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Bench-Scale Study for the Bioremediation of Chlorinated Ethylenes at Point Mugu Naval Air Weapons Station, Point Mugu California, IRP Site 24

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Bench-Scale Study for the Bioremediation of Chlorinated Ethylenes at Point Mugu Naval Air Weapons Station, Point Mugu California, IRP Site 24 AN ABSTRACT OF THE THESIS OF Matthew Thomas Keeling for the degree of Master of Science in Civil Engineering presented on November 23, 1998. Title: Bench-Scale Study for the Bioremediation of Chlorinated Ethylenes at Point Mugu Naval Air Weapons Station, Point Mugu California, IRP Site 24. Abstract approved. s Semprini Laboratory scale microcosm studies were conducted using site specific groundwater and aquifer solids to assess the feasibility of stimulating indigenous microorganisms in-situ to biologically transform Trichloroethylene (TCE) and its lesser chlorinated daughter products dichloroethylene (DCE) and vinyl chloride (VC). Three different treatments were conducted to determine the best approach for biologically remediating TCE under site specific conditions: anaerobic reductive dechlorination, aerobic cometabolism and sequential anaerobic/aerobic stimulation. Studies were conducted in batch serum bottles containing aquifer solids, groundwater and a gas headspace. Long-term (302 days) TCE anaerobic reductive dechlorination studies compared lactate, benzoate and methanol as potential anaerobic substrates. Site characteristic sulfate concentrations in the microcosms averaged 1,297 mg/L and TCE was added to levels of 2.3 mg/L. Substrates were added at one and a half times the stoichiometric electron equivalent of sulfate. Nutrient addition and bioaugmentation were also studied. Both benzoate and lactate stimulated systems achieved complete sulfate-reduction and prolonged dechlorination of TCE to VC and ethylene. Dechlorination was initiated between 15 to 20 days following lactate utilization and sulfate-reduction in the presence of approximately 300 mg/L sulfate. Benzoate amended microcosms did not initiate dechlorination until 120 to 160 days following the complete removal of available sulfate. After 302 days of incubation lactate and benzoate amended microcosms completely transformed TCE to VC with 7 to 15% converted to ethylene. Re-additions of TCE into both systems resulted in its rapid transformation to VC. The dechlorination of VC to ethylene was very slow and appeared to be dependent on VC concentration. Hydrogen addition at 10-3 and 10-4 atmospheres had no effect on the transformation of VC. Rapid methanol utilization resulted in its nearly stoichiometric conversion to methane and carbon dioxide without significant sulfate-reduction or dechlorination occurring. Nutrient addition slightly enhanced dehalogenation with lactate but inhibited it with benzoate. Bioaugmentation with a TCE dechlorinating culture from a previous benzoate amended Point Mugu microcosm effectively decreased lag-times and increased overall dechlorination. Aerobic cometabolism studies evaluated methane, phenol and propane as cometabolic growth substrates. Methane and phenol amended microcosms were able to remove only 50 to 60% of the added TCE after four stimulations, while propane utilizers were unable to cometabolize any TCE. Primary substrate utilization lag-times of 4 to 5 days, 0 to 0.5 days and 40 to 45 days were observed for methane, phenol and propane, respectively. Cometabolism of VC was possible in the presence of methane. Complete removal of 210 pg/L VC was achieved after 2 stimulations with methane under strictly aerobic conditions. Methane utilization and VC oxidation required nitrate addition, indicating that the system was nitrate limited. A sequential anaerobic/aerobic microcosm study failed to achieve methane utilization and VC transformation likely due to oxygen being utilized to re-oxidize reduced sulfate in the system. Bench-Scale Study for the Bioremediation of Chlorinated Ethylenes at Point Mugu Naval Air Weapons Station, Point Mugu California, IRP Site 24. by Matthew Thomas Keeling A THESIS submitted to Oregon State University in partial fulfillment of the requirements for the degree of Master of Science Presented November 23, 1998 Commencement June 1999 Master of Science thesis of Matthew Thomas Keeling presented on November 23, 1998 Major 'rofessor, repres ivil Engineering Chair of of Departmen of Civil, Construction, and Environmental Engineering Dean of Graduat chool I understand that my thesis will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my thesis to any reader upon request. Redacted for privacy Matthew Thomas Kee ing, Author ACKNOWLEDGEMENTS I would like to take this opportunity to recognize the people who have contributed in one way or another to this work and my life. First of all I would like to thank my advisor Dr. Lewis Semprini for giving me a reason to come to Oregon State University. His preeminent work in the field of chlorinated solvent remediation was the impetus for my interest in this field. Without his technical and financial