.................................................................................................................................. iv DEDICATION ............................................................................................................................... vi ACKNOWLEDGEMENTS .......................................................................................................... vii LIST OF TABLES .......................................................................................................................... x LIST OF FIGURES ....................................................................................................................... xi Chapter One: Expanded Distributions of Three Etheostoma Darters (subgenus Nothonotus) within the Upper Ohio River Watershed ........................................................................................ 1 1.1 Introduction ......................................................................................................................... 3 1.2 Study Area ........................................................................................................................... 5 1.3 Methods ............................................................................................................................... 5 1.4 Results ................................................................................................................................. 9 1.5 Discussion .......................................................................................................................... 16 1.6 Summary/Conclusions ....................................................................................................... 23 Literature Cited .......................................................................................................................... 24 Chapter Two: The genetic population structure of Etheostoma camurum (Bluebreast Darter) in the upper Ohio River watershed ................................................................................................... 29 2.1 Introduction ....................................................................................................................... 30 2.2 Methods............................................................................................................................. 35 2.3 Results ............................................................................................................................... 43 2.4 Discussion ......................................................................................................................... 59 2.5 Conclusions ....................................................................................................................... 73 Literature Cited ......................................................................................................................... 75 Chapter Three: Development of an aquatic environmental DNA (eDNA) method for detection of darters (subgenus Nothonotus) in the upper Ohio River watershed ............................................. 83 3.1 Introduction ....................................................................................................................... 84 3.2 Methods............................................................................................................................. 90 3.3 Results ............................................................................................................................. 100 3.4 Discussion ....................................................................................................................... 114 3.5 Conclusions ..................................................................................................................... 118 Literature Cited ....................................................................................................................... 121