AN ABSTRACT OF THE DISSERTATION OF Brian S. Dugovich for the degree of Doctor of Philosophy in Zoology presented on May 28, 2019. Title: Investigating Variation in Immunity and Infection Risk in Wild Ungulates. Abstract approved: ______________________________________________________ Anna E. Jolles Brian P. Dolan In the midst of the sixth mass extinction, understanding wildlife disease spillover is critical to maintaining protected wildlife areas. Studying ecoimmunology and wildlife disease ecology helps to understand immune and disease traits in an ecological context, which is invaluable in preventing pathogen spillover between livestock and wildlife. To investigate this interface, tests of innate immunity were validated in various ungulates and desert bighorn sheep, and then infectious spillover diseases in desert bighorn sheep and African buffalo were examined. In ungulates at Wildlife Safari in Oregon, innate immunity varied between species and strong positive correlations were found between innate immunity measures. In general, ecoimmunology study results are robust to the choice of innate immune defense assay. In desert bighorn sheep in California, an ELISA was developed that could measure natural antibody levels from blood samples, and differences in natural antibody levels and a bacterial killing assay were found between Peninsular and Mojave desert bighorn sheep. Spillover of Mycoplasma ovipneumoniae from domestic animals to desert bighorn sheep can lead to region-wide extinction. Disease prevalence, connectivity, genetic data, and immunophenotype were utilized to show that better connected populations resisted pathogen invasion. Finally, in Kruger National Park, South Africa, African buffalo calves infected with foot-and-mouth disease virus reinfected previously exposed animals to become carriers. Other risk factors for carrier status were identified: low body condition score, younger age, and being male. Identifying these attributes during risk assessment of stray buffalo could be used to help avoid foot-and-mouth disease spillover to cattle on the park border. ©Copyright by Brian S. Dugovich May 28, 2019 All Rights Reserved INVESTIGATING VARIATION IN IMMUNITY AND INFECTION RISK IN WILD UNGULATES by Brian S. Dugovich A DISSERTATION submitted to Oregon State University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Presented May 28, 2019 Commencement June 2020 Doctor of Philosophy dissertation of Brian S. Dugovich presented on May 28, 2019. APPROVED: Major Professor, representing Zoology Chair of the Department of Integrative Biology Dean of the Graduate School I understand that my dissertation will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my dissertation to any reader upon request. Brian S. Dugovich, Author ACKNOWLEDGEMENTS This work would not have been possible without the help of a diversity of organizations and incredible people. The Dolan Lab funded my first year of PhD studies and the Wildlife Safari Project through the Department of Biomedical Sciences at the Carlson College of Veterinary Medicine. Special thanks to Benji Alcantar, Kirsten Thomas, Sarah Cannizzo, and the rest of the Wildlife Safari staff for facilitating animal immobilization and sampling. I am grateful to Benji providing me a place to stay in Roseburg. Becca Sullivan, Lucie Crane, and Claire Behnke were invaluable for helping with fieldwork and immunoassays and camping by the South Umpqua. I would also like to thank Heather Broughton for helping on the futile task to validate the LPA. The desert bighorn project was made possible by funds from the Epps Lab through the Department of Fisheries and Wildlife at Oregon State University and the Jolles Lab through the Department of Biomedical Sciences. I am grateful to Heather Broughton, Rhea Hanselmann, Abby Sage, Rachel Crowhurst, Brian Henrichs, Lindsay Hunter, and Daniella Dekelaita for assisting on captures and running immunoassays and Ben Gonzalez and Lora Konde of the California Department Fish and Wildlife for mentorship and integrating our research team into the desert bighorn sheep captures. Additional thanks go to the Sweeney Granite Mountains Desert Research Center for providing housing and laboratory space and the Department of Fisheries and Wildlife for letting the Golden Falcon fly. I also greatly appreciate Tyler Creech and Brandon Nickerson providing their DBH genetic data for analysis. The Morris Animal Foundation made my fieldwork in South Africa possible by graciously funding me for two years. Much gratitude is given to SANParks for approving and overseeing the study. Sincere appreciation is given to Peter Buss, Michele Miller, Jenny Hewlett and the rest of the KNP Veterinary Wildlife Services team for tirelessly