Reductive Dechlorination Activity Research Articles

Groundwater Bioremediation through Reductive Dechlorination in a Permeable Bioelectrochemical Reactor

A new membrane-less bioelectrochemical reactor configuration was developed for contaminated groundwater remediation. The new bioelectrochemical reactor configuration was inspired by the utilisation of a permeable reactive barrier (PBR) configuration with no separation membrane. The corresponding reactive zones were created by using graphite granules and mixed metal oxide (MMO) electrodes to stimulate the reductive and oxidative biological degradation of chlorinated aliphatic hydrocarbons. In the present study, the PBR-like bioelectrochemical reactor has been preliminarily operated with synthetic contaminated groundwater, testing the reductive dechlorination activity on cis-dichloroethylene (cisDCE). Moreover, to assess the effects of competing anions presence for the electron donor (i.e., the cathode), the synthetic wastewater contained sulphate and nitrate anions. In the PBR-like reactor operation, nearly all cisDCE was removed in the initial sampling port, with only VC detected as the observable RD product. During the same biotic test of the PRB reactor, the presence of both the reductive dechlorination and anions reduction was confirmed by the complete nitrate reduction in the cathodic chamber of the PRB reactor. On the contrary, sulphate reduction showed a lower activity; indeed, only 25% of the influent sulphate was removed by the PRB reactor.

Integrated Advanced Molecular Tools Predict In Situ cVOC Degradation Rates: Field Demonstration.

Chlorinated volatile organic compound (cVOC) degradation rate constants are crucial information for site management. Conventional approaches generate rate estimates from the monitoring and modeling of cVOC concentrations. This requires time series data collected along the flow path of the plume. The estimates of rate constants are often plagued by confounding issues, making predictions cumbersome and unreliable. Laboratory data suggest that targeted quantitative analysis of Dehalococcoides mccartyi (Dhc) biomarker genes (qPCR) and proteins (qProt) can be directly correlated with reductive dechlorination activity. To assess the potential of qPCR and qProt measurements to predict rates, we collected data from cVOC-contaminated aquifers. At the benchmark study site, the rate constant for degradation of cis-dichloroethene (cDCE) extracted from monitoring data was 11.0 ± 3.4 yr-1, and the rate constant predicted from the abundance of TceA peptides was 6.9 yr-1. The rate constant for degradation of vinyl chloride (VC) from monitoring data was 8.4 ± 5.7 yr-1, and the rate constant predicted from the abundance of TceA peptides was 5.2 yr-1. At the other study sites, the rate constants for cDCE degradation predicted from qPCR and qProt measurements agreed within a factor of 4. Under the right circumstances, qPCR and qProt measurements can be useful to rapidly predict rates of cDCE and VC biodegradation, providing a major advance in effective site management.

Insights into the influence on 2,4,6-trichlorophenol microbial reductive dechlorination process by exposure to microplastics

As a newly emerging pollutant, the adverse effects of ubiquitous microplastic on ecosystems and living have attracted increasing attention, however, the microplastic intervention influencing anaerobic biotransformation behaviors of chlorinated aromatics remains poorly understood. Here, the significantly altered 2,4,6-trichlorophenol microbial reductive dechlorination activities were found by single- and multi-exposure to microplastics. Single contact with five different types of microplastics showed diverse dechlorination performance, that PET was the most promoted ( k T of 0.037 h -1 ) and PP was the largest inhibited ( k T of 0.01 h -1 ) compared to control ( k T of 0.023 h -1 ). The microplastic size was negatively correlated with dechlorination activities, with 1.8~1.9 times decreased k T as the particle size decreased from 1000μm to 75μm. The higher the concentration, the more promoted effects by PET and the more inhibited effects by PP on dechlorination performance, with the respective k T of 0.052 h -1 and 0.006 h -1 under 2.0 g/L condition. Bio-enrichment on hydrophilic surfaces of microplastics (e.g. PET) was speculated to be responsible for the favored dechlorination activity, while adsorption of chlorophenols posed little influence. However, the overall inhibited 2,4,6-TCP dechlorination performance was found under all conditions by extending exposure cycles. The obvious bacterial succession that the abundance of some absolute anaerobic fermentator ( Dysgonomonas ) or dechlorinator ( Desulfitobacterium ) was greatly decreased, while, some facultative anaerobic genera ( Comamonas , Pseudomonas ) that held microplastic degradation potential were enriched. The increased positive correlations among networks were found and the more simplified molecule network as the size of microplastics became larger. The study provided significant theoretical support for understanding anaerobic biotransformation process of CAs influenced by the microplastic intervention. • Altered microbial reductive dechlorination activities by microplastic intervention • PET mostly promoted and PP largest inhibited the dechlorination by single contact • Microplastic size was negatively correlated with the dechlorination activities • Serious inhibited dechlorination performance was found by multi-cycle exposure • Obvious population succession and increased positive correlations of eco-networks

