Search

Trichloroethylene (TCE) is a widespread environmental contaminant following decades of use as an industrial solvent, improper disposal, and remediation challenges. Consequently, TCE exposure continues to constitute a risk to human health. Despite epidemiological evidence associating exposure with adverse birth outcomes, the effects of TCE and its metabolite S -(1, 2-dichlorovinyl)-L-cysteine (DCVC) on the placenta remain undetermined. Flexible and efficient macronutrient and energy metabolism pathway utilization is essential for placental cell physiological adaptability. Because DCVC is known to compromise cellular energy status and disrupt energy metabolism in renal proximal tubular cells, this study investigated the effects of DCVC on cellular energy status and energy metabolism pathways in placental cells. Human extravillous trophoblast cells, HTR-8/SVneo, were exposed to 5-20 μM DCVC for 6 or 12 h. After establishing concentration and exposure duration thresholds for DCVC-induced cytotoxicity, targeted metabolomics was used to evaluate overall energy status and metabolite concentrations from energy metabolism pathways. The data revealed glucose metabolism perturbations including a time-dependent accumulation of glucose-6-phosphate+frutose-6-phosphate (G6P+F6P) as well as independent shunting of glucose intermediates that diminished with time, with modest energy status decline but in the absence of significant changes in ATP concentrations. Furthermore, metabolic profiling suggested that DCVC stimulated compensatory utilization of glycerol, lipid, and amino acid metabolism to provide intermediate substrates entering downstream in the glycolytic pathway or the tricarboxylic acid cycle. Lastly, amino acid deprivation increased susceptibility to DCVC-induced cytotoxicity. Taken together, these results suggest that DCVC caused metabolic perturbations necessitating adaptations in macronutrient and energy metabolism pathway utilization to maintain adequate ATP levels.

This study evaluated the biological removal of trichloroethylene (TCE) by Rhodococcus opacus using airlift bioreactor under continuous operation mode. The effect of inlet TCE concentration in the range 0.12-2.34 g m -3 on TCE removal has studied for 55 days. During the continuous bioreactor operation, a maximum of 96% TCE removal was obtained for low inlet TCE concentration, whereas the highest elimination capacity was 151.2 g m -3 h -1 for the TCE loading rate of 175.0 g m -3 h -1 . The carbon dioxide (CO 2 ) concentration profile from the airlift bioreactor revealed that the degraded TCE has primarily converted to CO 2 with a fraction of organic carbon utilized for bacterial growth. The artificial neural network (ANN) based model was able to successfully predict the performance of the bioreactor system using the Levenberg-Marquardt (LM) back propagation algorithm, and optimized biological topology is 3:12:1. The prediction accuracy of the model was high as the experimental data were in good agreement (R 2  = 0.9923) with the ANN predicted data. Overall, from the bioreactor experiments and its ANN modeling, the potential strength of R. opacus in TCE biodegradation is proved., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest between authors or any institute or funding agencies., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In this study, the YH consortium, an ethene-producing culture, was used to evaluate the effect of vitamin B 12 (VB 12 ) on trichloroethene (TCE) dechlorination by transferring the original TCE-reducing culture with or without adding exogenous VB 12 . Ultra-high performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS) was applied to detect the concentrations of VB 12 and its lower ligand 5,6-dimethylbenzimidazole (DMB) in the cultures. After three successive VB 12 starvation cycles, the dechlorination of TCE stopped mostly at cis-dichloroethene (cDCE), and no ethene was found; methane production increased significantly, and no VB 12 was detected. Results suggest that the co-cultured microbes may not be able to provide enough VB 12 as a cofactor for the growth of Dehalococcoides in the YH culture, possibly due to the competition for corrinoids between Dehalococcoides and methanogens. The relative abundances of 16 S rRNA gene of Dehalococcoides and reductive dehalogenase genes tceA or vcrA were lower in the cultures without VB 12 compared with the cultures with VB 12 . VB 12 limitation changed the microbial community structures of the consortia. In the absence of VB 12 , the microbial community shifted from dominance of Chloroflexi to Proteobacteria after three consecutive VB 12 starvation cycles, and the dechlorinating genus Dehalococcoides declined from 42.9% to 13.5%. In addition, Geobacter, Clostridium, and Desulfovibrio were also present in the cultures without VB 12 . Furthermore, the abundance of archaea increased under VB 12 limited conditions. Methanobacterium and Methanosarcina were the predominant archaea in the culture without VB 12 ., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Identifying co-metabolic TCE oxidation in polluted groundwater is challenging due to lack of indicative by-products. This challenge may theoretically be resolved if the oxidation process can be characterized by a distinct dual isotope enrichment. In