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INVESTIGATION OF FRESHWATER MUSSEL PHYSIOLOGY AND REPRODUCTIVE BIOLOGY TO INFORM CONSERVATION by ANDREA KAY FRITTS (Under the Direction of Robert B. Bringolf) ABSTRACT Filter feeding freshwater mussels (order Unionoida) fulfill an important ecological niche, but a suite of anthropogenic perturbations have made them the most imperiled faunal group in North America. Threats to mussels include habitat degradation, pollution, and alterations to natural flow regimes. The Apalachicola- Chattahoochee-Flint River Basin is a watershed under increasing pressure from human populations and is also home to a diverse assemblage of aquatic organisms including five federally listed mussel species. The federal recovery plan for these species outlines specific research objectives that will better equip scientists and managers with the necessary tools to protect and restore populations of these species. This dissertation has focused on addressing some of these critical data gaps by investigating the ecological relevance of the sodium chloride glochidia viability test, conducting host determination trials for the Purple Bankclimber mussel (Elliptoideus sloatianus), and developing nonlethal methods for assessing the physiological response of mussels to various stress events. Freshwater mussels are characterized by a unique lifecycle in which the glochidia larvae must attach to a vertebrate host to metamorphose into a juvenile mussel. This larval stage is used in toxicity testing to evaluate the effects of contaminants on freshwater mussels and for the derivation of water quality criteria. My results indicated that the viability of glochidia as measured by the sodium chloride test is an ecologically relevant measure of the health of glochidia. The discovery of Gulf Sturgeon (Acipenser oxyrinchus desotoi) as the primary hosts for Purple Bankclimber mussels has supplied important information for the preservation and management of wild mussel populations as well as providing the necessary data to initiate captive propagation. Changes in tissue glycogen and hemolymph chemistry parameters are potential biomarkers for monitoring stress in freshwater mussels. The factors of discharge, size, sex, and species were most commonly found to affect the biological responses in our models and we recommend that future research into the effects of drought and stress should include the use of alanine aminotransferase, aspartate aminotransferase, bicarbonate, and calcium. Combined, these data will allow scientists and managers alike to advance the conservation of these intriguing and ecologically important animals. INDEX WORDS: Glochidia, Unionidae, Elliptoideus sloatianus, Apalachicola- Chattahoochee-Flint River Basin, Biomarker INVESTIGATION OF FRESHWATER MUSSEL PHYSIOLOGY AND REPRODUCTIVE BIOLOGY TO INFORM CONSERVATION by ANDREA KAY FRITTS B.S., University of Wisconsin-River Falls, 2007 M.S. Missouri State University, 2009 A Dissertation Submitted to the Graduate Faculty of The University of Georgia in Partial Fulfillment of the Requirements for the Degree DOCTOR OF PHILOSOPHY ATHENS, GEORGIA 2013 © 2013 ANDREA KAY FRITTS All Rights Reserved INVESTIGATION OF FRESHWATER MUSSEL PHYSIOLOGY AND REPRODUCTIVE BIOLOGY TO INFORM CONSERVATION by ANDREA KAY FRITTS Major Professor: Robert B. Bringolf Committee: James T. Peterson Mary C. Freeman C. Rhett Jackson Electronic Version Approved: Maureen Grasso Dean of the Graduate School The University of Georgia August 2013 DEDICATION This dissertation would not have been possible without the unending support from Mark Fritts: fellow biologist, terrific cook, and wonderful husband. Our hours together in the rivers of Georgia and our countless dinner conversations about research projects, experimental design, and delectable recipes have made the past four years not only memorable, but also incredibly fun! I look forward to our continued exploration of the natural world, with you and I side by side. iv ACKNOWLEDGEMENTS When I first visited the University of Georgia and the lab of Dr. Robert Bringolf in January 2009, I realized very quickly that this was the place where I wanted to pursue my doctoral degree. Since the very first moment that I met Dr. Bringolf, he has been the type of advisor that every graduate student hopes for. Supportive and kind, yet one who challenges his students to jump out of their comfort zones and to grow as a scientist. His unending encouragement and passion for science has produced many wonderful experiences and memories that I will cherish as I leave Georgia and embark on new research adventures. Special thanks go out to Bob Ratajczak, Colin Shea, and Jason Wisniewski. Bob was always willing to lend a hand in the field or lab and he made sure that we were equipped with all of the peculiar supplies that we needed. Colin and Jason taught