Effect of polyhydroxyalkanoates on the microbial reductive dechlorination of polychlorinated biphenyls and competing anaerobic respirations in a marine microbial culture

The effect of polyhydroxyalkanoates (PHAs) with different composition on the reductive dechlorination activity of a polychlorinated biphenyls (PCBs) dechlorinating marine microbial community and on the activity of sulfate-reducing (SRB) and methanogenic bacteria (MB), were investigated in marine sediment microcosms and compared with the main monomer, 3-hydroxybutyric acid (3HB). Despite PHAs were fermented more slowly than 3HB, all electron donors stimulated constantly sulfate-reduction, methanogenesis and, only transiently, PCB reductive dechlorination. No relevant differences were observed with different compositions of PHAs. According to electron balances, the majority of the supplied electrons (50 %) were consumed by SRB and to less extent by MB (9–31 %), while a small percentage (0.01 %) was delivered to OHRB. In the studied conditions PHAs were confirmed as potential slow‑hydrogen releasing compounds in marine environment but their fermentation rate was sufficiently high to mainly stimulate the competitors of organohalide respring bacteria for electron donors.

Dehalogenation of Chlorinated Ethenes to Ethene by a Novel Isolate, "Candidatus Dehalogenimonas etheniformans".

Dehalococcoides mccartyi strains harboring vinyl chloride (VC) reductive dehalogenase (RDase) genes are keystone bacteria for VC detoxification in groundwater aquifers, and bioremediation monitoring regimens focus on D. mccartyi biomarkers. We isolated a novel anaerobic bacterium, "Candidatus Dehalogenimonas etheniformans" strain GP, capable of respiratory dechlorination of VC to ethene. This bacterium couples formate and hydrogen (H2) oxidation to the reduction of trichloro-ethene (TCE), all dichloroethene (DCE) isomers, and VC with acetate as the carbon source. Cultures that received formate and H2 consumed the two electron donors concomitantly at similar rates. A 16S rRNA gene-targeted quantitative PCR (qPCR) assay measured growth yields of (1.2 ± 0.2) × 108 and (1.9 ± 0.2) × 108 cells per μmol of VC dechlorinated in cultures with H2 or formate as electron donor, respectively. About 1.5-fold higher cell numbers were measured with qPCR targeting cerA, a single-copy gene encoding a putative VC RDase. A VC dechlorination rate of 215 ± 40 μmol L-1 day-1 was measured at 30°C, with about 25% of this activity occurring at 15°C. Increasing NaCl concentrations progressively impacted VC dechlorination rates, and dechlorination ceased at 15 g NaCl L-1. During growth with TCE, all DCE isomers were intermediates. Tetrachloroethene was not dechlorinated and inhibited dechlorination of other chlorinated ethenes. Carbon monoxide formed and accumulated as a metabolic by-product in dechlorinating cultures and impacted reductive dechlorination activity. The isolation of a new Dehalogenimonas species able to effectively dechlorinate toxic chlorinated ethenes to benign ethene expands our understanding of the reductive dechlorination process, with implications for bioremediation and environmental monitoring. IMPORTANCE Chlorinated ethenes are risk drivers at many contaminated sites, and current bioremediation efforts focus on organohalide-respiring Dehalococcoides mccartyi strains to achieve detoxification. We isolated and characterized the first non-Dehalococcoides bacterium, "Candidatus Dehalogenimonas etheniformans" strain GP, capable of metabolic reductive dechlorination of TCE, all DCE isomers, and VC to environmentally benign ethene. In addition to hydrogen, the new isolate utilizes formate as electron donor for reductive dechlorination, providing opportunities for more effective electron donor delivery to the contaminated subsurface. The discovery that a broader microbial diversity can achieve detoxification of toxic chlorinated ethenes in anoxic aquifers illustrates the potential of naturally occurring microbes for biotechnological applications.

