Anaerobic dechlorination and redox activities after full-scale Electrical Resistance Heating (ERH) of a TCE-contaminated aquifer

The effects of Electrical Resistance Heating (ERH) on dechlorination of TCE and redox conditions were investigated in this study. Aquifer and groundwater samples were collected prior to and after ERH treatment, where sediments were heated to approximately 100 degrees C. Sediment samples were collected from three locations and examined in microcosms for 250 to 400 days of incubation. Redox activities, in terms of consumed electron acceptors, were low in unamended microcosms with field-heated sediments, although they increased upon lactate-amendment. TCE was not dechlorinated or stalled at cDCE with field-heated sediments, which was similar or lower compared to the degree of dechlorination in unheated microcosms. However, in microcosms which were bioaugmented with a mixed anaerobic dechlorinating culture (KB-1) and lactate, dechlorination past cDCE to ethene was observed in field-heated sediments. Dechlorination and redox activities in microcosms with field-heated sediments were furthermore compared with controlled laboratory-heated microcosms, which were heated to 100 degrees C for 10 days and then slowly cooled to 10 degrees C. In laboratory-heated microcosms, TCE was not dechlorinated and redox activities remained low in unamended and lactate-amended sediments, although organic carbon was released to the aqueous phase. In contrast, in field-heated sediments, high aqueous concentrations of organic carbon were not observed in unamended microcosms, and TCE was dechlorinated to cDCE upon lactate amendment. This suggests that dechlorinating microorganisms survived the ERH or that groundwater flow through field-heated sediments carried microorganisms into the treated area and transported dissolved organic carbon downstream.     

The need for bioaugmentation after thermal treatment of a TCE-contaminated aquifer: Laboratory experiments

A microcosm study was conducted to evaluate the need for bioaugmentation after a thermal treatment to anaerobically dechlorinate trichloroethene (TCE) to ethene. The microcosms were either: heated to 100 degrees C and slowly cooled to simulate thermal remediation while bioaugmenting when the declining temperature reached 10 degrees C; or kept at ambient groundwater temperatures (10 degrees C) and bioaugmented for comparison. Aquifer samples from three sediment locations within a TCE-polluted source zone were investigated in duplicate microcosms. In biostimulated (5 mM lactate) and heated microcosms, no conversion of TCE was observed in 4 out of 6 microcosms, and in the remaining microcosms the dechlorination of TCE was incomplete to cDCE (cis-dichloroethene). By comparison, complete TCE dechlorination to ethene was observed in 4 out of 6 heated microcosms that were bioaugmented with a highly enriched dechlorinating mixed culture, KB-1, but no electron donor, and also in 4 of 6 microcosms that were augmented with KB-1 and an electron donor (5 mM lactate). These data suggest that electron donor released during heating, was capable of promoting complete dechlorination coincident with bioaugmentation. Heated microcosms demonstrated less methanogenesis than unheated microcosms, even with elevated H2 concentrations and addition of KB-1, which contains methanogens. This suggests that the heating process suppressed the native microbial community, which can decrease competition with the bioaugmented culture and increase the effectiveness of dechlorination following a thermal treatment. Specifically, cDCE removal rates were four to six times higher in heated than unheated bioaugmented microcosms. This study confirms the need for bioaugmentation following a laboratory thermal treatment to obtain complete dechlorination of TCE.     

Quantification of biotransformation of chlorinated hydrocarbons in a biostimulation study: added value via stable carbon isotope analysis

