, last accessed March 7, 2020). I have been studying the conservation biology of wild pandas since 1984. After over thirty-five years of multidisciplinary research with my team members and collaborators, I realize that our findings do not support the above pessimistic view: evolutionary dead-end or evolutionary cul-de-sac (Wei et al., 2015a, 2015b, 2015c). First, giant pandas have evolved different levels of adaptations to the bamboo diet, such as morphology (pseudothumbs), behavior (optimal foraging strategies), physiology (low metabolism), genetics/genomics (pseudogenization in umami receptor gene) and metagenomics (unique microbiome). Second, giant pandas have high reproductive rates and positive population growth. Third, giant pandas still maintain high genetic diversity and evolutionary potential. Last but not least, the plentiful bamboo resources in panda habitats should not be a limiting factor for population expansion. These findings strongly indicate that the giant panda is not a relic species or an “evolutionary cul-de-sac”, and address the pessimistic perception of panda’s future prospects and destiny prevalent in some sectors of the scientific community and the public. This adorable creature has a bright future. To respond to great concerns about investment in panda conservation and understand whether the investment translates into substantial returns for humanity, we estimated the values of the ecosystem service that panda and its habitat provided. Human existence relies on the vast number of provisioning, regulating and cultural ecosystem services provided by nature, such as the provision of clean water, breathable air, energy, food, timber, and places for recreation (Costanza et al., 1997; Daily, 1997). A global assessment of these services reached a total value of US $125 to US $145 trillion/year, greatly overshadowing the world’s collective Gross World Product (GWP) of US $75.2 trillion for 2011 (Costanza et al., 2014). To conserve pandas, the Chinese

Epilogue  Hope for the Giant Panda

government has established 67 nature reserves. The estimated ecosystem service value from pandas and their reserves fall between US $2.6 ~ 6.9 billion/year in 2010. Protecting the panda as an umbrella species and the habitat that supports it yields significant societal benefits, about 10 ~ 27 times the cost of maintaining the current reserves, motivating expansion of panda reserves and investments in natural capital in China. With these new knowledge at hand, we believe policy makers and society in general will take note and question reluctance to invest in panda conservation and species conservation (Wei et al., 2018). However, life is not always full of sunshine. Our research indicates that the endangerment of giant pandas is closely related to environmental pressures brought by growing human populations throughout the Chinese history (Zhu et al., 2013a). Deforestation, road and settlement construction, land reclamation, mineral exploitation, and large water engineering projects are now primary risk factors for giant pandas: they cause habitat loss/fragmentation and amplify the severity of natural disturbances, making giant panda populations more prone to local extinction. Small populations are particularly vulnerable to such threats (Zhu et al., 2010a, 2010b, 2011b). More recently, looking over the period ranging between 1985 and 2010, we found evidence that areas regularly affected by intense flooding (a type of natural disturbance mainly caused by intense precipitation and terrain characteristics) tend to be used by people rather than areas that pandas use more intensively (Ameca et al., 2019). Giant pandas are able to cope with these stochastic disturbances due to intrinsic adaptive capacities gained over evolutionary time. However, human responses to these disturbances (e.g., for mitigation) could reduce panda habitat resilience and quality, and hence cause declines in giant panda populations. We also find other threats such as diseases (panda worms, canine distemper virus, canine adenovirus, canine parvovirus) and environmental pollution (perfluorooctanesulfonate, perfluorooctanoate, organochlorine pesticides, polychlorinated biphenyls and brominated flame retardants) (Dai et al., 2006; Hu et al., 2008; Qin et al., 2010; Zhang et al., 2011b, 2015). Other researchers even find that future climate changes would impact the panda’s survival (Tuanmu et al., 2013; Li et al., 2015). In general, the endangerment status of the giant panda should be attributed to extrinsic factors rather than intrinsic ones, and the role of humans in threatening the viability of panda populations is unquestionable.

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To solve the problem above, Chinese government has implemented several laws and the Nature Reserve Management Regulations over the past decades. National conservation measures for giant pandas or closely related national initiatives have been taken to save giant pandas, including the ban of poaching, the panda habitat protection project, panda nature reserve network, natural forest protection project, and the Grain to Green project. These endeavors have led to significant conservation achievements: poaching has been eradicated; 67 giant panda nature reserves (and the beginnings of a new Giant Panda National Park) have been established protecting approximately 54% of panda habitats with over 67% of individuals; the fourth national survey showed a population increase and habitat expansion. Our recent investigation on panda reserve system reveals that pandas are well protected inside reserves and most indices of disturbance are decreasing (Wei et al., 2020b). At present, the Chinese government is implementing panda reintroduction in the Xiaoxiangling Mountains. One wild-rescued panda and three captive-bred individuals have been successively reintroduced to restore the isolated small population with high risk of extinction. Fortunately, the released individuals have begun to integrate into the local population and the female panda “Luxin” has successfully bred and produced one offspring in Liziping Reserve where she was reintroduced. Further, several habitat corridors such as Nibashan Corridor in the Daxiangling Mountains, Huangtuliang Corridor and Tudiling Corridor in the Minshan Mountains are under construction. These science-based practical conservation measures and the corresponding achievements foretell a bright future for the giant panda.