Microbial Chain Elongation and Subsequent Fermentation of Elongated Carboxylates as H2-Producing Processes for Sustained Reductive Dechlorination of Chlorinated Ethenes

In situ anaerobic groundwater bioremediation of trichloroethene (TCE) to nontoxic ethene is contingent on organohalide-respiring Dehalococcoidia, the most common strictly hydrogenotrophic Dehalococcoides mccartyi (D. mccartyi). The H2 requirement for D. mccartyi is fulfilled by adding various organic substrates (e.g., lactate, emulsified vegetable oil, and glucose/molasses), which require fermenting microorganisms to convert them to H2. The net flux of H2 is a crucial controlling parameter in the efficacy of bioremediation. H2 consumption by competing microorganisms (e.g., methanogens and homoacetogens) can diminish the rates of reductive dechlorination or stall the process altogether. Furthermore, some fermentation pathways do not produce H2 or having H2 as a product is not always thermodynamically favorable under environmental conditions. Here, we report on a novel application of microbial chain elongation as a H2-producing process for reductive dechlorination. In soil microcosms bioaugmented with dechlorinating and chain-elongating enrichment cultures, near stoichiometric conversion of TCE (0.07 ± 0.01, 0.60 ± 0.03, and 1.50 ± 0.20 mmol L-1 added sequentially) to ethene was achieved when initially stimulated by chain elongation of acetate and ethanol. Chain elongation initiated reductive dechlorination by liberating H2 in the conversion of acetate and ethanol to butyrate and caproate. Syntrophic fermentation of butyrate, a chain-elongation product, to H2 and acetate further sustained the reductive dechlorination activity. Methanogenesis was limited during TCE dechlorination in soil microcosms and absent in transfer cultures fed with chain-elongation substrates. This study provides critical fundamental knowledge toward the feasibility of chlorinated solvent bioremediation based on microbial chain elongation.

Water content as a primary parameter determines microbial reductive dechlorination activities in soil

Organohalide-respiring bacteria (OHRB) remove halogens from a variety of organohalides, which have been utilized for in situ remediation of different contaminated sites, e.g., groundwater, sediment and soil. Nonetheless, dehalogenation activities of OHRB and consequent remediation efficiencies can be synergistically affected by water content, soil type and inoculated/indigenous OHRB, which need to be disentangled to identify the key driving parameter and to elucidate the underlying mechanism. In this study, we investigated the impacts of water content (0–100%), soil type (laterite, brown soil and black soil) and inoculated OHRB (Dehalococcoides mccartyi CG1 and a river sediment culture) on reductive dechlorination of perchloroethene (PCE) and polychlorinated biphenyls (PCBs), as well as on associated microbial communities. Results suggested that the water content as a primary rate-limiting parameter governed dechlorination activities in environmental matrices, particularly in the soil, possibly through mediation of cell-to-organohalide mobility of OHRB. By contrast, interestingly, organohalide-dechlorinating microbial communities were predominantly clustered based on soil types, rather than water contents or inoculated OHRB. This study provided knowledge on the impacts of major parameters on OHRB-mediated reductive dechlorination in groundwater, sediment and soil for future optimization of in situ bioremediation of organohalides.

Impact of Fixed Nitrogen Availability on Dehalococcoides mccartyi Reductive Dechlorination Activity.