this work, we aimed to explore the carbon and chlorine isotope effects associated with TCE oxidation by a variety of oxygenases. These included pure strains and enrichment cultures of methane, toluene and ammonia oxidizers, as well as experiments with crude extracts. Isotope effects determined for TCE oxidation by toluene and ammonia oxidizers were mostly in line with expected values for epoxidation mechanism (ϵ 13 C -11.0 ± 0.7 to -24.8 ± 0.2‰ and ϵ 37 Cl +0.9 ± 0.5 to +1.0 ± 0.4‰), whereas, the methanotrophs resulted in distinctively different isotope effects (ϵ 13 C -2.4 ± 0.4 to -3.4 ± 0.8‰ and ϵ 37 Cl -1.8 ± 0.2 to -2.9 ± 0.9‰). It is suggested that in TCE oxidation by methanotrophs, substrate binding rather than bond cleavage is rate limiting, leading to this unexpected isotope effect. On the environmental level, our results imply that the oxidative process can be differentiated if catalyzed by toluene and ammonia oxidizers or by methanotrophs. Additionally, the oxidative process can be distinguished from the reductive one. However, using dual isotope analysis in the field may result in an under-estimation of the overall co-metabolic process if methanotrophs are to be excluded due to low isotope effects., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Electrochemical removal of chlorinated ethenes in groundwater plumes may potentially overcome some of the challenges faced by current remediation technologies. So far, studies have been conducted in simplified settings of synthetic groundwater and inert porous matrices. This study is a stepwise investigation of the influence of field-extracted groundwater, sandy sediment and groundwater aquifer temperatures on the removal of an aged partially degraded contamination of tetrachloroethylene (PCE) at a typical groundwater flow rate. The aim is to assess the potential for applying electrochemistry at contaminated sites. At a constant current of 120 mA, pH and conductivity were unaffected downgradient the electrochemical zone. Major groundwater species were reduced and oxidized. Some minerals deposited, others dissolved. Hydrogen peroxide, a strong oxidant, was formed in levels up to 5 mg L -1 with a limited distribution into the sandy sediment. Trichloromethane was formed, supposedly by oxidation of organic matter in the sandy sediment in the presence of chloride. The more realistic the settings, the higher the PCE removal, bringing concentrations down to 7.8 ± 2.3 μg L -1 . A complete removal of trichloroethylene and cis-1,2-dichloroethylene was obtained. The results suggest that competing reactions related to the natural complex hydrogeochemistry are insignificant in terms of affecting the electrochemical degradation of PCE and chlorinated intermediates., Competing Interests: Declaration of competing interest None., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Different pH conditions have been demonstrated to affect the activities of dechlorinating populations participating in the successive dechlorination of trichloroethylene to ethylene. However, the mechanism of the effect of pH conditions on the assembly of dechlorinating populations and their relations to the structure, function, and dynamics of the microbiome are unclear. In this study, we evaluated the effects of pH on microbiomes assembled in anaerobic trichloroethylene-dechlorinating reactors under neutral (pH 7.2), acidic (pH 6.2), and alkaline (pH 8.2) conditions. The results revealed that among the reactors, the acidic reactor had the highest efficiency for dechlorination without accumulation of dechlorinated metabolites, even at high loading rates. The results of high-throughput sequencing of the 16S rRNA gene indicated that the microbiomes in the 3 reactors underwent varied dynamic succession. The acidic reactor harbored a higher degree of complex microbes, dechlorinator diversity, and abundance of the Victoria subgroup of Dehalococcoides (1.2 ± 0.1 × 10 6  cell/mL), which were approximately 10-10 2 -fold higher than those at neutral and alkaline conditions. The pH settings altered species-species connectivity and complexity of microbial interaction networks, with more commensal interactions in the dechlorinators of the acidic reactor. As predicted, abundances of several functional gene categories were in strong linearity with pH values, and the microbiome possessed significantly more abundant functions in the acidic reactor (P < 0.001), such as potentially stimulating hydrogen production, cobalamin synthesis, cobalt transport, transport and metabolism of amino acids and secondary metabolites, cell motility, and transcription. All results of microbiomic analyses consistently revealed the observed superior dechlorination process and suggested an association of the reductive dechlorination process with the pH-dependent microbiome. The results of this study provide a new insight into the trichloroethylene dechlorination with regards to pH, and they will be useful for improving bioremediation and management of trichloroethylene-contaminated sites., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Deteriorated environmental conditions during the bioremediation of trichloroethene (TCE)-polluted groundwater cause decreased treatment efficiencies. This study assessed the effect of applying immobilized Clostridium