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English Pages 118 [127] Year 1999
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Four Decades of Vector Biology at the University of Notre Dame A Scientific Perspective Karamjit S. Rai
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Disclaimer Some images in the original version of this book are not available for inclusion in the netLibrary eBook. Copyright 1999 by University of Notre Dame Press Notre Dame, IN 46556 All Rights Reserved Manufactured in the United States of America Library of Congress CataloginginPublication Data Rai, Karamjit S., 1931– Four decades of vector biology at the University of Notre Dame : a scientific perspective / Karamjit S. Rai. p. cm. Includes bibliographical references. ISBN 0268028532 (alk. paper) 1. Mosquitoes as carriers of disease. 2. Aedes aegypti— Physiology. 3. Aedes aegypti—Genetics. 4. Aedes albopictus— Physiology. 5. Aedes albopictus—Genetics. I. Title. R A640.R35 1999 614.4'323—dc21 9914432 The paper used in this publication meets the minimum requirements of the American National Standard for Information Sciences—Permanence of Paper for Printed Library Materials, ANSI Z39.481984.
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Contents Dedication and Acknowledgments
ix
Introduction
1
1. Isolation and Characterization of Mutations and Genetic Mapping in Aedes aegypti
4
a. Morphological Mutations
4
b. Allozymes
6
c. Molecular Markers
7
2. Female Monogamy and Matrone: Sex and Physiological Mutations
7
3. Cytology: Genome Structure, Organization, and Evolution
8
a. Comparative Karyotypes
8
b. Giemsa CBanding Patterns: Heterochromatin Differentiation
9
c. Genome Size Variation
10
i. Interspecific Variation
10
ii. Intraspecific Variation
10
d. Intra and Interspecific Variation in Repetitive DNA Amount and Organization
11
e. Chromosome Mapping of Repetitive DNA
12
4. Genetics of Chromosomal Rearrangements
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5. Genetic Control: Evolution of the Concept
14
6. Seminar in Vector Genetics
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7. Genetic Control: Principles and Mechanisms
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a. SterileInsect Technique
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b. Chromosomal Translocations
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8. Genetic Control: Applications a. The Delhi Project: WHO/ICMR
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b. The Mombasa Project: USAID
26
9. Population Genetics
28
a. Allozymes
29
b. Molecular Markers
30
10. The Aedes albopictus Saga: Physiological Characteristics, Population Genetics, and Temporal Changes in the New World
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a. Photoperiodic Sensitivity
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b. ColdHardiness
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c. Allozyme Differentiation
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i. U.S. Populations
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ii. Native Populations
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iii. Geographic Origin of U.S. and Brazilian Populations
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iv. Temporal Changes in Genetic Structure
36
d. Ribosomal DNA (rDNA) Intergenic Spacer (IGS) Length Variation
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i. Population Genetic Survey
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ii. Temporal Variation in rDNA Spacer Length
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iii. Inheritance of rDNA Spacer Region
38
iv. Chromosome Mapping and Copy Number of rRNA Genes
38
e. Mitochondrial DNA Restriction Fragment Length Variation
25
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i. Intraspecific Variation
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ii. Interspecific Variation
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iii. Dynamics of mtDNA Haplotypes in Laboratory Cage Populations
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f. Reproductive Differentiation: Cytoplasmic Incompatibility in Aedes albopictus
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g. Competition between U.S. Strains of Ae. albopictus and Ae. aegypti
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h. Vector Competence to Dengue1 Virus i. Oral Susceptibility and Horizontal Transmission
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ii. Vertical Transmission
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iii. Midgut Basal Lamina Thickness and Virus Dissemination
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iv. Inheritance of Midgut Infection/Disseminated Infection
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v. Status of Ae. albopictus in U.S.
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11. Vector Competence and Arboviral Ecology
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12. Endocrinology and Reproductive Physiology
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13. Research and Training Funding
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14. Special VBL Resources and Facilities
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International Reference Centre (IRC)
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University of Notre Dame's Environmental Research Center (UNDERC)
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Mosquito Data Bank of the University of Notre Dame (MODABUND)
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St. Joseph County Mosquito Surveillance Program
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Indiana State Department of Health and the Laboratory for Arbovirus Research and Surveillance—University of Notre Dame (UNDLARS)
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Externship/Internship in Arbovirology for D.V.M. Students
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15. Linkages of the VBL with Tropical Institutions
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16. Vector Biology Courses and Visiting Scientist Program
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Epilogue
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Appendices 1. Vector Biology Faculty, University of Notre Dame, 1957–1999
42
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2. Postdoctoral Research Associates in Vector Biology/Parasitology since 1960
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3. Predoctoral Students in Vector Biology/Parasitology since 1961
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4. Undergraduate Students in Vector Biology since 1962
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5. Visiting Scientists/Professors and Guest Lecturers Relevant to Vector Biology at the University of Notre Dame
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References
105
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Dedication and Acknowledgments This book is dedicated, with affection, to the late George B. Craig, Jr., a colleague, a collaborator, and a friend for nearly four decades. Craig was likened by his graduate students to the indomitable and legendary Vince Lombardi, the coach of the Green Bay Packers. In a recent symposium entitled "George B. Craig, Jr.: His Impact on Entomology" held at the Annual Meeting of the Entomological Society of America, Nashville, Tennessee, December 1997, several of his former postdoctorals and graduate students provided memorable reminiscences about George Craig, the scientist and the man. His first graduate student, Robert VandeHey, remarked that "George frequently challenged our opinions and stimulated our thoughts" and that he ''took great pride in introducing us to the professional world of mosquito research" (personal communication). Another early student, Sister Mary Gerald Leahy, who did outstanding work on evolutionary relationships between Aedes aegypti and Ae. albopictus (Leahy and Craig, 1967), remarked that "George broadened us by both relating to us his impressions from his frequent trips and by engaging visitors from far and near in lively discussions by his leading, astute questions." George not only trained scores of scholars, he literally changed lives by the glow in his eyes, his very gifted intellect, his very quick and sharp wit and even sharper tongue, and by the breadth of his interests from academia to sports. He once remarked to the author that he owed his exceptional creativity, in part, to gout, which results in uric acid accumulation, and that Charles Darwin had the same affliction. He was honored by the University of Notre Dame and his peers in numerous ways. His election in 1983 as a Fellow of the National Academy of Science was a truly singular recognition for him. Notre Dame bestowed the Clark Chair on George in 1974. "Craig's students were devoted to their mentor even before they became students," wrote a local correspondent of the South Bend Tribune (Wayne Felda, South Bend Tribune, January 1, 1996) at the
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time of Craig's death. He was a naturalist par excellence; he loved to do creative science. Morton S. Fuchs, our colleague for some thirty years, remarked that Craig, like Barbara McClintock, possessed "a feeling for the Organism" (Fuchs, 1996). Several generations of students who studied under George in the laboratory and accompanied him to the University of Notre Dame Environmental Research Center (UNDERC)in Michigan's Upper Peninsula during mid to late spring and summer each year for "Aedes boot camp" have been immensely enriched as a result. Dr. Robert Novak, a former postdoctoral at the Vector Biology Laboratory and the 1996 President of the American Mosquito Control Association, remarked that "Professor Craig's death ends a truly magnificent era in mosquito biology" (Novak, 1996). George Craig introduced the author, a botanist by early training, to the beautiful "word" of mosquitoes and over the years profoundly influenced his interests, which became a lifelong passion. George's enthusiasm and commitment to this "world" were both inspiring and infectious. It is largely due to the breadth of his vision and catholicity of his person that, although at times we argued rather passionately and interpreted certain events and observations differently, we never allowed such mundane differences to affect our friendship and cordial relationship. The author also wishes to express grateful thanks to several other members of the Vector Biology Laboratory at the University of Notre Dame, in particular, Mort Fuchs, Paul Grimstad, Ted Crovello, and Ron Hellenthal, for numerous shared interests and courtesies. In a sense, we grew up together professionally. This distinctive group of individuals was able to create the multifaceted and interdisciplinary infrastructure of the Vector Biology Laboratory at Notre Dame, an enterprise much larger than the sum of their individual contributions. The comraderie, the working relationship created, and the give and take have been most fulfilling both professionally and personally. In academia how does one create anything, let alone leave a legacy, without intensely devoted postdoctorals and graduate and undergraduate students who are driven to the pursuit of excellence? The Vector Biology Laboratory has had more than its share of such
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colleagues, too numerous to mention here. We salute them all. They are listed in appendices 2, 3, and 4. This work has gone through what at times seemed all unending cycle of revisions. In addition, assembling and collating information about hundreds of former graduate and undergraduate students and postdoctorals going back forty years would certainly not have been possible were it not for a highly efficient secretary, Katie Merz, who possesses all the right genes for a wonderfully cheerful disposition and who never got tired of all the running around involved in preparing the manuscript. I also thank Paul Grimstad and Mort Fuchs for carefully reading the manuscript and suggesting several improvements. Nevertheless, I alone remain responsible for any shortcomings and errors of omission and commission. Finally, it is with great personal pride that I express my sincere gratitude to my wife, Gurmit, for her quiet inspiration and understanding, to our children, four sons and a daughter, all ardent Domers, and to their Families for their love, affection, and all their support and encouragement over the years. The University's commitment to the Vector Biology Laboratory has remained steadfast and strong throughout its existence. My sincere thanks for this to the Dean of the College of Science, Frank Castellino; to Vice Presidents James Merz and the late Robert Gordon; to Provost Nathan Hatch; and to several chairpersons of the Department of Biological Sciences, in particular, Ted Crovello, Mort Fuchs, Jack Duman, and the late Robert Gordon and Ralph Thorson. I thank Jim Langford, Director of the Notre Dame Press, for his quick and enthusiastic acceptance of this work for publication. The generous funding provided by several national and international agencies, particularly the National Institutes of Health, the U.S. Agency for International Development, and the World Health Organization, and by the University of Notre Dame in support of the original research mentioned in this book is gratefully acknowledged.
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Introduction On a global basis, the parasitic and viral diseases of man rank among the most important public health problems existing today. Arthropodborne diseases such as malaria, trypanosomiasis, filariasis, yellowfever, dengue, and the viral encephalitides afflict more than a billion humans on a yearly basis (Warren, 1988). Furthermore, several newly emerging infectious diseases pose an additional threat. The global warming, if real, is likely to exacerbate mosquito transmitted diseases. During the last forty years, the field of vector biology has evolved from a largely descriptive science to one of considerable sophistication, addressing fundamental as well as applied aspects of the abovementioned human diseases. Considerable emphasis has been placed on studying the biology, including genetics, field ecology, reproductive physiology, epidemiology, and more recently, molecular biology, of several important insect species that are vectors of the diseases mentioned above. Since its founding by the late Professor George B. Craig, Jr., in 1957, the Vector Biology Laboratory of the University of Notre Dame has played an important role in shaping the field into what it is today. In terms of scholarly productivity, national and international recognition, grant support, training, placement of graduate students and postdoctorals, and the overall quality of the published work, this group has few peers. Indeed, the case can perhaps be made that this group historically ranks among the best in the world in the broad area of vector biology spanning the last four decades. Two examples, one substantive and one metaphorical, may be cited as a mark of the recognition extended to UND's VBL by the community of its peers the world over. Professor James B. Kitzmiller, who is affectionately regarded as the "father" of the field of mosquito genetics not only in the United States but globally, donated his entire collection of more than 10,000 numbered and catalogued reprints involving a unique collection of articles and monographs dealing with genetics and embryology to the University of Notre Dame. This collection goes back to 1900, thus covering the
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very early days of genetics. This was a magnificent gift which will doubtlessly continue to enrich several generations of vector biologists at the University of Notre Dame. All tile famous names are among the authors of these reprints: Morgan, Muller, Sturtevant, Bridges, deVries, Holmes, Castle, Davenport, Dobzhansky, Wallace, Goldschmidt, Blakeslee, Emerson, Shull, and many others. Jim Kitzmiller also donated his collection of some 25,000 anopheline chromosome slides, including a box of slides from Professor Phineas W. Whiting. This box contains the original slides from which Prof. Whiting described the Culex chromosomes in 1917. The second example involves Professor Roger Wood of the University of Manchester, U.K., who has done pioneering work on the genetics of insecticide resistance and the sexratio distortion in the yellow fever mosquito, Aedes aegypti. In early 1970s I invited him for a seminar at VBL; my laboratory at the time was also involved in work on the cytogenetics of sexratio distortion. I introduced him to the group. He started his presentation by saying that each morning before beginning work on mosquitoes in his laboratory, he bows three times in the direction of Notre Dame and that his students follow the same ritual. This introduction just about brought the house down. However, there is perhaps a kernel of truth in this lighthearted metaphor. The VBL has at times been referred to as the "Mecca" of vector biology, where various "Mullahs" of the profession often flock to pay their "obeisance." The number and the stature of the scholars who have visited the VBL over the years is impressive indeed. Such visits have been highly beneficial to VBL personnel as well and have certainly enhanced the interdisciplinary nature of the program. This book provides a scientific perspective, in a historical context, on the origin, growth, and contributions of UND's VBL. Scientifically speaking, a detailed technical account of these contributions is available in more than five hundred publications by members of the VBL. Space limitations do not allow a detailed account, or even a mention, of each of the contributions during the last forty years; only the critical ones, chosen somewhat arbitrarily and subjectively by the author, are included here. Furthermore, although vector ecol
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ogy and related disciplines have constituted all important component of the VBL, the emphasis in this presentation is on vector genetics, genetic control, and population genetics. In the historical context, these disciplines represent the core, perhaps the "birthright," of VBL and provide whatever legitimacy exists for VBL's "claim to fame." Also, as the recent molecular orientation testifies, these disciplines represent all ongoing continuum. The account that follows is largely, though not entirely, chronological. The faculty involved in vector biology studies at Notre Dame since 1957 are listed in appendix 1. The postdoctorals, graduate students, undergraduate research participants, and visiting scholars are listed in appendices 2, 3, 4, and 5, respectively. These lists speak for themselves. As is obvious, individuals trained at VBL are currently occupying highly visible and leadership roles both in academia and government institutions in the U.S. and several countries around the globe. This, above all else, is the legacy of VBL. Historically, in the late '50s and early '60s, around the time the VBL was established, entomology was largely a descriptive discipline. In the field of medical entomology, the emphasis was on classical taxonomy, morphology, ecology, and management of arthropod pests of humans and domestic animals. The word "genetics" hardly existed in the entomological lexicon. There were essentially no geneticists whatsoever on entomological faculties in any of the major universities. In a sense, it is incredibly ironic that, although the infrastructure of genetics in the early part of the twentieth century was established largely through work with the venerable fruit fly, Drosophila melanogaster (all insect species and not infrequently a pest at that), early entomologists by and large did not regard such work as falling within the domain of entomology until other insect taxa began to be used in similar studies (Rai, 1973). It was in this context that the early work at the VBL began to be recognized as trailblazing and pioneering. One example of this among many may be mentioned: Craig and Rai were invited in 1966 to write a review article on the rather general theme of "Genetic approaches in entomology" for the Annual Review of Entomology (Thomas Mittler, personal communication, January 21, 1966). Nevertheless, for various
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reasons, in part because it was perceived as a bit premature, such a review was not written. Early history of Drosophila genetics is replete with historical accounts of "The Fly Room" at Columbia University (Sturtevant, 1965). The "changing of the guard" at the University of Notre Dame and the recent molecular transformation of the field provide the rationale for a perspective on what may be referred to as "The Mosquito Room" at Notre Dame. This account briefly describes the evolution of the VBL at UND, beginning with formal genetics and construction of linkage maps of Aedes aegypti, and moving on to cytology and cytogenetics, genetic control, population genetics first using allozymes and later various molecular markers, vector competence and arboviral ecology, endocrinology and reproductive physiology utilizing several mosquito taxa at various levels of taxonomic organization, and related activities. An integral part of such growth and evolution has been the role that computers played in providing deeper insights on various aspects of vector biology, simulation, modeling, and literature and data bank creation at UND. The major motivation for this perspective is to reflect on the past forty years and to attempt to weave the mystique of the place with a brief account of the major participants and the flavor of the science performed.
1— Isolation and Characterization of Mutations and Genetic Mapping in Aedes aegypti a— Morphological Mutations George B. Craig, Jr., was a gregarious individual, possessing unbounded enthusiasm for the field of entomology in general and
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mosquitoes in particular. The founding of what was subsequently named the Vector Biology Laboratory took place in 1957 when George Craig joined the Department of Biology at UND as an assistant professor. This happened shortly after the appearance of a seminal publication by James Kitzmiller (1953), "Mosquito genetics and cytogenetics," and less than ten years following the establishment of a Section on Medical Entomology in 1948 within the then parent organization, the American Association of Economic Entomologists (Steelman, 1989). Craig's Ph.D. mentor was the famous William R. Horsefall at the University of Illinois. At UND, aided by his first graduate student, Robert VandeHey, Craig began isolating morphological mutations through singlepair crosses, to characterize the same and to estimate the level of overall genetic variability in Aedes aegypti (Craig and VandeHey, 1962; VandeHey and Craig, 1962). This mosquito species is easy to colonize, it has a short generation span, its eggs can be stored for long periods, and the adults mate readily in small cages (Craig, 1965). The method they used of mating individual females and males in single pairs through the F2 generation was timeconsuming and labor intensive. Nevertheless, this project yielded several mutants, a study of the formal genetics of which, particularly the transmission pattern, came to be known as the Mosquito Genetics Project in the then Department of Biology (later, Department of Biological Sciences) at UND. This project was originally housed in the basement of the rather antique "Pipeshop," a maintenance facility on the campus, since dismantled. Some of the more interesting mutations that were thus isolated and characterized helped to establish a strong beginning of the formal genetics of Aedes aegypti and the groundwork for subsequent studies. This included the yellow larva (Craig and Gillham, 1959), sexratio distorter (Craig and VandeHey, 1960), bronze body (Bhalla and Craig, 1967), white eyes (Bhalla, 1968), and others. Ae. aegypti was the first mosquito species thus studied and quickly became a model mosquito species for genetic studies patterned after early work with D. melanogaster. Genetic analysis of the socalled "maleproducing" factor, Distortor, whereby males possessing this gene produce predominantly male progeny, set the
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stage for subsequent interest in using appropriate genes for genetic control purposes (Hickey and Craig, 1966). William Hickey later became the president of the well known Saint Mary's College in Notre Dame, Indiana, and attained national recognition as an astute administrator. More or less simultaneously, the dynamics of certain genes, e.g., yellowlarvae, began to be probed at the population level in laboratory cage studies. This work evaluated temporal changes in gene frequencies, given different proportions of the wildtype and mutant phenotypes to initiate breeding populations (Adhami, 1904 Ph.D. thesis). By 1967, more than 80 morphological mutants affecting almost all the body parts, including wings, antennae, legs, body segments, body size, body color, and so on, had been characterized; of these, 28 were assigned to the three linkage groups in Ae. aegypti (Craig and Hickey, 1967). Furthermore, the genetic basis of some important biological characteristics of Aedes, such as autogeny (O'Meara and Craig, 1969; O'Meara, 1972) began to be rigorously analyzed. Virtually all of this work was accomplished before the widespread use of gel electrophoresis fruitfully exploited by Hubby and Lewontin (1966). b— Allozymes Rapid utilization of Hubby and Lewontin's methodology ushered in the use of biochemical and molecular techniques and the subsequent founding of the domain of population genetics in mosquito studies. Beginning in the mid to late '60s, electrophoretic analyses of the protein products of enzyme loci (Hubby and Lewontin, 1966) provided a plethora of new isozyme markers. As a result, by 1979 some 200 genes were characterized and mapped on the three linkage groups (Munstermann and Craig, 1979). In time, efforts were also made to initiate formal genetic studies with mosquito species other than Ae. aegypti. These included Aedes albopictus (BattMiriam and Craig, 1966), Aedes mascarensis (Hartberg and Craig, 1974), Aedes atropalpus (Munstermann, 1980), and later, Aedes triseriatus (Matthews and Craig, 1980; Munstermann et al., 1982; Matthews and Munstermann, 1983).
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c— Molecular Markers Over the last decade, construction of saturated linkage maps has been made possible through the availability of large numbers of molecular markers generated through Polymerase Chain Reaction (PCR) amplification of small amounts of genomic template DNA. Severson et al. (1993) were the first to construct a linkage map of Ae. aegypti using 50 Restriction Fragment Length Polymorphism (RFLP) markers. More recently, a comparative linkage map for Ae. albopictus has been constructed using RFLP loci common to both these species (Severson et al., 1995). The resulting linkage maps in the two species are comparable in length. Antolin et al. (1996) and Mutebi et al. (1997) have constructed linkage maps of Ae. aegypti and Ae. albopictus, respectively, using Single Strand Conformation Polymorphism (SSCP) analysis of Random Amplified Polymorphic DNA (RAPD) markers. Studies in formal genetics and linkage map construction have, therefore, been ongoing and represent a continuum.
2— Female Monogamy and Matrone: Sex and Physiological Mutations As the formal genetic studies of Ae. aegypti progressed, new insights were obtained concerning several important facets of Aedes biology. For example, it was demonstrated that unlike D. melanogaster, the female Ae. aegypti mate only once in their lifetime. Such monogamy was shown to result from the transfer of a pheromone, termed Matrone, produced by the accessory glands in males (Craig, 1967; Fuchs et al., 1968). Gwadz (1972) established neurohormonal regulation of female sexual receptivity, and Nijhout and Craig (1971) showed involvement of a sexual pheromone to bring about reproductive isolation in Stegomyia mosquitoes. Also, the genetic bases of
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some of the more interesting sex phenotype abnormalities, such as gynandromorphs, in which one part of the body of an individual is of one sex and the other part of another sex, were established using appropriate genetic markers on each of the three chromosomes (Rai and Craig, 1963). The sexual mosaics or gynandromorphs result from fertilization of an egg by two sperm, one maledetermining (M) and the other femaledetermining (m). Interestingly, this is fundamentally different from the genetic basis of gynander formation in D. melanogaster. Intersexes represent another interesting sexphenotype abnormality. Morphologically, such individuals are neither typical females nor males, but somewhat intermediate. These result from relatively high temperature exposure to larvae containing an autosomal recessive gene, ix, in the homozygous state (Craig and Hickey, 1967), and from several other stimuli (e.g., Motara and Rai, 1977).