Biostimulation to promote reductive dechlorination is widely practiced, but the value of adding an exogenous nitrogen (N) source (e.g., NH4+) during treatment is unclear. This study investigates the effect of NH4+ availability on organohalide-respiring Dehalococcoides mccartyi (Dhc) growth and reductive dechlorination in enrichment cultures derived from groundwater (PW4) and river sediment (TC) impacted with chlorinated ethenes. In PW4 cultures, the addition of NH4+ increased cis-1,2-dichloroethene (cDCE)-to-ethene dechlorination rates about 5-fold (20.6 ± 1.6 versus 3.8 ± 0.5 μM Cl- d-1), and the total number of Dhc 16S rRNA gene copies were about 43-fold higher in incubations with NH4+ ((1.8 ± 0.9) × 108 mL-1) compared to incubations without NH4+ ((4.1 ± 0.8) × 107 mL-1). In TC cultures, NH4+ also stimulated cDCE-to-ethene dechlorination and Dhc growth. Quantitative polymerase chain reaction (qPCR) revealed that Cornell-type Dhc capable of N2 fixation dominated PW4 cultures without NH4+, but their relative abundance decreased in cultures with NH4+ amendment (i.e., 99 versus 54% of total Dhc). Pinellas-type Dhc incapable of N2 fixation were responsible for cDCE dechlorination in TC cultures, and diazotrophic community members met their fixed N requirement in the medium without NH4+. Responses to NH4+ were apparent at the community level, and N2-fixing bacterial populations increased in incubations without NH4+. Quantitative assessment of Dhc nitrogenase genes, transcripts, and proteomics data linked Cornell-type Dhc nifD and nifK expression with fixed N limitation. NH4+ additions also demonstrated positive effects on Dhc in situ dechlorination activity in the vicinity of well PW4. These findings demonstrate that biostimulation with NH4+ can enhance Dhc reductive dechlorination rates; however, a "do nothing" approach that relies on indigenous diazotrophs can achieve similar dechlorination end points and avoids the potential for stalled dechlorination due to inhibitory levels of NH4+ or transformation products (i.e., nitrous oxide).

Targeted detection of Dehalococcoides mccartyi microbial protein biomarkers as indicators of reductive dechlorination activity in contaminated groundwater

Dehalococcoides mccartyi (Dhc) bacterial strains expressing active reductive dehalogenase (RDase) enzymes play key roles in the transformation and detoxification of chlorinated pollutants, including chlorinated ethenes. Site monitoring regimes traditionally rely on qPCR to assess the presence of Dhc biomarker genes; however, this technique alone cannot directly inform about dechlorination activity. To supplement gene-centric approaches and provide a more reliable proxy for dechlorination activity, we sought to demonstrate a targeted proteomics approach that can characterize Dhc mediated dechlorination in groundwater contaminated with chlorinated ethenes. Targeted peptide selection was conducted in axenic cultures of Dhc strains 195, FL2, and BAV1. These experiments yielded 37 peptides from housekeeping and structural proteins (i.e., GroEL, EF-TU, rpL7/L2 and the S-layer), as well as proteins involved in the reductive dechlorination activity (i.e., FdhA, TceA, and BvcA). The application of targeted proteomics to a defined bacterial consortium and contaminated groundwater samples resulted in the detection of FdhA peptides, which revealed active dechlorination with Dhc strain-level resolution, and the detection of RDases peptides indicating specific reductive dechlorination steps. The results presented here show that targeted proteomics can be applied to groundwater samples and provide protein level information about Dhc dechlorination activity.

The lower base of corrinoid small molecules regulates reductive dehalogenase enzyme function in Dehalococcoides species

