‘Boom’ call variation in Cercopithecus mona as a possible proxy for genetic relatedness Thesis Presented in partial fulfillment of the requirement for the degree master of arts in the Graduate School of The Ohio State University. By Kaitlyn Rose Werling, B.A. Graduate Program in Anthropology The Ohio State University 2015 Thesis Committee: W. Scott McGraw, Advisor Dawn Kitchen Debbie Guatelli-Steinberg Mary E. Glenn Copyright by Kaitlyn Rose Werling 2015 Abstract The mona monkey population on the island of Grenada was introduced over 350 years ago during the trans-Atlantic slave trade. The source population in Africa has yet been determined. Variation in vocal ability is traditionally argued to be primarily genetic with minimal plasticity due to learned variations. If true, then similarities in vocal behaviors among populations can be used as a proxy for genetic material to estimate relatedness. I tested whether acoustic similarities in loud calls could help ascertain from which mainland African population the Grenadian Cercopithecus mona originated. Cercopithecus mona is one of nine guenon species known to emit loud call vocalizations known as ‘boom’ calls. In C. mona, these calls occur at low frequencies (Hz). They are also audibly short, tonal, double-phased calls. Booms are typically produced as responses to perceived threats, for territorial defense, or to initiate group movement. The lead males of mixed sex groups always give the boom calls. Boom call duration and bandwidth were analyzed in 18 boom calls from two Cercopithecus mona populations in Nigeria. They were then compared with previously analyzed calls recorded in Cameroon (n=19), Benin (n=17), and on Grenada (n=16). Boom calls from Nigeria had an average bandwidth of 358 Hz (range ~ 118-774 Hz), and duration of 121 ms. They are most similar to calls recorded in Cameroon. Calls from Benin and Grenada are distinctly similar, further supporting the hypothesis that mona monkeys on Grenada are descendants of a population from Benin. ii To my parents, Mark and Sara Werling for their unending support in all of my endeavors, iii Acknowledgements I wish to thank the members of my committee for their feedback and support with this project. I would also like to thank the Governments of Grenada, Nigeria and Cameroon, and the Office National du Bois and Mission Forestier Allemand du GTZ in Benin. I especially thank Thomas Struhsaker for granting access to boom calls from Cameroon, Reiko Matsuda Goodwin for access to the boom calls from Benin, Mary Glenn and Keith Bensen for access to calls from Grenada, and Carrie Vath and Fatsuma Olaleru for granting me access to recordings made in Nigeria. I also thank Matthew Medler and the Cornell Lab of Ornithology, Yoon Kim, Mark Hubbe, Rebekah Dickens, Amy Remer, Kena Worsham, May Patiño, as well as the many assistants who helped in the field. I am grateful to Ken Ayoob, the HSU College of Arts, Humanities, and Social Sciences, and the HSU Department of Anthropology for support. The Grenada research was funded by Humboldt State University, Windward Island Research and Education Foundation, National Geographic Society, Yerkes National Primate Research Center, and The Rockefeller University. Research in Benin was funded by NSF (SBR-9528348), Wenner-Gren Foundation for Anthropological Research (Gr. 9562), Leopold Schepp Foundation, and Primate Conservation, Inc. iv Vita May 2010.................................................Rio Americano High School May 2013……………………………….B.A. Anthropology, Humboldt State University August 2013 to present………………...Graduate Student, Department of Anthropology, The Ohio State University Publications Werling K, Ramsier M, Matsuda Goodwin R, Patiño M, Bensen KJ, and Glenn ME. (2015) Loud call variation in Cercopithecus mona: a proxy for genetic relatedness? American Journal of Physical Anthropology 156 (Suppl. 60): 322. Werling K, Worsham K, Patiño M, Ramsier M, and Glenn ME. (2013). Copulations calls of Cercopithecus mona in the wild. American Journal of Physical Anthropology 150(Suppl. 56):288. Patiño M, Werling K, Worsham K, Matsuda Goodwin R, Glenn ME, and Ramsier M. (2012) A new analysis of boom call variation among populations of mona monkeys (Cercopithecus mona) in Grenada, Benin, and Cameroon. American Journal of Primatology 74 (Suppl. 1): 38. Fields of Study Major Field: Anthropology Specialization: Primatology v Table of Contents Abstract…………………………………………………………………………………....ii Dedication………………………………………………………………………………...iii Acknowledgments………………………………………………………………………..iv Vita………………………………………………………………………………………...v List of Tables…………………………………………………………………………….vii List of Figures…………………………………………………………………………...viii Introduction………………………………………………………………………………..1 Methods and Materials…………………………………………………………………….6 Results……………………………………………………………………………………11 Discussion and Conclusion..