3— Cytology: Genome Structure, Organization, and Evolution a— Comparative Karyotypes The author (K. S. Rai) joined the Mosquito Genetics Project as its first postdoctoral research associate on December 1, 1960, and immediately initiated cytological and cytogenetic studies of Ae. aegypti. At the time, even the chromosomes number of this species was in dispute; one report suggested 2n=4 (Carter, 1918) and the other, 2n=6 (Sutton, 1942). Professor Osmond Breland (1959) of the University of Texas at Austin published ''preliminary observations" on the use of a simple, yet highly instructive method of using the socalled "squash technique" for studies with mosquito chromosomes. This technique had long been developed and championed by plant cytologists, beginning with the incomparable Barbara McClintock (1929). Rai had used it extensively in his Ph.D. disser
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tation research in plant genetics at the University of Chicago. The extension of this technique to mosquitoes, therefore, was relatively quick and highly productive (Rai, 1963a, 1966a). As a result, comparative karyotypes were established for more than 80 species belonging to 12 genera in the family Culicidae and related families in the author's laboratory (Rai et al., 1982). Indepth analyses of genome structure, organization, and evolution among mosquito taxa at various levels have remained a major focus of Rai's laboratory for almost four decades (Rai and Black, 1999). Because of the success of this effort, studies of mosquito chromosomes quickly became a popular enterprise in several laboratories in the U.S. and elsewhere. This work revealed that although the chromosome number stayed the same in all species examined at UND (2n=6), the individual karyotypes had undergone a 4.5fold change in total chromosomal length and structural repatterning to make for distinctive karyotypes and considerable chromosomal evolution (Rai et al., 1982; Rao and Rai, 1987a; Matthews and Munstermann, 1994). b— Giemsa CBanding Patterns: Heterochromatin Differentiation The application of the Giemsa Cbanding procedures to mosquito karyotypes that soon followed greatly improved the resolving power of chromosomal analysis. Motara and Rai (1977) reported two distinct types of heterochromatin, constitutive and facultative, in Aedes mosquitoes. The latter was shown to possess a unique ability of differential expression under certain specific genetic backgrounds. These studies also revealed that karyotypic differentiation in mosquitoes involved changes in amounts, expression, and location of heterochromatin associated with concomitant changes in length and morphology of individual chromosomes (Motara and Rai, 1978). Dr. Motara is currently serving as the Dean of Students at Rhoades University in South Africa and has become a recognized and an astute educator in the country. Rao and Rai (1987a) extended Giemsa Cbanding methodology to study 43 dipteran species belonging to 17 genera of the nematocerous families Tipulidae, Dixidae, Chaoboridae, and Culicidae.
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Dixa recens, Eucorethra underwoodi, and Mochlonyx velutinus have diploid numbers of eight; the rest have six chromosomes. The data indicated that Dixidae evolved from primitive Tipulidae, which then gave rise to Chaoboridae, which in turn gave rise to Culicidae. Anophelinae and Culicinae evolved along separate lines after arising from the same ancestor. During the last two decades, much progress has been made in studies of molecular aspects of mosquito genomes with particular emphasis on their molecular size, structure, organization, and evolution. c— Genome Size Variation i— Interspecific Variation Through the use of quantitative cytophotometry of Feulgenstained primary spermatocytes, haploid genome sizes were established for 36 species belonging to 12 genera of mosquitoes and other closely related taxa in the superfamily Culicoidea. These results showed an 8fold variation in haploid DNA amounts and a 4.5fold variation in total chromosomal lengths among the various taxa examined (Rao and Rai, 1987a, 1990). Furthermore, linear regression analysis of a relatively large data set involving 28 species belonging to 11 genera showed a significantly positive correlation between total chromosomal length and haploid genome size (Rao and Rai, 1987b). ii— Intraspecific Variation Haploid (1C) nuclear DNA amounts were also determined for 47 geographic populations of Aedes albopictus from 18 countries, including the continental U.S. and Hawaii, in order to determine the variation at the intraspecific level (Kumar and Rai, 1990a). Overall, the haploid DNA amounts varied nearly 3fold from 0.62 ± 0.02 (mean ± S.E.M.) picograms (pg) in a population from Koh Samui, Thailand, to 1.66 ± 0.08 pg in the original invading population found in Harris County, Texas (Sprenger and Wuithiranyagool, 1986). Within the continental U.S. populations, the values ranged from 1.03 ± 0.03 pg in the Chambers County, Texas population to
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1.66 ± 0.08 pg in the Harris County, Texas population (Rao and Rai, 1987a; Kumar and Rai, 1990a). There has been much debate concerning the Functional aspects of genome size variation; DNA content has been found to be correlated with a variety of cellular and organismic attributes. We found a significant positive correlation between genome size and development rate in 10 different strains of Aedes albopictus (Ferrari and Rai, 1989). d— Intra and Interspecific Variation in Repetitive DNA Amount and Organization Repetitive DNA sequences comprise a large proportion of the eukaryotic genome. The evolutionary history of the eukaryotic genome reflects the turnover of repetitive sequences, and speciation may be a consequence of such turnover. The mechanism(s) for such evolutionary turnover is not yet clearly understood. However, insights concerning such genome processes are vital. Using DNA reassociation kinetics, we have shown that the observed variation in nuclear DNA content is due mainly to differences in the amount of repetitive sequences; a linear correlation was found between genome size and proportion of the genome represented by repetitive DNA sequences. Such sequences comprise 20–84% of the genome among the various mosquito species and strains of Ae. albopictus examined. The amount of fold back, highly repetitive, and middle repetitive DNA increased with genome size. Also, interestingly, two distinctly different types of genome organization were observed in the same family, Culicidae. DNA repeat sequences in Anopheles quadrimaculatus follow the socalled longperiod interspersion pattern while all the culicine species examined showed a shortperiod interspersion pattern (Black and Rai, 1988). Further, the copy number of nine highly repetitive DNA sequences cloned from Aedes malayensis of the Aedes scutellaris subgroup and of eight such sequences from the OAHU strain of Ae. albopictus from the Aedes albopictus subgroup varied extensively, among several species in the two subgroups (McLain et al., 1986) and among 15 strains of Ae. albopictus (McLain et al., 1987).
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e— Chromosome Mapping of Repetitive DNA The chromosome location and genomic organization of five of McLain et al.'s (1987) clones (viz, H85, H61, H76, H19, and H115) were studied using in situ and Southern hybridization techniques. Preparations of meiotic and mitotic chromosomes showed that the sequences homologous to H85 DNA were dispersed throughout tile length of all three pairs of chromosomes in Ae. albopictus, Ae. seatoi, Ae. flavopictus of the Ae. scutellaris group, and the closely related species, Ae. aegypti belonging to the Ae. aegypti group, Dotblot hybridization revealed that the sequences homologous to H85 DNA were present in 12 species of mosquitoes examined, belonging to 6 genera in subfamilies Culicinae and Anophelinae, as well as in Mochlonyx velutinus, the presumptive ancestor of the Culicids (Kumar and Rai, 1991a). An analysis of chromosomal localization and genomic organization of three other cloned repetitive DNA fragments (H19, H61, and H76) showed that the sequences homologous to these fragments were also dispersed throughout the genome in Ae. albopictus, Ae. flavopictus, Ae. seatoi, Ae. alcasidi (Ae. albopictus subgroup) and Ae. polynesiensis, Ae. katherinensis (Ae. scutellaris subgroup), Ae. aegypti, Haemagogus equinus, Tripteroides bambusa, and Anopheles quadrimaculatus (Kumar and Rai, 1991b). The dispersion pattern of one of the clones, H115, was distinctive. This family of sequences was located at the intercalary position on both homologues of chromosome 1 in three strains of Ae. albopictus (OAHU, KOH SAMUI, SAVANNAH), Ae. seatoi, Ae. flavopictus, Ae. polynesiensis, Ae. alcasidi, Ae. katherinensis, and Ae. aegypti. The H115 sequences were widely conserved in Culicidae and were found in Hg. equinus and Tp. bambusa, besides Aedes and Anopheles species. DNA sequence analysis of a 284 bp fragment showed that the H115 insert contained two perfect inverted repeats and numerous perfect direct repeats. The occurrence of inverted repeats with potential to form intrastrand palindromic structure suggested that the H115 family of sequences might be involved in chromatin condensation (Kumar and Rai, 1992). Additional details concerning genome structure, organization, and evolution and vari
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ous interrelationships are included in two recent reviews (Kumar and Rai, 1993; Rai and Black, 1999).
4— Genetics of Chromosomal Rearrangements Soon after the methodology for chromosomal and cytogenetic studies in Aedes aegypti was well delineated, a concerted effort began in Rai's laboratory around early 1962 to isolate and genetically characterize chromosomal rearrangements, following radiation exposure of adult males, with particular emphasis on the reciprocal translocations and inversions. This became a major research focus during the decades of the '60s and '70s. The morphological mutants isolated and characterized by Craig and coworkers and the subsequent construction of marker stocks (in which each chromosome was marked by at least one mutation e.g., the "oldfaithful" RED stock with genes for red eyes (re), spot abdomen (s), and black tarsi (blt) on chromosomes 1, 2, and 3, respectively, provided the substrate for the studies aimed at isolating heritable chromosomal rearrangements. Reciprocal translocations are chromosome rearrangements involving the exchange of pieces of nonhomologous Chromosomes. In Ae. aegypti more than 150 reciprocal translocations were induced in a widely used laboratory stock, ROCKEFELLER (ROCK), and two field stocks, one from Delhi, India, and another from Mombasa, Kenya. These translocations were cytogenetically analyzed for their break points, fertility, fecundity, and transmission characteristics (Rai and McDonald, 1967; Rai et al., 1974; Hallinan et al., 1977). Studies were also undertaken on the competitive mating ability of males heterozygous for the more promising of these translocations in laboratory and field population cages (Rai and McDonald, 1971), and attempts were made to produce homozygotes for such translocations (Lorimer et al., 1972). Some of these translocations were
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also used to correlate the three genetic linkage groups with the three physical chromosomes in the species (McDonald and Rai, 1970). Computer simulations using the available data in Ae. aegypti indicated the potential role of various types of sexlinked and autosomal translocations for genetic control under various release strategies (McDonald and Rai, 1971). Chromosomal inversions which involve rotation of a segment of a chromosome by 180° also received considerable attention (McGivern and Rai, 1972, 1974). In time, much effort was expended on evaluating the feasibility of using the appropriate translocations for genetic control of field populations. Also, extensive studies were undertaken to establish the cytogenetic basis of radiation and chemicallyinduced fertility and fecundity so as to provide an understanding of the underlying mechanism of the socalled sterilemale technique in Ae. aegypti (Rai, 1963b, 1964a, b).
5— Genetic Control: Evolution of the Concept The development of insecticide resistance among several important mosquito vectors of human disease Such as Ae. aegypti, Culex fatigans, and Anopheles species in the '50s and '60s was recognized as a major public health problem (Brown and Pal, 1971). The traditional method of control—use of synthetic organic insecticides—in most cases proved ineffective and Undesirable, not only because of the relatively rapid development of resistance on the part of targeted insects, but also because of the effects on the nontarget population and the environmental pollution that ensues from the use of such chemicals. Such observations were largely instrumental in the emerging realization of the paramount need to develop alternative types of control methods and to evaluate them under field conditions. In simplest terms, genetic control involves any hereditary manipulation to suppress populations and is generally based on utiliti
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zation of individuals of a particular species to control populations of that species. Thus, nothing alien is introduced into the ecological niche of a pest population. Hence, maintenance of the environment in the original untreated from is likely to be assured, at least in theory. Understandably, mechanisms for genetic control for any species can emerge only from. a thorough investigation of the genetic biology of that species. Also, acquisition of detailed knowledge concerning the population dynamics and the ecology of the candidate vector species, particularly under field conditions, was recognized to be a necessary prerequisite for successful application of genetic control. The VBL had already become a leading center of research on various aspects of biology and genetics of Ae. aegypti. An important objective of this work had always been to use the information thus gained for possible genetic control purposes. As a result, development of genetic control methodology became a major mission of the VBL soon after its founding, and Ae. aegypti emerged as the prototype species among mosquitoes in which to rigorously test the new concept. This effort was nurtured with exceptional commitment by two major international agencies, the World Health Organization, Geneva, Switzerland, and the International Atomic Energy Agency, Vienna, Austria. An important mission of these agencies is to manage mosquito transmitted diseases through the development and application of appropriate method(s) by the former and the SterileInsect Technique (SIT) in particular by the latter agency. As a result, both these agencies played a vital role in providing appropriate forums for discussion of the issues involved and possible solutions of the same in the form of various group meetings, panels, seminars, and symposia in the '60s and early '70s and the sponsorship of subsequent field trials. Following are some of the more noteworthy activities sponsored by these two United Nations agencies: World Health Organization (WHO): WHO Scientific Group on Genetics of Vectors and Insecticide Resistance, 5–9 August 1963, Geneva, Switzerland (G. B. Craig, Jr., Invited Participant). WHO Techn. Rep. Ser. 268:40 pages, 1964.
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WHO Seminar on the Ecology, Biology, Control, and Eradication of Aedes aegypti, 16–20 August 1965, Geneva, Switzerland (G. B. Craig, Jr., and K. S. Rai, Invited Participants). Bull. WHO 36:519–702, 1967. WHO Informal Meeting of the Coordination Group on the Genetic Control of Insects of Public Health Importance, 5–9 June 1967, Washington, D.C. (G. B. Craig, Jr., Invited Participant). Bull. WHO 38:421–438, 1968. WHO Scientific Group on the Cytogenetics of Vectors of Disease of Man, Geneva, Switzerland, 31 October to 6 November 1967 (K. S. Rai, Invited Participant). WHO Techn. Rep. Ser. 398:41 pages, 1968. International Atomic Energy Agency (IAEA): Feasibility study on the application of the SterileMale Technique for the control of the filariasis vector, Culex fatigans, in Ceylon. IAEA Special Publication WP/5/283:38 pages, 1966b (K. S. Rai, author). IAEA Study Group on Effects of Radiation on Meiotic Systems, Vienna, Austria, 8–11 May 1967 (K. S. Rai, Invited Participant). STI/PUB/173:223 pages, 1968. IAEA Panel on SterileMale Technique for Eradication or Control of Harmful Insects, Vienna, Austria, 27–31 May 1968 (K. S. Rai, Invited Participant). STI/PUB/224:142 pages, 1969a. Isotopes in Entomology: Report to the Govt. of Brazil. IAEA Special Publication WP/5/483:38 pages, 1969b (K. S. Rai, author). IAEA symposium on the Sterility Principle for Insect Control or Eradication, IAEA, Vienna, Austria, 14–18 September 1970 (K. S. Rai, Invited Participant). STI/PUB/265:542 pages (1971). Historically, the involvement of Professors Craig and Rai in the above deliberations, beginning in 1963, helped lay the foundations first for the development of the theory and subsequently for the practice of genetic control of mosquitoes. On an invitation from
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the IAEA, the author (K. S. Rai) served as a consultant in Ceylon (now Sri Lanka) during May—September 1966 to advise the agency and the government of Ceylon on the feasibility of the use of the SterileInsect Technique for the control of the common house mosquito, Culex fatigans, the major vector of filariasis in the country. Although SIT was almost exclusively an IAEA domain, the senior personnel of WHO were eager to be kept informed and, if possible, involved in this IAEA sponsored project. Mr. James W. Wright and Dr. Rajinder Pal, Chief and scientist/biologist, respectively, of Vector Biology and Control of the World Health Organization, invited the author. (Letters dated 18 February 1966, and 1 March 1966) to visit WHO headquarters in Geneva, Switzerland, on his way to Sri Lanka, which he did. The purpose was to discuss ongoing WHO efforts involving genetic control of mosquitoes through the use of smallscale, sterilemale releases and the use of cytoplasmic sterility at their research unit on filariasis in Rangoon, Burma. These discussions were formalized in the form of a written document; WHO wished to collaborate with us on such efforts. Again, following the completion of the IAEA assignment in Ceylon and on an invitation from WHO, Dr. Cutkomp, the Head of Insect Eradication at IAEA, and the author (K. S. Rai) visited the WHO headquarters in Geneva to apprise Mr. Wright and Dr. Pal of the recommendations made by the author to the IAEA on the feasibility of the application of the sterilemale technique for Cx. fatigans control in Ceylon. This was soon followed by a Scientific Group on the "Cytogenetics of Vectors of Disease of Man" convened by WHO in Geneva during 31 October–6 November 1967. At this meeting the author presented two seminal papers, one entitled "Genetics of chromosomal aberrations in insect vectors of disease" (Rai and McDonald, 1967) and another on "Manipulation of cytogenetic mechanisms for genetic control of vectors" (Rai, 1967). The latter paper formally proposed the use of chromosomal translocations for vector control. Extensive work done in the author's laboratory on the genetics of chromosomal rearrangements, particularly translocations (Rai, 1967; Rai and McDonald, 1967; Rai and Asman, 1968) formed the basis of this proposal.
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6— Seminar in Vector Genetics Another major event that shaped the future of studies dealing with genetic control of mosquitoes at the VBL in particular, and around the globe in general, was a six week Seminar in Vector Genetics held at UND during 17 June–20 July 1968, Under the joint sponsorship of the World Health Organization and the University of Notre Dame. Following extensive discussions between Mr. Wright, Dr. Pal, and Professors Craig and Rai, the WHO selected ten established scientists from Tanzania, Uganda, Thailand, Singapore, Pakistan, India, Argentina, Venezuela, and Upper Volta who were actively involved in studies of various aspects of vector biology and public health, and UND selected six individuals from North America to participate in this seminar. All the graduate students and postdoctorals in residence at the VBL also participated. The participants were housed in the student dormitories at UND. The expenses for travel and per diem foreign participants and the faculty from institutions other than UND were borne by the WHO. The list of the teaching faculty and the participants involved in this seminar reads like a ''Who's Who" in the ranks of the discipline of Mosquito Biology that subsequently evolved the world over. It included the following: Faculty: Dr. A. W. A. Brown, University of Western Ontario, London, Ontario, Canada Dr. Donald G. Cochran, Virginia Polytechnic Institute, Blackburn, Virginia Dr. George B. Craig, Jr., University of Notre Dame, Notre Dame, Indiana Dr. Robert E. Gordon, University of Notre Dame, Notre Dame, Indiana Dr. William A. Hickey, Saint Mary's College, Notre Dame, Indiana Dr. James B. Kitzmiller, University of Illinois, Urbana, Illinois Dr. Leo E. LaChance, Metabolism and Radiation Research Laboratory, United States Department of Agriculture (USDA), Fargo, North Dakota
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Dr. Karamjit S. Rai, University of Notre Dame, Notre Dame, Indiana Dr. Rajinder Pal, Vector Biology and Control, World Health Organization, Geneva, Switzerland Participants: Dr. Chan Kai Lok, Vector Control Unit, Ministry of Health, Singapore Dr. R. Carcarvall, Francisco Deiro 3390, Buenos Aires, Argentina Dr. E. R. Shahgudian, Institute of Parasitology, Tropical Medicine, and Hygiene, Teheran, Iran Dr. A. S. Nasir, Atomic Energy Agency, Lahore, Pakistan Dr. G. B. White, East African Institute of Malaria and VectorBorne Disease, Amani, Tanzania Dr. C. E. MachadoAllison, Central University of Venezuela, Institute of Tropical Zoology, Caracas, Venezuela Dr. L. G. Mukwaya, East African Virus Research Institute, Entebbe, Uganda Dr. Raymond Subra, Centre Muraz, BoboDiculasso, Haute Volta Dr. Evans Offori, Ghana, Metabolism and Radiation Research Laboratory; USDA, Fargo, North Dakota Dr. Han Heng Yap, Malaysia, Entomology Dept., University of Minnesota, St. Paul, Minnesota Dr. Demrong Boonyeon, Thailand, School of Public Health, Tulane, New Orleans, Louisiana Maj. James Willman, U.S. Army Medical Laboratory, St. Louis, Missouri Dr. S. L. Narang, India, Dept. Zoology, University Of Illinois, Urbana, Illinois; (currently Group Leader, USDA, Washington) Dr. S. Kanda, Japan, Dept. Zoology; Univ. Of Illinois, Urbana, Illinois Capt. Louis Rutledge, Dept. Entomology, Walter Reed Army Medical Center, Washington, D.C. Mr. Michael Dunn, UND (currently Northwestern University Medical School, Chicago, Illinois) Miss Suzanne Buechler, St. Mary's College, Notre Dame, Indiana Mr. Wenceslaus Kilama, Tanzania and University of Notre Dame (currently Director General, Medical Research, Tanzania)
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Dr. W. K. Hartberg, University of Notre Dame (currently Chairman, Biology Dept., Baylor University) Mr. Robert Gwadz, University of Notre Dame (currently Malaria Section, National Institute of Health) Mr. George O'Meara, University of Notre Dame (currently Professor, University of Florida) Mr. Paul T. McDonald, University of Notre Dame (currently Entomologist with a chemical company) Mr. James McGivern, University of Notre Dame (currently Professor of Genetics, Gannon University) Mr. Jeffrey Powell, University of Notre Dame (currently Professor of Biology, Yale University) Mr. Harold Smith, University of Notre Dame (currently M.D., practicing medicine) The period during 17 June–12 July 1968 consisted of extensive, indepth lecture presentations on basic and applied principles of vector genetics, mutagenesis, sterile male technique, insecticide resistance, and theories and practice of genetic control, and of indepth, handson experience in laboratories at the UND Biology Department. The laboratory instruction reenforced the theoretical concepts imparted in lectures. The academic program at UND was followed by a two week (1326 July 1968) Mosquito Safari, a tour of laboratories and practical demonstrations of vector control methodology at the following major public health facilities in the U.S.: 1) The National Communicable Disease Center, Atlanta, Georgia 2) Technical Development Laboratory, United States Public Health Service (USPHS), Savannah, Georgia 3) Laboratory of Insects Affecting Man, USDA, Gainesville, Florida 4) Florida Entomological Research Center, Vero Beach, Florida The field trip, Undertaken on a bus rented for the purpose, promoted a rather close camaraderie among the participants and the UND faculty. It also laid the foundation of an excellent working relationship for years to come, thereby enhancing the science of vec
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tor biology and control by making it a cohesive group. The sixweek enterprise of living and working together, on an almost daily basis, was highly synergistic. In retrospect, an equally important facet of this seminar for the future of the field of genetic control of vector populations was the opportunity it provided for extensive and informal discussions, throughout the duration of the seminar, between Dr. Pal and the author as to where and how to test, under field conditions, the concepts of genetic control developed at UND and elsewhere. The focus of these discussions was on the use of chromosomal translocations and the SIT for genetic control, and the concept of population replacement of vector by nonvector populations through the use of translocation homozygotes and cytoplasmic incompatibility as originally proposed by Curtis (1968a) and Laven (1967), respectively. The concrete idea of the founding of a WHOsponsored research unit at an appropriate location in either the continent of Africa or Asia, where mosquitotransmitted diseases represented major public health problems, emerged from such informal discussions. Soon, a consensus evolved that in view of the scientific infrastructure, availability of needed facilities, and a sufficiently, trained scientific work force, an appropriate location in India would represent tile preferred choice. To follow up this idea, the WHO agreed to discuss with the Government of India the desirability of establishing a "Research Unit on Genetic (Control of Mosquitoes" in New Delhi, India. These discussions soon bore fruit: a collaborative research unit under joint sponsorship of the World Health Organization and the Indian Council of Medical Research (WHO/ICMR) became operational in late 1969. A spacious building on the Ring Road in New Delhi was rented for the purpose. The scientific and administrative facilities needed to undertake a large research enterprise were soon put in place and the staff hired. Dr. Carrol Smith from the USDA was the first project leader and most of the leading practitioners of genetic control of vectors at the time served as consultants at various times. Besides the author, these included Drs. George Craig, George Davidson, and Chris Curtis (London School of Tropical Medicine and Hygiene); Hannes Laven (University of Mainz, Germany), Max Whitten (Commonwealth Scientific and
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Industrial Research Organisation, Australia), and numerous others. Two graduate students from Rai's laboratory at UND, Edward Hallinan and Nancy Lorimer, were also hired as longterm WHO consultants to conduct work with translocation releases in Delhi.