Key interactions between proteins and small molecules regulate numerous environmental reactions. Such is the case with reductive dehalogenases that dechlorinate toxic chlorinated solvents only in the presence of corrinoid small molecules. Previous studies have demonstrated that the lower bases attached to these small molecule cofactors affect the activity of reductive dehalogenases differently in corrinoid auxotrophic Dehalococcoides mccartyi strains, which prefer the 5,6‐dimethylbenzimidazole (DMB) lower base. In contrast, Geobacter sulfurreducens strain PCA produces a small molecule corrinoid with 5‐hydroxybenzimidazole (5‐OHBza) as the lower base, which fails to support dechlorination catalyzed by reductive dehalogenases. Here, we cloned and heterologously expressed a unique Geobacter lovleyi O‐methyltransferase gene implicated in the methylation of 5‐OHBza to 5‐methoxybenzimidazole (5‐OMeBza). Unlike 5‐OHBza, 5‐OMeBza does support D. mccartyi reductive dechlorination activity. We demonstrated using HPLC and LC‐MS that the G. sulfurreducens mutant expressed the O‐methyltransferase, as evidenced by the native 5‐OHBza‐cobamide being converted into the 5‐OMeBza‐cobamide. Next, we co‐cultured D. mccartyi and the recombinant G. sulfurreducens in the absence of exogenous vitamin B12, which supported D. mccartyi growth and the complete dechlorination of cis‐1,2‐dichloroethene to ethene. These studies demonstrated that the O‐methyltransferase catalyzes a critical step in lower base biosynthesis and provides a genetic basis for the observation that D. mccartyi reductive dechlorination activity occurs in co‐cultures with G. lovleyi but not with G. sulfurreducens. Our results emphasize the critical role of small molecule interactions directing protein function in the case of dechlorinating toxic chlorinated solvents.Support or Funding InformationThe authors acknowledge funding support from the NIH and SERDP.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Estimating bioaugmentation efficacy of TCE dechlorination using long-term field well-to-well tests in a highly recharged and TCE-contaminated aquifer

This study demonstrates a combined field method accurately assessing the extent of trichloroethylene (TCE) reductive dechlorination activity and the mass fraction of its by-products. A combined method of injecting a known concentration of 1,1,2-trichloro-2-fluoroethene (TCFE) as a TCE bio-surrogate and a data processing technique of forced mass balance (FMB), considering the sorption effect on the mass fraction of chloroethene was evaluated by performing soil column and field bioaugmentation tests. In the soil column test, the FMB resulted in the mass fraction of 6% TCE, 48.3% cis-1,2-dichloroethene, 18.5% vinyl chloride and 27.2% ethylene. In the field bioaugmentation test, TCFE showed equivalent dechlorination pathways of TCE. The mass fractions estimated by FMB were very similar to those observed in the soil column bioaugmentation tests: 4.5% TCFE, 57.1% 1,2-dichloro-1-fluoroethene, 12% 1-chloro-1-fluoroethene and 26.4% fluoroethene (FE). The FMB method gave ∼50% higher mass fraction for more chlorinated ethenes (i.e., TCFE) and ∼10% lower mass fraction of less chlorinated ethenes (i.e., FE) than those considering only the aqueous concentrations of chlorofluoroethenes. A combined method of TCFE and FMB that could accurately estimate both the extent of dechlorination activities and mass distribution of TCE reductive dechlorination would be highly useful.

Characterization of biochars derived from different materials and their effects on microbial dechlorination of pentachlorophenol in a consortium.

The properties of biochars derived from different raw materials (rice husk, bamboo, caragana, and garbage) and their effects on the microbial reductive dechlorination of pentachlorophenol (PCP) were investigated to understand how biochars influence the biotransformation of environmental pollutants. The results indicated that only caragana-derived biochar showed stable electron transfer activity for PCP dechlorination. Electro(chemical) analyses revealed that caragana biochar had the highest electrical conductivity (EC) (2.22 × 106 μS cm−1), while those of the other biochars were <1500 μS cm−1. The electron transfer capacities were within the ranges of 61.63–155.83 μmol e− g−1. Cyclic voltammetry analysis suggested that there were no obvious redox peaks for the biochars, while the Fourier transform infrared analysis showed similar transmission spectra with variable absorption intensity; this suggested that all biochars possessed similar structures and functional group classes and the enhancement of PCP dechlorination was not attributable to the redox reaction. Overall, the beneficial effects of caragana biochar on PCP dechlorination depended on the EC rather than the redox functional groups, possibly because high EC values enabled the highest electron transfer, and thus resulted in the greatest promotion of reductive dechlorination activity.