 

  panda “Luxin” and her cub

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  panda “Luxin” and her cub

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Great news from the International Union for Conservation of Nature (IUCN) is that the giant panda is no longer “Endangered” on its Red List. It is downlisted to “Vulnerable” based on our systematic assessment of population parameters and IUCN standardized criteria (IUCN, 2012; Swaisgood et al., 2016). Our assessment showed profound reasons for hope for species conservation everywhere, and a milestone of successful conservation to make. However, we should take this positive message with caution. The Chinese government and international community should maintain its focus and investment in panda conservation, and develop strategic plans to deal with current threats such as habitat fragmentation and small populations, otherwise it will return to the “Endangered” status once again (Swaisgood et al., 2018). What should we do next to protect the pandas’ evolutionary potential? First, habitat protection and restoration should be the top priority; Second, implement reintroduction programs and habitat corridor construction to rescue isolated small populations; third, reinforce long-term monitoring and scientific research to provide a robust baseline the science-based conservation approaches. My research team will continue to work to better enable China’s giant panda to prosper. We hope more people will properly appreciate the pandas’ evolutionary potential and understand threats wild panda populations are facing, motivating more devotion to panda conservation. We wish the giant panda a better future!

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Afterword Ⅰ

Upon reading the manuscript Hope for the Giant Panda: Scientific Evidence and Conservation Practice written by Fuwen, my heart was filled with gratification and excitement. I couldn’t help but think of the old days in the 1990s when Chinese Academy of Sciences (CAS) was implementing the reform of the science and technology system with indomitable spirit led by President ZHOU Guangzhao of CAS. Taking advantage of this tide of reform, Fuwen came all the way to Beijing from Sichuan and was enrolled in the Institute of Zoology, CAS for doctoral study. At that time, the institute was promoting the idea of combining “macro and micro” in terms of scientific research and graduate training, that is, applying microscopic methods to macroscopic scientific problems, which was approved and supported by the Vice President XU Zhihong. Since then, with the support of the relevant departments in the institute, several public experimental platforms have been established to facilitate researchers and graduate students who engaged in macro-biological researches. This preliminarily solved the problems on experiments for researchers and graduate students working on macrobiology in fields such as taxonomy, ecology and conservation biology. Fuwen majored in animal ecology and decided to continue pursuing ecology and conservation biology after obtaining his doctorate. For the past 20 years in the Institute of Zoology, Fuwen has devoted himself wholeheartedly to giant panda research with great perseverance. What is more commendable is that he never confines his academic thinking to the conventional views, but can go beyond some authoritative conclusions and

solve one mystery after another along his way of exploring, thus shedding light on the evolution of giant pandas. The book is an advanced popular science work. Fuwen accurately and vividly explains some profound theoretical knowledge of life science in easy-to-understand words, so that besides the adorable image of the giant panda, readers can take a glance at how this creature has lived under the surpassing splendors of stegodons and carved its own path in the long evolutionary history of eight million years. Although the giant panda is morphologically structured as a carnivore, it can adapt to a low-energy bamboo diet and continue to live and breed to this day. Those profound evolutionary theories and tedious technical terms are transformed into such eloquent and fascinating sentences under Fuwen’s pen. This book will ignite a strong desire for readers to explore the mysterious evolutionary history of giant pandas and their future prospects. I believe Fuwen and his team will not stop here. They will continue looking for new scientific issues, and keep solving the enigmas in the evolution of giant pandas and determining the best way to protect them. I wish Fuwen and his team even more vibrant and vigorous. Go forge ahead in your scientific research journey while keeping in mind “the battle is not yet over in the near future”.

Prof. WANG Zuwang Animal Ecologist, Institute of Zoology, Chinese Academy of Sciences

Afterword Ⅱ Dr. WEI Fuwen—colleague, collaborator, “Lao Pengyou” (old friend)—is probably the most prolific of all giant panda scientists, and has taken the most integrative approach to giant panda biology and conservation. I take great pride and enjoyment in my circa two decades of collaboration with Fuwen. What’s more, I can think of no one better suited to write a book synthesizing decades of scientific research on the giant panda applied to this emerging conservation success story. Fuwen (I will call him here by the name I use for my old friend) delivers a tour de force, an expansive and integrative personal and professional account of his scientific journey to reveal the mysteries of the panda. I remember clearly when I first met Fuwen. He was still young, as was I, just establishing himself in academia. I was attending the 1997 Giant Panda Technical Conference in Chengdu. Chinese science was not what it is today, still recovering from years of neglect during the Cultural Revolution. But when Fuwen took the stage and presented his elegant work on microhabitat analyses for giant and red pandas, I took notice. Here was a fresh face, with fresh ideas and rigorous quantitative approaches. I introduced myself, complimented him on his work, and we ended up chatting for some time about all the work that could be accomplished with pandas in the future. We stayed in touch for a few years, meeting for dinners in Beijing and corresponding by email, but it wasn’t until 2003 that we finally found the opportunity to work together. That year we were joined by two biological giants, our mentors Dr. HU Jinchu and Dr. Donald Lindburg, for a tour of several panda reserves. We spent two weeks visiting Tangjiahe, Wanglang, Si’er and other panda reserves. We drank a lot of “Bai Jiu” during those visits, a necessary ingredient to make friends in Chinese culture. Then, we piled in the vehicle and were off along the windy mountainous roads to the next reserve. Between the drink and the roads, I remember having a headache most of the time. But what I remember most was the four of us engaged in great conversation about what we could achieve with a new joint project. About the same age, Fuwen and I also connected on topic of interest to young men in their thirties. Yet this rather inauspicious beginning was the seed and catalyst for what was to become nearly two decades (and counting!) of collaboration