7— Genetic Control: Principles and Mechanisms The basic principles and theoretical consideration underlying genetic control of insects have been adequately emphasized by LaChance and Knipling (1962), Craig (1963), Rai (1967, 1996; Knipling et al., 1968), and others. In general, because of the propagation of sterility and/or other desirable genetic factors from one generation to the next, these methods are based on birth rather than death control. As mentioned earlier, several potentially useful mechanisms have been proposed for genetic control of mosquitoes. Of these, the following two have been extensively evaluated in field trials with considerable involvement of the VBL personnel. a— SterileInsect Technique (SIT) The SIT is based on the induction of sexual sterility in males through the use of radiation or chemical sterilants and on inundating natural populations with such males (Knipling, 1955, 1959). To a large degree, interest in methods of genetic control of insects is a byproduct of the successful application of the sterileinsect release methods to control the screwworm fly; Cochliomyia hominivorax, from the southeast and southwestern United States and the West Indian island of Curaçao in the mid1950s (Knipling, 1959). Rai (1963, 1964a, b), LaChance (1967), and Curtis (1971) provided important insights concerning the genetic basis of mutageninduced dominant lethality in insects. Depending on the time of their application, radiation and/or chemosterilants can inhibit and
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completely prevent the production of gametes in insects (Rai 1964b, LaChance 1967). However, when the gametes are produced after such treatment, "sexual sterility" usually results from the induction of dominant lethality in sperm and ova, caused by structural. chromosomal aberrations such as loss of chromosomes or chromosome parts, particularly those that result in formation of broken ends. Mitotic anomalies such as the breakagefusionbridge cycles and the genetic imbalance that ensue from the induction of such drastic rearrangements in a developing embryo result in its death before embryogenesis is completed. Such fertilized eggs do not hatch. Although SIT cannot technically be regarded as a type of genetic control because complete sterility is not inherited (NAS 1969; Rai et al., 1973), it is customary to include it under genetic methods. The dominant lethal mutations induced in sperm of irradiated or chemically, sterilized males and that result in male sterility are caused by drastic chromosomal aberrations and are hence genetic in nature. The application of this technique for mosquito control has been reviewed by Rai (1969) and by Asman et al. (1981). All early, trials involving SIT were based on releases of males sterilized with radiation. Unfortunately, measured in terms of reductions of the target populations, none of these trials were successful. The reasons for these failures are fairly well understood (Rai, 1969). In general, they have been ascribed to either the use of massive doses of radiation to sterilize males, or to releases of males possessing reduced fitness ensuing from a long history of laboratory colonization. b— Chromosomal Translocations The use of inherited sterility associated with reciprocal chromosomal translocations has attracted considerable attention as a potential method for pest control. Although the potential of this method was originally proposed by a Russian geneticist (Serebrokskii, 1940), it was not until almost three decades later that its use for a number of insect species, such as mosquitoes, tsetse flies, and house flies, was contemplated and progress made (Rai, 1967; Rai and Asman, 1968; Rai and McDonald, 1971; Curtis, 1968b). Because approximately 50% of the gametes formed as a result of
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adjacent segregation from a translocation heterozygote produce inviable zygotes, crosses involving normal individuals and translocation heterozygotes are usually semisterile. Furthermore, half the progeny surviving such crosses also inherit the same translocation and in turn pass it on to their offspring. Crosses among individuals heterozygous for the same translocations are expected to produce translocation homozygotes, translocation heterozygotes, and standard (chromosomally normal) progeny in a ratio of 1:2:1. Individuals homozygous for a translocation would be expected to show full fertility when mated to normal individuals with the standard chromosome arrangement because each gamete formed contains the full haploid complement. All the progeny from such mating, however, will be translocation heterozygotes and semisterile. Thus, for field releases of a single translocation, homozygotes are expected to be more efficient than heterozygotes in terms of numbers of individuals released. Twice the number of heterozygotes would be required to introduce as many translocated chromosomes (Lorimer et al., 1972). Homozygotes are also useful in obtaining stocks of multiple translocations in which the genome is rearranged even more drastically, thereby increasing the sterility of the progeny: This can be done by irradiating translocation homozygotes and assaying for other translocations that are then also made homozygous. Double translocation heterozygotes can also be produced by crossing individuals heterozygous for two different, single translocations (Hallinan et al., 1977). Such double heterozygotes are characterized by approximately 75% sterility. In a release program in which large numbers of individuals must be reared, homozygotes are much easier to mass produce than heterozygotes because of their full fertility. Also, raising homozygotes eliminates the culling problem often associated with heterozygotes because all progeny inherit the same chromosomal rearrangement. As mentioned earlier, heterozygotes produce 50% normal progeny that have to be culled unless the translocation is very tightly linked with the male determining locus (M), in which case most or all sons will be translocated and all daughters normal. Homozygotes have also been proposed as a vehicle for driving desirable genes, such as refractoriness to parasitic diseases, conditional
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lethals, and other alterations, into natural populations (Curtis, 1968a). Such genes could be tightly linked with the translocation breakpoints and with appropriate manipulation driven to fixation in the population along with the translocation. In Ae. aegypti more than 150 reciprocal translocations were induced and cytogenetically analyzed to determine their fitness for genetic control (Rai et al., 1970; Rai and McDonald, 1972; Rai et al., 1974; Hallinan et al., 1977). Attempts made to produce homozygotes for such translocations yielded five homozygote stocks.
8— Genetic Control: Applications a— The Delhi Project: WHO/ICMR The feasibility of SIT was evaluated in the early 1970s in Cx. fatigans in several villages in the vicinity of Delhi, India, under the auspices of the abovementioned Research Unit on Genetic Control of Mosquitos, established by WHO in collaboration with ICMR, the Indian Council of Medical Research (Patterson et al., 1975; Yasuno et al., 1978). In several of these smallscale feasibility trials, the sterile males were not fully competitive under field conditions, and immigration of fertile females from surrounding areas to experimental sites could not be effectively curtailed. Nevertheless, these studies helped to highlight the need for additional requirements/developments to effectively control target populations in future field trials. Field work done under the sponsorship of the WHO/ICMR research unit on evaluating the feasibility of genetic control of mosquitoes in New Delhi, India, during the early 1970s demonstrated the genetic incorporation and maintenance of a genetic marker and a malelinked translocation in a natural population and their maintenance for several generations following the termination of the field releases (Rai et al., 1973). This was the first demonstration of
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its type among any vector species. Such longterm survival and perpetuation of an introduced genetic mechanism over several generations is an essential prerequisite for successful application of genetic control. Unfortunately, some of the largescale field releases planned by WHO/ICMR in India against relatively large urban populations of Ae. aegypti and Cx. fatigans following fiveyear feasibility studies were prematurely terminated for strictly political reasons, which have been discussed in detail elsewhere (WHO, 1976). b— The Mombasa Project: USAID At about the time the WHO/ICMR research unit was established in New Delhi, the U.S. Agency for International Development (USAID) awarded a contract entitled "Ecological Studies on Aedes aegypti Preliminary to Genetic Control" to UND with Craig and Rai as coPrincipal Investigators. George Craig was one of the research directors of the Nairobibased International Center of Insect Physiology and Ecology (ICIPE) at the time. We opened a field station just north of Mombasa, Kenya, and named it "The Mosquito Biology Unit" or MBU, the latter being the Swahili word for mosquito. MBU was, in a way, an outreach of VBL, and was affiliated with the ICIPE. It received its funding exclusively from USAID. MBU was based in a spacious beachfront house on the shores of the Indian Ocean with two large laboratories, two insectories, a mass rearing facility, a radiation source, a LandRover vehicle, an animal room, offices, a library; and a thirty comfortable suite of rooms with cooking and laundry facilities as living quarters (Lorimer and Lorimer, 1975). In view of the elegance and the location of this unit on the shores of the Indian Ocean, with its clean private, sandy beach front and the graciousness of the resident hostess, Mary Ann Hausermann, MBU quickly became an attractive place to visit. Craig, Rai, and Crovello made numerous trips to plan and oversee the work; Paul Weinstein, the department chairman, visited the laboratory and wrote a report. The president of the university, Father Theodore Hesburgh, made an unannounced visit and celebrated mass on Christmas Eve in 1974. The field site located approximately 20 kilometers away, just off the main road to Nairobi, consisted of several small villages belong
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ing to the Rabai tribe. Three distinct types of populations of Ae. aegypti with little gene flow among them in the Rabai areas and other locations in East Africa were recognizable. They are the dark forestdwelling form, the peridomestic form breeding in manmade containers outside the houses, and the highly domestic populations living inside houses in close association with man (Tabachnick et al., 1979). The domestic form, which is important as a disease vector, was the focus of our work in Mombasa preliminary to and as a part of our field releases for genetic control. For a major part of the Mombasa MBU operations, Dr. Walter Hausermann served as the resident project leader, and at different times, Drs. John Peterson, Paul McDonald, and Nancy Lorimer worked on various aspects of the project. The first two had earlier worked in Craig's laboratory and the other two in Rai's laboratory at the VBL. The use of single and double translocation heterozygotes for genetic control of Ae. aegypti was evaluated in villages around Mombasa, Kenya (Peterson et al., 1977; McDonald et al., 1977). The targets were small village populations. Substantial (60–70%) sterility), was introduced into native females during and immediately following these releases. However, no population suppression could be documented as a consequence of infiltration of Ae. aegypti spp. formosus (the jungle form) into the otherwise domestic habitat of the type form Ae. aegypti. The concept of population replacement through the release of translocation homozygotes was also evaluated in Mombasa. However, the results were negative because of the lack of competitiveness of the released strain under field conditions and because of behavioral differences in oviposition preference between the homozygous strain released and the indigenous population (Lorimer et al., 1976). The releases of translocations stocks of Ae. aegypti in Mombasa were also terminated prematurely in the mid1970s because of the largescale reemergence of malaria and the need perceived by USAID to shift focus to work with Anopheles gambiae, under the presumption that Ae. aegypti was not an important vector species in Africa. Although the results of the field trials to date in Delhi and
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in Mombasa admittedly were not startling because of the reasons discussed, nevertheless they established the validity of the scientific principle of genetic control. In no case can the lack of optimal success be ascribed to a limitation or a negative feature of the concepts themselves. Furthermore, much was learned about the field biology, ecology, and dynamics of field populations as a result of the field releases both in India and East Africa. Recent developments in molecular genetics offer exciting possibilities for extension and possible future applications of genetic control. These are highlighted in Crampton and Eggleston, 1992; Crampton, 1992; Besansky et al., 1992; Curtis, 1992; Kidwell and Ribeiro, 1992; Collins, 1994; Besansky and Collins, 1994; and others. Currently, the focus of this work is on the production of competitive transgenic mosquito strains and on the isolation of appropriate transposable elements that could be used to drive disease refractory genes into vector populations.
9— Population Genetics Extensive work has been done in the field of population genetics at various levels of taxonomic organization at the VBL over the years. The objective has been to establish and characterize patterns and levels of genetic variation at the individual, population, species, and generic levels. Such studies are critical in understanding various facets of vectorborne disease, epidemiology, and vector control. Leonard Munstermann joined the VBL as a graduate student in 1970 and received his Ph.D. degree in 1979 with George Craig as his mentor. He rose through the ranks to Associate Faculty Fellow at UND. He has been a pioneer in this field: Leonard is an exceptionally creative individual whose contributions in mosquito population genetics and systematics are universally recognized. Besides, he is an accomplished photographer whose mosquito slides are used
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around the globe. Several postdoctorals and graduate students have collaborated with him or have followed in his footsteps. Among others, these include Walter Tabachnick, Jeff Powell, Annalisa Marchi, Stephen Saul, Tom Mathews, William Hawley; Phil Lounibos, David Taylor, and more recently, JohnPaul Mutebi in Craig's laboratory. Drs. Kelly McLain, Larry Hilburn, Larry Szymczak, Dorothy Pashley, James Ferrari, William Black, and Srinivas Kambhampati in Rai's laboratory have similarly made outstanding contributions to this field. Drs. Black and Kambhampati, in particular, have been major participants in elucidating the population genetic structure of Ae. albopictus from numerous locations in its native habitat as well as from several locations in the U.S. following its introduction in 1985. Furthermore, Tabachnick and Black (1995) and Munstermann and Conn (1997) have highlighted the roles of molecular and cladistic analysis in population genetic studies in vector biology. a— Allozymes The early work involved the use of allelic frequency differences at various polymorphic enzyme loci (allozymes) and delineation of genetic structure of sympatric populations of Ae. aegypti in East Africa (Munstermann, 1979; Tabachnick et al., 1979); Aedes species on the Kenya coast (Lounibos and Munstermann, 1981); the sibling species, Ae. triseriatus and Aedes hendersoni (Saul et al., 1977a, 1978; Mathews and Munstermann, 1983; Mathews, 1983; Munstermann, 1985); Ae. atropalpus complex (Munstermann, 1980; Szymczak et al., 1986); Ae. scutellaris group (Hilburn and Rai, 1981; Kambhampati and Rai, 1991a); three Stegomyia groups, aegypti, walbus, and scutellaris (Pashley and Rai, 1983a; Pashley et al., 1985); Ae. stimulans group (Eldridge et al., 1986); Sabethes cyaneus (Munstermann and Marchi, 1986); Nearatic Aedes species (Munstermann, 1988); Ae. albopictus (Black et al., 1987, 1988; Kambhampati et al., 1991); Culex species (Saul et al., 1977b; Corsaro and Munstermann, 1984); Anopheles minimus (Green et al., 1990); Anopheles balabacensis (Hii et al., 1991); and the Anopheles dirus complex (Green et al., 1992). In several aedine species, particularly Ae. aegypti and Ae. triseriatus (Munstermann et al., 1982), and Ae. scutellaris subgroup species (Pashley and Rai, 1983b), al
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lozyme loci have been mapped. In certain studies, Nei's genetic distances have been used to separate the populations into various groupings and sometimes to compare geographic distances with genetic distances. Such pairwise comparisons employing coefficients of genetic distances (Nei's D) indicated that the sympatric, domestic, and sylvan populations of Ae. aegypti along the Kenya coast were fairly discrete with restricted gene flow (Tabachnick et al., 1979). Rai's laboratory paid special attention to the evolutionary population genetics of the Ae. scutellaris group in the subgenus Stegomyia, during the mid70s to mid90s. This group is divided into two subgroups: the scutellaris subgroup consisting of some 34 species which are largely allopatric in distribution on the islands of the South Pacific; and the albopictus subgroup with 11 described species having a predominantly sympatric distribution in Southeast Asia (Rai et al., 1982). Because of their unique zoogeography, the Ae. scutellaris group species represent a major resource for evolutionary studies designed to address fundamental questions dealing with the onset of reproductive isolation and the patterns and rates of genetic differentiation as a function of their contrasting distribution patterns. Consequently, much progress has been made with various species in the two subgroups in the areas of (a) experimental hybridization (Hilburn and Rai, 1981; Dev and Rai, 1982, 1985; Sherron and Rai, 1983); (b) matechoice tests as indicators of the intensity of isolation indices (McLain et al., 1985; McLain and Rai, 1986); (c) chromosomal differentiation (Dev and Rai, 1984; Sherron and Rai, 1984); (d) population genetics (Hilburn and Rai, 1981; Pashley and Rai, 1983a; Pashley et al., 1985); and (e) molecular organization and evolution of the genomes (McLain et al., 1986, 1987; Black and Rai, 1988). Also, using calibration of known geologic events, e.g., the separation of two islands in the South Pacific, these studies have allowed estimates of the rates of genetic differentiation as a function of evolutionary time (Pashley et al., 1985; Kambhampati and Rai, 1991c). b— Molecular Markers During the last few years, a variety of molecular markers have been used for population genetic analysis, e.g., ribosomal RNA
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genes (Black et al., 1989a; Kambhampati et al., 1990), repetitive DNA sequences in the Ae. scutellaris subgroup (McLain et al., 1986) and in the albopictus subgroup (McLain et al., 1987), mtDNA among scutellaris group species (Kambhampati and Rai, 1991c) and Ae. albopictus populations (Kambhampati and Rai, 1991b), and Random Amplified Polymorphic DNA (RAPD) (Kambhampati et al., 1992a).
