Chlorinated aliphatic hydrocarbons (CAHs), such as tetrachloroethene (PCE) and trichloroethene (TCE), are ubiquitously pollutants in aquifer sediments and groundwater due to their heavy usage in industry and inappropriate disposal in the last century. Among about 1300 NPL (National Priorities List) sites, PCE and TCE are the two most frequently detected hazardous contaminants. Engineered bioremediation, including biostimulation and bioaugmentation, is a promising technology to clean those PCE and/or TCE contaminated sites. However, in many contaminated groundwater systems and hazardous waste sites, pH can be lower than 5 to 6. And release of HCl (strong acid) from anaerobic reductive dechlorination may lower the pH of groundwater. Besides, another main source of acidity comes from the fermentation of additive electron donors such as alcohols, organic acids and etc. Decreasing pH has been proved to be detrimental to the microbes that dechlorinated PCE or TCE. We intended to enrich and isolate microorganisms, which can perform anaerobic reductive dechlorination at low pH environments, by establishing microcosms, which will be beneficial to in situ bioremediation. We also screened some existing cultures for dechlorinating activity at low pH and determined the pH tolerance of consortium BDI, which had been successfully, applied for in situ bioremediation. Besides, this study investigated and explored the effects of solids on BDI consortium under low pH conditions. Generally, various dechlorinating pure cultures and consortium BDI show highest dechlorination rates and extent at circumneutral pH. Only Sulfurospirillum multivorans among tested cultures dechlorinated PCE to cDCE at pH 5.5. The screening efforts suggest that microbes capable of dechlorination below pH 5.5 are not common. It was observed that solids play an important role for enhancing microbial activities under low pH conditions. And BDI consortium can recover from up to 8 weeks exposure to low pH conditions, although the VC-to-ethene dechlorination step was affected.

​This volume provides a review of the past 10 to 15 years of intensive research, development and demonstrations that have been on the forefront of developing bioaugmentation into a viable remedial technology. This volume provides both a primer on the basic microbial processes involved in bioaugmentation, as well as a thorough summary of the methodology for implementing the technology. This reference volume will serve as a valuable resource for environmental remediation professionals who seek to understand, evaluate, and implement bioaugmentation.

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.

This book summarizes the current state of knowledge concerning bacteria that use halogenated organic compounds as respiratory electron acceptors. The discovery of organohalide-respiring bacteria has expanded the range of electron acceptors used for energy conservation, and serves as a prime example of how scientific discoveries are enabling innovative engineering solutions that have transformed remediation practice. Individual chapters provide in-depth background information on the discovery, isolation, phylogeny, biochemistry, genomic features, and ecology of individual organohalide-respiring genera, including Dehalococcoides, Dehalogenimonas, Dehalobacter, Desulfitobacterium and Sulfurospirillum, as well as organohalide-respiring members of the Deltaproteobacteria. The book introduces readers to the fascinating biology of organohalide-respiring bacteria, offering a valuable resource for students, engineers and practitioners alike.