me the mussels of the Flint River and graciously helped to collect mussels not only during the pleasant summer months but also during chilly winter sampling trips. Many other individuals have contributed to the successful completion of this dissertation research, including: Sandy Abbott, Brett Albanese, Chris Barnhart, Mike Bednarski, Shay Bush, Ben Carswell, Greg Cope, Julie Creamer, Zack DeWolf, Justin Dycus, Dewayne Fox, Mark Fritts, Pete Hazelton, Karen Herrington, Whitney Jacobs, Cecil Jennings, Jennifer Johnson, Rachel Katz, Kristen Kellock, Teresa Newton, Doug Peterson, Sandy Pursifull, Bernard Sietman, Channing St. Aubin, Jim Stoeckel, Brittany Trushel, Amos Tuck, Dan Watrous, Deb Weiler, Naeem Willett, and Jim Williams. My committee members, Dr. v Mary Freeman, Dr. Jim Peterson, and Dr. Rhett Jackson have also been terrific sources of support and constructive input over the course of this project. Owens and Williams Fish Farm were incredibly generous in donating a tremendous number of largemouth bass for an assortment of experiments. Lastly, a big thank you to my family, who were the first to instill a love of nature within me. My parents, Bill and Anna Crownhart, my siblings, Maria, Amy, John, and Rachel, and my sweet husband, Mark Fritts---you have all been incredible sources of support during my graduate school career. vi TABLE OF CONTENTS Page ACKNOWLEDGEMENTS .................................................................................................v LIST OF TABLES ...............................................................................................................x LIST OF FIGURES ......................................................................................................... xiii CHAPTER 1 BACKGROUND AND CHAPTER ORGANIZATION ...................................1 INTRODUCTION .......................................................................................1 REFERENCES ..........................................................................................10 2 ASSESSMENT OF TOXICITY TEST ENDPOINTS FOR THE GLOCHIDIA LARVAE OF FRESHWATER MUSSELS .............................16 ABSTRACT ...............................................................................................17 INTRODUCTION .....................................................................................18 METHODS ................................................................................................20 RESULTS ..................................................................................................26 DISCUSSION ............................................................................................29 REFERENCES ..........................................................................................35 3 CRITICAL LINKAGE OF IMPERILED SPECIES: GULF STURGEON AS HOST FOR THE PURPLE BANKCLIMBER MUSSELS.............................48 ABSTRACT ...............................................................................................49 INTRODUCTION .....................................................................................51 vii METHODS ................................................................................................54 RESULTS ..................................................................................................58 DISCUSSION ............................................................................................58 REFERENCES ..........................................................................................65 4 EVALUATION OF PHYSIOLOGICAL BIOMARKERS OF STRESS IN FRESHWATER MUSSELS ............................................................................80 ABSTRACT ...............................................................................................81 INTRODUCTION .....................................................................................83 METHODS ................................................................................................87 RESULTS ..................................................................................................93 DISCUSSION ............................................................................................95 REFERENCES ........................................................................................103 5 NON-LETHAL ASSESSMENT OF FRESHWATER MUSSEL RESPONSE TO CHANGES IN ENVIRONMENTAL FACTORS IN THE LOWER FLINT RIVER BASIN, GEORGIA, U.S.A. .................................................120 ABSTRACT .............................................................................................121 INTRODUCTION ...................................................................................123 METHODS ..............................................................................................127
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