Impacts of low-temperature thermal treatment on microbial detoxification of tetrachloroethene under continuous flow conditions

Coupling in situ thermal treatment (ISTT) with microbial reductive dechlorination (MRD) has the potential to enhance contaminant degradation and reduce cleanup costs compared to conventional standalone remediation technologies. Impacts of low-temperature ISTT on Dehalococcoides mccartyi (Dhc), a relevant species in the anaerobic degradation of cis-1,2-dichloroethene (cis-DCE) and vinyl chloride (VC) to nontoxic ethene, were assessed in sand-packed columns under dynamic flow conditions. Dissolved tetrachloroethene (PCE; 258 ± 46 μM) was introduced to identical columns bioaugmented with the PCE-to-ethene dechlorinating consortium KB-1®. Initial column temperatures represented a typical aquifer (15 °C) or a site undergoing low-temperature ISTT (35 °C), and were subsequently increased to 35 and 74 °C, respectively, to assess temperature impacts on reductive dechlorination activity. In the 15 °C column, PCE was transformed primarily to cis-DCE (159 ± 2 μM), which was further degraded to VC (164 ± 3 μM) and ethene (30 ± 0 μM) within 17 pore volumes (PVs) after the temperature was increased to 35 °C. Regardless of the initial column temperature, ethene constituted >50 mol% of effluent degradation products in both columns after 73–74 PVs at 35 °C, indicating that MRD performance was greatly improved under low-temperature ISTT conditions. Increasing the temperature of the column initially at 35 °C resulted in continued VC and ethene production until a temperature of approximately 43 °C was reached, at which point Dhc activity substantially decreased. The abundance of the vcrA reductive dehalogenase gene exceeded that of the bvcA gene by 1–2.5 orders of magnitude at 15 °C, but this relationship inversed at temperatures >35 °C, suggesting Dhc strain-specific responses to temperature. These findings demonstrate improved MRD performance with low-temperature thermal treatment and emphasize potential synergistic effects at sites undergoing ISTT.

Resilience and recovery of Dehalococcoides mccartyi following low pH exposure.

Bioremediation treatment (e.g. biostimulation) can decrease groundwater pH with consequences for Dehalococcoides mccartyi (Dhc) reductive dechlorination activity. To explore the pH resilience of Dhc, the Dhc-containing consortium BDI was exposed to pH 5.5 for up to 40 days. Following 8- and 16-day exposure periods to pH 5.5, dechlorination activity and growth recovered when returned to pH 7.2; however, the ability of the culture to dechlorinate vinyl chloride (VC) to ethene was impaired (i.e. decreased rate of VC transformation). Dhc cells exposed to pH 5.5 for 40 days did not recover the ethene-producing phenotype upon transfer to pH 7.2 even after 200 days of incubation. When returned to pH 7.2 conditions after an 8-, a 16- and a 40-day low pH exposure, tceA and vcrA genes showed distinct fold increases, suggesting Dhc strain-specific responses to low pH exposure. Furthermore, a survey of Dhc biomarker genes in groundwater samples revealed the average abundances of Dhc 16S rRNA, tceA and vcrA genes in pH 4.5-6 groundwater were significantly lower (P-value < 0.05) than in pH 6-8.3 groundwater. Overall, the results of the laboratory study and the assessment of field data demonstrate that sustained Dhc activity should not be expected in low pH groundwater, and the duration of low pH exposure affects the ability of Dhc to recover activity at circumneutral pH.

A Data Mining Approach to Predict In Situ Detoxification Potential of Chlorinated Ethenes.

