University of Massachusetts Amherst ScholarWorks@UMass Amherst Masters Theses Dissertations and Theses December 2020 In-vitro Propagation and Fish Assessments to Inform Restoration of Dwarf Wedgemussel (Alasmidonta Heterodon) Jennifer Ryan University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/masters_theses_2 Part of the Aquaculture and Fisheries Commons, and the Terrestrial and Aquatic Ecology Commons Recommended Citation Ryan, Jennifer, "In-vitro Propagation and Fish Assessments to Inform Restoration of Dwarf Wedgemussel (Alasmidonta Heterodon)" (2020). Masters Theses. 990. https://doi.org/10.7275/18786186 https://scholarworks.umass.edu/masters_theses_2/990 This Open Access Thesis is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Masters Theses by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. IN-VITRO PROPAGATION AND FISH ASSESSMENTS TO INFORM RESTORATION OF DWARF WEDGEMUSSEL (ALASMIDONTA HETERODON) A Thesis Presented by JENNIFER ELAINE RYAN Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment Of the requirements for the degree of MASTER OF SCIENCE September 2020 Environmental Conservation Wildlife, Fish and Conservation Biology IN-VITRO PROPAGATION AND FISH ASSESSMENTS TO INFORM RESTORATION OF DWARF WEDGEMUSSEL (ALASMIDONTA HETERODON) A Thesis Presented by JENNIFER ELAINE RYAN Approved as to style and content by: ____________________________________ Allison H. Roy, Chair ____________________________________ David L. Perkins, Member ____________________________________ Peter D. Hazelton, Member ____________________________________ Curtice R. Griffin . Environmental Conservation ACKNOWLEDGMENTS First and foremost, I would like to thank my advisor, Allison Roy for accepting me as a student in her lab, for her continued support throughout this process, and for fostering such a strong community in her lab. I would also like to thank my committee members, Peter Hazelton and David Perkins for their feedback and encouragement these last two years. I would like to acknowledge my funding source, the Science Support Partnership Program with the US Fish and Wildlife Service and the US Geological Survey. In particular, I would like to thank Heather Galbraith, Barry Baldigo and Barbara St. John White from the U.S. Geological Survey. I would also like to thank Melissa Grader and Susi von Oettingen from the U.S. Fish and Wildlife Service for supplying the data that allowed me to complete my research. I would also like to thank members of the Richard Cronin Aquatic Resource Center (CARC), specifically Timothy Warren, for aiding me in the setup for all my experiments and braving the cold water to collect gravid mussels. The assistance Tim provided throughout all my experiments was invaluable and I am deeply grateful for his help. I would also like to thank Ayla Skorupa and Virginia Martell for welcoming me into the mussel lab with open arms. I am thankful to have worked alongside such intelligent and driven women who were willing to answer my endless questions about graduate school and CARC before and throughout my time at UMass. This experience would have been much more exhausting if not for their presence. I would also like to thank my technicians, Kara Duprey, Sarah Endyke and Sam Rode for their enthusiasm for the project, their endless positivity and flexibility throughout the field season and iii beyond. Their work was instrumental in the completion of this project. Thank you to Ethan Nedeau (Biodrawversity, LCC) for collecting gravid dwarf wedgemussels. I want to thank Monte McGregor and Julieann Jacobs with the Kentucky Department of Fish and Wildlife Resources Center for Mollusk Conservation taught me how to successfully propagate freshwater mussels using in-vitro methods. Without their training, guidance and willingness to share, my ability to conduct successful in-vitro propagation would not have been possible. Thank you for taking precious time out of your busy schedule to train me and foster a passion for this very specific line of research. I also want to thank Robert Wick and Claire Walsh who offered me their endless support to identify the fungal contamination plaguing my in-vitro dishes. Their curiosity, interest, and dedication to figuring out a big piece of the puzzle helped in my research as well as future research for freshwater mussel propagation. Thank you to the fisheries and mussel biologists throughout the Northeast took time out of their busy field season to help with data collection. Specifically, I would like to thank Brian Keleher and Steven Mattocks from Massachusetts Division of Fisheries and Wildlife; Ross Shramko and Jeanette Bowers of New