10— The Aedes albopictus Saga: Physiological Characteristics, Population Genetics, and Temporal Changes in the New World The subgenus Stegomyia of the genus Aedes contains approximately 110 described species divided into seven groups, including the scutellaris group which, as mentioned earlier, is subdivided into the scutellaris and the albopictus subgroups consisting of 34 and 11 species, respectively (Rai et al., 1982). Aedes albopictus belonging to the Ae. albopictus subgroup is a remarkably successful colonizing species with a wide distribution and an important vector of pathogens causing human disease, particularly dengue and dengue hemorrhagic fevers. From its presumed origin in Southeast Asia it has expanded its range from Madagascar in the west to Hawaii in the east during the last approximately one hundred years. Within the past thirty years, the species has colonized the Solomon and Santa Cruz Islands in the South Pacific (see Hawley, 1988, and Rai, 1991, for references). In a grant application to the NIH in October 1983 entitled ''Genetic Differentiation of the Aedes Albopictus Subgroup," Rai predicted that "Introduction and subsequent establishment of Ae. albopictus in the continental United States cannot be discounted in view of the well documented success of this species to move through commerce routes and to colonize new regions. . . . For the
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continental United States, the potential public health importance of this species becomes even greater when one realizes that this species is considerably more cold tolerant than Ae. aegypti. Thus, if it were introduced to the United States, it might become established in more northern (Midwestern) states where Ae. aegypti does not exist. In Japan and China its distribution reaches just about 40°N latitude and in the Oriental region (North India, West Pakistan, Nepal) it can withstand long periods of freezing." Both these predictions were borne out by subsequent events. Relatively large populations of Ae. albopictus were found throughout Harris County, Texas, in August 1985 (Sprenger and Wuithiranyagool, 1986). From there it spread relatively quickly; the species is currently found to be widely distributed in some twentyfour states in the United States extending as far south as Brownsville, Texas, and south of Miami County, Florida, and north throughout the Midwest from Kansas City, Missouri, and Chicago, Illinois, to Baltimore, Maryland in the east. Using data on the distribution of the species in north Asia, Nawrocki and Hawley (1987) estimated that the 0°C isotherm is the northern limit of the overwintering range and the 5°C isotherm limits the maximum northward expansion of the species during summers. In the U.S., this is much farther north than the related Ae. aegypti can colonize and unquestionably puts more of the midwestern and eastern parts of the country potentially in contact with Ae. albopictus. The detection of Ae. albopictus in Harris County, Texas, presumably very shortly after its successful introduction into the continental U.S., provided a rare opportunity to study various aspects of the biology, population genetics, and vector competence of a species in the process of rapidly colonizing a new continent. Interest in research on Ae. albopictus consequently increased dramatically following its introduction and spread into other areas of the Americas from the mid1980s to the present. The ongoing work on the genetics of this species underway in Rai's laboratory since the early 80s was followed in Craig's laboratory in the mid80s. Again, these studies set the pace for the rest of the country. It should be emphasized that Ae. albopictus is a primary vector of dengue virus in rural areas of Southeast Asia. However, the species has adapted itself in
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creasingly to urban habitats both in its native habitat, e.g., in Singapore, and in the U.S., e.g., Chicago, Houston, and New Orleans among other cities, thus enhancing the potential for urban epidemics should dengue virus be introduced and spread. Among all the viruses which Ae. albopictus can vector, dengue poses the most serious threat in the Americas, followed by its potential for La Crosse virus transmission in the midwestern U.S. Dengue virus activity has remained at a relatively high level in recent years, with epidemics involving all four dengue serotypes occurring in various parts of the Americas (Gubler, 1988). Furthermore, many cases of suspected dengue are imported into the continental United States every year. Perhaps more ominously, in view of the demonstrated ability of the species to transovarially transmit this virus, Ae. albopictus has the potential to effectively import the virus into the U.S. and initiate an epidemic without an infected human reservoir population. This made it all the more imperative to genetically characterize the U.S. populations, to decipher their origin, and to determine their vector competence. This was done using the following parameters; the first two in Craig's laboratory and the remaining in Rai's laboratory: a— Photoperiodic Sensitivity Hawley et al. (1987) examined seventeen strains of Ae. albopictus for photoperiodicallyinduced embryonic diapause by exposing females in the laboratory to either long or shortday conditions. The strains tested from the U.S. (Texas, Louisiana, Florida, Tennessee, and Indiana) and from northern Asia (Beijing, Tokyo, and Korea) showed a photoperiodicallyinduced egg diapause, i.e., when exposed to the shortday lengths, the females laid eggs which either failed to hatch or gave low hatches. However, none of the strains tested from tropical Asia (Mauritius, Madagascar, and northern Taiwan) (Hawley et al., 1987) and none of the six from Brazil (Hawley and Craig, 1989) showed any photoperiodic response. b— ColdHardiness Exposure of embryonated eggs to subfreezing temperatures in the laboratory (10°C for a 24hour period after they were cold
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conditioned at 5°C for two weeks), for eggs laid by females of six North American and two Japanese strains subjected to longday photoperiods, caused mortality of 22% or less. Conversely, similar treatment of eggs of two Ae. albopictus strains from tropical Malaysia and of a laboratory strain of Ae. aegypti resulted in nearly 100% mortality, whereas a subtropical strain from Taiwan exhibited intermediate coldhardiness. In addition, the degree of coldhardiness of the eggs of North American Ae. albopictus strains was similar to those of the Japanese strains (Hawley et al., 1989). These studies on photoperiodic sensitivity and coldhardiness suggested all Asian origin of U.S. populations. c— Allozyme Differentiation Studies on allozyme variation addressed three questions concerning the Ae. albopictus populations in the U.S.: (a) what is the breeding structure of the newly established populations? (b) what is the geographic origin of the US populations? and (c) what is the nature of changes in breeding structure that accompany colonization of a new environment? Questions (a) and (b) above were answered by comparing the breeding structure of the U.S. populations with that of populations from several native habitats. i— U.S. Populations To characterize the breeding structure of the newly established U.S. populations of Ae. albopictus, we analyzed allele frequencies at seven enzymatic loci in 17 populations. The collections were made during the summer and fall of 1986 from several locations within the cities of Houston, Texas, and New Orleans, Louisiana, and one each from Memphis, Tennessee, Jacksonville, Florida, and Evansville and Indianapolis, Indiana. Unique alleles and relatively high levels of heterozygosity were detected in the NEW ORLEANS, HOUSTON, and INDIANAPOLIS strains, indicating relatively large introductions into these cities. Low heterozygosities and no unique alleles were observed in other populations, indicative of population bottlenecks ensuing from a relatively small number of founding individuals or through insecticidal control efforts at these locations. Populations from the various cities were genetically distinct; how
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ever, partitioning of variance in allele frequencies indicated that the amount of differentiation among collections within a city was 3 to 4 times as large as the variance among cities suggestive of extensive local differentiation (Black et al., 1987). ii— Native Populations In a subsequent study, an analysis of 11 populations from Malaysia and Borneo indicated that (a) the average expected genic heterozygosities in the native populations were remarkably similar to those of the Houston and New Orleans strains, and (b) extensive local differentiation observed in U.S. populations was not a consequence of the colonization event, but rather an attribute of the natural breeding structure of the species probably due to the patchy distribution of the breeding sites and the low dispersal capacity of the adults (Black et al., 1988). iii— Geographic Origin of U.S. and Brazilian Populations To genetically trace the geographic origin of the U.S. and the Brazilian populations, we conducted a worldwide survey of genetic variation at eight polymorphic loci in 57 populations from nine countries. The results indicated that populations from the various regions were sufficiently genetically distinct from one another and that the populations from within a region were sufficiently genetically similar to one another to enable a strong inference regarding the origin of the U.S. and the Brazilian populations. A discriminant analysis of allele frequencies separated the populations from the various countries into nine nonoverlapping clusters; the U.S., Japanese, Chinese, and Brazilian populations formed closely placed, but distinct, clusters. The probability of assigning a population to the correct country was greater than 98%. The U.S. and the Brazilian populations were closest in terms of genetic distance to the Japanese populations. Based on discriminant and genetic distance analyses, we concluded that the U.S. and Brazilian Ae. albopictus originated in Japan. Furthermore, the data indicated that the U.S. population(s) were introduced with virtually all of their genetic variation intact, significantly enhancing the probability of longterm establishment (Kambhampati et al., 1991).
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iv— Temporal Changes in Genetic Structure Species that colonize a new environment may undergo drastic and rapid reduction in genetic variation levels due to the loss of lowfrequency alleles. Any such changes are detectable only after several generations have been completed in the new environment and are related to the breeding structure of a founding population(s). Although our studies showed that the breeding structure of the newly established U.S. populations did not differ substantively from that of the native Asian populations, we wished to determine if changes in the breeding structure have or were taking place in subsequent generations. We therefore undertook a survey of temporal variation at 10 loci in 17 populations from locations in and around Houston, Texas, between 1986 and 1988. The results revealed significant but nondirectional changes in allele frequencies. The variance in allele frequencies remained unchanged as did the levels of variation within and among populations, indicating that the breeding structure of Ae. albopictus in the U.S. did not differ substantially from that in the native habitat, either immediately following colonization or after several generations in the new habitat. The fact that no significant decrease in genetic variation was observed over time is also suggestive of the absence of a bottleneck. The results also suggested that there was considerable movement of populations within the U.S., presumably by the eggs moved in used tires (Reiter and Sprenger, 1987), subsequent to establishment in Houston, Texas, which must have resulted in considerable gene flow. A rough estimate of effective population sizes indicated an increase between 1986 and 1988. A large founder population, gene flow; and a rapid increase in population size would be expected to impede the loss of genetic variation during founder events (Kambhampati et al., 1990). d— Ribosomal DNA (rDNA) Intergenic Spacer (IGS) Length Variation Molecular genetic surveys of multigene families have received considerable attention in recent years. Of the many multigene families, the genes coding for ribosomal RNA (rDNA) have been used widely in population genetic analyses. We undertook studies on this important multigene family at the population and species levels.
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i— Population Genetic Survey We conducted a worldwide survey of variation in the rDNA gene family in 17 populations of Ae. albopictus at four levels: in individuals, in populations within a city, among cities, and among countries, with the objective of determining the patterns of divergence in individual components of the rDNA cistron. We also compared these patterns in newly established U.S. populations with those from the native range (Black et al., 1989a). Three clones of ribosomal cistrons were isolated from a genomic library and mapped. These clones were subcloned into plasmids and used to probe for intraspecific variation. The 18S and 28S ( & ) coding regions were similar in size among all 17 populations of Ae. albopictus, while extensive variation was found in the intergenic spacer (IGS) within and among populations. Individual mosquitoes in a given population carried a unique set of spacers, and most of the variation in spacer length existed at the population level with 65% of the variance arising within populations. This differs markedly from the situation in An. albimanus (Beach et al., 1989) and Drosophila melanogaster (Coen et al., 1982; Williams et al., 1985), where spacer diversity exists mainly among, rather than within, populations. Due to the large amount of intrapopulational variation, samples from the U.S. were not similar to any one foreign population examined. Consequently, this part of the genome could not be used as a tool to determine the origin of populations. ii— Temporal Variation in rDNA Spacer Length To better understand the sources of the extensive intrapopulation variation in IGS length, studies were undertaken to determine (a) the shortterm changes in IGS frequencies in field populations and (b) the mode of inheritance of the IGS length (Kambhampati and Rai, 1991a). Results of studies on temporal variation in the rDNA IGS length in eight different populations from Texas over a threeyear period indicated that the high level of variation observed by Black et al. (1989a) was being maintained, with random fluctuations in the frequencies of the various size repeats. Statistically significant variation was detected among populations at each sampling
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date; significant but nondirectional changes were observed within populations over time. iii— Inheritance of rDNA Spacer Region Singlepair crosses were made between individuals with differing IGS lengths. The results indicated that the IGS is inherited codominantly in a Mendelian fashion. Progeny of single pair crosses contain spacer sizes from both parents in some combination, although often one and sometimes two IGS genotypes out of a possible four predominate. In addition, new spacer sizes, presumably arising from recombination within the IGS region or in the flanking regions of the rDNA locus, were also observed (Kambhampati and Rai, unpublished). The rate of generation of new IGS length classes was estimated to be 102 per generation, which is 10 times greater than that reported for Drosophila spp. (Coen et al., 1982). The results of the inheritance and field studies indicated that (a) the rate of generation of new length variants is high and (b) once generated, these new spacers are subject to population level phenomena resulting in a high turnover rate. The results help explain some of the variation observed in natural populations. iv— Chromosome Mapping and Copy Number of rRNA Genes To physically map rRNA genes, in situ hybridization using 3Hlabeled 18S and 28S rDNA probes from Ae. albopictus was performed on the mitotic chromosomes of 20 species of mosquitoes, including three albopictus subgroup species (Ae. pseudalbopictus, Ae. flavopictus, and Ae. seatoi). In all but one species examined, the rRNA genes were localized to a single site per haploid genome; in Ae. triseriatus the rRNA genes were located on two chromosomes (Kumar and Rai, 1990a). The presence of the rDNA locus on different chromosomes and at different locations in different species suggested that considerable chromosomal repatterning has taken place over evolutionary time. Dotblot hybridization was also used to estimate copy number of rRNA genes (Kumar and Rai, 1990a). The results indicated a striking 26fold difference among the various species in rDNA copy number. In the Ae. albopictus subgroup species, it ranged from about
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180 copies in Ae. seatoi to over 1000 copies in Ae. albopictus and Ae. flavopictus. e— Mitochondrial DNA Restriction Fragment Length Variation Mitochondrial DNA (mtDNA) is a sensitive tool to determine lineages and evolutionary history of a species. It is maternally inherited, has a relatively simple organization, often evolves 5 to 10 times faster than single copy nuclear DNA, and there is no recombination among genomes of different maternal clones. We undertook studies on inter and intraspecific variation in mtDNA. i— Intraspecific Variation We surveyed 17 populations of Ae. albopictus from 10 countries, including six from the U.S., for mtDNA restriction fragment length polymorphisms. We wished to determine if mtDNA, because of its inherent advantages, can provide further insights into the breeding structure of Ae. albopictus and provide a more precise indication of the origin of the U.S. populations. Of the 18 enzymes used, 6 did not reveal any restriction sites. The remaining 12 yielded a total of 1,060 fragments for the 17 populations. The mtDNA size was estimated to be about 17.5 kb. Only three populations showed the presence of novel mtDNA haplotypes: MAURITIUS with HaeIII and HpaII, HONG KONG with EcoRI and HinfI, and SINGAPORE with HaeIII. The mean F value (i.e., proportion of shared sites) was 0.995. A value of 1 means complete identity (Kambhampati and Rai, 1991b). The low level of mtDNA polymorphism observed in Ae. albopictus is in contrast to most other animals and insects. Consequently, mtDNA analyses could not be used to trace the origin and movement of populations of the species in the continental United States. However, the results suggested that most of the range expansion in this species had taken place relatively recently, possibly as a result of humanaided transport. ii— Interspecific Variation To reconstruct the phylogeny of the species in the Ae. scutellaris group, studies using mtDNA RFLP analysis were undertaken. There
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was extensive variation among the seven species of the scutellaris subgroup and four species in the albopictus subgroup; all average proportion of 0.338 restriction fragments were shared among them (Kambhampati and Rai, 1991c). The phylogeny that resulted from the mtDNA RFLP analysis, not surprisingly, was incongruent with those derived from allozymes, morphology, and biogeography. The presumed monophyly of the species in the scutellaris subgroup and those in the albopictus subgroup was also not reflected in the mtDNA analysis. iii— Dynamics of mtDNA Haplotypes in Laboratory Cage Populations A laboratory cage study was undertaken to determine (a) if mtDNA haplotypes in Ae. albopictus are selectively neutral and (b) the potential of cytoplasmic incompatibility (CI) to bring about population replacement using mtDNA haplotype as a genetic marker. Item (a) was undertaken in response to published studies which purported to show that mtDNA haplotypes are not selectively neutral in Drosophila pseudoobscura (Macrae and Anderson, 1990) and Drosophila stimulans (Nigra and Prout, 1990), which if true has implications for the use of mtDNA in phylogenetic and population genetic studies. Our results based on frequency estimates of two mtDNA haplotypes in three replicate cages over 10 generations, each of bidirectionally compatible and unidirectionally incompatible matings, indicated that in Ae. albopictus, (1) one mtDNA haplotype does not increase At the expense of the other if matings are bidirectionally compatible, and (2) in cages with unidirectionally incompatible strains, the mtDNA haplotypes carried by the females that lay inviable eggs was completely replaced in two generations, suggesting the CI can be used for populations replacement (Kambhampati et al., 1992b). f— Reproductive Differentiation: Cytoplasmic Incompatibility in Aedes albopictus Endosymbiontmodulated cytoplasmic incompatibility (CI) has been reported for a number of insect species, including several from the genus Aedes. Infection of the gonads by rickettsia, e.g., Wolbachia spp., has been implicated in cases of insect CI. In preliminary
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studies we found that matings between Ae. albopictus females from the island of Mauritius and males from any other geographic region resulted in inviable eggs. The reciprocal crosses produce viable eggs. In subsequent studies, crosses were made among six U.S. and Asian strains of Ae. albopictus (Kambhampati et al., 1993). In interpopulation crosses involving MAURITIUS females and males of any other five populations, the eggs were inviable. All other inter and intrapopulation crosses, as well as the reciprocal crosses, yielded egg hatches ranging from 65% to 99%. The maternal mode of inheritance of the incompatibility factor was confirmed in backcrosses involving three different populations (Kambhampati et al., 1993). Transmission electron microscope studies on ovarian tissues of MAURITIUS, INDIANAPOLIS, and OAHU females indicated that Wolbachia or Wlike microorganisms were present in the ovaries of all individuals of all populations. [Thus the observed incompatibility, was not a result of matings between infected males and uninfected females.] In all attempt to ''cure" the mosquitoes of the Wolbachia infection, INDY males were treated with tetracycline and mated to MAURITIUS females. This resulted in a partial restoration of compatibility with an egg hatch of 28%. Although the results are suggestive of endosymbiont involvement in CI, only a partial restoration of compatibility after antibiotic treatment to restore compatibility suggests a complex situation. CI may play, a role in speciation and can potentially be used for regulation of pest populations and for introduction of genes into insect hosts that harbor populations of Wolbachia. g— Competition between U.S. Strains of Ae. albopictus and Ae. aegypti Mosquito abatement workers in HOUSTON and NEW ORLEANS have observed that the introduction of Ae. albopictus was accompanied by a decline in Ae. aegypti populations. A series of laboratory studies on competition between U.S. strains of the two species was undertaken. The results did not suggest that the U.S. Ae. albopictus populations are inherently more competitive in the laboratory, than Ae. aegypti (Black et al., 1989b). It seems likely, that the densities of the two species in the U.S. are fluctuating indepen
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dently of each other. Alternatively, the presumed replacement of Ae. aegypti by Ae. albopictus may ensure from (a) a greater ability of the latter species to develop eggs than the former after they ingested small blood meals, and (b) the greater longevity of adult Ae. albopictus females than that of Ae. aegypti under conditions of starvation (Klowden and Chambers, 1992). h— Vector Competence to Dengue1 Virus i— Oral Susceptibility and Horizontal Transmission Eight geographic strains of Ae. albopictus from North America and Asia and one North American strain of Ae. aegypti were tested for their vector competence with a Fiji strain of dengue serotype 1 virus. Thc Asian strains were derived from regions with known recent dengue nonendemic, endemic, and epidemic history. All strains tested, except the HOUSTON strain, showed high susceptibility to midgut infection. However, there were marked differences in percent disseminated infection and transmission rates among the various strains (Boromisa et al., 1987). Strains from dengue endemic/epidemic regions of Asia showed significantly higher susceptibility to disseminated infection and oral transmission than those of the U.S. and Japan (nonendemic). Among the U.S. strains, mosquitoes of the OAHU strain were nearly twice as likely to develop disseminated infections as were individuals of any of the other strains tested. Overall, the three North American strains were comparable to the strain of Ae. aegypti from Houston, Texas in their ability to transmit DEN1 virus. However, the detailed profiles of vector competence among the three North American strains tested showed considerable differences between the HOUSTON strain and the MEMPHIS/NEW ORLEANS strains, more or less paralleling allozyme differentiation among the NEW ORLEANS and HOUSTON strains (Black et al., 1987). Based on these studies, the North American strains were more similar to a northern Asian strain (TOKYO) than to the three Malaysian (southern Asia) strains, supporting the hypothesis that the Ae. albopictus populations introduced into the United States had a northern Asian origin.
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ii— Vertical Transmission Five geographical strains of Ae. albopictus were compared for their ability to transmit vertically a DEN1 virus isolate from Jamaica. The OAHU strain of Ae. albopictus and a strain of Ae. aegypti from the U.S. were included as controls. The offspring of orally infected females were assayed individually for vertical infection. Vertical transmission rates among strains ranged from 11 to 41% and filial infection rates of strains ranged from 0.5 to 2.9%. Filial infection rates of individual positive families within strains ranged from 1.4 to 17.4% (Bosio et al., 1992). These rates were higher than those previously recorded for Ae. albopictus. The observed differences in rates of vertical transmission among the strains were not statistically significant, however, because 95% of the measured variation was attributed to families within strains. The most significant source of variation in vertical transmission of this DEN1 virus by these Ae. albopictus strains/populations was at the individual level. iii— Midgut Basal Lamina Thickness and Virus Dissemination Strain differences in midgut basal lamina thickness, assessed by measurement in transmission electron micrographs, and disseminated infection rates of DEN1 virus were compared among three laboratory strains of Ae. albopictus (Thomas et al., 1993). Mean basal lamina thicknesses for the NEW ORLEANS and HOUSTON strains were significantly greater than those for the OAHU strain, which exhibited a higher disseminated infection rate than the former two. Although basal lamina thicknesses among the F1 progeny of reciprocal crosses of the OAHU and HOUSTON strains were intermediate between the parental strains, they were too variable to be useful as markers in genetic studies. Measurements of basal laminae among individuals of the NEW ORLEANS strain, with disseminated or nondisseminated infections, failed to demonstrate a role for basal lamina thickness as a modulator of DEN1 virus dissemination across the midgut epithelium of Ae. albopictus, in contrast to the demonstration of this phenomenon, apparently of con
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siderable importance, in the Ae. triseriatus—La Crosse virus system (Grimstad and Walker, 1991). iv— Inheritance of Midgut Infection/Disseminated Infection We examined the inheritance patterns of these two traits prior to establishing extreme phenotypes through selection. The F1 progenies were obtained by reciprocally crossing adults of OAHU with those of NEW ORLEANS and HOUSTON following infectious blood meals prepared according to the methods of Boromisa et al. (1987), which results in titres of 8.0–8.5 log10 of DEN1 virus/ml. Indirect immunofluorescent assays (IFA) of midgut and head squashes of all bloodfed adults were used to establish midgut and disseminated infection rates among the progeny. The midgut infection rates did not show much variance in the progenies of the control and F1 crosses, reflecting the lack of much heterogeneity among the parental strains. However, the disseminated infection rates of the F1 progeny in most cases approximated those of the maternal strains used in the hybrid crosses. For example, crosses involving OAHU as the female parent had higher disseminated midgut infection rates than the reciprocal crosses where OAHU was used as the male parent. The data from the eight sets of backcrosses involving OAHU and NEW ORLEANS did not show any obvious segregation patterns for either trait and do not allow any definitive conclusions to be made about their mode of inheritance at this time. However, generally higher rates of both midgut and disseminated infections were observed when either NEW ORLEANS was used as the female parent or the F1 parent received the NEW ORLEANS female—determining sex chromosome. This suggests a possible linkage of susceptibility for these traits with this chromosome (Thomas and Rai, unpublished). v— Status of Ae. albopictus in U.S. Craig complained loudly that the federal government issued warnings about the Ae. albopictus infestations in the U.S. but took little action to contain it. He had a running battle with the Centers
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for Disease Control and Prevention (CDC) about the inadequacy of the funding it provided and its overall efforts. This was one issue where Craig and Rai interpreted the potential for future damage differently. Craig "emerged as the leading proponent of an aggressive response to the Asian tiger mosquito" (Hall, 1987a). Zealot that he was, he pushed CDC and state authorities to wipe out the species from the U.S. by whatever means and whatever funding it required. Rai, on the other hand, emphasized that the U.S. strains of Ae. albopictus were "no more effective at transmitting dengue fever than Ae. aegypti which has been in the country for three centuries," that "Ae. albopictus is here to stay," and that "it has already become part of our national fauna and we may have to learn to live with it" (Hall, 1987a). Rai also emphasized that ''insecticides, whether you spend $1 million or $100 million, will be ineffective" against a species which has already moved to the treehole habitats at most locations in the U.S. (Hall, 1987b).