Stable carbon isotope analysis of chlorinated aliphatic compounds was performed at an in situ biostimulation pilot test area (PTA) at a site where 1,2-dichloroethane (1,2-DCA) and trichloroethene (TCE) were present in groundwater. Chlorinated products of TCE reductive dechlorination (cis-dichloroethene (cDCE) and vinyl chloride (VC)) were present at concentrations of 17.5 to 126.4 micromol/L. Ethene, a potential degradation product of both 1,2-DCA dihaloelimination and TCE reductive dechlorination was also present in the PTA. Emulsified soybean oil and lactate were added as electron donors to stimulate anaerobic dechlorination in the PTA. Stable carbon isotope analysis provided evidence that dechlorination was occurring in the PTA during biostimulation, and a means of monitoring changes in dechlorination efficiency over the 183 day monitoring period. Stable carbon isotope analysis was also used to determine if ethene production in the PTA was due to dechlorination of TCE, 1,2-DCA, or both. Fractionation factors (alpha) were determined in the laboratory during anaerobic biotransformation of 1,2-DCA via a dihaloelimination reaction in four separate enrichment cultures. These alpha values (as well as the previously published ranges of alpha for the dechlorination of TCE, cDCE and 1,2-DCA) were used, along with isotopic values measured during the pilot test, to derive quantitative estimates of biotransformation during the pilot test. Dechlorination was found to account for 10.7 to 35.9%, 21.9 to 74.9%, and 54.4 to 67.8% of 1,2-DCA, TCE and cDCE concentration loss respectively in the PTA. Stable carbon isotope analysis indicates that dechlorination of 1,2-DCA, TCE and cDCE were all significant processes during the pilot test, while ethene production during the pilot test was dominated by 1,2-DCA dihaloelimination. This study demonstrates how stable carbon isotope analysis can provide more conservative estimates of the extent of biotransformation than do conventional protocols. In addition, in a complex mixed plume such as this, compound specific isotope analysis is shown to be one of the few methods available for clarifying dominant biotransformation pathways where breakdown products are non-exclusive (i.e. ethene).     

Microbial PCB dechlorination in dredged sediments and the effect of moisture

Evidence of reductive dechlorination of polychlorinated biphenyls (PCBs) in sediments was investigated in Hudson River sediments dredged and encapsulated in 1978 at Moreau, NY. The effect of different moisture contents in dredged sediments on dechlorination and dechlorinating microorganisms was also determined using PCB-spiked sediments in which the moisture level was adjusted by simulating a dewatering process. The congener pattern of PCBs indicated that the dechlorination in the dredged sediments was far less advanced than that in the river sediments collected from the general area of the dredged site (Ft. Edward site). Dechlorination in encapsulated sediments at the Moreau site appeared to have stopped soon after dredging. When microorganisms eluted from the encapsulated sediments were inoculated in clean sediments spiked with Aroclor 1242, an extensive dechlorination was observed, indicating that the encapsulated sediments still harbored dechlorinating microorganisms. However, the same inoculum failed to further dechlorinate residual congeners in the dredged sediments. On the other hand, an inoculum obtained in 1990 from the dredged site in the Hudson River dechlorinated the residual congeners further. In simulated dredged sediments, the maximum level of dechlorination was lower at reduced moisture contents. The population size of dechlorinating microorganisms, as determined by the most probable number (MPN) technique, was also smaller at the lower moisture levels. There was a significant correlation between the maximum extent of dechlorination and the specific death rate of dechlorinating populations. These results indicate that the underlying mechanism of the moisture-dependent maximum dechlorination is the moisture-dependence of the death rate of dechlorinating microorganisms.     

The reactive transport of trichloroethene is influenced by residence time and microbial numbers

The dechlorination rate in a flow-through porous matrix can be described by the species specific dechlorination rate observed in a liquid batch unless mass transport limitations prevail. This hypothesis was examined by comparing dechlorination rates in liquid batch with that in column experiments at various flow rates (3-9-12 cm day(-1)). Columns were loaded with an inoculated sand and eluted with a medium containing 1mM trichloroethene (TCE) for 247 days. Dechlorination in the column treatments increased with decreasing flow rate, illustrating the effect of the longer residence time. Zeroth order TCE or cis-DCE degradation rates were 4-7 folds larger in columns than in corresponding batch systems which could be explained by the higher measured Geobacter and Dehalococcoides numbers per unit pore volume in the columns. The microbial numbers also explained the variability in dechlorination rate among flow rate treatments marked by a large elution of the dechlorinating species' yield as flow increased. Stop flow events did not reveal mass transport limitations for dechlorination. We conclude that flow rate effects on reactive transport of TCE in this coarse sand are explained by residence time and by microbial transport and that mass transport limitations in this porous matrix are limited.     