and friendship. Some of the product of that fruitful collaboration is detailed here in this book. One of the things that strikes me most about Fuwen and his work is his collaborative spirit and integrative approach to biological research, driven by his curiosity and hunger for knowledge. If Fuwen doesn’t know how to do something (e.g., a scientific discipline or methodology), he seeks out those that do. He enlists their help and he absorbs their ideas. I was honored to be one of his main collaborators in the field of ecology, but he enlisted so many others along the way—geneticists, endocrinologists, chemists, nutritional ecologists, economists, and many more. These productive collaborations have allowed his research on pandas to soar above all others’. That he has finally put the sum total of this accumulated wisdom down in print is fortunate for the rest of us. Frankly, it has been difficult to keep up with Fuwen and learn about all of his new findings. Fuwen came of age academically in the shadow of pessimism. People had little hope for the giant panda; Dr. George Schaller even penned a book called The Last Panda. There was indeed reason for pessimism at the time. Panda numbers were declining precipitously and although some protections were put in place, they seemed insufficient to stave off extinction. That all began to change—thankfully—when people like Fuwen really started to put their shoulders into conservation efforts. Too numerous to enumerate here, a plethora of new policies and protections issued forth from the Chinese government and scientists and conservationists globally rallied to protect the panda. It is one of conservation’s greatest success stories in the making. There is still much work to do to keep the panda on the path to recovery, but it is reassuring and hope-inspiring that things have begun to turn around for this most charismatic of species. This book, better than any other single written product, reveals the previously unknown biological mysteries of this enigmatic species and catalogues how science has contributed to the panda’s recovery. It is destined to become a classic.

Dr. Ronald Swaisgood Animal Behaviorist and Ecologist, San Diego Zoo Global

Afterword Ⅲ I first met Fuwen in 1999 when he came to the lab that I had recently established at Cardiff University to work with my then postdoc and long-time collaborator Benoit Goossens. Benoit had worked with Fuwen a few years previously on giant panda ecology and they were keen to see whether it was possible to produce reliable DNA profiles for giant panda feces, which would potentially greatly assist the Chinese authorities with population census estimates and reveal new and exciting facets of giant panda reproductive biology and demography. I think it was Fuwen’s first long stay at an English-speaking lab, but he settled in quickly and became both a valued lab member and a good friend (we are almost exactly the same age), with whom I have had the pleasure to work with for more than 20 years. I vividly remember Fuwen accepting giving a departmental seminar in our school, despite his newly developing spoken English skills, and holding a packed room enthralled for over an hour with many stories and scientific discoveries of this fascinating species. It is a seminar that is still talked about in Cardiff 20 years later. Armed with this new DNA profiling knowledge, Fuwen returned to Beijing and started to establish a new lab working in this area and we began to collaborate on many different genetic studies of the giant and red panda and other endangered mammals in China. These projects have resulted in more than 20 publications and collaborations with Fuwen and his colleagues and students (such as LI Ming, YANG Guang, ZHANG Baowei, HU Yibo, ZHU Lifeng, LIU Zhijin, MA Tianxiao) and including a long and productive collaboration with his former student ZHAN Xiangjiang. These collaborations have been intense, hard work and very

rewarding and has allowed me to experience the extraordinary transformation in Chinese science from I first visited Prof. ZHOU Kaiya in 1993 to the present day, and seen in microcosms at the Institute of Zoology, CAS in Beijing, which has transformed dramatically from when I first visited nearly 20 years ago into one of the world’s powerhouse zoological research institutes today. It has also allowed me to travel extensively across China, often in Fuwen’s company, and experience its natural wonders, cultural diversity and fabulous cuisine, so that I now feel China is a second home, so much so that I spent a sabbatical at the Institute in 2018. During all this time, my discussions and collaboration with Fuwen have remained a constant, and I was delighted when he was elected to the Chinese Academy of Sciences – just deserves for an amazing career in ecology and conservation biology. Long may the research continue! This book should appeal to those interested in conservation biology in its most general sense. This is because the giant panda offers an excellent and very approachable example of how the use of modern scientific methods can unravel the most apparently impenetrable of life-styles. The life history of this species, its many complexities and apparent paradoxes, are rapidly becoming understood, thanks in no small part to the research efforts of the world leader on this species, Prof. WEI Fuwen.

Prof. Michael Bruford Conservation Geneticist, Cardiff University, UK

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