11— Vector Competence and Arboviral Ecology After getting his Ph.D. at the University of Wisconsin under the direction of Professor Gene DeFoliart, Dr. Paul Grimstad joined the Vector Biology Laboratory in 1974 as a postdoctoral research associate to work with George Craig. DeFoliart's group had incriminated the treehole mosquito, Aedes triseriatus, as the major vector of La Crosse (LAC) virus in Wisconsin. Craig's laboratory had earlier conducted a series of studies showing the genetic basis of susceptibility of Ae. aegypti to the bird malarial parasite, Plasmodium gallinacium (Kilama and Craig, 1969), and to the filarial worms, Foleyella seasonalis (Terwedo, 1972) and Waltonella flexicauda (Terwedow and Craig, 1977). Also, Rodriguez and Craig (1973) and Paige and Craig (1975) had shown susceptibility of several East
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African populations of Ae. aegypti to Brugia pahangi. Kilama (1976) similarly showed such variation to Wuchereria infections. With this as the backdrop, Craig was eager to demonstrate the genetic basis of arboviral susceptibility in mosquitoes. However, he recognized a major paradox and called attention to the same: "The situation in arbovirus research is astonishing. Single genes for aphids and leaf hopper transmission of plant viruses have been known since the early 30s. In spite of the strong evidence that genes will also control mosquito susceptibility to arboviruses, there is no proven case in the literature" (Craig, 1975). In this context, Grimstad's addition to the VBL in 1974 was expected to fill an important niche. However, although considerable progress has been made, the goal of establishing the genetic basis of susceptibility of any of the mosquito transmitted viruses has remained elusive. This likely is due to the complexity of viral transmission by the vector species, involving several seemingly independent steps and barriers such as ingestion, replication, and dissemination of the virus from the midgut to the hemocoel, the salivary glands, and finally the ability to transmit. In all probability, these interrelated steps are controlled by different genetic factors and their interaction with the intrinsic environment of the specific vector population and numerous extrinsic environmental factors may also come into play. Also, and perhaps even more importantly, the premature termination of both the WHO/ICMRsponsored field releases in Delhi and of USAIDsponsored work in Mombasa during the mid70s brought about a shift in focus of both Craig's and Rai's research efforts. An epidemic of St. Louis encephalitis in the United States in 1975 with more than 2,300 cases and several hundred deaths was the trigger for a change in George Craig's research focus. This epidemic occurred throughout the central U.S., but it was most serious in the Midwest, particularly in Illinois, Indiana, and Michigan. Craig declared, "As the only medical entomologist then resident in Indiana, it behooved me to stay and tend to the home fires for a little while rather than worry about what is going on in Africa" (Craig, 1976). At least some ten mosquitoborne viruses cause central nervous system (CNS) infection and often overt disease such
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as encephalitis; the symptoms of CNS infection include high fever, headache, nausea and vomiting, frequent disorientation, stupor, and sometimes coma. In severe cases, these viral infections of the CNS cause inflammation of the brain (i.e., encephalitis). There is no treatment, only protective therapy to lower the fever and treat other severe symptoms. Considerable work has been done over the years at the VBL on several important mosquitotransmitted arboviruses particularly important in the Midwest. These include the following: (1) St. Louis encephalitis (SLE) virus maintained in North American bird species and transmitted to man by the common house mosquito, Culex pipiens, and other species (Cx. tarsalis, Cx. nigripalpus, Cx. salinarius). (2) Eastern equine encephalitis (EEE) virus, found in discrete foci in northern Indiana and southwest Michigan with 50–80% mortality of humans and horses that contract the disease. There are sporadic cases of this disease during most late summers and falls in two to ten counties in and around Indiana and Michigan. (3) The La Crosse (LAC) virus, first isolated in 1964 from brain tissue of a young female who died in 1960 in a La Crosse, Wisconsin, hospital. This virus is most active in the midwestern United States (Grimstad, 1988). As mentioned earlier, LAC virus is transmitted in the Midwest by Ae. triseriatus, the treehole mosquito, and is maintained in nature in a cycle involving select small mammals, e.g., chipmunks and squirrels. Two other recent emerging diseases in the Midwest are caused by infection with Jamestown Canyon (JC) virus (Grimstad et al., 1986) and the Cache Valley (CV) virus (Neitzel and Grimstad, 1991). Jamestown Canyon virus has been implicated since 1980 in a growing number of severe central nervous system infections in North America. Human antibody prevalence ranges from 18% in Michigan's Lower Peninsula to 42% in its Upper Peninsula (Grimstad, pers. comm.). Firsttrimester CV virus infections have the potential of affecting the developing fetus and may be a cause of miscarriages in women as well as birth defects in newborns (Calisher and Sever, 1995). Most of the studies dealing with determination of the particular mosquito species as the vectors and the epidemiology of the disease are being conducted under the
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direction of Dr. Paul Grimstad. These studies have led to the important concept that the differential ability to transmit a particular viral pathogen call be a characteristic of individuals or populations as well as species. Also, extensive serological surveys have been conducted on the incidence of LAC and SLE virus infections in human populations in Indiana (Grimstad et al., 1984). Infection of more than 12% for residents of individual counties was detected for both of these viruses. Moreover, significant differences among various geographic strains of mosquitoes were detected in their ability for oral infection, disseminated infection, and transmission ability (Boromosa et al., 1987).
12— Endocrinology and Reproductive Physiology Professor Morton S. Fuchs, who joined the then Department of Biology and the VBL in 1960, has been a leader in studies dealing with the reproductive physiology of mosquitoes. He has provided extraordinary leadership in fostering rapid growth and evolution of the department toward a more cellular and molecular orientation. Fuchs served as the chairman of the Department of Biology from 1975 to 1981, of the Department of Microbiology from 1981 to 1984, and of the combined (biology and microbiology) Department of Biological Sciences from 1984 to 1993. The work in Dr. Fuch's laboratory over the past thirty years has been directed toward understanding the endocrinological and molecular parameters of ovarian development in mosquitoes. The longrange rationale is that such knowledge can be exploited to develop specific biochemical strategies designed to interrupt the mosquito's reproductive cycle and thereby could possibly be used for effective population control. Ovarian development in many insects is a
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complex process regulated by the interaction and participation of the endocrine system. Using mosquitoes as a model, Fuch's group (and others) have found that at least three different hormones are required for successful oocyte maturation in mosquitoes. At the molecular level, these hormones may manifest their modes of action via a combination of transcriptional control as well as posttranslocational modifications. The major focus of his laboratory has been to unequivocally establish the molecular basis for this hormonal regulation, to identify specific hormones involved, and to establish their mode of action. Shortly after adult eclosions the cells which comprise the putative ovary in Ae. aegypti grow and differentiate into primary follicles (approximately 50 per ovary). These follicles consist of an oocyte, surrounded by follicular epithelial cells and seven nurse cells. This process is complete by approximately 40–4751606.FXP60 hours postadult emergence. At this point, the ovaries are said to have reached their "resting stage." In an autogenous mosquito such as Ae. aegypti, the ovaries will remain in this state until a bloodmeal is ingested. The ingested blood is hydrolyzed by proteases in the midgut and the products transported via the hemolymph to the fat body, where they are resynthesized into yolk proteins called vitellin. Vitellin is then released by the fat body, after which it is sequestered by the oocytes and is now called vitellogenenin. Thus, oocyte maturation is completed after vitellogenesis, and fertilization can then occur. Work performed in Fuch's laboratory (and others) has led to a complex endocrinological picture clearly showing that juvenile hormone (JH), 20OH ecdysone, and numerous peptide hormones are required for successful oocyte maturation. Previtellogenic development which results in "resting stage" ovaries is regulated by JH secreted from the Corpus Allatum (CA) shortly after adult eclosion (Gwadz, 1973; Hagedorn, 1989). This hormone also "primes" the fat body for subsequent vitellin synthesis (Ma et al., 1988). After bloodfeeding the ovary releases 20OH ecdysone, which in turn induces the fat body to synthesize vitellin. In addition to JH and 20OH ecdysone, head or brain factors also appear to be involved. These other factors, all peptides, either regulate JH and/or 20OH
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ecdysonesynthesis (Meola and Lea, 1972; Feinsod and Spielman, 1980; Rossignol et al., 1981) or proteolytic enzyme synthesis (Borovsky et al., 1991).
13— Research and Training Funding Through the past four decades, faculty in the VBL have received support from major U.S. federal funding agencies, such as the National Institute of Health (NIH), the Environmental Protection Agency, the National Science Foundation, and the United States Department of Agriculture; from the Indiana State Department of Health and the St. Joseph County (Indiana) Health Department; through individual research grants from private foundations like the John D. and Catherine T. MacArthur Foundation and the Wellcome Trust; from international funding bodies, including the World Health Organization; and from the University of Notre Dame. Craig received continuous funding from 1958 to the time of his death; he had been awarded an NIH merit grant in 1988 for the period 1/88–12/98. On his death, NIH named Craig's longtime collaborator, Paul Grimstad, as principal investigator to oversee the remaining three years of the grant. Grimstad also served as replacement mentor for four graduate students "orphaned" by George's death and Rai and Fuchs mentored one each. In addition, the Vector Biology and Parasitology programs have been supported by three major training grants awarded by the NIH for almost thirty consecutive years. One grant, entitled "Mosquito Biology: Genetic, Organismal, Environmental" (5T01 AI00378 TMP), with Dr. K. S. Rai as the program director, provided funding for the period of July 1, 1969 to September 30, 1974. A second, entitled "Parasitology: Biochemical, Developmental, Genetic" (5T01 AI00400 TMP), with Dr. Paul P. Weinstein, the former chief of Parasitic Disease at NIH, as the program director, covered the
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period July 1, 1970, to December 30, 1975. When NIH, due to a change in national policy, withdrew support from all training grants in these areas, both these grants automatically came to a close at the end of their respective award periods. With the reintroduction of a policy of support once again, a third training grant, entitled "Experimental Parasitology and Vector Biology" (5 T32 AI07030 TMP), with Dr. Weinstein again as the program director, was awarded by NIH for a period of five years for five postdoctoral trainees a year commencing July 1, 1976. A supplement to this grant was awarded in 1979 to support three predoctoral trainees a year. This latter grant (5 T32 AI07030 TMP) was renewed twice for an additional ten years (July 1, 1981 to June 30, 1986, and September 1, 1988 to August 31, 1993) to support the training of five postdoctoral and three predoctoral trainees a year. Following one year's interruption, this grant was renewed again for a fiveyear period (July 1, 1994 to June 30, 1999) with funding for three predoctoral trainees. Dr. Weinstein served as the program director for this training grant during 1976–1986, and Dr. Rai served as the program director during 1988–1998. Dr. Collins became the program director in June 1998. These training grants, running for the longest period in the country, have been extraordinarily successful in terms of bringing faculty and students with disparate interests into interdisciplinary collaboration and synergistic interaction.
14— Special VBL Resources and Facilities International Reference Centre (IRC). The VBL was designated as the WHO's International Reference Centre for Aedes in the mid60s and continues in this capacity on an ongoing basis. This designation recognized the pioneering contributions of the VBL personnel in the field of Aedes genetics. The functions of the Reference Centre include consultation, dissemination of information,
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and strain maintenance. Over the years, this laboratory has maintained over 150 strains of mosquitoes, including 22 species, geographic populations, mutant strains, etc. During the past four decades, more than 1,000 requests for standard strains, inbred lines, and genetically marked strains have been filled. It is unquestionably an unparalleled international germ plasm resource for Aedes mosquitoes. Currently, it houses one of the world's largest reference collections of Anopheles species and genetic strains as well. University of Notre Dame's Environmental Research Center (UNDERC). The central mission of UND Environmental Research Center (UNDERC) has focused on basic and applied research and educational programs that explore solutions to global environmental problems based on an understanding of undisturbed natural habitats and biological diversity. UNDERC encompasses 2,975 hectares (approximately 11.5 square miles) on both sides of the state line between Wisconsin and Michigan's Upper Peninsula in Vilas County (Wisconsin) and Gogebic County (Michigan). It includes a land area of 2,485 hectares and thirty lakes and bogs with a combined surface area of 490 hectares. Among the aquatic habitats that lie wholly on the property are nine dystrophic bogs, many permanent ponds and small lakes, and several marsh habitats. Scientific investigations conducted at UNDERC have considered the chemical and biological impact of acidification of aquatic habitats, natural processes that lead to the production of the "greenhouse" gases in aquatic habitats, and manipulation of biological processes to control the effect of nonpoint source nutrient additions to aquatic environments. Studies of the epidemiology of several arthropodborne zoonosis arc also under way. California serogroup viruses circulate in local deer and small rodents. Tickborne Lyme Arthritis occurs in nearby sites in Vilas County. There are ample opportunities for studying wildlife disease cycles that are maintained by biting flies and arthropod ectoparasites. During May–June, many vernal ponds exist on the property. Mosquito populations in many of these ponds have been surveyed annually for more than twenty years by VBL students and faculty. UNDERC maintains a winterized laboratory building, a classroom, and housing for up to sixty students and re
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searchers. Field vehicles and numerous small boats are maintained for teaching and research operations. More than 140 scientific papers have been published based on research at UNDERC, nearly 100 of these since 1980. Since 1976, a tenweek summer field course provides training in aquatic and environmental biology. Beginning in 1992, a graduatelevel course in field vector parasitology has also been offered at UNDERC. It has been developed specifically for the parasitology/vector biology training grant participants and emphasizes sampling, field collection methods, and host, vector, and parasite identification techniques. The insect fauna of UNDERC is rich and varied. Systematic survey of the aquatic insects is ongoing. Thirtyone mosquito species have been found to date, most of them members of the Aedes communis or Aedes excrucians groups. Treeholebreeding Aedes, pitcherplant breeding Wyeomyia, and aquatic reedbreeding Coquillettidia are all abundant. There are also locally intense populations of black flies (Simulium), deer flies (Chrysops), and horse flies (Hybomitra and Tabanus). Mosquito Data Bank of the University of Notre Dame (MODABUND). This computerized mosquito literature data bank was established by professor Theodore J. Crovello, who was a member of the VBL faculty from 1966 to 1984 and is currently Dean of Graduate Studies, California State University, Los Angeles. The purpose of MODABUND was to provide students with maximum information about mosquitoes, both in terms of literature retrieval and raw data. There are more than 34,000 references contained in MODABUND organized around approximately fifty subject categories (Crovello, 1972). For each reference, MODABUND contains the following data: (a) author(s); (b) date; (c) title; (d) citation; and (e) subject number. MODABUND can be searched for any combination of author, date, key words in the title, or deep index subject numbers. This service has been available to anyone interested in the same. As for our own students, particularly the incoming ones, this was an efficient method to present them with tailormade computer printouts of the past literature in each fold. Even with the current availability of several online computer databases for literature re
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trieval and other websites, the MODABUND facility, focusing exclusively on mosquitoes, still provides a useful service. However, in time it may largely be supplanted. St. Joseph County Mosquito Surveillance Program. The St. Joseph County Mosquito Surveillance Program is an annual activity of the VBL. The laboratory contracts with the St. Joseph County (Indiana) Health Department to provide basic medical entomology services to the county. These services include adult mosquito surveillance with a grid of sixteen light traps and with biting collections; larval mosquito surveillance using a variety of collecting methods; tick surveillance to detect Rocky Mountain Spotted Fever foci in the county; serological screening of deer sera for arboviral antibody (herd immunity) presence; and special ecological investigations of selected mosquito species. Particular effort is currently given to studies of the biology and control of Coquillettidia perturbans, the likely sector of Eastern equine encephalitis virus in the county. A special larval sampling method was developed for this species for surveillance for larval sites and for insecticide tests. Teknar, Abate, and Dursban were tested against larval Coq. pertubans; plans are made to text Altosid also. Since the death of George Craig, Jr., Paul Grimstad directs the SJCMSP; a postdoctoral research associate acts as field supervisor, and graduate and undergraduate students are field workers. Members of the VBL clearly benefit from the activities of the SJCMSP by gaining practical, applied field experience in mosquito research and control. Indiana State Department of Health and the Laboratory for Arbovirus Research and Surveillance—University of Notre Dame (UNDLARS). An ongoing program is maintained to follow activity of arboviruses in Indiana. Viruses monitored include SLE, EEE, western equine encephalitis (WEE), and numerous bunyaviruses. Each summer, more than 5,000 birds are bled and tested for antibodies against SLE, EEE, and WEE viruses. In addition, mosquitoes are trapped and tested for infectious virus as needed. This early warning system has been useful in averting epidemics by alerting control agencies (M. Sinsko, pers. comm.).
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Externship/Internship in Arbovirology for D.V.M. Students. This program was initiated in 1985 by Grimstad in response to the request of the Subcommittee on Veterinary Arbovirology of the American Committee on Arthropodborne Virus to place veterinary students in arbovirus laboratories and field studies.
15— Linkages of the VBL with Tropical Institutions The VBL faculty have a long history of overseas involvement. We have trained doctoral students from Tanzania, Malaysia, India, Taiwan, Ghana, Philippines, Sri Lanka, Thailand, China, Kenya, Uganda, Mexico, Panama, Brazil, and Nigeria. We have had visiting faculty and postdoctorals from India, Uganda, Malaysia, Ghana, and South Africa. Moreover, faculty of the VBL have been active as consultants to various national and international agencies, including USPHS, USAID, WHO, IAEA, and several universities and commercial organizations. Our graduate students and postdoctoral fellows have had ample opportunities to participate in field research activities of these agencies and at our research stations in the tropical environment abroad. This provides an excellent opportunity for individuals interested in combining field and laboratory research in their work. Over the years, the VBL has been actively involved in several field projects abroad, i.e., in Ceylon (IAEA), Brazil (IAEA), India (WHO), Tanzania (WHO), and Kenya (USAID). Dr. Craig was involved in the effort to eradicate Ae. aegypti in the southern U.S. and in particular in Mississippi (USPHS). Professor Rai has served as a consultant for feasibility studies for genetic control of Culex fatigans with radiationsterilized males in Sri Lanka and Brazil and of Ae. aegypti with chromosome translocations in India and East Africa. Professor Craig was a senior advisor and reviewer for the Board on Science and Technology for International Development (BOSTID)
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of the National Academy of Sciences since its inception. He also served as chairman of the BOSTID Working Group on Manpower Needs and Career Opportunities in the Field Aspects of Vector Biology in Washington, D.C., in 1983. Craig was also one of the founding directors of the International Centre for Insect Physiology and Ecology (ICIPE), Nairobi, Kenya. As mentioned earlier, UND maintained a laboratory, and field project near Mombasa, Kenya, from 1970 through 1977 to study genetic control of Ae. aegypti in villages under a grant from USAID. Five postdoctorals were stationed there; two, Drs. Nancy Choitz and Paul McDonald, worked in the laboratory of Professor Rai and received their Ph.D. degrees under his direction. Drs. Walter Hausermann, Dean Fanara, and Jack Paterson worked on field biology and ecology under Dr. George Craig. Professor Rai served as a member of the WHO Advisory Group for Genetic Control of Culicine Mosquitoes in New Delhi, India, from 1970 to 1975. He also served as research director in Delhi and two of his graduate students (Nancy Lorimer and Edward Hallinan) evaluated the feasibility of the use of the chromosomal translocations for genetic control of Ae. aegypti at the Delhi unit. This work constituted part of their doctoral dissertations at UND. The WHO/ICMR research unit was incorporated into the Malaria Research Centre, Delhi, India, in 1976. Professors Frank Collins and Nora Besansky have been engaged in collaborative research projects whose aim is to better describe population structure of the major malaria vectors in Africa. Most of this work involves the use of microsatellites, mtDNA sequence, and the sequence of nuclear regions (usually introns) as the population genetic markers. The key people doing the work have been Nora Besansky, postdoc Tovi Lehmann, Ms. Luna Kamau, a Kenya Ph.D. student at Kenyatta University being supervised by William Hawley, and Frederic Simard, a Ph.D. student being supervised by Didier Fontenille of ORSTOM in Dakar, Senegal. The initial objectives were to assess the performance of microsatellite and DNA sequence markers as population genetic tools and then use these markers to define deme size and rates of gene flow in the Savanna karyotype of Anopheles gambiae. Some collections made in western
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Kenya and coastal Kenya in the mid1980s are being examined to look at effective population size in An. gambiae from these two locations. The work has since been extended to An. arabiensis in collaboration, with Mario Coluzzi of Rome and Yeya Toure of Bamako, Mali, and includes studies of other cytotypes in the An. gambiae taxon—namely, the Bamako and Mopti types. Through Coluzzi, Besansky has been able to obtain specimens from several parts of West Africa in addition to Mali and Senegal. Collins is also collaborating (mainly contributing data and primers in advance of publication) with Harold Townson of Liverpool, whose Ph.D. student is doing a study of the population structure of An. arabiensis in Tanzania and Malawi. Besansky also has been particularly interested in defining rates of gene flow among species in the An. gambiae complex, particularly between An. gambiae and An. arabiensis. In addition to the above, Frank Collins is a collaborator on a recently funded WHO TDR grant to John Vulule of KEMRI (Kenya Medical Research Institute). Vulule has documented permethrin resistance in An. gambiae from western Kenya, and Collins is collaborating on the effort to determine the molecular basis of this resistance and then use that knowledge to determine rates of spread in the resistanceconferring alleles in a population now under pressure from a large permethrin impregnated bed net study. Nora Besansky has recently started to assemble a set of microsatellite markers for the African malaria vector An. funestus, work now being done by her postdoc Steve Sinkins. The work will also tie in with research by John Vulule and William Hawley in Kenya and by Didier Fontenille, who will provide specimens from several sites in West Africa as well as Madagascar. Frank Collins also collaborates with Cathy Walton of the University of Leeds, who is studying the population genetics of the An. dirus complex in Southeast Asia. Most of the above links with the field are essentially links with collaborators who are either based overseas or who do field work during transmission seasons. Professor David Severson has an NIHfunded field project in Trinidad and Tobago in West Indies in collaboration with the Ministry of Health. The objective is to assay population genetics of Ae.