Assessing chlorinated ethene degradation in a large scale contaminant plume by dual carbon-chlorine isotope analysis and quantitative PCR

The fate of chlorinated ethenes in a large contaminant plume originating from a tetrachloroethene (PCE) source in a sandy aquifer in Denmark was investigated using novel methods including compound-specific carbon and chlorine isotope analysis and quantitative real-time polymerase chain reaction (qPCR) methods targeting Dehaloccocoides sp. and vcrA genes. Redox conditions were characterized as well based on concentrations of dissolved redox sensitive compounds and sulfur isotopes in SO(4)(2-). In the first 400 m downgradient of the source, the plume was confined to the upper 20 m of the aquifer. Further downgradient it widened in vertical direction due to diverging groundwater flow reaching a depth of up to 50 m. As the plume dipped downward and moved away from the source, O(2) and NO(3)(-) decreased to below detection levels, while dissolved Fe(2+) and SO(4)(2-) increased above detectable concentrations, likely due to pyrite oxidation as confirmed by the depleted sulfur isotope signature of SO(4)(2-). In the same zone, PCE and trichloroethene (TCE) disappeared and cis-1,2-dichloroethene (cDCE) became the dominant chlorinated ethene. PCE and TCE were likely transformed by reductive dechlorination rather than abiotic reduction by pyrite as indicated by the formation of cDCE and stable carbon isotope data. TCE and cDCE showed carbon isotope trends typical for reductive dechlorination with an initial depletion of (13)C in the daughter products followed by an enrichment of (13)C as degradation proceeded. At 1000 m downgradient of the source, cDCE was the dominant chlorinated ethene and had reached the source δ(13)C value confirming that cDCE was not affected by abiotic or biotic degradation. Further downgradient (up to 1900 m), cDCE became enriched in (13)C by up to 8 ‰ demonstrating its further transformation while vinylchloride (VC) concentrations remained low (<1 μg/L) and ethene was not observed. The correlated shift of carbon and chlorine isotope ratios of cDCE by 8 and 3.9 ‰, respectively, the detection of Dehaloccocides sp genes, and strongly reducing conditions in this zone provide strong evidence for reductive dechlorination of cDCE. The significant enrichment of (13)C in VC indicates that VC was transformed further, although the mechanism could not be determined. The transformation of cDCE was the rate limiting step as no accumulation of VC occurred. In summary, the study demonstrates that carbon-chlorine isotope analysis and qPCR combined with traditional approaches can be used to gain detailed insight into the processes that control the fate of chlorinated ethenes in large scale plumes.     

Comparison between acetate and hydrogen as electron donors and implications for the reductive dehalogenation of PCE and TCE

Bioremediation by reductive dehalogenation of groundwater contaminated with tetrachloroethene (PCE) or trichloroethene (TCE) is generally carried out through the addition of a fermentable electron donor such as lactate, benzoate, carbohydrates or vegetable oil. These fermentable donors are converted by fermenting organisms into acetate and hydrogen, either of which might be used by dehalogenating microorganisms. Comparisons were made between H2 and acetate on the rate and extent of reductive dehalogenation of PCE. PCE dehalogenation with H2 alone was complete to ethene, but with acetate alone it generally proceeded only about half as fast and only to cis-1,2-dichloroethene (cDCE). Additionally, acetate was not used as an electron donor in the presence of H2. These findings suggest the fermentable electron donor requirement for PCE dehalogenation to ethene can be reduced up to 50% by separating PCE dehalogenation into two stages, the first of which uses acetate for the conversion of PCE to cDCE, and the second uses H2 for the conversion of cDCE to ethene. This can be implemented with a recycle system in which the fermentable substrate is added down-gradient, where the hydrogen being produced by fermentation effects cDCE conversion into ethene. The acetate produced is recycled up-gradient to achieve PCE conversion into cDCE. With the lower electron donor usage required, potential problems of aquifer clogging, excess methane production, and high groundwater chemical oxygen demand (COD) can be greatly reduced.     

