Kinetics of Dehalobacter and Dehalococcoides Strains and Their Effects on Competition for Growth

Title of Document: KINETICS OF DEHALOBACTER AND DEHALOCOCCOIDES STRAINS AND THEIR EFFECTS ON COMPETITION FOR GROWTH SUBSTRATES. YenJung Lai, Doctor of Philosophy, 2010 Directed By: Associate Professor, Jennifer G. Becker, Department of Environmental Science and Technology The chlorinated solvents tetrachloroethene (PCE) and trichloroethene (TCE) are common groundwater contaminants. Reductive dechlorination of PCE and TCE at contaminated sites is commonly carried out by dehalorespiring bacteria that utilize these compounds as terminal electron acceptors, but often results in the accumulation of cis-1,2-dichloroethene (cDCE) and vinyl chloride (VC), rather than non-toxic ethene. This project focused on evaluating how interactions among dehalorespiring populations that may utilize the same electron acceptors, electron donors and/or carbon source may affect the extent of PCE dechlorination in situ. These interactions may be particularly important if both Dehalococcoides ethenogenes (Dhc. ethenogenes) and Dehalobacter restrictus (Dhb. restrictus) are present because these bacteria utilize the same electron donor (H2) and both respire PCE and TCE. However, unlike Dhc. ethenogenes, Dhb. restrictus cannot dechlorinate PCE beyond cDCE. Therefore, the outcome of the population interactions may determine the extent of detoxification achieved. Monod kinetic parameter estimates that describe chlorinated ethene and electron donor utilization by Dhc. ethenogenes and Dhb. restrictus at non-inhibitory substrate concentrations were obtained in batch assays. Substrate inhibition effects on both populations were also evaluated. Highly chlorinated ethenes negatively impacted dechlorination of the lesser chlorinated ethenes in both populations. In Dhc. ethenogenes, cometabolic transformation of VC was also inhibited by the presence of other chlorinated ethenes. PCE and TCE dechlorination by Dhb. restrictus was strongly inhibited by VC. The microbial interactions between Dhc. ethenogenes and Dhb. restrictus was investigated using reactors and mathematical models under engineered bioremediation and natural attenuation conditions. Under engineered bioremediation conditions, Dhc. ethenogenes became the dominant population, and the modeling predictions suggested that the inhibition of Dhb. restrictus by VC was a key factor in determining this outcome. Dechlorination rates by Dhb. restrictus appeared to be affected very little by low acetate concentrations under natural attenuation conditions, giving it an advantage over Dhc. ethenogenes, which requires relatively high acetate concentrations. This study highlighted that substrate interactions among dehalorespiring bacteria can influence their performance and contaminant fate under common bioremediation scenarios. A better understanding of the factors affecting the outcomes of these microbial interactions was achieved, which should aid in the design of successful bioremediation strategies. Kinetics of Tetrachloroethene-Respiring Dehalobacter and Dehalococcoides Strains and Their Effects on Competition for Growth Substrates