Microbial Community Production and Tolerance in Heavy Metals Polluted Sediment

University of Montana ScholarWorks at University of Montana Graduate Student Theses, Dissertations, & Professional Papers Graduate School 2002 Microbial community production and tolerance in heavy metals polluted sediment Christian D. Frazar The University of Montana Follow this and additional works at: https://scholarworks.umt.edu/etd Let us know how access to this document benefits ou.y Recommended Citation Frazar, Christian D., "Microbial community production and tolerance in heavy metals polluted sediment" (2002). Graduate Student Theses, Dissertations, & Professional Papers. 6877. https://scholarworks.umt.edu/etd/6877 This Thesis is brought to you for free and open access by the Graduate School at ScholarWorks at University of Montana. It has been accepted for inclusion in Graduate Student Theses, Dissertations, & Professional Papers by an authorized administrator of ScholarWorks at University of Montana. For more information, please contact [email protected]. Maureen and Mike MANSFIELD LIBRARY The University ofjMONTANA Permission is granted by the author to reproduce this material in its entirety, provided that tliis material is used for scholarly purposes and is properly cited in published works and reports. ** Please check "Yes" or "No" and provide signature** Yes, I grant permission X T No, I do not grant permission ____ Author's Signature. Date 9 V-/' X X Any copying for commercial purposes or financial gain may be undertaken only with the author's explicit consent. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Microbial Community Production and Tolerance in Heavy Metals Polluted Sediment By Christian D. FVazar B. A. Colby College, WatervUle, ME, 1999 Presented in partial fulfillment of the requirements for the degree of Master of Science The University of Montana August 2002 Apfn^ved by: /f ur!<A Cfhairman, Boaid of Examiners Dean, Graduate School - f Z -o Date Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number: EP37678 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed. a note will indicate the deletion. UMT DiwdfWion Publishing UMI EP37678 Published by ProQuest LLC (2013). Copyright in the Dissertation held by the Author. Microform Edition © ProQuest LLC. All rights reserved. This work is protected against unauthorized copying under Title 17, United States Code ProQuest ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. u Frazar, Christian D. MS., May 2002 Microbiology Measurement of Heavy Metal Tolerance and Biomass Production in Sediment Microbial Communities Director: Dr. James E. Gannon With the aid of four large floods in the late 1800’s and early 1900’s, mining wastes from the Butte - Anaconda region of Montana established a heavy metal gradient from the headwaters of the Clark Fork River down stream over 200 Ion. Over time microbial communities have been exposed to different metal concentrations dependant upon their place in the gradient. Microbial secondary productivity and biomass were measured seasonally and were not impaired relative to the metal concentrations in the sediment. Productivity levels were similar to those reported by other researchers in uncontaminated systems. These broad measures of community parameters suggest that microbial communities have adapted to the heavy metal concentrations. One mechanism for microbial adaptation to heavy metals is increased tolerance. Mine tailings and other mining wastes are routinely washed into the Clark Fork River from the floodplain and interact with the microbial communities. Four sites along the Clark Fork River and one uncontaminated tributary were stressed using increasing volumes of mine tailings and microbial productivity was measured using ['^C]leucine incorporation. The sites with the greatest previous metal exposure exhibited the greatest tolerance to the mine tailings. Microbial metal tolerance followed the metals gradient with sites furthest from the source exhibiting the lowest tolerance. These data suggest that the use of mine tailings to stress microbial communities may be a more realistic way to measure microbial tolerance to heavy metals than using conventional metal salt assays. In addition community productivity is still influenced by bank tailings that wash into river sediments. 'I Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. Ill ACKNOWLEDGMENTS I would like to thank Dr. James Gannon for the opportunity to work in his lab and for his advice and encouragement. I would also like to thank my committee members Dr. George Card and Dr. Johnnie Moore for their advice and guidance. Kevin Ferris, Janine Hall, Justin Harris, Philip Ramsey, and Nate Stevens assisted me in various ways including sample collection, geochemical analysis and general laboratory procedures. Their support and help was much appreciated. I would also like to thank Dr. Matthias Rillig for his patience and assistance with my statistical questions. Finally, I would like to thank Sarah Banner and my parents for their patience and support. \ Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. IV TABLE OF CONTENTS Chapter Page 1 General Introduction 1 2 Seasonal Microbial Productivity and Biomass Along a Gradient of 9 Mining Contamination in the Claik Foric River of Western Montana Abstract 9 Introduction 10 Materials and Methods 11 Results 14 Discussion 19 Conclusions 26 Acknowledgements 26 References 27 3 Measurement of Heavy Metal Tolerance in Sediment Microbial 31 Communities using IVOne Tailings Abstract 31 Introduction 32 Materials and Methods 34 Results 38 Discussion 43 Conclusions 49 Acknowledgements 50 References 51 4 General Discussion 54 General References 58 \ Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. V LIST OF FIGURES Chanter Figure Descriotion Page 2 1 Map of study sites 12 2 Seasonal biomass and productivity at CFRC and RCES 16 3 Seasonal biomass and productivity at CFGC and BHNB 17 4 Seasonal biomass and productivity at SBMC and LBGJ 18 5 Plot of Isotope Dilution 19 6 Comparison of mean biomass measurements for study sites 20 3 1 Map of study sites 35 2 Mine tailings inhibit microbial productivity 39 3 Linear regression of metal tolerance 40 4 Isotope dilution 41 5 Mine tailings result in lower sample pH 42 6 Both pH and mine tailings play a role in productivity 43 inhibition 7 Metal tolerance decreases with increasing distance from 45 the metal source \ Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. VI LIST OF TABLES Chaoter Table Description Page 2 1 List of test and reference sites 12 2 Concentrations of heavy metals in sediment samples 18 3 Bacterial carbon production at study sites 24 3 1 Sample composition for metal tolerance studies 37 2 Metal concentrations in sediment and tailings samples 44 \ Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. CHAPTER 1 GENERAL INTRODUCTION TO HEAVY METAL EFFECTS ON MICROBIAL COMMUNITIES: BACTERIAL PRODUCnVHY Over a century of mining activities have left a legacy of hazardous waste throughout many rivers and lakes in the United States. There are an estimated 550,000 abandoned hardrock mines nationwide and 12,000 miles of rivers polluted with heavy metals (38). Mining in the Butte, Montana region began in the mid-1860’s. By the 1890’s over 4,500 metric tons of sulfide ores were being mined daily from Butte Hill. Soon after mining reached these levels one of the world’s largest smelters was built in nearby Anaconda, MT. Early mining and smelting activities in the Butte - Anaconda region resulted in the accumulation of large volumes of mining wastes. Over 100 million metric tons of tailings and other mining wastes were routinely discarded directly to the land surface or to the headwaters of Silver Bow Creek. A series of large floods in the late 1800’s and early 1900’s helped to establish a heavy metals gradient extending over 200km downstream. The floodplain of Silver Bow Creek currently holds vast quantities of mine tailings and heavy-metal laden sediment, which still enter the creek during rainstorms and other disturbances. Extensive fish kills have been documented as a result of the sudden influx of heavy metals into the river following thunderstorms, demonstrating that the mining wastes still impact the river today (27). Silver Bow Creek becomes the Clark Fork River several miles downstream. The Clark Fork River Basin is now the largest superfund complex, covering an area approximately l/5‘^ the size of Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. 2 Rhode Island (37). Metals of concern in this contaminated system include various species of arsenic, cadmium, copper, iron, lead and zinc. Several studies have explored the effects of heavy metals, including metal tolerance, on sediment microbial

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