butyricum (a hydrogen-producing bacterium) in silica gel on enhancing the reductive dechlorination efficiency of TCE with the slow polycolloid-releasing substrate (SPRS) supplement in groundwater. The responses of microbial communities with the immobilized system (immobilized Clostridium butyricum and SPRS amendments) were also characterized by the metagenomics assay. A complete TCE removal in microcosms was obtained within 30 days with the application of this immobilized system via reductive dechlorination processes. An increase in the population of Dehalococcoides spp. was observed using the quantitative polymerase chain reaction (qPCR) analysis. Results of metagenomics assay reveal that the microbial communities in the immobilized system were distinct from those in systems with SPRS only. Bacterial communities associated with TCE biodegradation also increased in microcosms treated with the immobilized system. The immobilized system shows a great potential to promote the TCE dechlorination efficiency, and the metagenomics-based approach provides detailed insights into dechlorinating microbial community dynamics. The results would be helpful in designing an in situ immobilized system to enhance the bioremediation efficiency of TCE-contaminated groundwater., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Dehalococcoides is able to completely dehalogenate tetrachloroethene (PCE) and trichloroethene (TCE) to ethene (ETH). However, the dechlorination efficiency of Dehalococcoides is low and result in the accumulation of toxic intermediates. In this study, Shewanella oneidensis MR-1 (S. oneidensis MR-1) was added to the Dehalococcoides-containing culture and the complete TCE to ETH dechlorination was shortened from 24 days to 16 days. Dehalococcoides-targeted 16S rRNA gene and two model reductive dehalogenase (RDase) genes (tceA and vcrA), responsible for dechlorinating TCE to vinyl chloride (VC) and VC to ETH respectively, were characterized. Results showed that S. oneidensis MR-1 has no effect on the cell growth while the RDase genes expression was up-regulated and the RDase activity of Dehalococcoides was elevated. The mRNA abundance of vcrA increased approximately tenfold along with the increased concentration of vitamin B 12 (cyanocobalamin). Interestingly, the addition of S. oneidensis MR-1 increased the concentration of vitamin B 12 by affecting the microbial community structure. Therefore, the addition of S. oneidensis MR-1 might have a positive effect on regulating the activity of RDase of functional microorganisms and uptake of vitamin B 12 , and further provided a practical vision of chloroethene dechlorination by the Dehalococcoides-containing culture., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

ISCO using activated sodium persulphate is a widely used technology for treating chlorinated solvent source zones. In sensitive areas, however, high groundwater sulphate concentrations following treatment may be a drawback. In situ biogeochemical transformation, a technology that degrades contaminants via reduced iron minerals formed by microbial activity, offers a potential solution for such sites, the bioreduction of sulphate and production of iron sulphides that abiotically degrade chlorinated ethenes acting as a secondary technology following ISCO. This study assesses this approach in the field using hydrochemical and molecular tools, solid phase analysis and geochemical modelling. Following a neutralisation and bioaugmentation, favourable conditions for iron- and sulphate-reducers were created, resulting in a remarkable increase in their relative abundance. The abundance of dechlorinating bacteria (Dehalococcoides mccartyi, Dehalobacter sp. and Desulfitobacterium spp.) remained low throughout this process. The activity of iron- and sulphate-reducers was further stimulated through application of magnetite plus starch and microiron plus starch, resulting in an increase in ferrous iron concentration (from

The effectiveness of using gamma poly-glutamic acid (γ-PGA) as the primary carbon and nitrogen sources to bioremediate trichloroethene (TCE)-contaminated groundwater was studied in this pilot-scale study. γ-PGA (40 L) solution was injected into the aquifer via the injection well (IW) for substrate supplement. Groundwater samples were collected from monitor wells and IW and analyzed for TCE and its byproducts, geochemical indicators, dechlorinating bacteria, and microbial diversity periodically. Injected γ-PGA resulted in an increase in total organic carbon (TOC) (up to 9820 mg/L in IW), and the TOC biodegradation caused the formation of anaerobic conditions. Increased ammonia concentration (because of amine release from γ-PGA) resulted in the neutral condition in groundwater, which benefited the growth of Dehalococcoides. The negative zeta potential and micro-scale diameter of γ-PGA allowed its globule to distribute evenly within soil pores. Up to 93% of TCE removal was observed (TCE dropped from 0.14 to 0.01 mg/L) after 59 days of γ-PGA injection, and TCE dechlorination byproducts were also biodegraded subsequently. Next generation sequence (NGS) analyses were applied to determine the dominant bacterial communities. γ-PGA supplement developed reductive dechlorinating conditions and caused variations in microbial diversity