Reference Diatom Assemblage Response to Transplantation into a Stream Receiving Treatment for Acid Mine Drainage in Southeastern Ohio A thesis presented to the faculty of the College of Arts and Sciences of Ohio University In partial fulfillment of the requirements for the degree Master of Science Jonathon B. Gray November 2011 © 2011 Jonathon B. Gray. All Rights Reserved. 2 This thesis titled Reference Diatom Assemblage Response to Transplantation into a Stream Receiving Treatment for Acid Mine Drainage in Southeastern Ohio by JONATHON B. GRAY has been approved for the Program of Environmental Studies and the College of Arts and Sciences by Morgan L. Vis Professor of Environmental and Plant Biology Howard Dewald Interim Dean, College of Arts and Sciences 3 ABSTRACT GRAY, JONATHON B., M.S., November 2011, Environmental Studies Reference Diatom Assemblage Response to Transplantation into a Stream Receiving Treatment for Acid Mine Drainage in Southeastern Ohio Director of Thesis: Morgan L. Vis Acid mine drainage (AMD) is a prevalent legacy of coal mining within Appalachia. Streams receiving AMD effluent are drastically altered both chemically and biologically. Hewett Fork, a stream in southeastern Ohio, is one such affected stream. Although treatment methods have reduced acidity considerably downstream, the ability of Hewett Fork to sustain a biological community compared to those found in reference conditions remains unclear. To assess this, tiles colonized with diatom assemblages from an unimpacted stream were transplanted into Hewett Fork along a stream health gradient, from poor to good, and sampled after one, three, and six weeks in the treated stream. Chlorophyll a concentrations and species diversity metrics were calculated to compare reference assemblages to transplanted assemblages. Results suggested that after an initial one week acclimation period, assemblages at the uppermost and lowermost sites along the reach were relatively similar to those found in reference conditions, while sites within the middle region continued to show signs of impairment, although the factor(s) causing this impairment remain unknown. These findings suggest that although treatment has been effective on a site-specific basis, the expected linear-response to treatment may not be achieved due to underlying factors that are inhibiting reference-like biological communities from reestablishing within the affected stream reach. 4 Approved: _____________________________________________________________ Morgan L. Vis Professor of Environmental and Plant Biology 5 ACKNOWLEDGMENTS First and foremost, I would like to thank Dr. Morgan Vis for the uncountable hours she has invested in my success as a graduate student. She was always there to provide me with the encouragement and advice that I needed whenever uncertainty reared its ugly head. I am most grateful for the opportunity to join her lab. It is also imperative that I thank my thesis committee members, Drs. Kelly Johnson and Brian McCarthy for their assistance in the development of my research project as well as their continued support as my project moved forward. I must also acknowledge Nate Smucker for his role in showing me the ropes upon arriving in the lab and for continuing to put up with my inquiries into…everything. Also, Justin Pool for taking me out into the world of southeastern Ohio and sharing with me the sage-like advice of an experienced fellow tile scraper. Furthermore, I would like to thank Lisa DeRose, Dr. Daryl Lam, Mariah Thrush, and Lauren Fuelling for assisting me in sampling the wilds of Hewett Fork; we came, we saw, we sampled! And I can’t forget my lab mates, Emily Johnston, Eric Salomaki, Lauren F. (as previously mentioned) for making our little office into so much more than a workspace; you made it feel like a home. Not to mention the entirety of the Environmental Studies Program and the Department of Environmental and Plant Biology. Thanks to the Phycological Society of America for awarding me the Croasdale Fellowship so that I might attend Diatom Camp, as well as Drs. Edlund and Potapova for introducing me to diatom ecology. And for that matter, thank you diatoms! Thank you to all of the following funding sources for their generosity and support: Graduate Student Senate Original Work and Travel Grants, the Voinovich School of Leadership and Public 6 Affairs (especially Jen Bowman!), the Department of Environmental and Plant Biology, and the OU Research and Creativity Expo. Finally, there is no way any of this would have been possible without the support of my wife, Mindy. Is it any wonder that the last ten years of my life have been the best years thus far? I love you, Minders. 7 TABLE OF CONTENTS Page Abstract............................................................................................................................... 3 Acknowledgments............................................................................................................... 5 List of Tables ...................................................................................................................... 9 List of Figures................................................................................................................... 10 Introduction....................................................................................................................... 12 Materials and Methods...................................................................................................... 20 Results............................................................................................................................... 30 Discussion......................................................................................................................... 37 Conclusions and Future Directions................................................................................... 43 References......................................................................................................................... 44 Appendix A: Diatom Growth Forms and the Representative Genera, with Descriptions of Growth Form and Potential Costs/Benefits of that Growth Form. ................................... 71 Appendix B: Acid Mine Drainage Diatom Index of Biotic Integrity (AMD-DIBI) Component Metrics from Zalack et al. (2010).................................................................. 73 Appendix C: Habitat Characteristics at the Time of Initial Tile Colonization, Transplantation, and 1-, 3-, and 6-Weeks After Transplantation. N/A in Water Velocity Represents Below Detectable Levels of Measurement, yet Definite Flow was Visible... 75 Appendix D: Water Chemistry for Each Site at 1, 3, and 6 Weeks After Transplantation. ........................................................................................................................................... 77 Appendix E: Number of Cumulative (n = 3) Taxa Identified at Each Site for Week 1 Sampling. .......................................................................................................................... 80 Appendix F: Bray-Curtis Dissimilarity Scores for Each Site and Sampling Time Using Pooled Replicates (n = 3). Mean is the Mean Dissimilarity Between Clear Creek and Hewett Fork Sites.............................................................................................................. 94 Appendix G: Bray-Curtis Dissimilarity Scores for Each Site and Sampling Time, Rounded to Four Decimal Places...................................................................................... 95 Appendix H: Number of Cumulative (n = 3) Taxa Identified at Each Site for Week 3 Sampling. .......................................................................................................................... 98 Appendix I: Number of Cumulative (n = 3) Taxa Identified at Each Site for Week 6 Sampling ......................................................................................................................... 112 Appendix J: Number of Cumulative (n = 3) Taxa Identified at Each Site from the Comparative Study of Clear Creek, HF039 and HF090................................................. 129 8 Appendix K: Bray-Curtis Dissimilarity Scores for Clear Creek and the Most Downstream (HF039) and Upstream (HF090) Sites on Hewett Fork After a 6-Week Colonization Period Using Pooled Replicates (n = 3). Mean is the Mean Dissimilarity Between Clear Creek and Hewett Fork Sites.................................................................. 142 2 Appendix L: The χ 0.05,1 Values from Comparisons of Growth Forms Between Clear Creek and HF039, and Clear Creek and HF090 After a 6-Week Colonization Period... 143 9 LIST OF TABLES Page Table 1: Mean and range (in parentheses) historical water chemistry data for Hewett Fork for months June - September, with n = number of samples available for each parameter from www.watersheddata.com ................................................................ 52 Table 2: Mean and range (in parentheses) historical biodiversity data for Hewett Fork for sampling months June – September, with n = the number of samples available for each parameter from www.watersheddata.com and Pool (2010). N/A represents no data available due to limited samples available (in the case of data ranges) or a lack of samples for that site .............................................................................................. 53 Table 3: Mean water chemistry (n = 3; range in parentheses) from each site for the duration