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The purpose of this study is to compare different approaches to modeling the reductive dechlorination of chlorinated ethenes in the anaerobic region of an upward flow constructed wetland by microbial consortia. A controlled simulation experiment that compares three different approaches to modeling the degradation of chlorinated ethenes in wetland environments is conducted and investigates how each of the modeling approaches affect simulation results. Concepts like microbial growth in the form of a biofilm and spatially varying contaminant concentrations bring the validity of the CSTR assumption into question. These concepts are incorporated into the different modeling approaches to evaluate the CSTR assumption. Model simulations show that spatially varying contaminant concentrations have a significant effect on contaminant effluent concentrations. Additionally, the significance of the incorporation of a biofilm concept depends on the time characteristics of both diffusive mass transport and reaction kinetics.
Chlorinated solvents and their daughter products are the most common contaminants of groundwater at industrial and military facilities in the United States. Limitations of conventional technologies have intensified efforts to find alternative methods to remediate contaminated sites to regulatory goals set by CERCLA. Natural attenuation of chlorinated solvents is a promising alternative to traditional pump and treat methods but has not been well understood or widely accepted. This modeling study investigated the ability of TCE to completely degrade under various aquifer conditions and rate order constants. It also examined a case study of a former landfill site at Moody AFB. We found unusually high flow of ground water by advection or dispersion inhibits the complete degradation of TCE. High concentrations of sulfate or nitrate inhibit the creation of methanogenic conditions and therefore inhibit reductive dechlorination of TCE. We also found an electron donor co-contaminant a critical factor for the complete destruction of TCE because it creates anaerobic conditions. The model illustrated a possible explanation for the lack of down gradient contaminants at the landfill site may be the coupling of reductive dechlorination and cometabolism naturally attenuation the contaminants.
Contamination by chlorinated ethenes is widespread in groundwater aquifers, sediment, and soils worldwide. The overarching objectives of my research were to understand how the bacterial genus Dehalococcoides function optimally to carry out reductive dechlorination of chlorinated ethenes in a mixed microbial community and then apply this knowledge to manage dechlorinating communities in the hydrogen-based membrane biofilm reactor (MBfR). The MBfR is used for the biological reduction of oxidized contaminants in water using hydrogen supplied as the electron donor by diffusion through gas-transfer fibers. First, I characterized a new anaerobic dechlorinating community developed in our laboratory, named DehaloR^2, in terms of chlorinated ethene turnover rates and assessed its microbial community composition. I then carried out an experiment to correlate performance and community structure for trichloroethene (TCE)-fed microbial consortia. Fill-and-draw reactors inoculated with DehaloR^2 demonstrated a direct correlation between microbial community function and structure as the TCE-pulsing rate was increased. An electron-balance analysis predicted the community structure based on measured concentrations of products and constant net yields for each microorganism. The predictions corresponded to trends in the community structure based on pyrosequencing and quantitative PCR up to the highest TCE pulsing rate, where deviations to the trend resulted from stress by the chlorinated ethenes. Next, I optimized a method for simultaneous detection of chlorinated ethenes and ethene at or below the Environmental Protection Agency maximum contaminant levels for groundwater using solid phase microextraction in a gas chromatograph with a flame ionization detector. This method is ideal for monitoring biological reductive dechlorination in groundwater, where ethene is the ultimate end product. The major advantage of this method is that it uses a small sample volume of 1 mL, making it ideally suited for bench-scale feasibility studies, such as the MBfR. Last, I developed a reliable start-up and operation strategy for TCE reduction in the MBfR. Successful operation relied on controlling the pH-increase effects of methanogenesis and homoacetogenesis, along with creating hydrogen limitation during start-up to allow dechlorinators to compete against other microorgansims. Methanogens were additionally minimized during continuous flow operation by a limitation in bicarbonate resulting from strong homoacetogenic activity.
Trichloroethene (TCE) is a ubiquitous soil and groundwater contaminant. The most common bioremediation approach for TCE relies on the process of reductive dechlorination by Dehalococcoides mccartyi. D. mccartyi uses TCE, dichloroethene, and vinyl chloride as electron acceptors and hydrogen as an electron donor. At contaminated sites, reductive dechlorination is typically promoted by adding a fermentable substrate, which is broken down to short chain fatty acids, simple alcohols, and hydrogen. This study explored microbial chain elongation (MCE), instead of fermentation, to promote TCE reductive dechlorination. In MCE, microbes use simple substrates (e.g., acetate, ethanol) to build medium chain fatty acids and also produce hydrogen during this process. Soil microcosm using TCE and acetate and ethanol as MCE substrates were established under anaerobic conditions. In soil microcosms with synthetic groundwater and natural groundwater, ethene was the main product from TCE reductive dechlorination and butyrate and hydrogen were the main products from MCE. Transfer microcosms using TCE and either acetate and ethanol, ethanol, or acetate were also established. The transfers with TCE and ethanol showed the faster rates of reductive dechlorination and produced more elongated products (i.e., hexanoate). The microbial groups enriched in the soil microcosms likely responsible for chain elongation were most similar to Clostridium genus. These investigations showed the potential for synergistic microbial chain elongation and reductive dechlorination of chlorinated ethenes.