and dominant bacterial species. The dominant four groups of bacterial communities including dechlorinating bacteria, vinyl chloride degrading bacteria, hydrogen producing bacteria, and carbon biodegrading bacteria., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Trichloroethylene is an industrial solvent and common environmental pollutant. Despite efforts to ban trichloroethylene, its availability and usage persist globally, constituting a hazard to human health. Recent studies reported associations between maternal trichloroethylene exposure and increased risk for low birth weight. Despite these associations, the toxicological mechanism underlying trichloroethylene adverse effects on pregnancy remains largely unknown. The trichloroethylene metabolite S-(1,2-dichlorovinyl)-L-cysteine (DCVC) induces mitochondrial-mediated apoptosis in a trophoblast cell line. To gain further understanding of mitochondrial-mediated DCVC placental toxicity, this study investigated the effects of DCVC exposure on mitochondrial function using non-cytolethal concentrations in placental cells. Human trophoblasts, HTR-8/SVneo, were exposed in vitro to a maximum of 20 μM DCVC for up to 12 h. Cell-based oxygen consumption and extracellular acidification assays were used to evaluate key aspects of mitochondrial function. Following 6 h of exposure to 20 μM DCVC, elevated oxygen consumption, mitochondrial proton leak and sustained energy coupling deficiency were observed. Similarly, 12 h of exposure to 20 μM DCVC decreased mitochondrial-dependent basal, ATP-linked and maximum oxygen consumption rates. Using the fluorochrome TMRE, dissipation of mitochondrial membrane potential was detected after a 12-h exposure to 20 μM DCVC, and (±)-α-tocopherol, a known suppressor of lipid peroxidation, attenuated DCVC-stimulated mitochondrial membrane depolarization but failed to rescue oxygen consumption perturbations. Together, these results suggest that DCVC caused progressive mitochondrial dysfunction, resulting in lipid peroxidation-associated mitochondrial membrane depolarization. Our findings contribute to the biological plausibility of DCVC-induced placental impairment and provide new insights into the role of the mitochondria in DCVC-induced toxicity., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Parkinson's disease (PD) is characterised pathologically by degeneration of the dopaminergic (DA) neurones of the substantia nigra pars compacta (SNpc) and the presence of α-synuclein containing Lewy body inclusions. Trichloroethylene (TCE) has been suggested as a potential environmental chemical that may contribute to the development of PD, via conversion to the neurotoxin, 1-Trichloromethyl-1,2,3,4-tetrahydro-β-carboline (TaClo). We investigated the effect of an 8 week exposure to TCE or TaClo on wild type and, as an experimental model of PD, A30P mutant α-synuclein overexpressing mice using a combination of behaviour and pathology. TCE or TaClo exposure caused significant DA neuronal loss within the SNpc in both wild type and transgenic mice. Cell numbers were lower in A30P animals than wild type, however, no additive effect of TCE or TaClo exposure and A30P overexpression was found. TCE or TaClo did not appear to lead to acceleration of motor or cognitive deficits in either wild type or A30P mutant mice, potentially because of the modest reductions of DA neuronal number in the SNpc. Our results do however suggest that TCE exposure could be a possible factor in development of PD like changes following exposure., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Trichloroethylene (TCE) was negative for developmental toxicity after inhalation and oral gavage exposure of pregnant rats but fetal cardiac defects were reported following drinking water exposure throughout gestation. Because of the deficiencies in this latter study, we performed another drinking water study to evaluate whether TCE causes heart defects., Methods: Groups of 25 mated Sprague Dawley rats consumed water containing 0, 0.25, 1.5, 500, or 1,000 ppm TCE from gestational day 1-21. TCE concentrations were measured at daily formulation, when placed into water bottles each day and when water bottles were removed from cages. Four additional mated rats per group were used for plasma measurements. At termination, fetal hearts were carefully dissected fresh and examined., Results: All TCE concentrations were >90% of target when initially placed in water bottles and when bottles were placed on cages. All dams survived with no clinical signs. Rats in the two higher dose groups consumed less water/day than other groups but showed no changes in maternal or fetal weights. The only fetal cardiac observation was small (<1 mm) membranous ventricular septal defect occurring in all treated and water control groups; incidences were within the range of published findings for naive animals. TCE was not detected in maternal blood, but systemic exposure was confirmed by detecting its primary oxidative metabolite, trichloroacetic acid, although only at levels above the quantitation limit in the two higher dose groups., Conclusions: Ingesting TCE in drinking water ≤1,000 ppm throughout gestation does not cause cardiac defects in rat offspring., (© 2019 The Authors. Birth Defects Research published by Wiley Periodicals, Inc.)

