ROCK POOL MOSQUITO ECOLOGY OF THE SOUTHERN APPALACHIAN MOUNTAINS 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 Corey Allen Day Director: Dr. Brian Byrd Professor, School of Health and Human Sciences Committee Members: Dr. Thomas Martin, Department of Biology Dr. Joseph Pechmann, Department of Biology Dr. James Vonesh, Virginia Commonwealth University May 2020 ACKNOWLEDGEMENTS First, I wish to acknowledge my advisor and mentor throughout my undergraduate and graduate studies, Dr. Brian Byrd. He has gone well beyond any reasonable expectations to support and guide me. I deeply appreciate his guidance and the multitude of opportunities he has offered me, even when it was inconvenient to him and wholly unrequired. I owe him more than I can likely ever repay. I must also extend my gratitude to Dr. Thomas Martin, Dr. Joseph Pechmann, and Dr. James Vonesh. While their guidance as committee members certainly shaped and influenced this thesis, each of them also played crucial roles as classroom instructors. My participation in their thoughtfully designed courses prepared me with the knowledge and skills that I needed to conduct this research. I would also like to extend my thanks to my peers, who would be better described as friends, for their support and companionship during these years as a graduate student. I must acknowledge Kaylin Lewandowski, Ellie Armstrong, Miranda Pavey, and Justin Graham for their help with data collection for this thesis, but just as importantly I want to thank them and my other lab mates for making my learning experience more enjoyable with their personalities and passions for learning. Finally, I want to thank my family. So many of you have played immeasurable roles in my life, but unfortunately I cannot name everyone here. Specifically I will thank my parents, Mark and Kelly Day, as well as my fiancé Dakota Little, for their unwaivering love and support, and for constantly pushing me to pursue my interests even when we did not understand where they would take me. ii TABLE OF CONTENTS ROCK POOL MOSQUITO ECOLOGY OF THE SOUTHERN APPALACHIAN MOUNTAINS ............................................................................................................................ i ACKNOWLEDGEMENTS ....................................................................................................... ii LIST OF TABLES ...................................................................................................................... v LIST OF FIGURES ...................................................................................................................vi ABSTRACT ............................................................................................................................ vii INTRODUCTION ...................................................................................................................... 1 AIM 1: DETERMINE THE ANNUAL PHENOLOGY OF ROCK POOL MOSQUITOES ........5 Methods ..................................................................................................................................6 Study Site ............................................................................................................................6 Sampling Method ................................................................................................................6 Statistical Methods ..............................................................................................................9 Results .................................................................................................................................. 11 Abiotic Data ...................................................................................................................... 11 Wing Lengths of Mosquitoes Collected from Rock Pools .................................................. 11 Species Composition and Abundance ................................................................................ 12 Discussion ............................................................................................................................. 15 Abiotic Data ...................................................................................................................... 15 Mosquito Wing Lengths .................................................................................................... 15 Mosquito Species Compositions at Bull Pen and Bull Sluice ............................................. 16 Overall Abundances of Invertebrate Taxa .......................................................................... 18 Conclusion ............................................................................................................................ 19 AIM 2: ANALYZE THE FITNESS OF AUTOGENOUS AEDES ATROPALPUS REARED AT DIFFERENT TEMPERATURES .............................................................................................. 31 Methods ................................................................................................................................ 32 Experimental Design ......................................................................................................... 32 Statistical Methods ............................................................................................................ 34 Results .................................................................................................................................. 37 Comparisons of Size-Fecundity Relationships ................................................................... 37 Population Growth Correlates ............................................................................................ 37 iii Finite Population Growth Rates ......................................................................................... 38 Discussion ............................................................................................................................. 39 Effect of Temperature on Size-Fecundity Relationship ...................................................... 39 Effect of Temperature on Finite Population Growth Rate................................................... 39 Conclusion ............................................................................................................................ 43 REFERENCES ......................................................................................................................... 51 iv LIST OF TABLES Table 1A. Bull Pen pool measurements taken on 12 January 2019. ............................................. 8 Table 1B. Bull Sluice pool measurements taken on 15 January 2019. ......................................... 8 Table 2. Summary statistics for monthly water chemistry data at Bull Pen. Mean values and range are reported for each sampling date. ................................................................................. 21 Table 3. Summary statistics for monthly water chemistry data at Bull Sluice. Mean values and range are reported for each sampling date. ................................................................................. 22 Table 4. Results of generalized linear mixed-effect models comparing mean conductivity and pH among pools at BP and BS across the entire study period with pool as a random effect. ............ 22 Table 5. Average wing lengths of female Aedes japonicus japonicus collected as pupae at Bull Pen. ........................................................................................................................................... 23 Table 6. Average wing lengths of female Aedes atropalpus collected as pupae at Bull Sluice. .. 23 Table 7. Mean abundances of mosquitoes per pool at Bull Pen (BP). ........................................ 26 Table 8. Mean abundances of mosquitoes per pool at Bull Sluice (BS). .................................... 27 Table 9. List of taxa and the numbers of individuals observed during the sampling period ........ 29 Table 10. Linear regression of wing length (independent) and fecundity (dependent) and linear equations for functions relating fecundity and wing length (w) .................................................. 46 Table 11. Summary statistics for males that emerged from the temperature treatments. Values shown are grand means, with ranges based on means of the individual replicates. ..................... 46 Table 12. Tukey’s HSD contrasts of MANOVA with male data. .............................................. 47 Table 13. p-values of univariate contrasts of MANOVA for males and females with Bonferroni corrections. Each contrast had two degrees of freedom. ............................................................. 47 Table 14. Summary statistics for females that emerged from the temperature treatments. Values shown are grand means, with ranges based on means of the individual replicates. ..................... 48 Table 15. Tukey’s HSD contrasts of MANOVA with female data. ........................................... 48 Table 16. Tukey’s HSD contrasts of average 휆′ (finite population growth rate). ........................ 50 v LIST OF FIGURES Figure 1. Map of the northern Chattooga river highlighting the headwaters and the two rock pool sampling sites. .............................................................................................................................7 Figure 2. Rock pools 5