FEASIBILITY STUDY FOR RESTORATION OF FRESHWATER MUSSELS (VILLOSA IRIS AND LAMPSILIS FASCIOLA) INTO THE UPPER OCONALUFTEE RIVER IN NORTH CAROLINA A thesis presented to the faculty of the Graduate School of Western Carolina University in partial fulfillment of the requirements for the degree of Master of Science in Biology. By Rachael Elizabeth Finigan Director: Dr. Thomas Martin Associate Professor Department of Biology Committee Members: Dr. Karen Kandl, Biology Mr. Michael LaVoie, Eastern Band of Cherokee Indians Natural Resources March 2019 ACKNOWLEDGEMENTS I would like to thank my advisor Dr. Thomas Martin for his contributions to this project and dealing with my seemingly endless questions. Also, thank you to the rest of my committee, Dr. Karen Kandl and Mr. Michael LaVoie, for their advice throughout this process. Thank you to Dr. Jeremy Hyman for reading this thesis. Thank you to my field partner, Michelle Ruigrok, for helping me through long field days and keeping me laughing. Thank you to Rachael Hoch, Luke Etchison, and Stephania Maehrlein at the North Carolina Wildlife Resources Commission for their help with tagging, installation, and advice. Thank you to numerous people from the Eastern Band of Cherokee Indians Natural Resources for helping build enclosures and assisting with installation, including Caleb Hickman, Dallas Bradley, Josh Parrish, Nick Reed, and Aaron Ducker. Thank you to many others who helped with field work or provided advice, including Ethan Fite, Sierra Kincaid, Erin Clingerman, Steve Fraley, Jason Mays, Ezra Gardiner, and Mollie Cashner. Finally, thank you to TVA for funding this project, making it all possible. ii TABLE OF CONTENTS List of Tables ................................................................................................................................. iv List of Figures ..................................................................................................................................v Abstract ......................................................................................................................................... vii Introduction ......................................................................................................................................1 Methods............................................................................................................................................8 Study Site and Animal Collection ...............................................................................................8 Experimental Design ...................................................................................................................9 Data Collection .........................................................................................................................12 Analysis .....................................................................................................................................13 Results ............................................................................................................................................15 Initial Length, Survival, and Growth ........................................................................................15 Water Quality, Urbanization, and Snorkel Surveys ..................................................................22 Tags ...........................................................................................................................................27 Discussion ......................................................................................................................................30 Survival and Growth ................................................................................................................30 Tags ..........................................................................................................................................36 Conclusions ..............................................................................................................................37 Works Cited ...................................................................................................................................39 iii LIST OF TABLES Table 1. Locations of each site described by river, GPS coordinates, and river kilometers. ...........9 Table 2. ANOVA summary for tests of differences in initial length of V. iris and L. fasciola .....16 Table 3. ANOVA summary for tests of differences in maximum length of V. iris .......................19 Table 4. ANOVA summary for tests of differences in maximum length of L. fasciola ................21 Table 5. ANCOVA summary table of specific conductivity (µs/cm) ...........................................23 Table 6. ANCOVA summary table of dissolved oxygen (mg/L) ..................................................24 Table 7. ANCOVA summary table of Temperature (°C) ..............................................................25 Table 8. ANOVA summary for analysis of velocity (m/s) ............................................................26 Table 9. ANOVA summary for tests of differences in tags ...........................................................28 iv LIST OF FIGURES Figure 1. Mussel silo developed by Dr. M. Christopher Barnhart at Missouri State University and modified by the Virginia Department of Game and Inland Fisheries (VDGIF) (Barnhart et al. 2007) ......................................................................................... 11 Figure 2. Hallprint shellfish tags (left), queen bee tags (right top of mussel), laser etched tags (right bottom of mussel) on L. fasciola (top) and V. iris (bottom) ................................11 Figure 3. Initial length (mm) of L. fasciola (left) and V. iris (right) at site 1 (red circle), site 2 (green triangle), site 3 (blue square), and control (purple cross) ..................................16 Figure 4. Percent survival of L. fasciola (circle) and V. iris (triangle) at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from May to November. The purple arrow shows experiment installation month, when no measurements were taken. .............................................................................................................................17 Figure 5. Average length (mm) of V. iris at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from March to November. Initial measurements were taken in March and the purple arrow shows experiment installation month, when no measurements were taken. May data was removed from the analysis ..........................18 Figure 6. Average growth in length (mm) of V. iris at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from March to November. Initial measurements were taken in March and the purple arrow shows experiment installation month, when no measurements were taken. May data was removed from the analysis ....................................................................................................................19 Figure 7. Average length (mm) of L. fasciola at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from March to November. Initial measurements were taken in March and the purple arrow shows experiment installation month, when no measurements were taken. May data was removed from the analysis .....................20 Figure 8. Average growth in length (mm) of L. fasciola at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from March to November. Initial measurements were taken in March and the purple arrow shows experiment installation month, when no measurements were taken. May data was removed from the analysis ....................................................................................................................21 Figure 9. Specific conductivity (µs/cm) of sites 1 (red circle), 2 (green triangle), and 3 (blue square) plotted against USGS gauge specific conductivity (µs/cm) .............................23 Figure 10. Dissolved oxygen (mg/L) of sites 1 (red circle), 2 (green triangle), and 3 (blue square) plotted against USGS gauge dissolved oxygen (mg/L) .................................................24 Figure 11. Temperature (°C) of sites 1 (red circle), 2 (green triangle), and 3 (blue square) plotted against USGS gauge temperature (°C) ..........................................................................25 Figure 12. Velocity (m/s) from May to November at sites 1 (red circle), 2 (green triangle), and 3 (blue square) ..................................................................................................................26 Figure 13. Urbanization criteria scores at sites 1, 2, 3, and control ...............................................27 Figure 14. Average tag condition of queen bee tags (red circle), Hallprint tags (green triangle), and laser tags (blue square) of L. fasciola and V. iris ...................................................28 v Figure 15. Average Hallprint (A), queen bee (B), and laser tag (C) condition of L. fasciola and V. iris at sites 1 (red circle), 2 (green triangle), 3 (blue square), and control (purple cross) from May to November. ................................................................................................29 Figure 16. Percent survival of A. viridis (left), L. fasciola (middle), and V. iris (right) at sites 1 (red circle), 2 (green triangle), 3 (blue