Jersey Department of Environmental Protection; Brian Eltz, Michael Beauchene and Laura Saucier from Connecticut Department of Energy and Environmental Protection; and Scott Decker, Benjamin Nugent, Melissa Doperalski and Jason Carrier from New Hampshire Fish and Game as well as all of their field technicians who helped gather data, plan and execute fish surveys in the summer of 2019. iv I would like to give a special thank you to all of the Roy lab members, past and present who offered advice and support, as well as a community I can to take with me for the rest of my career. I would also like to thank my younger sister, Alyssa, who showed me the ropes of graduate school at UMass and who welcomed me with open arms. Finally, I would like to thank my parents, Trudy and Robert Ryan who fostered a love and appreciation of the outdoors, especially freshwater ecosystems. The life they gave me allowed me to pursue my dreams and for that, I am forever grateful. v ABSTRACT PROPAGATION AND FISH ASSESSMENTS TO INFORM RESTORATION OF DWARF WEDGEMUSSEL (ALASMIDONTA HETERODON) SEPTEMBER 2020 JENNIFER E. RYAN, B.S., STATE UNIVERSITY OF NEW YORK AT POSTDAM M.S., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Allison H. Roy The dwarf wedgemussel (Alasmidonta heterodon) is a federally endangered freshwater mussel that once ranged from New Brunswick to North Carolina, but now only exists in isolated populations throughout its diminished range. Laboratory propagation in conjunction with augmentation or reintroduction is considered a critical component of dwarf wedgemussel restoration. My thesis aimed to I) develop in-vitro propagation techniques including methods to minimize microbial contamination for two Alasmidonta species and II) compile and assess critical fish assemblage information at existing dwarf wedgemussel presence and absence locations to inform future restoration activities. For the first objective (Chapter 2) I assessed three methods of contamination mitigation (media change frequency, concentration of the antifungal Amphotericin B, and method of antifungal replenishment). Across all experiments, higher levels of contamination severity had a negative impact on transformation success. In my experiments, dishes that had media changes every other day (vs. those changed daily and every 3 days) had the highest contamination and the lowest glochidia transformation success. Treatments with vi the lowest (0 µg/mL) concentration of the antifungal, Amphotericin B, and got a low- dose replenishment of Amphotericin B (vs frequent media changes) had the highest transformation success. The fungus was identified as Candida parapsilosis, a common fungus found in aquatic and human environments; future propagation efforts should use mitigation methods that are specific to the fungal contaminant. In the assessment of host fish near dwarf wedgemussel locations (Chapter 3), two fish repeatedly emerged as indicators of dwarf wedgemussel occurrence and abundance: the tessellated darter (Etheostoma olmstedi) and brown trout (Salmo trutta). Tessellated darter was positively related to dwarf wedgemussel which supports the importance of tessellated darter as a host fish in the wild. The negative relationship between dwarf wedgemussel and brown trout may indicate a difference in habitat between the species or brown trout’s predation upon wild hosts. Results from this project will be used in conjunction with habitat and genetic information to inform future restoration plans in the northeast and add to the growing body of literature on in-vitro propagation of freshwater mussels. vii TABLE OF CONTENTS Page ACKNOWLEDGEMENTS ............................................................................................... iii ABSTRACT ....................................................................................................................... vi LIST OF TABLES ............................................................................................................. xi LIST OF FIGURES ......................................................................................................... xiii CHAPTER 1. INTRODUCTION .........................................................................................................1 1.1. Study Species – The Dwarf Wedgemussel (Alasmidonta heterodon) ....................1 1.2. Freshwater Mussel Propagation and Host Fish for Restoration .............................4 1.3. Study Objectives .....................................................................................................6 2. ASSESSING METHODS FOR MITIGATING CONTAMINATION IN FRESHWATER MUSSEL IN-VITRO PROPAGATION