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aegypti relative to vector control efforts using organophosphates. His team will be looking at the population structure (1) just before spraying, (2) soon after spraying, and (3) long after sprays have been terminated, using RFLP markers. Collins and Hawley, the latter a former associate faculty fellow at UND, will collaborate in these studies. David Severson has also established a field project at Kenya Medical Research Institute (KEMRI) to look for vector competence of field populations of An. gambie to Plasmodium falciparum and correlative relationships with microsatellites and RFLP. Professors Nora Besansky and William Black, the latter a former postdoctoral research associate in Rai's laboratory, taught a WHOsponsored course in population genetics at ICIEPE in Nairobi, Kenya, during June 1998. Rev. Tom Streit, Research Assistant Professor, is currently involved in studies on the epidemiology of filariasis in Haiti under the auspices of CDC. His field research deals with evaluating the effectiveness of a relatively new drug, Ivermectin, against filariasis. This drug is used in the U.S. to treat heartworm disease in dogs, a disease closely related to filariasis. Dr. V. P. Sharma, who received his postdoctoral training in Rai's laboratory from 1964 to 1966, is the Director of the Malaria Research Centre, a major government enterprise in Delhi, India. He maintains close contact with the VBL, has sent students for training in vector genetics at UND, and has supplied the VBL with field strains of Ae. albopictus and Ae. aegypti from India. The Centre has several field stations in India, whose facilities are available to our students. The Director of the Assam field unit in East India, Dr. Vas Dev, is a 1984 Notre Dame Ph.D. recipient. The VBL has had special ties with the Florida Medical Entomology Laboratory (FMEL), a branch of the University of Florida. Professor Craig was an adjunct professor at FMEL. Over the years, our students have attended training sessions on Tropical Medical Entomology at Vero Beach in early January each year. At a recent FMEL workshop on mosquito ecology, more than half of the sixty participants had trained at Notre Dame at some time in their career.
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In addition, we have had an active ongoing collaboration with the Harris County Mosquito Abatement District in Houston, Texas, resulting in joint publications. Dr. Dan Sprenger, the entomologist at the district who first discovered Ae. albopictus in Houston, has supplied eggs of Ae. albopictus from twelve locations in Houston on a regular basis for our studies on temporal changes in genetic structure of that species after initial colonization. Other Notre Dame graduates or postdoctoral students currently working overseas include: Dr. Moosa A. Motara, Professor of Zoology and Dean of Students, Rhodes University, Grahamstown, South Africa Dr. W. L. Kilama, Director, National Institute of Medical Research, Amani, Tanzania Dr. Robert Copeland, Malariologist, U.S. Army Medical Research Service, Nisuma, Kenya Dr. Raymond Beach, CDC Malaria Research Team, Costa Rica
16— Vector Biology Courses and Visiting Scientist Program There are approximately forty courses taught by various faculty in UND's Department of Biological Sciences which are available to the students enrolled in Vector Biology. Of these, the following are particularly relevant: Course
Title
Instructors of Record
Bios 406
General Entomology
Craig (now Collins)
Bios 513
Medical Veterinary Entomology
Craig (now Collins)
Bios 506
Cytogenetics
Rai
Bios 515
Vector Genetics
Craig, Rai, and Severson
(table continued on next page)
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(continued) Course
Title
Instructors of Record
Bios 520
Arbovirology
Grimstad
Bios 520
Immunology
Diffley, Müller
Bios 536
Advanced Virology
Fraser
Bios 554
Biological Research Applications of Computers
Hellenthal
Bios 562
Aquatic Insects
Craig, Hellenthal
Bios 569
Practicum on Aquatic Biology
Staff
Bios 574
Topics in Evolutionary and Systematic Biology
Feder, Besansky, Collins
Bios 577
Topics in Genetics/Molecular Biology
Rai and Staff
Bios 663–665
Methods in Cellular and Molecular Biology Staff
Special mention must be made of the course entitled Vector Genetics (Bios 515). Beginning in fall 1968, this new, graduatelevel course began to be taught by Craig and Rai on a regular basis. This course was modeled on the WHO Seminar in Vector Genetics taught in 1967. It was the first course of its kind in the country. It constitutes the core course for graduate students and postdoctorals. It provides analyses of various aspects of the genetic biology of principal mosquito species that cause human disease. Areas stressed include formal genetics, cytogenetics, population genetics, evolutionary molecular genetics, and manipulation of genetic mechanisms for controlling vector populations. An introduction to computer simulation of pest populations is also included. Diverse vector species have also often been utilized to provide laboratory experience in genetic mapping, chromosomal analyses, insect sterilization, insecticide resistance, and capacity to transmit disease. Several faculty members and visiting scientists often participate in this course. Several scientists from all parts of the world have visited the VBL during the last forty years to present lectures to the VBL personnel and to meet with specific groups (appendix 5). These visiting scientists, many of whom have attained international reputation for their research in various aspects of insect biology and control, supplement
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the training provided by the resident faculty. Such formal and informal contacts with a wide variety of viewpoints help to stimulate student interest and to emphasize the interdisciplinary nature of the overall program.
Epilogue The death of George Craig in December 1995 left a void. However, the addition in 1997 of Frank Collins from CDC to fill the Clark Chair formerly held by Craig, Nora Besansky from CDC, David Severson from the University of Wisconsin, and Thomas Streit, one of Craig's former students, has greatly strengthened the program, particularly in molecular vector biology, and has broadened it to include multifaceted molecular genetic studies of the important malaria vector An. gambiae and related species in the An. gambiae complex. A major effort is underway, directed by Frank Collins, to physically map and subsequently clone and sequence the gene for encapsulation, which encapsulates and kills the malaria parasite early in sporogony, and the gene for malarial parasite refractoriness. Collins's laboratory is also working on isolating appropriate transposable DNA elements which could potentially be used to replace vector by nonvector populations. Nora Besansky is working to elucidate molecular evolutionary relationships among An. gambiae and An. arabiensis, which show sympatric distribution throughout tropical Africa in particular, and among other sibling species in the complex as well. DNA sequence variation from multiple loci, individuals, and geographical regions is being used to characterize population structure and to seek insights on the origin of behavioral differences among them. David Severson is a leader in quantitative and population genetics of Ae. aegypti and is actively employing molecular DNA markers, e.g., RFLP to map genes that confer susceptibility
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to filarial parasites Brugia malayi and Dirofilaria immitis and to the malarial parasite Plasmodium falcipaum. Finally, Tom Streit is actively involved in establishing a major laboratory in epidemiology and biology of several viral infections particularly common in the Midwest, in addition to his major field project in Haiti on filariasis control.
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Appendix 1— Vector Biology Faculty, University of Notre Dame, 1957–1999 Name Yrs. at N.D. Research Interests Yr. and Institution of Ph.D. George B. Craig, Jr.
1957–1995
Medical Entomology, Vector Genetics, Aquatic Insects
1956, Univ. of Illinois
Karamjit S. Rai
1960–1999
Vector Genetics, Cytogenetics, Molecular Evolutionary Genetics
1960, Univ. of Chicago
Theodore J. Crovello
1966–1984
Population Dynamics, Use of Computers
1966, Univ. of California, Berkeley
Morton S. Fuchs
1966–
Reproductive Biochemistry, Endocrinology
1967, Michigan State Univ.
Paul R. Grimstad
1976–
Arbovirology, Epidemiology, Emerging Infections
1973, Univ. of Wisconsin, Madison
Ronald A. Hellenthal
1977–
Research applications of computers
1977, Univ. of Minnesota, Twin Cities
Frank H. Collins
1997–
Molecular Vector Biology and Genetics
1981, Univ. of California, Davis
Nora J. Besansky
1997–
Molecular Evolutionary Genetics
1990, Yale University
David W. Severson
1997–
Population and Quantitative Genetics
1983, Univ. of Wisconsin, Madison
Thomas G. Streit
1997–
Arbovirology
1994, Univ. of Notre Dame
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Appendix 2— Postdoctoral Research Associates in Vector Biology/Parasitology since 1960 Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Rai, Karamjit S.
Univ. of Chicago, Ph.D., 1960
1960–62
Craig
Professor of Biology, Univ. of Notre Dame, Notre Dame, IN
BatMiriam, Mariassa
Hebrew Univ., Ph.D., 1963
1962–64
Craig
Prof. Emeritus, Sackler Med. School, Univ. of Tel Aviv, Israel
McClelland, G. A. H.
London School of Tropical Medicine, Ph.D., 1962
1962–63
Craig
Professor of Entomology, U.C. Davis, Davis, CA
Trebatosky, Sr. Alice
Univ. of Notre Dame, Ph.D., 1967
1967–68
Craig
Professor, Dept. of Biology, Pace Univ., New York
Hausermann, Walter
Basle, Switzerland, Ph.D.
1968–70
Craig
CIBAGEISYSA Centre De Recherches Agricoles, Aubin, Switzerland
Rodriguez, Paul
Univ. of Rhode Island, Ph.D., 1970
1970–73
Craig
Professor of Genetics, Div. of Life Sciences, Univ. of Texas—San Antonio, San Antonio, TX
(table continued on next page)
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Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Hacker, Carl S.
Rice Univ., Ph.D., 1968
1970–73
Craig
Assist. Professor of Ecology, Univ. of Texas, Health Science Center, Houston, TX
von Ende, Carl
Cornell Univ., A.B., 1967
1970–74
Craig / Ross
Assoc. Prof., Dept. of Biol. Sci., Northern Illinois Univ., DeKalb, IL
Fanara, Dean
U.C. Riverside, Ph.D.
1971–73
Craig
R&D International/Clorox, Pleasanton, CA
Trpis, Milan
Charles Univ., Prague, C.Sc., RNDr., Ph.D., 1971–74 1960
Craig
Professor of Medical Entomology, Dept. of Immunology & Infectious Diseases, Johns Hopkins Univ., MD
Igbokwe, Emmanual C.
Queen's Univ., Canada, Ph.D., 1970
1972–75
Craig
Science and Science Technology, Southern Univ. Shreveport, Shreveport, LA
Lounibos, Philip
Harvard Univ., Ph.D., 1974
1973–77
Craig
Florida Medical Entomology Lab, Univ. of Florida, Vero Beach, FL
(table continued on next page)
Page 66
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Grimstad, Paul
Univ. of Wisconsin, Ph.D., 1973
1974–76
Craig
Assoc. Professor of Biology, Dept. of Biological Sciences, Univ. of Notre Dame, Notre Dame, IN
Saul, Steven
Univ. of Rochester, Ph.D., 1970
1974–77
Craig
Assoc. Professor of Entomology, Dept. of Entomology, Univ. of HawaiiManoa, Honolulu, HI
Fish, Durland L.
Univ. of Florida, Ph.D., 1976
1976–80
Craig
Research Scientist, Dept. of Epidemiology & Public Health, Yale School of Medicine, New Haven, CT
Novak, Robert
Univ. of Illinois, Ph.D., 1976
1976–78
Craig
Illinois National Historical Survey, Champaign, IL
Sims, Steve
Unknown
1978–80
Craig
Location unknown
Nasci, Roger
Univ. of Massachusetts, Ph.D., 1983
1979–80
Craig
Research Entomologist, Dept. of VectorBorne Infect. Diseases, Centers for Disease Control, Fort Collins, CO
(table continued on next page)
Page 67
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Walker, Edward
Univ. of Massachusetts, Ph.D., 1984
11/83–3/86
Craig
Assoc. Professor, Dept. of Entomology, Michigan State Univ., East Lansing, MI
Beier, John
Johns Hopkins Univ., Sc.D., 1980
7/80–6/82
Craig
Assoc. Professor, Dept. of Trop. Med., School of Public Health & Trop. Med., Tulane Univ., New Orleans, LA
Munstermann, Leonard E. Univ. of Notre Dame, Ph.D., 1979
9/80–6/81
Craig
Research Scientist, Dept. of Epidemiology & Public Health, Yale Univ., New Haven, CT
Haramis, Linn
7/81–4/83
Craig
Entomologist/Program Manager, Arbovirus Surveillance and Vector Control Programs, Illinois Dept. of Public Health, Div. of Environmental Health, Springfield, IL
Ohio State Univ., Ph.D., 1981
(table continued on next page)
Page 68
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Rowton, Jr., Edgar
Univ. of Georgia, Ph.D., 1981
9/81–9/83
Craig
Research Entomologist, Walter Reed Army Inst., Washington, D.C.
Hawley, William
Univ. of Oregon, Ph.D., 1984
10/84–9/85
Craig
Research Entomologist, Malaria Branch, Centers for Disease Control, Atlanta, GA
Cully, Jack
Univ. of New Mexico, Ph.D., 1984
1989–90
Craig
Assist. Unit Leader—Wildlife, Dept. of Entomology, Kansas State Univ., Manhattan, KS
Blackmore, Mark
Univ. of Utah, Ph.D., 1989
3/91–8/93
Craig
Assist. Professor, Dept. of Biology, Valdosta State Univ., Valdosta, GA
Lord, Cynthia
Princeton Univ., Ph.D., 1991
2/92–8/93
Craig
Florida Medical Entomology Lab, Univ. of Florida, Vero Beach, FL
Brooks, Daniel
St. George Univ., M.S.; Ph.D.
1978–79
Crovello
Dept. of Zoology, Univ. of Toronto, Toronto, Ontario, Canada
(table continued on next page)
Page 69
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Uznanski, Richard
Univ. of Nebraska, Lincoln, Ph.D., 1976
1978–79
Crovello
Location unknown
Thompson, Steven R.
Oregon State Univ., Ph.D., 1966
1966–68
Fuchs
Assoc. Professor of Genetics, Ithaca College, Ithaca, NY
Kang, Sukkee
Univ. of Notre Dame, Ph.D., 1973
1973–74
Fuchs
Professor Emeritus and Head, Dept. of Biology, Univ. of Seoul, South Korea
France, Kenneth
Univ. of California, Davis, Ph.D.
1976–78
Fuchs
Johnson & Johnson, Venezuela
Kelly, Thomas
Univ. of Illinois, UrbanaChampagn, Ph.D. 1976–78
Fuchs
Deceased
Schlaeger, Dorothy
Univ. of Notre Dame, Ph.D., 1973
1980–83
Fuchs
VocationReligious Formation, Mt. St. Francis, Colorado Springs, CO
Whisenton, LaVern
Univ. of Notre Dame, Ph.D., 1980
8/80–6/82
Fuchs
Assoc. Professor, Dept. of Biology, Millersville Univ. of Pennsylvania, Millersville, PA
(table continued on next page)
Page 70
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Bang, Sookie S.
Univ. of California, Davis, Ph.D., 1981
1982–85
Fuchs
Assoc. Professor, Dept. of Biology, South Dakota School of Mines & Technology, Rapid City, SD
Powell, James
Ph.D., 1983 Chemistry
1983–88
Fuchs
Professor, Ancilla College, Donaldson, IN
Smith, Timothy
Ph.D.
1983–84
Fuchs
Dept. of Pharmacology, Univ. of Nebraska, Omaha
Thomas, William R.
Mississippi State Univ., Ph.D., 1982
9/83–8/85
Fuchs
Merck Pharmaceuticals, Sumneytown Pike, West Point, PA
Hollander, Andrew
Univ. of Massachusetts, Ph.D., 1983
10/83–9/85
Fuchs
Business Process Consultant, Office of Information Technologies, Univ. of Notre Dame, Notre Dame, IN
Hilliard, Richard
Texas A&M Univ., Ph.D., 1983
10/84–6/86
Fuchs
Director of Research Compliance, Graduate School Research Division, Univ.of Notre Dame, Notre Dame, IN
(table continued on next page)
Page 71
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Rivers, Elissa
Univ. of Minnesota, Ph.D., 1985
1989–90
Fuchs
Deceased
Burks, Charles
Univ. of Missouri, Ph.D., 1991
7/91–8/93
Fuchs
U.S. Grain Market Research Lab., USDA, Manhatten, KS
Blackmore, Carina
Univ. of Notre Dame, Ph.D., 1996
4/96–9/96
Grimstad
Temporary Assist. Professor, Valdosta State Univ., Valdosta, GA
Costero, Adriana
McGill Univ., Canada, Ph.D., 1994
9/96–
Grimstad
Postdoctoral Fellow, Univ. of Notre Dame, Notre Dame, IN
Garvin, Mary
Univ. of Florida, Ph.D., 1996
9/96–
Grimstad
Postdoctoral Fellow, Univ. of Notre Dame, Notre Dame, IN
Wallace, John
Michigan State Univ., Ph.D., 1997
6/97–
Grimstad
Postdoctoral Fellow, Univ. of Notre Dame, Notre Dame, IN
Mescher, Alma Louise Sr.
Univ. of Notre Dame, Ph.D., 1963
6/64–8/64
Rai
Professor, St. Mary's of the Woods, Terre Haute, IN
Dhillon, T.S.
Univ. of Chicago, Ph.D., 1960
1964–65
Rai
Professor Emeritus, Dept. of Botany, Univ. of Hong Kong, Hong Kong
(table continued on next page)
Page 72
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Sharma, V. P.
Univ. of Allahabad, Ph.D.
1965–67
Rai
Director, Malaria Research Centre, Delhi, India
Hartberg, Warren Keith
Univ. of Notre Dame, Ph.D., 1968
5/68–7/68
Rai
Chmn., Dept. of Biology and Professional Biology, Baylor Univ., Waco, TX
McDonald, Paul
Univ. of Notre Dame, Ph.D., 1970
1970–74
Rai
Project Manager for Insecticides, Uniroyal Chemical Co., Bethany, CT
Brat, Ved
Oxford Univ., Ph.D.
1971–73
Rai
President, Bratton Biotech, Bethesda, MD
Lorimer, Nancy
Univ. of Notre Dame, Ph.D., 1975
1973–75
Rai
Entomologist, USDA Forest Research Service, St. Paul, MN
Marchi, Annalisa
Univ. of Cagliari, Italy, Ph.D.
1976–77
Rai
Univ. of Cagliari, Cagliari, Italy
Hilburn, Larry R.
Texas Tech Univ., Ph.D., 1978
1978–80
Rai
Professor of Biology, Rhodes Univ., Memphis, TN
Chang, HoChi
Univ. of Nebraska, Lincoln, Ph.D., 1979
1979–80
Rai
Location unknown
Pashley, Dorothy P.
Univ. of Texas, Ph.D., 1980
1980–82
Rai
Professor, Dept. of Entomology, Louisiana State Univ., Baton Rouge, LA
(table continued on next page)
Page 73
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position McLain, Denson Kelly
Emory Univ., Ph.D., 1982
1983–85
Rai
Assoc. Professor, Biology Dept., Georgia Southern Univ., Statesboro, GA
Boromisa, Robert
Univ. of Michigan, M.P.M., 1983
1985–86
Rai
Location unknown
Ferrari, James A.
Univ. of Calif., Riverside, Ph.D., 1985
1985–86
Rai
Assoc. Professor, Dept. of Biology, California State Univ., San Bernardino,CA
Black, William
Iowa State Univ., Ph.D., 1985
1985–87
Rai
Assoc. Professor, Dept. of Microbiology, Univ. of Colorado, Fort Collins, CO
Kambhampati, Srinivas
Simon Fraser Univ., Canada, Ph.D., 1988
1987–92
Rai
Assoc. Professor, Dept. of Entomology, Kansas State Univ., Manhattan, KS
Kumar, Arun
Banaras Hindu Univ., India, Ph.D., 1982
1988–93
Rai
Assist. Professor, Indian Institute of Science, Bangalore, India
(table continued on next page)
Page 74
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Thomas, Rex
Univ. of Oklahoma, Ph.D., 1986
1990–91
Rai
Staff Scientist/Medical Entomologist, Paravax,Inc., Fort Collins, CO
Wu, Wheikuo
Yale Univ., Ph.D., 1989
1990–92
Rai
Senior Fellow, Dept. of Lab. Medicine, Univ. of Washington, Seattle, WA
Sinkins, Steven
Univ. of London, Ph.D., 1996
1997–
Besansky
Research Associate, Univ. of Notre Dame, Notre Dame, IN
Mukabayire, Odette
Univ. of La Sapeinza, Rome, Ph.D., 1993
1997–
Collins
Research Associate, Univ. of Notre Dame, Notre Dame, IN
(table continued on next page)
Page 75
Appendix 2, con't. Name Prior Institution, Degree and Date Years in Residence Mentor at Notre Dame Current Position Ranson, Hilary
Univ. of Wales, Ph.D., 1996
1997–
Collins
Visiting Scholar, Univ. of Notre Dame, Notre Dame, IN
Ke, Zhaoxi (Peter)
Univ. of Arizona, Ph.D., 1992
1997–
Collins
Research Associate, Univ. of Notre Dame, Notre Dame, IN
Sarkar, Abbimanyu
Univ. of Maryland, Ph.D., 1998
1997–
Collins
Research Associate, Univ. of Notre Dame, Notre Dame, IN
*
Some of the addresses and years listed above may not be current and/or complete.
Page 76
Appendix 3— Predoctoral Students in Vector Biology/Parasitology since 1961 Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position VandeHey, Robert
St. Norbert College, B.A., 1946; Univ. of Notre Dame, M.S., 1957
Ph.D., 1961
Craig
Professor, Dept. of Biology, St. Norbert College, De Pere, WI
Leahy, Sr. Mary Gerald
USC, B.A., 1945; Catholic Univ., M.A., 1947 Ph.D., 1962
Craig
Carondolet Center, Los Angeles, CA
Mescher, Alma Louise Sr.
Immaculate Heart College, B.A., 1942; Marquette Univ., M.S., 1953
Ph.D., 1963
Craig
Professor, St. Mary's of the Woods, Terre Haute, IN
Adhami, Usman
Aligarh Muslim Univ., India, M.S., 1960
Ph.D., 1964
Craig
Professor of Genetics, Aligarh Muslim Univ., Dept. of Zoology, Aligarh, India
Hickey, William A.