Use of statistical tools to evaluate the reductive dechlorination of high levels of TCE in microcosm studies

A large, multi-laboratory microcosm study was performed to select amendments for supporting reductive dechlorination of high levels of trichloroethylene (TCE) found at an industrial site in the United Kingdom (UK) containing dense non-aqueous phase liquid (DNAPL) TCE. The study was designed as a fractional factorial experiment involving 177 bottles distributed between four industrial laboratories and was used to assess the impact of six electron donors, bioaugmentation, addition of supplemental nutrients, and two TCE levels (0.57 and 1.90 mM or 75 and 250 mg/L in the aqueous phase) on TCE dechlorination. Performance was assessed based on the concentration changes of TCE and reductive dechlorination degradation products. The chemical data was evaluated using analysis of variance (ANOVA) and survival analysis techniques to determine both main effects and important interactions for all the experimental variables during the 203-day study. The statistically based design and analysis provided powerful tools that aided decision-making for field application of this technology. The analysis showed that emulsified vegetable oil (EVO), lactate, and methanol were the most effective electron donors, promoting rapid and complete dechlorination of TCE to ethene. Bioaugmentation and nutrient addition also had a statistically significant positive impact on TCE dechlorination. In addition, the microbial community was measured using phospholipid fatty acid analysis (PLFA) for quantification of total biomass and characterization of the community structure and quantitative polymerase chain reaction (qPCR) for enumeration of Dehalococcoides organisms (Dhc) and the vinyl chloride reductase (vcrA) gene. The highest increase in levels of total biomass and Dhc was observed in the EVO microcosms, which correlated well with the dechlorination results.     

Comparison of an assay for Dehalococcoides DNA and a microcosm study in predicting reductive dechlorination of chlorinated ethenes in the field

The study aims to compare the detection of 16S rRNA gene of Dehalococcoides species and the microcosm study for biotransformation in predicting reductive dechlorination of chlorinated ethenes in ground water at hazardous waste sites. A total of 72 ground water samples were collected from 12 PCE or TCE contaminated sites in the United States. The samples were analyzed and used to construct microcosms in the laboratory. The results showed that the presence of Dehalococcoides DNA was well associated with dechlorination to ethene in the field. Nearly half of the wells where Dehalococcoides DNA was detected had ethene as a dechlorination end product. In comparison, for ground water samples of 16 wells where ethene was detected, ethene was produced in 11 of the corresponding microcosms. For most microcosms, during two years of incubation, dechlorination was less extensive than that observed in the field.     

The study on the dechlorination of OCDD with Pd/C catalyst in ethanol-water solution under mild conditions

Dechlorination of octachlorodibenzo-p-dioxin (OCDD) was carried out in ethanol-water (v/v=1:1) solution of NaOH in the presence of Pd/C catalysts with the use of H(2). The substrate was dechlorinated with Pd/C under mild conditions (atmospheric pressure and <100 degrees C) to give a chlorine-free product, dibenzo-p-dioxin (DD), in high yields. After reaction of 3h at 50 degrees C, 95.9% OCDD was degraded to low dechlorinated congeners and the yield of DD was 77.4%. We have also studied the dechlorination selectivity of chlorine atoms on the different substituted positions and postulated the dechlorination pathway of OCDD. For OCDD, the 2-position has higher reactivity than 1-position, but the difference is very small. From the distribution statistics of the intermediates during the reaction, we postulate that the steric effect plays an important role during the reaction and affect the dechlorination pathway of OCDD.     

Synergic effect of calcium oxide and iron (III) oxide on the dechlorination of hexachlorobenzene

Dechlorination of hexachlorobenzene (HCB) was achieved by a mixture of commercial CaO and alpha-Fe2O3 (CaO/alpha-Fe2O3) in closed systems at temperatures of 300 degrees C and 350 degrees C, which exhibited a synergic effect compared to CaO or alpha-Fe2O3 alone, and the dechlorination efficiency was dramatically enhanced. When CaO and alpha-Fe2O3 coexisted, HCB was dechlorinated by about 98% after 0.5 h reaction at 300 degrees C. All TeCB-, TrCB-, and DCB-isomers were detected, showing the presence of more than one dechlorination pathway. The pathway to form 1,2,4-TrCB must be a major pathway because the greatest amount of 1,2,4-TrCB was detected. There existed discrepancy of the material balance between the starting and dechlorinated materials. It implies that besides dechlorination other decomposition processes may be present. The mechanism of synergic effect of calcium and iron oxides was investigated.     