……………………………………………………………..16 References………………………………………………………………………………..19 Appendix A: Map of field location in Benin…………...………………………………..26 Appendix B: Map of field locations in Nigeria………………………………………….28 Appendix C: Map of field location in Cameroon………………………………………..30 Appendix D: Map of field location in Grenada………………………………………….32 vi List of Tables Table 1. Sample size and source location of genetic material…………………………….3 Table 2. Sample size and location of boom vocalizations……………………………….11 Table 3. Tests of normality, call duration and bandwidth…………….…………………13 Table 4. ANOVA, boom call duration…….……………………………………………..14 Table 5. Post-hoc LSD multiple comparison, boom call duration……………………….15 Table 6. Independent-Samples Kruskal-Wallis test, unit bandwidth…………………….15 Table 7. Post-hoc pairwise comparison, boom call bandwidth on Grenada (GD) Benin (BJ), Cameroon (CM), and Nigeria (NG)……………..………………………….16 vii List of Figures Figure 1. Geographic map of endemic and introduced populations of C. mona………….2 Figure 2. Spectrogram of Cercopithecus mona boom call on Grenada…………………...7 Figure 3. Analysis chart of boom call unit………………..………………...……………12 viii INTRODUCTION Cercopithecus mona (mona monkey) is a species of arboreal monkey endemic to West Africa. The original range of Cercopithecus mona spanned forested areas from eastern Ghana to central Cameroon (Booth 1955, 1956; Struhsaker 1970; Oates 1988). Mona monkeys are generalists in diet and habitat, thriving in most forest types including seasonally dry forests and mangrove swamps (Glenn 1996; Matsuda Goodwin 2007). Approximately 300 years ago, during the transatlantic slave trade, mona monkeys were introduced to the island nations of São Tomé and Príncipe in the Gulf of Guinea and Grenada in the West Indies (Glenn et al., 1999 unpub. report). Sailors often brought back exotic animals from their travels to sell to colonists or to keep as pets (Eaden 1931; Sade and Hildreth 1965; McGuire 1974; Denham 1982, 1987). São Tomé and Príncipe were trans-shipping points during the Atlantic slave trade. It is hypothesized that Cercopithecus mona was first introduced to São Tome and from there, a single, possibly pregnant female Cercopithecus mona was taken to Grenada where she escaped and founded the present island population (Horsburgh et al., 2002). 1 Figure 1. Geographic map of endemic and introduced populations of C. mona The ability of Cercopithecus mona to be a generalist has allowed them to successfully survive these island introductions, despite experiencing extreme genetic bottlenecks (Glenn and Bensen 2013). The most recent extreme bottleneck occurred in 2004. Hurricane Ivan destroyed Grenada, leveling nearly all of the Grand Etang National Park and Forest Reserve along with most of the forested and urban sections of the island. A census to determine how many monkeys remain on the island has not been conducted post-Ivan. In order to understand the effect of the genetic bottlenecks on variation in the Grenada mona monkey genome, Horsburgh et al., (2002) examined the mitochondrial DNA control region. This was used as a measure of intraspecific variability. Tissue samples were collected during field seasons on Grenada between 1992 and 1994, and on São Tomé and Príncipe in 1998 (Glenn, 1996; Glenn et al., unpub. report). No fresh 2 tissue samples were available for mainland populations. Skins collected by naturalists during the 1900s were used (samples made available by the Powell-Cotton Museum, Kent, and the Museum of Natural History, London) (Table 1). Table 1. Sample size and source location of genetic material. Country n Grenada 21 São Tomé 12 Príncipe 4 Cameroon 9 Benin 1 Total 47 Mitochondrial DNA was extracted from samples, amplified using the Polymerase Chain Reaction (PCR), and then sequenced (Horsburgh et al., 2002). To estimate genetic variation of the three island populations and mainland Africa populations, Horsburgh and associates calculated a diversity index using Nei’s number. Only shorter chains of DNA could be extracted from the African Cercopithecus mona samples. Therefore, longer and shorter sequences of DNA were indexed separately (Horsburgh et al., 2002). Due to degradation of the mainland African genetic samples, the colonizing source of the São Tomé population cannot be determined. The results of Horsburgh et al.,’s analysis (2002) suggest that mona monkeys on Grenada were brought there from São Tomé. We do not know, however, from which mainland Africa population the Grenada lineage stemmed before their arrival to São Tomé. As viable genetic samples from mainland Africa are unavailable, a proxy to 3 determine relatedness is needed. Vocalization recordings, however,