Despite advances in physicochemical remediation technologies, in situ bioremediation treatment based on Dehalococcoides mccartyi (Dhc) reductive dechlorination activity remains a cornerstone approach to remedy sites impacted with chlorinated ethenes. Selecting the best remedial strategy is challenging due to uncertainties and complexity associated with biological and geochemical factors influencing Dhc activity. Guidelines based on measurable biogeochemical parameters have been proposed, but contemporary efforts fall short of meaningfully integrating the available information. Extensive groundwater monitoring data sets have been collected for decades, but have not been systematically analyzed and used for developing tools to guide decision-making. In the present study, geochemical and microbial data sets collected from 35 wells at five contaminated sites were used to demonstrate that a data mining prediction model using the classification and regression tree (CART) algorithm can provide improved predictive understanding of a site's reductive dechlorination potential. The CART model successfully predicted the 3-month-ahead reductive dechlorination potential with 75.8% and 69.5% true positive rate (i.e., sensitivity) for the training set and the test set, respectively. The machine learning algorithm ranked parameters by relative importance for assessing in situ reductive dechlorination potential. The abundance of Dhc 16S rRNA genes, CH4, Fe(2+), NO3(-), NO2(-), and SO4(2-) concentrations, total organic carbon (TOC) amounts, and oxidation-reduction potential (ORP) displayed significant correlations (p < 0.01) with dechlorination potential, with NO3(-), NO2(-), and Fe(2+) concentrations exhibiting precedence over other parameters. Contrary to prior efforts, the power of data mining approaches lies in the ability to discern synergetic effects between multiple parameters that affect reductive dechlorination activity. Overall, these findings demonstrate that data mining techniques (e.g., machine learning algorithms) effectively utilize groundwater monitoring data to derive predictive understanding of contaminant degradation, and thus have great potential for improving decision-making tools. A major need for realizing the predictive capabilities of data mining approaches is a curated, open-access, up-to-date and comprehensive collection of biogeochemical groundwater monitoring data.

Catalytic hydrodechlorination of monochloroacetic acid in wastewater using Ni-Fe bimetal prepared by ball milling.

Monochloroacetic acid (MCA) is a chemically stable and biologically toxic pollutant. It is often generated during the production of the pesticide dimethoate. Conventional wastewater treatment processes have difficulty degrading it. In this work, the dechlorination effects of Ni-Fe bimetal prepared using ball milling (BM) technology for the high concentrations of MCA in wastewater were examined. The MCA in aqueous solution was found to be degraded efficiently by the Ni-Fe bimetal. However, S-(methoxycarbonyl) methyl O, O-dimethyl phosphorodithioate (SMOPD) in wastewater, a by-product of the dimethoate production process, significantly inhibited the reductive dechlorination activity of Ni-Fe bimetal. Increasing the reaction temperature in the MCA wastewater enhanced the reduction activity of the Ni-Fe bimetal effectively. Oxygen was found to be unfavorable to dechlorination. Sealing the reaction to prevent oxidation was found to render the degradation process more efficient. The process retained over 88% efficiency after 10 treatment cycles with 50 g/L of Ni-Fe bimetal under field conditions.

Bench-scale evaluation of aerosol delivery for biostimulation and bioaugmentation in the vadose zone.

Aerosol delivery was evaluated for distributing biostimulation and bioaugmentation amendments in vadose zones. This technique involves transporting amendments as micron-scale aerosol droplets in injected gas. Microcosm experiments were designed to characterize reductive dechlorination of trichloroethene (TCE) under unsaturated conditions when delivering components as aerosols. Delivering amendments and/or microbes as aqueous aerosols resulted in complete dechlorination of TCE, similar to controls operated under saturated conditions. Reductive dechlorination was achieved with manual injection of a bioaugmentation culture suspended in soybean oil into microcosms. However, aerosol delivery of the culture in soybean oil induced little reductive dechlorination activity. Overall, the results indicate that delivery as aqueous aerosols may be a viable option for delivery of amendments to enhance vadose zone bioremediation at the field-scale.

Identification of a reductive tetrachloroethene dehalogenase in Shewanella sediminis.

The genome sequence of psychrophilic Shewanella sediminis revealed the presence of five putative reductive dehalogenases (Rdhs). We found that cell extracts of pyruvate/fumarate-grown S. sediminis cells catalysed reduced methyl viologen-dependent reductive dechlorination of tetrachloroethene (PCE) to trichloroethene (TCE) at a specific activity of approximately 1 nmol TCE min(-1) (mg protein)(-1). Dechlorination of PCE followed Michaelis-Menten kinetics with an apparent Km of 120 μM PCE. No PCE dechlorination was observed with heat-denatured extract or when cyanocobalamin was omitted from the growth medium; however, the presence of PCE in the growth medium increased PCE transformation rates. Analysis of mutants carrying in-frame deletions of all five Rdhs encoding genes showed that only deletion of Ssed_3769 resulted in the loss of PCE dechlorination activity suggesting that Ssed_3769 is a functional Rdh. This is the first study to show reductive dechlorination activity of PCE in a sediment-dwelling Shewanella species that may be important for linking the flux of organohalogens to organic carbon via reductive dehalogenation in marine sediments.