Quantifying chlorinated ethene degradation during reductive dechlorination at Kelly AFB using stable carbon isotopes

Stable isotope analysis of chlorinated ethene contaminants was carried out during a bioaugmentation pilot test at Kelly Air Force Base (AFB) in San Antonio Texas. In this pilot test, cis-1,2-dichloroethene (cDCE) was the primary volatile organic compound. A mixed microbial enrichment culture, KB-1, shown in laboratory experiments to reduce chlorinated ethenes to non-toxic ethene, was added to the pilot test area. Following bioaugmentation with KB-1, perchloroethene (PCE), trichloroethene (TCE) and cDCE concentrations declined, while vinyl chloride (VC) concentrations increased and subsequently decreased as ethene became the dominant transformation product. Shifts in carbon isotopic values up to 2.7 per thousand, 6.4 per thousand, 10.9 per thousand and 10.6 per thousand were observed for PCE, TCE, cDCE and VC, respectively, after bioaugmentation, consistent with the effects of biodegradation. While a rising trend of VC concentrations and the first appearance of ethene were indicative of biodegradation by 72 days post-bioaugmentation, the most compelling evidence of biodegradation was the substantial carbon isotope enrichment (2.0 per thousand to 5.0 per thousand) in ?13C(cDCE). Fractionation factors obtained in previous laboratory studies were used with isotope field measurements to estimate first-order cDCE degradation rate constants of 0.12 h(-1) and 0.17 h(-1) at 115 days post-bioaugmentation. These isotope-derived rate constants were clearly lower than, but within a factor of 2-4 of the previously published rate constant calculated in a parallel study at Kelly AFB using chlorinated ethene concentrations. Stable carbon isotopes can provide not only a sensitive means for early identification of the effects of biodegradation, but an additional means to quantify the rates of biodegradation in the field.     

Dechlorination of PCE in the presence of Fe0 enhanced by a mixed culture containing two Dehalococcoides strains

A mixed culture capable of supplying its energy requirements by the oxidation of zero-valent iron (Fe0) and concomitant reduction of chlorinated ethenes was established. The culture contained Dehalococcoides species as determined by polymerase chain reaction (PCR) with genus specific primers. The use of a newly designed ARDRA procedure and subsequent sequencing revealed the presence of two Dehalococcoides strains, one closely related to Dehalococcoides ethenogenes strain 195, a bacterium respiring with chlorinated ethenes, and one closely related to strain CBDB1 a chlorobenzene and dioxin dehalogenating anaerobe. The mixed culture was used to study dechlorination of tetrachloroethene (PCE) to ethene in the presence of Fe0. Whereas abiotic transformation of PCE by Fe0 led to incomplete dechlorination, the mixed culture mediated fast and complete dechlorination of PCE to ethene with Fe0 as electron donor. Compared to cultures with hydrogen added as electron donor, cultures with Fe0 as electron donor showed the same or higher rates of PCE dechlorination. Growth of the Dehalococcoides strains in the mixed culture is linked to the presence of Fe0 as electron donor and PCE as electron acceptor demonstrating that Dehalococcoides spp. play a pivotal role in the dechlorination of chlorinated ethenes in Fe0 systems.     

Enhancing polychlorinated biphenyl dechlorination in fresh water sediment with biostimulation and bioaugmentation

Polychlorinated biphenyls (PCBs) are toxic compounds ubiquitously distributed in ecosystems. Microbial attenuation of these contaminants is a potential means of remediation. Two promising microbial PCB remediation technologies, biostimulation and bioaugmentation, were investigated in different sediments. Biostimulation experiments in which electron donor was supplied (H2 via elemental iron, Fe(0)) resulted in only a marginal improvement in the dechlorination of amended 2,3,4,5-tetrachlorobiphenyl (2,3,4,5-CB), likely because of an inadequate population of indigenous H2-utilizing dechlorinators. Extensive dechlorination was observed, however, after bioaugmenting microcosms with a PCB-dechlorinating enrichment culture. Dechlorination of 2,3,4,5-CB began prior to the 20th day of incubation and proceeded to 2-chlorobiphenyl. This extensive dechlorination activity was maintained in both sediments over 70d at 10 and 25 degrees C. This research demonstrates that although past studies of biostimulation were promising, a great deal must be known about the PCB-dechlorinating organisms present before successful biostimulation is expected. Bioaugmentation, however, appears to be a promising PCB remediation strategy and should be further pursued.     