Univ. of Notre Dame, M.S., 1959
Ph.D., 1965
Craig
President Emeritus, St. Mary's College, Notre Dame, IN
Schoenig, Fr. Enrique (Heinrich?)
Univ. of Notre Dame, M.S., 1952
Ph.D., 1966
Craig
Dept. of Biology, Univ. of San Carlos, Cebu City, Philippines (Deceased)
(table continued on next page)
Page 77
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Trebatoski, Sr. Alice
Alverno College, B.S., 1958; Univ. of Notre Ph.D., 1967 Dame, M.S., 1964
Craig
Professor, Dept. of Biology, Pace Univ., New York
Bhalla, Satish
E. Punjab Univ., M.S., 1956; Univ. of Kansas, M.S., 1963
Ph.D., 1967
Craig
Location unknown
Hartberg, Warren Keith
Wabash College, A.B., 1963; Univ. of Notre Ph.D., 1968 Dame, MSc, 1965
Craig
Chmn., Dept. of Biology and Professional Biology, Baylor Univ., Waco, TX
O'Meara, George F.
Univ. of Notre Dame, B.S., 1964; M.A., 1967 Ph.D., 1969
Craig
Professor, Florida Medical Entomology Lab., Univ. of Florida, Vero Beach, FL
Gwadz, Robert
Univ. of Notre Dame, B.S., 1962
Ph.D., 1970
Craig
Staff Fellow, Primate Malaria Unit, NIH, GA
Quinn, Thomas
Univ. of Notre Dame, B.S., 1969
M.S., 1970
Craig
Professor of Medicine, Johns Hopkins Univ.,Baltimore, MD
Kilama, Wenceslaus
Univ. of Notre Dame, M.S., 1967; M.A., 1968
Ph.D., 1971
Craig
Director, National Inst. Medical Research, Salaam, Tanzania
(table continued on next page)
Page 78 Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Ramalingam, S.
Madras Univ, India, B.S., M.S.
Ph.D., 1973
Craig
Curator of Parasitology, Univ. of Alberta, Canada
Ph.D., 1973
Craig
Dean of the College of Science, Central Univ.of Venezuela, Caracas, Venezuela
Terwedow, Henry
Univ. of Notre Dame, B.S., 1968; Montclair Ph.D., 1973 State Univ., M.A., 1969
Craig
Biostatistician Epidemiologist, MA General Hospital, Molecular Neurogenetics Unit, Charlestown, MA
O'Flynn, Sr. Ignatius
Univ. of Notre Dame, M.S., 1972
Ph.D., 1974
Craig
St. Ursula's School, Collymore Rock, Barbados, West Indies
von Ende, Carl
Cornell Univ., A.B., 1967
Ph.D., 1975
Craig / Ross
Assoc. Professor, Dept. of Biological Sciences, Northern Illinois Univ.
Petersen, John
Columbia Univ., M.S., 1971
Ph.D., 1976
Craig
Assoc. Professor, Panama Canal College, Panama
Beach, Raymond
Univ. of Notre Dame, B.S., 1970
Ph.D., 1976
Craig
Centers for Disease Control, Malaria Research Team, Costa Rica
MachadoAllison, Carlos Central Univ. of Venezuela, M.S.
(table continued on next page)
Page 79
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Sinsko, Michael
Eastern Illinois Univ., M.S., 1972
Ph.D., 1976
Craig
Sr. Medical Entomologist, Indiana State Board of Health, Indianapolis, IN
Shroyer, Donald
Purdue Univ., M.S., 1974
Ph.D., 1977
Craig
Medical Entomologist, Indian River Mosquito Abatement, Vero Beach, FL
Munstermann, Leonard
Univ. of Minnesota, M.A., 1968
Ph.D., 1979
Craig
Yale Univ., Arbovirus Research Unit
Huang, SioMei Lein
Taipai, Taiwan
M.S., 1979
Craig
Location unknown
McCombs, Susan
Univ. of Notre Dame, M.S., 1980
M.S., 1980
Craig
Assist. Entomologist,Dept. of Entomology, Univ. of Hawaii—Manoa, Honolulu, HI
Peloquin, John J.
Univ. of Notre Dame, B.S., 1975
M.S., 1980
Craig
Research Biochemist, U.C. Irvine, Dept. of Molecular Biology, Irvine, CA
Durso, Stephan
Univ. of Notre Dame, B.S., 1979
M.S., 1982
Craig
Director, Palm Springs Mosquito Abatement District, Palm Springs, CA
(table continued on next page)
Page 80
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Rohrer, William H.
St. Mary's College, Minnesota, B.A., 1982
Ph.D., 1985
Craig
Research Assist., Merck Co. Foundation, Rutgers Univ., Biomedical Engineering, Piscataway, NJ
Berry, William
Univ. of Notre Dame, B.S., 1981
M.S., 1983
Craig
Environmental Scientist, U.S. Atomic Energy Commission, Moscow, ID
Taylor, David B.
Univ. of Utah, M.S., 1979
Ph.D., 1983
Craig
Entomologist, USDA Screwworm Lab, Lincoln, NB
Pumpuni, Charles
Univ. of Ghana, B.S., 1982
M.S., 1986; Ph.D., 1989
Craig
Entomologist, Univ. Maryland Medical School, Baltimore, MD
Leiser, Lorraine
Purdue Univ., B.S., 1979
Ph.D., 1986
Craig
Research Associate, USDA Screwworm Lab, Lincoln, NE
Paulson, Sally
Miami Univ., M.S., 1981
Ph.D., 1987
Craig
Associate Professor of Entomology, Virginia Polytech Univ., Blacksburg, VA
(table continued on next page)
Page 81
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Copeland, Robert
Louisiana State Univ., M.S., 1981
Ph.D., 1987
Craig
International Centre of Insect Physiology & Ecology, Nairobi, Kenya
Hanson, Scott
Andrews Univ., B.S., 1986
M.S., 1988; Ph.D., 1991
Craig
Research Entomologist, Medical School, Univ. of Tokyo, Japan
Wesson, Dawn
Univ. of Illinois at Chicago, M.S., 1985
Ph.D., 1990
Craig
Asst. Professor, Department of Tropical Medicine, Tulane Univ., New Orleans, LA
Mutebi, JohnPaul
Makerere Univ. College, B.Sc., 1984
M.S., 1991 Ph.D., 1995
Craig
Univ. of Texas, Medical Branch, Dept of Pathology Research, Galveston, TX
Streit, Fr. Thomas
Univ. of Notre Dame, M. Div., 1985
M.S., 1991 Ph.D., 1994
Craig
Res. Asst. Professor, Dept. of Biological Sciences, Univ. of Notre Dame
Niebylski, Mark
Louisiana State Univ., M.S., 1988
Ph.D., 1992
Craig
Research Scientist, Rocky Mountain National Lab, Hamilton, MT
(table continued on next page)
Page 82
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Scoles, Glen
Cornell Univ., B.S., 1980
M.S., 1994 Ph.D., 1997
Craig
Postdoc, Dept. of Epidemiology & Public Health, Yale Univ., New Haven, CT
Kreuger, Betty
Univ. of Kentucky, B.S., 1993
M.S., 1996
Craig
Graduate Student, Univ. of Kentucky, Lexington, KY
Russo, Raymond
Northeast Missouri State Univ., M.A., 1971 Ph.D., 1977
Crovello
Assoc. Professor of Ecology, Ind. Univ.—Purdue Univ., Indianapolis, IN
Keller, Clifton
Washington Univ., M.A.
Ph.D., 1979
Crovello
Director, Cedar Lake, MI
Hauser, Larry A.
Emporia State Univ., M.S.
M.A., 1982
Crovello
Supervisor, Marion Merrill Dow, Inc., BristolMyers Parmaceutical, Evansville, IL
McMillan, Jack D.
Indiana Univ. of Pennsylvania, M.S.
Ph.D., 1984
Crovello
Asst. Data Manager, Washington State Department of Natural Res., Olympia, WA
Carter, Joan Smith
Goshen College, B.A., 1968
M.S., 1971
Fuchs
Head, Math and Science Program, Durham Tech. Community College, Durham, NC
(table continued on next page)
Page 83
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Whisenton, LaVern
Morningside College, B.S.
Ph.D., 1980
Fuchs
Assoc. Professor, Dept. of Biology, Millersville State Univ., Millersville, PA
Hiss, Edwin
Univ. of Notre Dame, B.S., 1966
Ph.D., 1971
Fuchs
Administrative Manager, Washington Univ., Dept. of Chemistry, St. Louis, MO
Schlaeger, Sr. Dorothy
Univ. of Notre Dame, M.S., 1969
Ph.D., 1973
Fuchs
VocationReligious Formation, Mt. St. Francis, Colorado Springs, CO
Kang, Suk Hee
Univ. of Wisconsin, M.S.
Ph.D., 1973
Fuchs
Professor Emeritus and Head, Dept. of Biology, Univ. of Seoul, South Korea
Pomato, Nicholas
Univ. of Notre Dame, B.S., 1967
Ph.D., 1974
Fuchs
Dept. Head, Biochemistry, Bionetics Research Institute, Rockville, MD
Fong, WangFun
Chinese Univ. of Hong Kong, B.S.
Ph.D., 1975
Fuchs
Location unknown
Thompson, Stephen
Unknown
Ph.D., 1977
Fuchs
Location unknown
(table continued on next page)
Page 84
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Masler, E. Peter
Univ. of New Hampshire, M.S.
Ph.D., 1978
Fuchs
Cornell Univ., Ithaca, NY
Sundland, Barry
Univ. of Missouri, St. Louis, B.A.
M.S., 1979
Fuchs
Family Practice Physician, Aurora, CO
Fortunato, Paul J.
Penn. State Univ., B.S.
M.S., 1979
Fuchs
Medical Lab Technician/Clinical Microbiologist, Indiana Hospital, Indiana, PA
Banavaliker, Sujata
St. Xaviers College (Bombay), B.S., 1981
M.S., 1984
Fuchs
Deceased
Lee, Seung Koo
Sungkyunkwan Univ., B.S., 1983
M.S., 1986, Ph.D., 1992
Fuchs
Postdoctoral Fellow, Medical School, Dept. of Biochemistry, Stanford Univ., Stanford, CA
Elton, Tami
Univ. of Notre Dame, B.S., 1993
M.S., 1996
Fuchs/Craig
Technician, Dept. of Biological Sciences, Univ. of Notre Dame, Notre Dame, IN
Boromisa, Robert
Univ. of Michigan, M.P.H., 1980
Ph.D., 1985
Grimstad
Wadesworth Ctr. For Labs & Research, Griffin Laboratories, Albany, NY
(table continued on next page)
Page 85
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Zhang, Mingbao
Medical Univ. of HarbinChina, B.S.
Ph.D., 1993
Grimstad
Wadesworth Ctr. For Labs & Research, Griffin Laboratories, Albany, NY
Blackmore, Carina
Swedish Univ., D.V.M., 1989
Ph.D., 1996
Grimstad
Temporary Assist. Professor, Valdosta State Univ., Valdosta, GA
Nowicki, Wendi
Manchester College, B.S.
Ph.D., 1996
Grimstad
Bayer, Inc., Elkhart, IN
Liu, Hope
Virginia Polytech Univ., B.S., 1994
M.S., 1996
Grimstad/Craig
Graduate Student, Virginia Polytech Univ., Blacksberg, VA
Heard, Phillip
Univ. of Oklahoma, M.D., 1987
Ph.D., 1997
Grimstad
Epidemiologist, Maryland Dept. of the Environment
Anderson, Justin
Ripon College, B.A., 1995
In progress
Grimstad
Graduate Student, Univ. of Notre Dame, Notre Dame, IN
Brockus, Catherine
Central Michigan Univ., B.S., 1984
In progress
Grimstad
Graduate Student, Univ. of Notre Dame, Notre Dame, IN
Meece, Jennifer
Western Illinois Univ., M.S., 1995
In progress
Grimstad
Graduate Student, Univ. of Notre Dame, Notre Dame, IN
(table continued on next page)
Page 86
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Asman, Monica
Univ. of Notre Dame, M.S., 1960
Ph.D., 1966
Rai
Retired Entomologist/Dir. Univ. CA; St. Francis Center, Redwood City, CA
Akhtaruzzaman, M.
Dacca Univ., M.Sc.
Ph.D., 1967
Rai
Professor and Head, Dept. of Botany, Univ. of Dacca, Bangladesh
McDonald, Paul
Yale Univ., B.A., 1964
Ph.D., 1970
Rai
Project Manager for Insecticides, Uniroyal Chemical Co., Bethany, CT
Mehra, Romesh
Agra Univ., India, M.Sc.
M.Sc., 1970
Rai
Dept. of Biology, Indiana Univ., South Bend, IN
Mukunnemkeril, George M.
DePaul Univ., M.S.
Ph.D., 1973
Rai
Dept. of Biology, Greensboro College, Greensboro, IL
McGivern, James
Iona College, B.S.
Ph.D., 1973
Rai
Assoc. Professor, Dept. of Biology, Gannon Univ., Erie, PA
Lorimer, Nancy L.
Indiana Univ., B.S.
Ph.D., 1975
Rai
Staff Entomologist, USDA Forest Research Service, St. Paul, MN
(table continued on next page)
Page 87
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Hallinan, Edward
Unknown
Ph.D., 1976
Rai
Director, Cytogenetics, St. Luke's Hospital, Bethlehem, PA
Motara, Moosa A.
Rhodes Univ., S. Africa, M.S., 1972
Ph.D., 1978
Rai
Dean of Students, Rhodes Univ., Grahamstown, S. Africa
Soderquist, Ann
Vanderbilt Univ., M.Div.
Ph.D., 1980
Rai
Pastor, Minister, Nashville, TN
Szymczak, Larry
Indiana Univ., South Bend, B.A., 1975
Ph.D., 1981
Rai
Assoc. Professor, Dept. of Biology, Chicago State Univ., Chicago, IL
Sherron, David A.
N. Carolina State Univ., M.S., 1979
Ph.D., 1983
Rai
High School Biology Teacher, Houston Independent School Dist., Houston, TX
Dev, Vas
Panjab Univ., India, M.S., 1978
Ph.D., 1984
Rai
Sr. Research Officer, Malaria Research Centre, Berhampur, India
Greenlee, Judith
Indiana Univ., South Bend, B.A., 1979
Ph.D., 1984
Rai
Assoc. Professor, Pasadena City College, Dept. of Life Sciences, Pasadena, CA
(table continued on next page)
Page 88
Appendix 3, con't. Name Prior Institution, Degree and Date Notre Dame Degree and Date Mentor at Notre Dame Current Position Knaak, Christian
Univ. of Delaware, B.A., 1983
M.S., 1985
Rai
Research Assoc., Louisiana State Univ., Baton Rouge, LA
Rao, P. Nageshwara
Univ. of Calcutta, M.S., 1980
Ph.D., 1985
Rai
Assoc. Professor, Univ. of California, Med. Sch., Los Angeles
Bosio, Chris
Bucknell Univ., B.S., 1989
M.S., 1993
Rai
Research Asst., Dept. of Entomology, Colorado State Univ., Fort Collins, CO
Taafe, Christine
Univ. of Notre Dame, B.S., 1993
M.S., 1997
Rai/Craig
Technician, Administrative Asst., VBL, Univ. of Notre Dame, Notre Dame, IN
Rohr, Cherise
Yale Univ., M.P.H., 1994
In progress
Besansky
Graduate Student, Univ. of Notre Dame, Notre Dame, IN
Dymbrowski, Kirk
Univ. of Illinois at Chicago, B.A., 1996
In progress
Collins
Graduate Student, Univ. of Notre Dame, Notre Dame, IN
*
Some of the addresses and years listed above may not be current and/or complete.
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Appendix 4— Undergraduate Students in Vector Biology since 1962 Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Keeley, Larry L.
B.S., 1962
Purdue Univ., Ph.D.
Craig
Professor, Dept. of Entomology, Texas A&M, College Station, TX
Berry, Richard
B.S., 1964
Ohio State Univ., Ph.D.
Craig
Medical Entomologist, Ohio Dept. of Health, Columbus, OH
Truman, James W.
B.S., 1965
Harvard Univ., Ph.D., 1970
Craig
Professor, Dept. of Zoology, Univ. of Washington, Seattle, WA
Powell, John
B.S., 1966
Univ. of Michigan, M.D.
Craig
Dermatologist, St. Louis, MO
Dunn, Michael
B.S., 1967
Northwestern Univ., M.D.
Craig
Office of Secretary of Defense, OASD (Health Affairs), Dayton, OH
Quinn, Thomas
B.S., 1969
Northwestern Univ., M.D.
Craig
Professor of Medicine, Johns Hopkins Univ., Baltimore, MD
Lounibos, Philip
B.S., 1969
Harvard Univ., Ph.D., 1974
Craig
Florida Medical Entomology Lab, Univ. of Florida, Vero Beach, FL
(table continued on next page)
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Appendix 4, con't. Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Powell, Jeffrey
B.S., 1969
U.C. Davis, Ph.D., 1972
Craig
Professor, Ecology, Evolution & Systematics, Yale Univ.
Beach, Raymond
B.S., 1970
Univ. of Notre Dame, Ph.D., 1976
Craig
Centers for Disease Control, Atlanta, GA
Nijhout, Frederick
B.S., 1971
Harvard Univ., Ph.D.
Craig
Professor, Dept. of Zoology, Duke Univ.
Murtaugh, Michael
B.S., 1973
Ohio State Univ., Ph.D., 1980
Craig
Prof., Univ. of Minnesota, Dept. of Vet. Pathology, St. Paul, MN
Connor, William
B.S., 1973
Cornell Univ., Ph.D.
Craig
V.P. and Actuary, Prudential Ins. Co. of America, Newark, NJ
Paige, Christopher
B.S., 1974
Cornell Univ., Ph.D., 1981
Craig
Location unknown
Burkot, J. Thomas
B.S., 1975
Univ. of St. Thomas, MBA
Craig
Financial Planner, Metlife Securities, Inc., Wexford, PA
Peloquin, John
B.S., 1975
Univ. of Notre Dame, M.S., 1980
Craig
Research Biochemist, U.C. Irvine, Dept of Molecular Biology, Irvine, CA
Piotrowski, Joseph
B.S., 1975
Hahnemann Univ., M.D.
Craig
Surgeon, Sante Fe, NM
(table continued on next page)
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Appendix 4, con't. Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Landry, Steven
B.S., 1977
Univ. of Wisconsin, Ph.D.
Craig
Health Advisor, USAID, Office of Health, Washington, D.C.
Berges, Ann
B.S., 1977
Oregon State Univ., M.S.
Craig
Teacher, Florence, KY
Taylor, David B.
B.S., 1977
Univ. of Notre Dame, Ph.D., 1983
Craig
Research Scientist, USDA, ARL, Univ. of NebraskaLincoln
Sullivan, Kathleen M.
B.S., 1978
Univ. of California, San Diego, Ph.D.
Craig
Assoc. Prof., Univ. of Miami, Dept. of Biology, Coral Gables, FL
McKiernan, Margaret
B.S., 1979
Univ. of Notre Dame, M.S., 1981
Craig
Argonne National Lab., Argonne, IL
Durso, Stephen
B.S., 1979
Univ. of Notre Dame, M.S., 1982
Craig
Entomologist, Corona, CA
Berry, William
B.S., 1981
Univ. of Notre Dame, M.S., 1983; Iowa Craig State Univ., Ph.D.
(table continued on next page)
Environmental Scientist, Compliance Specialist, Jason Assoc., Idaho Falls, ID
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Appendix 4, con't. Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Skahan, Kenneth J.
B.S., 1981
Ohio State Univ., M.D.
Craig
Infectious Disease Specialist, Univ. of Cincinnati Medical Center, Cincinnati, OH
Gimnig, John E.
B.S., 1991
Unknown
Craig
Research Assist., U.C. Davis, Dept. of Entomology, Davis, CA
Peterson, Rebecca
B.S., 1996
Unknown
Craig/Rai
Location unknown
Kogge, Stephen N.
B.S., 1971
Univ. of Notre Dame, Ph.D., 1976
Fuchs
Computer Engineer, Univ. of Maryland, Dept. of Computer Science, College Park, MD
Zielinski, Henry
B.S., 1972
Loyola Univ., Chicago, M.D.
Fuchs
Surgical Partner, Marietta, GA
Philip, Carter
B.S., 1967
Univ. of Notre Dame, Ph.D., 1971
Rai
Professor of Microbiology, North Carolina State Univ., Raleigh, NC
Smith, Harold
B.S., 1969
Unknown
Rai
Physician, Homer, AK
Westenkirschner, David
B.S., 1973
Medical College of Ohio, M.D.
Rai
Assoc. Director, Pediatric ICU, Riley Hospital for Children, Indianapolis, IN
(table continued on next page)
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Appendix 4, con't. Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Podlasek, Stanley, J.
B.S., 1975
Univ. of Notre Dame, M.S., 1977
Rai
Physician, Washington, D.C.
Baum, James
B.S., 1976
Unknown
Rai
Research Assoc., Univ. of Georgia, Dept. of Genetics, Athens, GA
Malko, Eileen
B.S., 1976
Univ. of Pennsylvania, Ph.D.
Rai
Research Scientist, Univ. of Pennsylvania, Philadelphia, PA
Murphy, Michael
B.S., 1982
Univ. of TexasAustin, Ph.D.
Rai
Assist. Prof., Albany Medical College, Dept. of Pharmacology, Albany, NY
Herman, Greg
B.S., 1984
Jefferson Medical College, M.D.
Rai
Family Physician, Keller Army Hospital, Dept. of Primary Care, West Point, NY
Pompa, Patty A.
B.S., 1990
Unknown
Rai
Cytogenetics Technician, Southwest Genetics, San Antonio, TX
Burgun, Stephen J.
B.S., 1991
SUNYSyracuse, M.D.
