<I>Trichomonas Gallinae</I> in Wild Birds
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University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 8-2018 Investigation of the potential role of bird baths in the transmission of Trichomonas gallinae in wild birds Kathryn Erin Purple University of Tennessee Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Recommended Citation Purple, Kathryn Erin, "Investigation of the potential role of bird baths in the transmission of Trichomonas gallinae in wild birds. " PhD diss., University of Tennessee, 2018. https://trace.tennessee.edu/utk_graddiss/5085 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Kathryn Erin Purple entitled "Investigation of the potential role of bird baths in the transmission of Trichomonas gallinae in wild birds." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the equirr ements for the degree of Doctor of Philosophy, with a major in Comparative and Experimental Medicine. Richard W. Gerhold Jr., Major Professor We have read this dissertation and recommend its acceptance: Debra L. Miller, Marcy J. Souza, Mary Katherine Tolbert Accepted for the Council: Dixie L. Thompson Vice Provost and Dean of the Graduate School (Original signatures are on file with official studentecor r ds.) Investigation of the potential role of bird baths in the transmission Trichomonas gallinae in wild birds A Dissertation Presented for the Doctor of Philosophy Degree The University of Tennessee, Knoxville Kathryn Erin Purple August 2018 ACKNOWLEDGEMENTS I would like to acknowledge and emphasize the unwavering support and guidance I have received from my advisor, Dr. Richard Gerhold, without whom none of this would be possible. I have grown as researcher with the direction and challenges from my entire committee: Drs. Marcy Souza, Deb Miller and Katie Tolbert. For her indispensable input and moral support, I want to thank my dear friend Dr. Sawsan Ammar. My personal support network has gone above and beyond to elevate my spirits and keep me focused. I am eternally grateful to my family, especially my mother, Debbie Purple, who has supported me through all of my toughest personal and academic challenges. To my husband, Matt Murphy, there is no one I would rather have by my side. Thank you for always knowing how to make me laugh and always knowing just what to say to lift my spirits. I also want to thank my son, Jonathan “Jack” George Murphy, whom we welcomed in the middle of my PhD for reminding me that there are more important things in life than doctorate degrees. And last, but not least, thank you to my canine babies, Hallie and Parker, who have loved me unconditionally through thick and thin. ii ABSTRACT Understanding the role of bird baths and shared water sources in the ecology and epidemiology of the deadly protozoan parasite Trichomonas gallinae is crucial to identify mitigation strategies for population-limiting epidemics. We evaluated multiple factors that influence the transmission of this pathogen including characterizing persistence in simulated bird baths, evaluating potential effects persistence may have on virulence, and investigating the molecular epidemiology of a concurrent outbreak. Trichomonads were thought to be extremely labile in the environment since their discovery over 200 years ago to 5 years ago when maximum recorded persistence in water was 20 min. We show that trichomonads persist up to 48 hr in simulated bird baths at 37°C with organic material (OM). We also measured persistence in a variety of conditions including, with and without organic material (4 and 16 hr, respectively), with artificially decreased dissolved oxygen (30 hr), and with exposure to UV light (4hr). Moreover, when cytopathic effect (CPE) of post-persistence trichomonads on cultured avian cells was compared to non-persistence control isolates, virulence changed significantly by treatment or persistence time. Post-UV treated persistence broad-winged hawk isolate destroyed significantly more of the cell monolayer than those from the OM treatment illustrating that persistence type can alter virulence. Currently, live animal or cell culture infection trials are the only tool to define virulence of an isolate; however, the more we understand the phylogenetics and epidemiology of Trichomonas spp, the better we can contribute phenotypic differences to genetic data. Using molecular techniques, we showed that circulating genotypes in subclinical hunter-killed doves differ from birds with lesions consistent with trichomonosis. Therefore, hunter-killed doves may not be the source of deadly strains circulating in other sympatric species. Combining laboratory work with the parasite, assessment of virulence on cell culture, and molecular techniques we have shown that the role of bird baths and water sources in transmission cannot be underestimated when addressing outbreaks and that molecular information will continue to improve our effects to mitigate the disease in wild birds. iii TABLE OF CONTENTS CHAPTER I INTRODUCTION ............................................................................................................. 1 ABSTRACT ............................................................................................................................................ 2 LITERATURE REVIEW .............................................................................................................................. 3 CHAPTER II TRICHOMONAS GALLINAE PERSISTENCE IN FOUR WATER TREATMENTS .......... 12 ABSTRACT .......................................................................................................................................... 13 SHORT COMMUNICATION ...................................................................................................................... 14 Acknowledgments ......................................................................................................................... 19 CHAPTER III PERSISTENCE OF TWO ISOLATES OF TRICHOMONAS GALLINAE IN SIMULATED BIRD BATHS WITH AND WITHOUT ORGANIC MATERIAL .............................................................. 20 ABSTRACT .......................................................................................................................................... 21 INTRODUCTION .................................................................................................................................... 22 MATERIALS AND METHODS ................................................................................................................... 23 Statistical analysis ......................................................................................................................... 24 RESULTS ............................................................................................................................................ 24 DISCUSSION ........................................................................................................................................ 27 Acknowledgments ......................................................................................................................... 27 CHAPTER IV ARTIFICIALLY DECREASED DISSOLVED OXYGEN INCREASES THE PERSISTENCE OF TRICHOMONAS GALLINAE IN WATER ............................................................ 29 ABSTRACT .......................................................................................................................................... 30 INTRODUCTION .................................................................................................................................... 31 RESULTS ............................................................................................................................................ 32 DISCUSSION ........................................................................................................................................ 35 MATERIALS AND METHODS ................................................................................................................... 38 Measuring DO levels with added OM ............................................................................................. 38 Measuring DO with added Oxyrase® ............................................................................................. 39 Measuring persistence of trichomonads in water with decreased DO ............................................. 39 Acknowledgements ....................................................................................................................... 40 CHAPTER V EVALUATION OF THE CYTOPATHIC EFFECT OF TRICHOMONAS GALLINAE BEFORE AND AFTER PERSISTENCE IN SIMULATED BIRD BATHS USING AVIAN FIBROBLASTS IN CELL CULTURE ........................................................................................................................... 41 ABSTRACT .......................................................................................................................................... 42 INTRODUCTION ...................................................................................................................................