Dechlorination and organohalide-respiring bacteria dynamics in sediment samples of the Yangtze Three Gorges Reservoir

Several groups of bacteria such as Dehalococcoides spp., Dehalobacter spp., Desulfomonile spp., Desulfuromonas spp., or Desulfitobacterium spp. are able to dehalogenate chlorinated pollutants such as chloroethenes, chlorobenzenes, or polychlorinated biphenyls under anaerobic conditions. In order to assess the dechlorination potential in Yangtze sediment samples, the presence and activity of the reductively dechlorinating bacteria were studied in anaerobic batch tests. Eighteen sediment samples were taken in the Three Gorges Reservoir catchment area of the Yangtze River, including the tributaries Jialing River, Daning River, and Xiangxi River. Polymerase chain reaction analysis indicated the presence of dechlorinating bacteria in most samples, with varying dechlorinating microbial community compositions at different sampling locations. Subsequently, anaerobic reductive dechlorination of tetrachloroethene (PCE) was tested after the addition of electron donors. Most cultures dechlorinated PCE completely to ethene via cis-dichloroethene (cis-DCE) or trans-dichloroethene. Dehalogenating activity corresponded to increasing numbers of Dehalobacter spp., Desulfomonile spp., Desulfitobacterium spp., or Dehalococcoides spp. If no bacteria of the genus Dehalococcoides spp. were present in the sediment, reductive dechlorination stopped at cis-DCE. Our results demonstrate the presence of viable dechlorinating bacteria in Yangtze samples, indicating their relevance for pollutant turnover.     

Continuous-flow column study of reductive dehalogenation of PCE upon bioaugmentation with the Evanite enrichment culture

A continuous-flow anaerobic column experiment was conducted to evaluate the reductive dechlorination of tetrachloroethene (PCE) in Hanford aquifer material after bioaugmentation with the Evanite (EV) culture. An influent PCE concentration of 0.09 mM was transformed to vinyl chloride (VC) and ethene (ETH) within a hydraulic residence time of 1.3 days. The experimental breakthrough curves were described by the one-dimensional two-site-nonequilibrium transport model. PCE dechlorination was observed after bioaugmentation and after the lactate concentration was increased from 0.35 to 0.67 mM. At the onset of reductive dehalogenation, cis-dichloroethene (c-DCE) concentrations in the column effluent exceeded the influent PCE concentration indicating enhanced PCE desorption and transformation. When the lactate concentration was increased to 1.34 mM, c-DCE reduction to vinyl chloride (VC) and ethene (ETH) occurred. Spatial rates of PCE and VC transformation were determined in batch-incubated microcosms constructed with aquifer samples obtained from the column. PCE transformation rates were highest in the first 5 cm from the column inlet and decreased towards the column effluent. Dehalococcoides cell numbers dropped from approximately 73.5% of the total Bacterial population in the original inocula, to about 0.5% to 4% throughout the column. The results were consistent with estimates of electron donor utilization, with 4% going towards dehalogenation reactions.     

