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Bacterial Characterization of Louisiana Groundwater Contaminated By Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2009 Bacterial characterization of Louisiana groundwater contaminated by DNAPL-containing chloroethanes and other solvents Kimberly Bowman Louisiana State University and Agricultural and Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Part of the Civil and Environmental Engineering Commons Recommended Citation Bowman, Kimberly, "Bacterial characterization of Louisiana groundwater contaminated by DNAPL-containing chloroethanes and other solvents" (2009). LSU Doctoral Dissertations. 3340. https://digitalcommons.lsu.edu/gradschool_dissertations/3340 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. BACTERIAL CHARACTERIZATION OF LOUISIANA GROUNDWATER CONTAMINATED BY DNAPL-CONTAINING CHLOROETHANES AND OTHER SOLVENTS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College In partial fulfillment of the Requirements for the degree of Doctor of Philosophy In The Department of Civil Engineering by Kimberly Bowman B.S., San José State University, 1993 May, 2009 ACKNOWLEDGEMENTS I would like to thank the members of my advisory committee Dr. William Moe, Dr. Fred Rainey, Dr. John Pardue, Dr. David Constant, and Dr. Maud Walsh for contributing their valuable time and guidance to this dissertation. In particular, I would like to express my gratitude to Dr. Moe for all of the time and energy he put into mentoring me, and for the exceptional patience and support he provided me in my pursuit of balance in academics and family. I would like to thank Brian Rash for his assistance with LIBSHUFF and rarefaction analysis and Ms. Cindy Henk for her assistance with microscopy. And I would like to acknowledge my graduate school peers: Jun Yan, Marilou Nabatilan, Jyoti Rao, Congna Li, Luis Espinoza, Gabriel Kassenga, Stephen Mbuligwe, Brian Rash, Ryan Callegan, and Manish Shukla for their friendship and help, and our student worker, Rachael Dupré, for her high quality work. My husband Dave, and my daughters Sierra and Zoey deserve immense gratitude for their unconditional love, patience, and support, as do other members of my family, including my parents, Pat and Jay Bainbridge and my in-laws, Phyllis and Jim Bowman. Finally, I would like to thank the state of Louisiana for the Economic Development Assistantship and Dr. William Moe for the financial assistance, and the Governor’s Biotechnology Initiative of the Louisiana Board of Regents and NPC Services, Inc for funding my research. ii TABLE OF CONTENTS Acknowledgements......................................................................................................................... ii List of Tables .................................................................................................................................. v List of Figures................................................................................................................................ vi Abstract......................................................................................................................................... vii Chapter 1: Introduction and Literature Review ............................................................................. 1 1.1 Introduction..................................................................................................................... 1 1.2 Biotransformation of Chlorinated Aliphatic Compounds............................................... 3 1.3 Bacterial Tolerance to Chlorinated Solvents .................................................................. 6 1.4 Microbially Mediated Hydrogen Production as a Source for Alternative Energy.......... 9 1.5 Background of the PPI Site........................................................................................... 10 1.6 Research Objectives...................................................................................................... 10 1.7 Organization of the Dissertation ................................................................................... 11 Chapter 2: Enumeration and Diversity of Bacteria in the DNAPL Source Zone of the PetroProcessors of Louisiana, Inc. Superfund Site....................................................................... 12 2.1 Introduction................................................................................................................... 12 2.2 Experimental Design..................................................................................................... 12 2.3 Results........................................................................................................................... 19 2.4 Discussion..................................................................................................................... 30 2.5 Conclusions................................................................................................................... 37 Chapter 3: Chlorinated Solvent Tolerance of Clostridium Species............................................. 39 3.1 Introduction................................................................................................................... 39 3.2 Experimental Design..................................................................................................... 39 3.3 Results........................................................................................................................... 42 3.4 Discussion..................................................................................................................... 51 3.5 Conclusions................................................................................................................... 54 Chapter 4: Rate and Yield of Hydrogen Production by Clostridium Isolates and Reference Strains ........................................................................................................................................... 56 4.1 Introduction................................................................................................................... 56 4.2 Experimental Design..................................................................................................... 58 4.3 Results........................................................................................................................... 62 4.4 Discussion..................................................................................................................... 64 4.5 Conclusions................................................................................................................... 69 Chapter 5: Further Characterization of Two Clostridium Isolates and Their Description as New Species .......................................................................................................................................... 70 5.1 Introduction................................................................................................................... 70 5.2 Experimental Design..................................................................................................... 70 iii 5.3 Results........................................................................................................................... 74 5.4 Discussion..................................................................................................................... 81 5.5 Conclusions................................................................................................................... 86 Chapter 6: Conclusions and Recommendations for Future Research.......................................... 87 6.1 Conclusions................................................................................................................... 87 6.2 Recommendations for Future Research........................................................................ 91 Literature Cited ............................................................................................................................. 93 Vita……………………………………………………………………………………………...111 iv LIST OF TABLES Table 2.1: Chlorinated solvent concentrations measured in groundwater from well W-1024-1 at the PPI site .................................................................................................................................... 20 Table 2.2: Geochemical parameters measured in groundwater from well W-1024-1 at the PPI site................................................................................................................................................. 20 Table 2.3: Phylogenetic summary of bacterial community based on 16S rRNA gene sequences amplified using universal bacterial primers.................................................................................. 23 Table 2.4: Phylogenetic summary of bacterial community based on 16S rRNA gene sequences from anaerobic isolates ................................................................................................................. 24 Table 2.5: LIBSHUFF comparisons of isolate and clone libraries ............................................. 30 Table 3.1: Hydrogen produced in the absence of chlorinated solvents......................................
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