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Distribution and Coinfection of Microparasites and Macroparasites in Juvenile Salmonids in Three Upper Willamette River Tributaries

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Distribution and Coinfection of Microparasites and Macroparasites in Juvenile Salmonids in Three Upper Willamette River Tributaries AN ABSTRACT OF THE THESIS OF Sean Robert Roon for the degree of Master of Science in Microbiology presented on December 9, 2014. Title: Distribution and Coinfection of Microparasites and Macroparasites in Juvenile Salmonids in Three Upper Willamette River Tributaries. Abstract approved: ______________________________________________________ Jerri L. Bartholomew Wild fish populations are typically infected with a variety of micro- and macroparasites that may affect fitness and survival, however, there is little published information on parasite distribution in wild juvenile salmonids in three upper tributaries of the Willamette River, OR. The objectives of this survey were to document (1) the distribution of select microparasites in wild salmonids and (2) the prevalence, geographical distribution, and community composition of metazoan parasites infecting these fish. From 2011-2013, I surveyed 279 Chinook salmon Oncorhynchus tshawytscha and 149 rainbow trout O. mykiss for one viral (IHNV) and four bacterial (Aeromonas salmonicida, Flavobacterium columnare, Flavobacterium psychrophilum, and Renibacterium salmoninarum) microparasites known to cause mortality of fish in Willamette River hatcheries. The only microparasite detected was Renibacterium salmoninarum, causative agent of bacterial kidney disease, which was detected at all three sites. I identified 23 metazoan parasite taxa in these fish. Nonmetric multidimensional scaling of metazoan parasite communities reflected a nested structure with trematode metacercariae being the basal parasite taxa at all three sites. The freshwater trematode Nanophyetus salmincola was the most common macroparasite observed at three sites. Metacercariae of N. salmincola have been shown to impair immune function and disease resistance in saltwater. To investigate if N. salmincola affects disease susceptibility in freshwater, I conducted a series of disease challenges to evaluate whether encysted N. salmincola metacercariae increase susceptibility of juvenile Chinook salmon to Flavobacterium columnare and Aeromonas salmonicida infection. These bacteria cause high mortality in juvenile hatchery salmonid populations in the Willamette River, and are a potential threat to wild juvenile salmonids. Juvenile Chinook salmon were first infected with N. salmincola through cohabitation with infected freshwater snails, Juga spp., then challenged through static immersion with either F. columnare or A. salmonicida. Cumulative percent mortality from F. columnare was higher in N. salmincola­ parasitized compared to non-parasitized juvenile Chinook salmon. In contrast, cumulative percent mortality from A. salmonicida did not differ between N. salmincola-parasitized and non-parasitized juvenile Chinook salmon. No mortalities were observed in the N. salmincola-only and control groups from either challenge. These results show that a high infection intensity (>200 metacercariae per posterior kidney) of encysted N. salmincola metacercariae does not cause mortality alone, but can increase susceptibility to certain bacterial infections in juvenile Chinook salmon. ©Copyright by Sean Robert Roon December 9, 2014 All Rights Reserved Distribution and Coinfection of Microparasites and Macroparasites in Juvenile Salmonids in Three Upper Willamette River Tributaries. by Sean Robert Roon A THESIS submitted to Oregon State University in partial fulfillment of the requirements for the degree of Master of Science Presented December 9, 2014 Commencement June 2015 Master of Science thesis of Sean Robert Roon presented on December 9, 2014 APPROVED: Major Professor, representing Microbiology Chair of the Department of Microbiology Dean of the Graduate School I understand that my thesis will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my thesis to any reader upon request. Sean Robert Roon, Author ACKNOWLEDGEMENTS I would like to express my sincere appreciation to my thesis committee: Dr. Jerri L. Bartholomew, Dr. Kym Jacobson, Dr. Michael L. Kent, and Dr. Martin Fisk for their feedback throughout my time at OSU. I specifically want to thank my major advisor, Dr. Jerri L. Bartholomew, for providing the opportunity to pursue an advanced degree and for her continued advice, support, and encouragement throughout this project. I am grateful to Dr. Kym Jacobson for her assistance with experimental design and snail care tips. I would like to thank Dr. Michael Kent for being willing to discuss parasites at any moment and providing insight and new directions to my project. I wish to thank all the groups that have supported me financially: The Department of Microbiology John L. Fryer Graduate Fellowship, the U.S. Army Corps of Engineers, and the Oregon Department of Fish and Wildlife Restoration and Enhancement Board. I am indebted to Luke Whitman, Brian Cannon, Brian Bangs, Paul Olmstead, Jeremy Romer, Fred Monzyk, Kurt Schroeder, and seasonal staff at the ODFW Corvallis Research Lab along with Matt Stinson and Ken Lujan at the U.S. Department of Fish and Wildlife for their assistance in fish collection, transport, and care. I would like to thank all the members of the Bartholomew lab, past and present, for their assistance, advice, and levity during the not so successful times. Thanks to Dr. Julie Alexander for teaching me the ways of statistics and providing sage advice throughout my time here. I want to thank everyone at ODFW Fish Health Services for their assistance and teaching me the fish health ropes. Specifically, I want to thank Craig Banner for his continued mentorship in the ways of fish parasites. I am grateful for Amy Long and Steve Whitlock for their critical reviews and professional advice. Thanks to John Rogers for his help with fish necropsy and general lab work. I would like to give a big thanks to all my friends for providing a bunch of laughs and a break from being a scientist. Lastly, I would like to thank my family, specifically my parents, for always supporting and encouraging me throughout my pursuits. CONTRIBUTION OF AUTHORS Dr. Jerri L. Bartholomew provided advice on experimental design, data interpretation, and manuscript editing throughout this thesis. Dr. Julie Alexander provided assistance with experimental design, data interpretation, and statistical analysis for all the research in this thesis. For the first manuscript of this thesis, Michelle Jakaitis, John Rogers, and Craig Banner provided field sampling, fish necropsy, and diagnostic assistance along with data collection. For the second manuscript of this thesis, Dr. Kym Jacobson provided assistance with experimental design and data interpretation. TABLE OF CONTENTS Page Chapter 1: Introduction .................................................................................................. 1 Disease ecology .................................................................................................. 1 Salmonids in the U.S. Pacific Northwest ............................................................ 4 Microparasites of Willamette River salmonids ...................................... 8 Parasite coinfection ........................................................................................... 15 References ......................................................................................................... 17 Chapter 2: Survey of microparasites and macroparasites in wild juvenile Chinook salmon and rainbow trout in three upper Willamette River tributaries ........................ 25 Abstract ............................................................................................................. 26 Introduction ....................................................................................................... 27 Methods ............................................................................................................. 29 Results ............................................................................................................... 35 Discussion ......................................................................................................... 39 Acknowledgements ........................................................................................... 46 References ......................................................................................................... 47 Chapter 3: Coinfection effects of Nanophyetus salmincola Chapin and two pathogenic bacteria on juvenile Chinook salmon, Oncorhynchus tshawytscha (Walbaum) ........... 68 Abstract ............................................................................................................. 69 Introduction ....................................................................................................... 69 Methods ............................................................................................................. 73 TABLE OF CONTENTS (Continued) Page Results ............................................................................................................... 80 Discussion ......................................................................................................... 81 Acknowledgements ........................................................................................... 85 References ......................................................................................................... 87 Chapter 4: Summary ..................................................................................................... 94 Synthesis of survey
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