Chlorinated ethenes such as tetrachloroethene (PCE) and trichloloethene (TCE) are among the most prevalent contaminants in soil, sediments and groundwaters. Currently, Insitu bioremediation via anaerobic reductive dechlorination has become a widely used technology for groundwater contaminated with chlorinated ethenes. To better understand the reductive dechlorination remediation process and the inter-relationships among the complex microbial communities that comprise it, a comprehensive biokinetic model was recently developed at Cornell University by Gretchen Heavner, a modification of an earlier Cornell model developed by Donna Fennell. The Heavner model uses specific biomasses based on quantitative PCR-based population data, and under some conditions can accurately predict kinetics of dechlorination, fermentation of electron donors, and competition for electron donors between dechlorinators and methanogens, and generation of methane. However, the platform used to run the model - STELLA® (High Performance Systems) - is cumbersome for simulation of long time-spans, limiting the model's utility. Furthermore, the model uses an empirical, "mRNA-tuning" technique to improve data fits at high PCE-loadings, which makes the model descriptive, rather than predictive, in such cases. Additionally, electron donor fermentation is not predicted well at high electron-donor feeding rates. The overall purpose of this thesis research was to address some of the limitations of the Heavner model. The STELLA® model was successfully converted to run in MATLAB® using Runge-Kutta 4th-order integration. The model fits at high-PCE and high electron-donor loadings were improved by utilizing the inhibitory effects of high PCE on dechlorination and methanogenesis, and by postulating additional pathways of butyrate's fermentation and acetate's hydrogenation to storage products. Model simulations indicate that by adding 2nd-order Haldane inhibition instead of mRNA tuning, the model revised in this thesis research predicts the dechlorination, methanogenesis and donor fermentation well over a broad range of PCE feeding rates. Moreover, when simulating donor fermentation at high-PCE-loadings, butyrate's fermentations and acetate's hydrogenation to storage products must be considered to obtain a mass balance between butyrate consumption and product formation.

This research focused on the enhanced reductive dechlorination of trichloroethene (TCE) and its surrogate, trichlorofluoroethene (TCFE), using two bioremediation methods in anaerobic conditions. Two anaerobic bioremediation studies were conducted to investigate the effects of microbial communities in the presence of different electron acceptors and donors during anaerobic reductive dechlorination of TCE and TCFE. The first study was conducted in the groundwater microcosm bottles, filled with groundwater and sediments collected from Richmond site, CA. Parallel reductive dechlorination of TCE and TCFE was evaluated in the presence of fumarate and its product, succinate, while active reduction of high background concentrations of sulfate (2.5 mM) occurred. Because sulfate was assumed as a favorable electron acceptor during reductive dechlorination of chlorinated aliphatic hydrocarbons (CAHs), all microcosms receiving TCE and TCFE with substrates showed enhanced reductive dechlorination activity and even no substrate addition microcosms generated biotransformation products. From the electron mass balance calculations, more than 87.5% of electrons went to sulfate reduction and less than 10% of available electrons involved in dechlorination after sulfate reductions. After amending varying concentrations of sulfate (0 2.5 mM), no inhibition was found between reductive dechlorination of TCE and sulfate reduction. The result indicated that reductive dechlorination could be directly competed with sulfate reduction for available electrons. The second study investigated the effectiveness of in situ push-pull tests to evaluate bioaugmentation in physical aquifer models (PAMs) using dehalogenating strains to reductively dechlorinate TCE to ethene and TCFE to FE in the TCE contaminated sediments. Complete reduction of TCE to ethene occurred in less than 14 days with repeated additions of TCE (13.0 to 46.0 mg/L) and TCFE (15.0 mg/L) was completely transformed to FE in under 24 days. Increased rate and extent of dechlorination in the bioaugmented PAM compared to the nonaugmented control PAM indicated successful transport of the bioaugmented culture through the PAM. Similar transformation rates and time course of TCE and TCFE also indicated that TCFE was a bioprobe for reductive dechlorination of TCE. TCE and TCFE were transformed to cisdichloroethene (c-DCE) and cis-dichlorofluoroethene (c-DCFE) respectively at two of the three sampling ports after 50 days of incubation in the nonaugmented PAM indicating reductive dechlorination activity of indigenous microorganisms. The results showed that it is possible to increase the rate and extent of reductive dechlorination of TCE and TCFE by bioaugmentation and that push-pull tests are effective tools for detecting and quantifying these processes in situ. The third study focused on numerical modeling of the second study. The objectives of this study were (1) to evaluate a simplified method for estimating retardation factors for injected solutes and bioaugmented microorganisms using "pushpull" test injection phase breakthrough curves, (2) to identify whether bioaugmented microorganisms have kept the same transformation capacity of Evanite culture using Michaelis-Menten kinetics by the values provided by Yu et al. (2005) and to verify in situ rates of TCFE reductive dechlorination rates of push-pull tests by numerical modeling, and (3) to investigate a reasonable answer for the nonuniform recovery of ethene and FE during the activity test and the push-pull test. The bioaugmented microorganisms were effectively transported through Hanford sediment. The estimated retardation factor was 1.33. A numerical simulation predicted cell transport in the PAM as far as port 5. This was qualitatively confirmed by cell counts obtained during bioaugmentation but, cells were distributed nonuniformly. The transport test indicated that TCE and TCFE transport was relatively retarded compared to coinjected bromide tracer (retardation factors ranged from 1.33-1.62 for TCE and from 1.44-1.70 for TCFE). The modeling simulation of Michaelis-Menten kinetics for the activity test was well matched for reductive dechlorination rates for TCE and less dechlorinated ethenes using the previous published values of kmax and Ks of chlorinated ethenes by Yu et al. (2005); the model match indicated that the bioaugmented microorganisms kept the same transformation capacity as the original source, Evanite culture (Yu et al., 2005) over 4 months in the PAM. A numerical simulation resulted in the simple first order FE production rate of 1 day' using STOMP code (2002) and the value of FE production rate was in the range of the transformation rates of TCFE during the activity test. The bioaugmented PAM has caused slow loss of injected CAHs during the activity test and the push-pull test.

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.