capturing buffalo every two months and making scores of sedated buffalo a tranquil affair and Louis Van Schalkwyk and Lin- Mari deKlerk-Lorist for many more mornings of buffalo capture. I would also like to thank Katherine Scott and Eva Perez-Martin for helping me with the FMDV data. My South Africa time will always be precious to me with lifetime memories made with the Jolles Lab buffalo team including Courtney Coon, Julie Rushmore, Erin Gorsich, Kath Potgieter Forssman, Hannah Tavalire, Claire Couch, Henri Combrink, Leigh Combrink, Caroline Glidden, Robert Spaan, Johannie Spaan, and Danielle Sisson. Many thanks go to the Department of Integrative Biology at Oregon State University for funding my post- fieldwork PhD studies through teaching assistantships and to Lindsay Biga and Devon Quick for teaching mentoring. Finally, I owe these amazing wildlife research opportunities, compelling challenges, fieldwork and lab work assistance, amazing discoveries, and exceptional mentorship to Anna Jolles, Brian Dolan, Clinton Epps, and Brianna Beechler. CONTRIBUTION OF AUTHORS Authors: * designate co-authors and ** designate co-principal investigators Chapters 1 – 4: Dr. Anna E. Jolles (A.E.J.) was my co-adviser, co-principal investigator on the Wildlife Safari project, co-principal investigator for the desert bighorn sheep project, and the principal investigator on the USDA-AFRI FMDV African buffalo project. Chapters 1 – 3: Dr. Brian P. Dolan (B.P.D) was my co-adviser, co-principal investigator on the Wildlife Safari project, and co-principal investigator for the desert bighorn sheep project. Chapters 2 – 3: Dr. Clinton W. Epps (C.W.E.) was co-principal investigator on the desert bighorn sheep project. Chapters 1 – 4: Dr. Brianna R. Beechler (B.R.B.) was a post-doctoral researcher and assistant professor of research who was involved in all projects. Chapter 1: B.P.D., A.E.J., B.R.B. conceived the project idea and B.P.D. and B.R.B designed the study’s methodology. Dr. Benji Alcantar’s role as a zoo veterinarian was responsible for ungulate capture at Wildlife Safari. Dr. Lucie L. Crane helped collect data and ran the TLR experiment. B.P.D. helped write the manuscript. Chapter 2: Dr. Melanie J. Peel (M.J.P.)* was co-author with me on this manuscript. B.P.D., B.R.B., A.E.S., C.W.E., and M.J.P. helped conceptualize the project. Investigation was led by Amy. L. Palmer (A.L.P.), Dr. Aleksandra E. Sikora (A.E.S.), and Dr. Ryszard A. Zielke. M.J.P., A.E.S., and B.P.D. designed the study’s methodology. The project was administrated by C.W.E. and B.P.D. and supervised by A.E.S., A.E.J., and B.P.D. B.R.B. and B.P.D. helped write the original draft with review and editing by A.E.S., A.E.J., and C.W.E. Chapter 3: A.E.J.**, C.W.E.**, B.R.B., and B.P.D. conceived the project’s idea and helped with writing. B.R.B., B.P.D., C.W.E., and Rachel S. Crowhurst designed methodology. Dr. Benjamin Gonzalez was responsible for desert bighorn sheep capture. Chapter 4: A.E.J., B.R.B., Bryan Charleston (B.C.), and Francois F. Maree (F.F.M.) conceived the project’s idea and A.E.J., B.R.B., B.C., F.M., Eva Perez-Martin, and Katherine Scott designed the methodology. B.R.B. was the project’s administrator. TABLE OF CONTENTS Page INTRODUCTION ......................................................................................................... 1 CHAPTER 1: MULTIPLE INNATE ANTIBACTERIAL IMMUNE DEFENSE ELEMENTS ARE CORRELATED IN DIVERSE UNGULATE SPECIES .... 5 Abstract .............................................................................................................. 6 Introduction ........................................................................................................ 6 Materials and Methods ....................................................................................... 9 Results ................................................................................................................ 16 Discussion .......................................................................................................... 17 Figures and Tables ............................................................................................. 19 CHAPTER 2: DETECTION OF BACTERIAL-REACTIVE NATURAL IGM ANTIBODIES IN DESERT BIGHORN SHEEP (OVIS CANADENSIS NELSONI) POPULATIONS .............................................................................. 23 Abstract .............................................................................................................. 24 Introduction ........................................................................................................ 24 Materials and Methods ......................................................................................