Rai
Clinical Instructor, Ohio State Univ., Dept. of Internal Medicine, Columbus, OH
(table continued on next page)
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Appendix 4, con't. Name Notre Dame Degree and Date Graduate Institution, Degree and Date Mentor at Notre Dame Current Position Anbar (Bozer), Marni
B.S., 1991
Rai
Masters Candidate, Univ. of Rochester, Dept. of Education, Rochester, NY
Villa, John F.
B.S., 1991
Syracuse HSC, M.D.
Rai
Physician, Florida Hospital, Orlando, FL
Biros, Daniel
B.S., 1992
Univ. of MinnesotaSt. Paul, D.V.M.
Rai
Veterinarian, Colorado State Univ., Vet. Teaching Hospital, Fort Collins, CO
Haas, David
B.S., 1993
N/A
Rai
Medical Student, St. Louis Univ. School of Medicine, St. Louis, MO
*
This is only a partial listing of the undergraduate research assistants in the VBL at ND. Some of the addresses listed above may be incorrect and/or incomplete.
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Appendix 5— Visiting Scientists/Professors and Guest Lecturers Relevant to Vector Biology at the University of Notre Dame A— Visiting Scientists/Professors Dr. Mariassa BatMiriam, Univ. of Tel Aviv, Israel. Visiting Prof. 1963–64. Dr. A. W. A. Brown, University of Western Ontario, London, Ontario, Canada. Visiting Prof. June 17–July 26, 1968. Dr. Donald G. Cochran, Virginia Polytechnic Inst., Blacksburg, VA. Visiting Prof. June 17–July 26, 1968. Dr. William Hickey, St. Mary's College, Notre Dame, IN. Visiting Prof. June 17–July 26, 1968. Dr. James. B. Kitzmiller, University of Illinois, Urbana, IL. Visiting Prof. June 17–July 26, 1968. Dr. Leo E. LaChance, Metabolism and Radiation Research Lab., USDA, Fargo, ND. Visiting Prof. June 17–July 26, 1968. Dr. Rajindar Pal, Div. of Vector Biology and Control, WHO, Geneva, Switzerland. Global vector control problems; Chemical control; Insecticide resistance and environmental pollution; Alternative methods of control: biological and genetic; The future of vector control. June 17–July 26, 1968; July 12–23, 1971; August 31, 1976. Dr. Reinhart Brust, Dept. of Entomology, Univ. of Manitoba, Winnepeg, Manitoba, Canada. Visiting Prof. 1971. Dr. William Z. Coker, Entomology Dept., Univ. of Ghana, Legon, Acra, Ghana. Visiting Prof. 1977; 1981; 1986. Dr. Terry Matthews, Dept. of Biology, Indiana Univ., N.W., Gary, IN. Visiting Prof. 1979–1985 (summers). Dr. James Long, Biology Dept., Sam Houston State Univ., Huntsville, TX. Visiting Prof. 1981; 1982 (6 mos). Dr. Chester L. Sutula, President, Agdra Inc., Elkhart, IN. Guest Assoc. Prof. 1982. Dr. Eddie W. Cupp, Medical Entomology, Cornell Univ., Ithaca, NY. Visiting Prof. 1982; 1983 (6 mos). Dr. B. N. Chowdiah, Professor of Zoology, Bangalore Univ., India. Visiting Professor. May 12–25, 1982. Dr. A. Balankura, Dept. of Biology, Ramkhamaeng Univ., Bangkok, Thailand. Visiting Professor. Oct. 1–Nov. 15, 1982.
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Dr. Bruce Eldridge, Entomology Dept., Oregon State Univ., Corvallis, OR. Visiting Prof. 1984 (6 mos). Dr. Louis G. Mukwaya, East African Virus Research Institute, Entebbe, Uganda. Visiting Prof. 1984 (6 mos). Dr. Moosa Motara, Dept. of Zoology and Entomology, Rhodes Univ., South Africa. Visiting Prof. Sept. 1–Nov. 30, 1984. Dr. Stanley N. Grove, Dept. of Biology, Goshen College, Goshen, IN. Visiting Assoc. Prof. 1985. Dr. Itam Sulaiman, Univ. of San Malaysia, Malaysia. Visiting Assoc. Prof. 1985. Dr. R. I. Sommerville, Dept. of Zoology, Univ. of Adelaide, Adelaide, Australia. Visiting Prof. Dr. Jaroslaw Krzywinski, University of Gdansk, Gdansk, Poland. Visiting Scientist. 1998. B— Guest Lecturers Dr. Peter Mattingly, British Museum, London, England. ''Problems in mosquito zoogeography." October 22–23, 1969. Dr. Max Whitten, Dept. of Entomology, CSIRO, Australia. "Application of genetics for sheep blowfly control in Australia." December 17, 1969. Dr. Jack C. Jones, Univ. of Maryland, College Park, MD. "Stereoscan studies with Aedes aegypti." April 14–15, 1970. Dr. J. Austin Kerr, Pan American Health Org. "Mosquitoborne disease around the world." April 15–16, 1970. Dr. Satish C. Bhalla, Univ. of Maryland, Baltimore, MD. "Chromosomal aberrations and sex linked lethals in Aedes aegypti." May 11, 1970. Dr. George D. Hanks, Indiana Univ., Northwest Campus, Gary, IN. "Meiotic Drive." May 27, 1970. Dr. Jack Seawright, USDA, laboratory on "Insects Affecting Man," Gainesville, FL. "Genetics of Aedes aegypti; Field testing of translocation heterozygotes of Aedes aegypti." June 15–17, 1970; November 18, 1971. Dr. H. J. Barr, Univ. of Wisconsin, Madison, WI. "Development and cytology of insect embryos genetically deficient for nucleolar organizers." July 22, 1970. Dr. J. Laarman, Netherlands Institute for Trop. Medicine, Leiden, Netherlands. "Genetics and behavior in Anopheles maculipennis." September 17, 1970. Dr. Gillian Winner, Univ. of Windsor, Windsor, Ontario, Canada. "Ap
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plication of electron microscopy to the study of the insect reproductive system." October 9, 1970. Dr. Ernest Boesiger, National Centre de la Recherche Scientifique, Gifsurvette, France. "Heterogeneity of natural populations"; "Genetical load"; "Polymorphism; Geographical differentiation of populations"; "Courtship and sexual selection in Drosophila"; "Inbreeding depression and heterosis in Drosophila and in Japanese quail''; "Developmental and genetical homeostasis." December 7–18, 1970. Dr. R. H. Baker, Pakistan Medical Research Center; Univ. of Maryland, Lahore, West Pakistan. "The genetics of Culex tritaeniorhynchus." December 16, 1970. Dr. Chas. R. Burnham, Dept. of Genetics, Univ. of Minnesota, St. Paul, MN. "Chromosomal interchanges: studies of chromosome pairing and related behavior." March 14–15, 1971. Dr. M. Aslamkhan, Pakistan Medical Research Centre, Lahore, Pakistan. "Endimology and transmission of filariasis in Pakistan"; "Genetics of vectorability of Wuchereria bancrofti in Pakistan"; "Genetics of Anopheles stephensi and the transmission of malaria in Pakistan." March 17, 1971. Dr. Ralph Barr, Univ. of California, Los Angeles. "Genetic control of Culex pipiens." April 28, 1971. Dr. Canute Khamala, Univ. College, Nairobi, Kenya, E. Africa. "Research on mosquitoes and biting flies in Kenya." May 12, 1971. Dr. Robert W. Gwadz, Dept. of Trop. & Public Health, Harvard School of Public Health, Boston, MA. "The effects of juvenile hormone on the development of Brugia pahangi in Aedes aegypti"; "Genetics of vectorial capacity for malaria and filariasis." July 6, 1971; July 17, 1972. Dr. Jean David, Laboratoire d'Entomologie Experimentale et de Genetique, Lyon, France. "Oviposition behavior in geographic races of Drosophila melanogaster." September 1, 1971. Dr. Roger Williams, School of Public Health, Columbia Univ., New York NY. "A quest for Africa's smallest game; arboviruses." October 5, 1971. Dr. G. G. Foster, CSIRO, Canberra, Australia. "Synthesis and use of compound chromosomes for genetic control of mosquitoes and other insect pests"; "Genetics of the Notch locus in Drosophila melanogaster." October 17–18, 1971. Dr. Manabu Sasa, Institute of Med. Science, Univ. of Tokyo & Fogarty International Centre, Tokyo, Japan. "Ecological and epidemiological aspects of various types of schistosomiasis and filariasis endemic
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in Asia"; "How to breed mites, mosquitoes and snails in the laboratory." October 18–20, 1971. Dr. William Horsfall, Dept. of Entomology, Univ. of Illinois, Urbana, IL. "Who's on first? (sex reversal in mosquitoes by thermal stress)." April 26–28, 1972. Dr. Gene R. DeFoliart, Dept. of Entomology, Univ. of Wisconsin, Madison, WI. "Ecology and transmission of California encephalitis viruses." May 2, 1972. Dr. Keith Hutcheson, Dept. of Biology, Univ. of Windsor, Windsor, Ontario, Canada. "Ultrastructural study of spermiogenesis in the normal and male producing strains of Aedes aegypti." June 26, 1972. Dr. Carl Hacker, Population Studies Module School of Public Health, Univ. of Texas, Houston, TX. "Predictive models as tools in studies of population dynamics of Culex." October 9, 1972. Dr. Keith W. Hartberg, Dept. of Biology, Southern College, Statesboro, GA. "Reproductive isolation and phylogeny of Stegomyia mosquitoes." March 15–16, 1973. Dr. E. D. Garber, Dept. of Biology, Univ. of Chicago, Chicago, IL. "Enzymes as taxonomic and genetic tools." June 11, 1973. Dr. Paul McDonald, ICIPEMBU, Mombasa, Kenya, E. Africa. "Production of translocations for genetic control of Aedes aegypti in East Africa." August 21–28, 1973. Dr. Reuben Olembo, Dept. of Botany, Univ. of Nairobi, Nairobi, Kenya. "Opportunities for genetical research in Africa." September 5–14, 1973. Dr. John D. O'Connor, Dept. of Biology, Univ. of California, Los Angeles. "Qualitative and quantitative analysis of ecdysones in biological samples." November 6, 1973. Dr. Philip Lounibos, Harvard Univ., Cambridge, MA. "Seasonal development and photoperiodism in Wheomyia smithii, the Pitcher Plant Mosquito." June 24–28, 1974. Dr. V. P. Sharma, Director, Malaria Research Centre, New Delhi, India. "Malaria in India: Epidemiology and Prospects of its control." September 22–25, 1984. Dr. Hewson Swift, Professor of Biology, Univ. of Chicago, Chicago, IL. "Cellular Ecology: Genome interaction between nucleus and cytoplasm"; "Cellular Selection: Cell competition in development"; "Cellular Evolution: Origin of Genome diversity." March 26–28, 1985. Dr. Lynn Margulis, Boston University. "Early Life on Earth"; "Sym
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biosis and Evolution"; "Origins of Cell Motility." October 23–25, 1988. Dr. Anthony J. Nappi, Loyola University. "Biochemical and Cellular Aspects of Insect Immunity." November 7, 1988. Dr. Bruce Christensen, University of Wisconsin. "Mechanisms of Incompatibility in Mosquitofilarial Worm Associations." January 20, 1989. Dr. William Z. Coker, University of Accra, Ghana. "Yellow Fever epidemic, Oyo State, Nigeria, 1987." November 22, 1989. Dr. Richard Williams, Universitat Karlsruhe, West Germany. "New methods to identify proteins involved in hostparasite communication." January 18, 1990. Dr. R. C. Kim, Dept. of Entomology, Pennsylvania State University. "Evolution of parasitehost associations: Parasitic arthropods versus vertebrate hosts." March 27, 1990. Dr. Bernd Heinrich, University of Vermont. "Thermoregulation in Bufferflies." September 11, 1990. Dr. Margaret Kidwell, University of Arizona. "Population genetics of the P transposable elements in Drosophila: A model for developing gene transfer systems in Insects." April 2, 1991. Dr. Theodore F. Tsai, CDC. Fort Collins, CO. "The epidemiology of domestic arboviral infections." April 16, 1991. Dr. David Le Sueur, Research Institute for Diseases in a Tropical Environment, Durban, South Africa. "The Anopheles gambiae complex and malaria: The scourge of Africa." September 3, 1991. Dr. Brian Sharp, Research Institute for Diseases in a Tropical Environment, Durban, South Africa. "The Anopheles gambiae complex and malaria: The scourge of Africa." September 3, 1991. Dr. Scott O'Neill, University of Illinois. "The mysterious tale of the giant killer sperm: Cytoplasmic incompatibility in insects." October 29, 1991. Dr. Anthony S. Fauci, "Immunopathogenic mechanisms of human immunodeficiency virus infections; AIDS: Consideration for the 1990's." November 11, 1991. Dr. Jeffrey Feder, University of Chicago. "Host Race Formation and Sympatic Speciation in Rhagoletis Fruit Flies." January 28, 1992. Dr. YunBo Shi, Carnegie Institute of Washington. "Gene Regulation during Xenopus laevis Metamorphosis." January 30, 1992. Dr. Elizabeth Dolliver Eldon, Baylor College of Medicine. "Genetic Molecular and developmental analysis of the Drosophila 18Wheeler Gene." February 4, 1992.
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Dr. Ronald Atlas, Univ. of Louisville. "Polymerase Chain Reaction for Environmental Monitoring." April 22, 1992. Dr. Gerald McLaughlin, Purdue University. "Molecular Epidemiology of Infectious Diseases"; "The discovery of New Bioactive Natural Products using benchtop bioassays." October 6–7, 1992. Dr. Stephen J. Gould, Harvard University: "The proper integration of development and evolution." December 9, 1992. Dr. Margaret Humphries, Harvard School of Public Health. "Yellow Fever and the South, 1878–1905." April 2, 1992. Dr. Ingrid Müller, University of Lausanne, Epalinges, Switzerland. "Role of T Lymphocytes in Experimental Murine Leischmaniasis." February 17, 1993. Dr. Bruce W. Stillman, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York. "Inheritance of DNA: 40 years after the double helix"; "Replication of DNA: viruses as model systems"; "Replication of chromosomes: Mechanisms and control." February 23–25, 1993. Dr. Jeremy Sternberg, Department of Zoology, University of Aberdeen, Aberdeen Scotland. "African Trypanosomes: We're Quite Happy to be Extracellular." September 27, 1994. Dr. Ann Elizabeth Eakin, Luce Faculty Candidate, Department of Biochemistry, University of Puerto Rico, School of Medicine, San Juan, PR. "Molecular Approaches to the Design of Drugs Targeted to Enzymes of Trypanosoma Cruzi, the Etiologic Agent of Chagas' Disease." February 13, 1995. Dr. Elizabeth L. Wilder, Harvard Medical School, Department of Genetics. "Analysis of Wingless Signaling and Function in the Patterning of the Drosophila Adult." February 27, 1995. Dr. Howard K. Schachman, University of California, Berkeley, Department of Molecular and Cell Biology. "Maintaining Integrity in Science: Roles of Government and Universities." March 1, 1995. Dr. Michael J. Wade, University of Chicago, Department of Ecology and Evolution. "The CoEvolution of all Endosymbiont (Wolbachia) and Its Flour Beetle Host (Tribolium Confusum)." March 7, 1995. Dr. John Donelson, University of Iowa. "New Mechanisms of Immune Evasion by African Trypanosomes." April 3, 1995. Dr. Timothy G. Geary, Senior Scientist, Animal Health Discovery Research, Upjohn Laboratories, Kalamazoo, Michigan. "The Physiology of Nematode FMR Famidelike Neuropeptides." April 4, 1995. Dr. Kathryn V. Anderson, Professor of Genetics, University of Califor
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nia, Berkeley. "Genetic Analysis of Embryonic Pattern Formation." April 10, 1995. Dr. Kathryn V. Anderson, Professor of Genetics, University of California, Berkeley. "Generating the DorsalVentral Pattern of the Drosophila Embryo: Extracellular Proteases and Localized Ligand Production." April 11, 1995. Dr. Kathryn V. Anderson, Professor of Genetics, University of California, Berkeley. "Generating the DorsalVentral Pattern of the Drosophila Embryo: A Conserved Cytoplasmatic Signaling Pathway." April 12, 1995. Dr. Richard Merritt, Department of Entomology, Michigan State University, East Lansing, Michigan. "Filter Feeding Ecology of Aquatic Insects." Also presented an informal lecture, "Effect of BTI on Black Flies and Nontargeted Organisms in the UP of Michigan." April 19, 1995. Dr. Anne E. Hershey, Department of Biology, University of Minnesota, Duluth, Minnesota. "Organic Matter Processing by Black Flies in Stream Ecosystems." September 26, 1995. Dr. Evelyn Fox Keller, Program of Science, Technology and Society, Massachusetts Institute of Technology: "Drosophila Embryology in Historical Context: Donald Poulson and Christiane NüssleinVolhard." October 3, 1995. Dr. Louis H. Miller, Chief, Laboratory of Malaria Research, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland. "In the Game Between Parasites and the Host Immune System, Why do Some Parasites Play for High Stakes?" October 4, 1995. Dr. Louis H. Miller, Chief, Laboratory of Malaria Research, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland. "A Superfamily of Malaria Receptor Proteins Involved in Pathogenesis." October 5, 1995. Dr. William Black, Department of Microbiology, Colorado State University. "New Molecular Methods for Genetic Mapping in Mosquitoes." October 6, 1995. Dr. Robert Etges, Freiburg, Germany "Leishmanolysin: A Novel GPIanchored Metzincin Proteinase of Leishmania promastigotes. Biochemical and Structural Characterization." October 31, 1995. Dr. Mitch Dushay, Center for Developmental Biology, University of Texas, Southwest Medical Center, Dallas, Texas. "The Role of Tumor Suppressor Genes in the Development of the Vertebrate Peripheral Nervous System." December 12, 1995. Dr. Takafumi Tsuboi, Department of Parasitology, Ehime University School of Medicine, Shigenobucho, Ehime JAPAN. "New Devel
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opments for a Malaria Transmission Blocking Vaccine." February 14, 1996. Dr. Robert G. McLean, Centers for Disease Control and Prevention, Fort Collins, Colorado, "Out of Africa: Hunting for Ebola Virus." April 12, 1996. Dr. Martin Heisenberg, Professor of Genetics, TheodorBoveriInstitut Für Biowissenschaften Würzburg University "Genes, Brains and the Biological Roots of the Mind." April 16, 1996. Dr. Martin Heisenberg, Professor of Genetics, TheodorBoveriInstitut Für Biowissenschaften Würzburg University. "Vision in Drosophila: Genetics of Microbehavior—12 Years later." April 17, 1996; "Drosophila, the Model Brain: What the Mushroom Bodies and Central Complex Do for the Fly." April 18, 1996. Dr. Frank H. Collins, Centers for Disease Control and Prevention, Chamblee, Georgia. "Anopheles Gambiae: Genetics, Genomics, and Malaria Control." July 9, 1996. Dr. Nora J. Besansky, Centers for Disease Control and Prevention, Chamblee, Georgia. "Evolutionary Relationships Between Anopheles gambiae Sibling Species: An Incestuous Affair?" July 16, 1996. Dr. Anthony A. James, Department of Molecular Biology and Biochemistry, University of California, Irvine. "Molecular Genetic Manipulation of Vector Mosquitoes." July 23, 1996. Dr. Eric Wieschaus, Nobel Laureate in Medicine (1995), Professor, Princeton University, "Embryonic Transcription and the Control of Development Pathways." September 12, 1996; "Of Flies and Men: Genes and Embryonic Development of the Fruit Fly." September 14, 1996. Dr. Richard Komuniecki, Department of Biology, University of Toledo. "The Role of the Pyruvate Dehydrogenase Complex in AerobisAnaerobic Transitions During Nematode Development." October 8, 1996. Dr. Douglas Fishkind, Department of Biological Sciences, University of Notre Dame. "ActinMyosin Dynamics in Cell Division." November 5, 1996. Dr. Scott O'Neill, Department of Entomology and Public Health, Yale University School of Medicine. "Wolbachia/Insect Interactions: Making a Living From Exploitation Rather than Bribery." January 16, 1997. Dr. Steve Pratt, Argonne National Laboratories, Argonne, Illinois. "Applications of XRays in Molecular Environmental Science." January 21, 1997.
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Dr. Vern B. Carruthers, Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri. "There's a Burglar Amongst Us: How the Ubiquitous Parasite Toxoplasma gondii Actively Enters Host Cells." February 11, 1997. Dr. Gard Otis, Department of Entomology, University of Guelph, Ontario, Canada. "Biodiversity in Asian Honeybees." February 18, 1997. Dr. David Severson, Department of Animal Health and Biomedical Sciences, University of Wisconsin, Madison. "Molecular Markers and Genome Analysis in Aedes aegypti." February 26, 1997. Dr. Peter M. Vitousek, Department of Biological Sciences, Stanford University. "Global Environmental Change: A Reality, Not a Controversy." April 14, 1997. Dr. Leonard E. Munstermann, Department of Epidemiology & Public Health, Yale University School of Medicine. "Mosquitoes, Mosquitoes, Mosquitoes: Mapping Applications in Insect Vectors." April 29, 1997. Dr. G. Arturo SanchezAzofeifa, Research Center on Sustainable Development, University of Costa Rica. "Are Protected Areas Conserving Biodiversity in the Tropics? New Perspectives From Remote Sensing and Geographic Information Systems." October 15, 1997. Dr. Kevin S. McCann, Division of Environmental Studies, University of California, Davis. "On the Dynamical Influences of Life History Strategies in Aquatic Communities." January 20, 1998. Dr. Jeffrey R. Powell, Dept. of Ecology and Evolutionary Biology, Yale University "Molecular Evolution and Phylogenetics of Drosophila." April 28, 1998. Dr. Greg Dwyer, Dept. of Ecology and Evolution, University of Chicago. "Theory and Experiment on the Ecology of Disease." February 5, 1998. *
This is not a complete list. The dates and names of some of the earlier visitors were impossible to obtain.
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