support this work would have not been possible. I also owe a special thanks to all of my fellow grad students and researchers: Adisorn (Merf) Tovanabootr, Young Kim, Pardi Jitnuyanont, and especially George Pon for his preliminary work with Point Mugu microcosms. Their patience with my questions and willingness to help me prepared the groundwork for this work. Jason Cole deserves special recognition for his unwavering technical support - Merryfield Lab will suffer in his absence. I would also like to thank my committee members Dr. Jack Istok, Dr. Sandra Woods and Dr. Richard Vong for taking an interest in my work. I can never thank enough my family without whose love, nurturing and support I would not be the person I am today. My brothers Gregg, David and John never lacked advice to give me, but were always understanding when I had to do things my own way. My mother and father deserve special credit for raising four kooky boys without going completely mad. My thanks and love goes out to them for being there whenever I needed them. I cannot emphasize enough the group of individuals who without their acquaintance at OSU this would be just another piece of paper: Alex 'the Exterminator' Dehger with her ability to kill anything with a single word or chlorinated phenol molecule. Kristin Frey with her contagious laughter and endless knowledge of the celluloid. Marcus 'the Highway Avenger' Quigly and his 25% larger brain. With Marcus there will always be a better way. Rob Murchison for his down home southern morals, deeply introspective mind and willingness to talk with anyone no matter how smelly. My soul-sister Cassandra Robertson for her environmental idealism, warm hugs, traffic stopping smile and beautiful curly hair. Darin Trobaugh, his ability to sew fleece ACKNOWLEDGEMENTS, CONTINUED jog bras forty-eight hours straight gooned on hard candy and coke will lead him to greatness. His openess to amoral behavior and general craziness gave us all the chance to change him forever may he never be the same. Mark 'Cookie Monster' Havighorst who needs to quit his day job and write political satire for the Wall Street Journal; his tremendous wit and knowledge of worldly things is wasted on me. Jason and Jessica "Is that a no wax shine?" Cole who gave waxing the kitchen floor a totally new look. You are both compulsive and I love you that way. Just stay off the butcher block for God sake. My life would not be complete without the company of David and Kelly Roberts. Nothing is better than good friends and family you are both. Oh yeah, and I cannot forget Joe 'hide your sister' Lotrario and Paul 'which way did he go' Kwon. May they both find what they're looking for. See you all at our next therapy session. A very special thanks goes out to Sandra Uesugi for her review of this overwritten piece of research, her disdain for extraneous comas,,,, and her wonderful company. Finally, to the person who started this entire sorted academic affair, Kate Gaynor. How can I ever forgive her for telling me about environmental engineering and giving me the research paper "Where's the Benzene?" Kate, you really suck. TABLE OF CONTENTS Page 1. INTRODUCTION 1 1.1 Site Background 1 1.2 Scope of Work 4 1.3 Acknowedgement 5 2. LITERATURE REVIEW 6 2.1 Sources & Significance of TCE Contamination 6 2.2 Chlorinated Aliphatic Hydrocarbon Transformation 8 2.3 Biostimulation 8 2.4 Effects of Redox Potential & Chloride Substitution 9 2.4.1 Redox Potential 10 2.4.2 Anaerobic Systems 11 2.4.3 Aerobic Systems 12 2.5 Metabolism & Cometabolism 12 2.6 Anaerobic Reductive Dechlorination 14 2.6.1 Reductive Dechlorination by Microbial Cultures 19 2.6.1.1 methanogenesis 19 2.6.1.2 acetogenesis 22 2.6.1.3 sulfate reduction 23 2.6.1.4 iron reduction 26 2.6.1.5 facultative organisms 26 TABLE OF CONTENTS (Continued) Page 2.6.2 Hydrogen as the Electron Donor for Dechlorination 27 2.6.2.1 interspecies hydrogen transfer & syntrophy 29 2.6.2.2 hydrogen partial pressure and competition between hydrogenotrophs 29 2.6.3 Transition Metal Coenzymes & Cofactors 33 2.6.4 Chlororespiration 34 2.7 Aerobic Cometabolism 39 2.7.1 Aerobic Microorganisms & Enzymes 41 2.7.2 Competitive Inhibition & Transformation Product Toxicity 42 2.8 Combined Anaerobic/Aerobic Transformation 45 2.9 In-Situ Transformation 46 2.9.1 Natural Attenuation 47 2.9.2 In-Situ Biostimulation 48 2.9.2.1 anaerobic systems 49 2.9.2.2 aerobic systems 50 2.10 Conclusions 52 3. ANAEROBIC STUDY 53 3.1 Materials & Methods 53 3.1.1 Microcosm Preparation 53 3.1.2 Analytical Methods 57 3.2 Results & Discussion 59 3.2.1 Anaerobic Reactions 61 3.2.2 Lactate Stimulation 63 3.2.2.1 lactate utilization 63 3.2.2.2 dechlorination 69 TABLE OF CONTENTS (Continued) Page 3.2.3 Benzoate Stimulation 71 3.2.3.1 dechlorination 75 3.2.3.2 augmentation 76 3.2.4 Methanol Stimulation 79 3.2.5 Effect of Added Nutrients 82 3.2.5.1 lactate 82 3.2.5.2 benzoate 85 3.2.6 Sulfide Inhibition 87 3.2.7 Hydrogen Effects 89 3.2.8 Acetate Accumulation & pH 93 3.2.9 Electron Equivalence Mass Balance 94 3.3 Summary & Conclusions 98 4.
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