Anaerobic abiotic transformations of cis-1,2-dichloroethene in fractured sandstone

A fractured sandstone aquifer at an industrial site is contaminated with trichloroethene to depths greater than 244 m. Field data indicate that trichloroethene is undergoing reduction to cis-1,2-dichloroethene (cDCE); vinyl chloride and ethene are present at much lower concentrations. Transformation of cDCE by pathways other than reductive dechlorination (abiotic and/or biotic) is of interest. Pyrite, which has been linked to abiotic transformation of chlorinated ethenes, is present at varying levels in the sandstone. To evaluate the possible role of pyrite in transforming cDCE, microcosms were prepared with groundwater, ～40 mg L^?1 cDCE + [¹⁴C]cDCE, and crushed solids (pure pyrite, pyrite-rich sandstone, or typical sandstone). During 120 d of incubation, the highest level of cDCE transformation occurred with typical sandstone (11–14% ¹⁴CO₂, 1–3% ¹⁴C-soluble products), followed by pyrite-rich sandstone (2–4% ¹⁴CO₂, 1% ¹⁴C-soluble products) and even lesser amounts with pure pyrite. These results indicate pyrite is not likely the mineral involved in transforming cDCE. A separate experiment using only typical sandstone compared the rate of cDCE transformation in non-sterilized, autoclaved, and propylene-oxide sterilized treatments, with pseudo-first order rate constants of 8.7, 5.4, and 1.0 yr^?1, respectively; however, transformation stopped after several months of incubation. Autoclaving increased the volume of pores, adsorption pore diameter, and surface area in comparison to non-sterilized typical sandstone. Nevertheless, autoclaving was less disruptive than chemical sterilization. The results provide definitive experimental evidence that cDCE undergoes anaerobic abiotic and biotic transformation in typical sandstone, with formation of CO₂ and soluble products.     

Variability in microbial carbon isotope fractionation of tetra- and trichloroethene upon reductive dechlorination

The variability of stable carbon isotope fractionation upon reductive dechlorination of tetra- and trichloroethene by several microbial strains was investigated to examine the uncertainties related to the in situ application of compound specific isotope analysis (CSIA) of chlorinated ethenes. Carbon isotope fractionation was investigated with a set of microorganisms representative for the currently known diversity of dehalorespirers: Dehalococcoides ethenogenes strain 195, Desulfitobacterium sp. strain Viet1, Desulfuromonas michiganensis and Geobacter lovleyi sp. strain SZ and compared to the previous reports using Sulfurospirillum spp. and Desulfitobacterium sp. strain PCE-S. Carbon isotope fractionation of tetrachloroethene (PCE) and trichlorethene (TCE) was highly variable ranging from the absence of significant fractionation to carbon isotope fractionation (epsilonC) of 16.7 and 3.5-18.9 for PCE and TCE, respectively. Fractionation of both compounds by D. ethenogenes strain 195 (PCE: epsilonC=6.0; TCE: epsilonC=13.7) was similar to the literature data for mixed cultures containing Dehalococcoides spp. D. michiganensis (PCE: no significant fractionation; TCE: epsilonC=3.5) and G. lovleyi sp. strain SZ (PCE no significant fractionation; TCE: epsilonC=8.5) generated the lowest fractionation of all studied strains. Desulfitobacterium sp. strain Viet1 (PCE: epsilonC=16.7) gave the highest enrichment factor for PCE.     

Quantifying the effects of fumarate on in situ reductive dechlorination rates

In situ methods are needed to evaluate the effectiveness of chemical amendments at enhancing reductive dechlorination rates in groundwater that is contaminated with the priority pollutant, trichloroethene (TCE). In this communication, a method that utilizes single-well, “push–pull” tests to quantify the effects of chemical amendments on in situ reductive dechlorination rates is presented and demonstrated. Five push–pull tests were conducted in each of five monitoring wells located in a TCE-contaminated aquifer at the site of a former chemical manufacturing facility. Rates for the reductive dechlorination of the fluorinated TCE-surrogate, trichlorofluoroethene (TCFE), were measured before (test 1) and after (test 5) three successive additions (tests 2–4) of fumarate. Fumarate was selected to stimulate the growth and activity of indigenous microorganisms with the metabolic capability to reduce TCFE and TCE. In three wells, first-order rate constants for the reductive dechlorination of TCFE increased by 8.2–92 times following fumarate additions. In two wells, reductive dechlorination of TCFE was observed after fumarate additions but not before. The transformation behavior of fumarate was also monitored following each fumarate addition. Correlations between the reductive dechlorination of TCFE and the reduction of fumarate to succinate were observed, indicating that these reactions were supported by similar biogeochemical conditions at this site.