Unexpected specificity of interspecies cobamide transfer from Geobacter spp. to organohalide-respiring Dehalococcoides mccartyi strains.

Dehalococcoides mccartyi strains conserve energy from reductive dechlorination reactions catalyzed by corrinoid-dependent reductive dehalogenase enzyme systems. Dehalococcoides lacks the ability for de novo corrinoid synthesis, and pure cultures require the addition of cyanocobalamin (vitamin B(12)) for growth. In contrast, Geobacter lovleyi, which dechlorinates tetrachloroethene to cis-1,2-dichloroethene (cis-DCE), and the nondechlorinating species Geobacter sulfurreducens have complete sets of cobamide biosynthesis genes and produced 12.9 ± 2.4 and 24.2 ± 5.8 ng of extracellular cobamide per liter of culture suspension, respectively, during growth with acetate and fumarate in a completely synthetic medium. G. lovleyi-D. mccartyi strain BAV1 or strain FL2 cocultures provided evidence for interspecies corrinoid transfer, and cis-DCE was dechlorinated to vinyl chloride and ethene concomitant with Dehalococcoides growth. In contrast, negligible increase in Dehalococcoides 16S rRNA gene copies and insignificant dechlorination occurred in G. sulfurreducens-D. mccartyi strain BAV1 or strain FL2 cocultures. Apparently, G. lovleyi produces a cobamide that complements Dehalococcoides' nutritional requirements, whereas G. sulfurreducens does not. Interestingly, Dehalococcoides dechlorination activity and growth could be restored in G. sulfurreducens-Dehalococcoides cocultures by adding 10 μM 5',6'-dimethylbenzimidazole. Observations made with the G. sulfurreducens-Dehalococcoides cocultures suggest that the exchange of the lower ligand generated a cobalamin, which supported Dehalococcoides activity. These findings have implications for in situ bioremediation and suggest that the corrinoid metabolism of Dehalococcoides must be understood to faithfully predict, and possibly enhance, reductive dechlorination activities.

Electron donor availability for microbial reductive processes following thermal treatment

Thermal treatment is capable of removing significant free-phase chlorinated solvent mass while potentially enhancing bioremediation effectiveness by establishing temperature gradients in the perimeter of the source zone and by increasing electron donor availability. The objectives of this study were to determine the potential for enhanced reductive dechlorination activity at the intermediate temperatures that establish in the perimeter of the heated source zone, and to evaluate the effect of electron donor competition on the performance of the microbial reductive dechlorination process. Microcosms, constructed with tetrachloroethene- (PCE-) and trichloroethene- (TCE-) impacted soils from the Great Lakes, IL, and Ft. Lewis, WA, sites were incubated at temperatures of 24, 35, 50, 70, and 95 °C for 4 months. Reductive dechlorination did not occur in microcosms incubated at temperatures above 24 °C even though mesophilic PCE-to- cis-1,2-dichloroethene dechlorinators were present in Ft. Lewis soil suggesting electron donor limitations. Five days after cooling the microcosms to 24 °C and bioaugmentation with the methanogenic, PCE-to-ethene-dechlorinating consortium OW, at least 85% of the initial PCE and TCE were dechlorinated, but dechlorination ceased prior to complete conversion to ethene. Subsequent biostimulation with hydrogen gas mitigated the dechlorination stall, and conversion to ethene resumed. The results of this study demonstrated that temperatures >35 °C inhibit reductive dechlorination activity at the Great Lakes and Ft. Lewis sites, and that the majority of reducing equivalents released from the soil matrix during heat treatment are consumed in methanogenesis rather than reductive dechlorination. These observations suggest that bioaugmenting thermal treatment sites with cultures that do not contain methanogens may allow practitioners to realize enhanced dechlorination activity, a potential benefit of coupling thermal treatment with bioremediation.