Trichloroethylene (TCE) is one of the most ubiquitous halogenated organic compounds of concerns of carcinogens in groundwater in Taiwan. Bioremediation has been recognized as a cost-effective approach in reducing TCE concentration. Five pilot-scale wells were constructed to monitor TCE concentrations in contaminated groundwater. With injection of EOS ® , TCE was effectively degraded to 42%-93% by the end of 175 days. The biostimulation with EOS ® was useful in establishing a micro-site anaerobic but with limited contribution. Dilution of the aquifer movement also caused the TCE reduction among injection and monitoring wells. The degradability was affected by the location and the proximity from the injection well. TCE concentrations found to be negatively correlated with the associated Dehalococcoides spp. and functional genes levels. Dhc concentration of 10 8 copies L -1 caused the initial 40% of TCE degradation. The well with the optimal degradation owned tceA of 10 9  cells L -1 . T-RFLP results indicate the wells with the superior TCE degradability also performed the highest Shannon index number (means the highest diversity), which occurred on the same day that Dhc levels started to enlarge. Desulfovibrio desulfuricans and Desulfuromonas chloroethenica were predominant species identified in the T-RFLP fingerprint profile. In brief, a variety of different factors including well locations, geochemical indicators, and microbial contribution were useful to explain the site-specific optimal TCE remediation approach. The consistence among TCE degradation, Dhc growing pattern, functional gene levels, and the dynamics of the microbial community structure present the novelty of this study., (Copyright © 2019. Published by Elsevier Ltd.)

Subsurface environments hold the largest reservoir of microbes in the biosphere. They play essential roles in transforming nutrients, degrading contaminants and recycling organic matter. Here, we propose a previously unrecognised fundamental microbial process that influences aquifer bioremediation dynamics and that applies to all microbial communities. In contrast to previous models, our proposed Piggyback-the-Persistent (PtP) mechanism occurs when viruses become more dominated by those exhibiting temperate rather than lytic lifestyles driven by persistent chemicals (in our case chlorinated-hydrocarbon pollutants) that provide long-term carbon sources and that refocus the aquifer carbon cycle, thus altering the microbial community. In this ultra-oligotrophic system, the virus:microbial ratio (VMR) ranges from below the detection limit of 0.0001 to 0.6, well below the common aquatic range of 3-10. Shortest-average-path network analysis revealed VMR and trichlorethene (TCE) as nodes through which ecosystem information and biomass most efficiently pass. Novel network rearrangement revealed a hierarchy of Kill-the-Winner (KtW), Piggyback-the-Winner (PtW) and PtP nodes. We propose that KtW, PtW and PtP occur simultaneously as competing strategies, with their relative importance depending on conditions at a particular time and location with unusual nutrient sources, such as TCE, appearing to contribute to a shift in this balance between viral mechanisms., (© FEMS 2019.)

Zero-valent iron nanoparticles (ZVIN) have found applications in many strategies for on-site soil and groundwater decontamination. A number of studies have reported the prospective utilization of ZVIN in the reduction of chlorinated organic compounds such as dense non-aqueous phase liquids (DNAPLs) in groundwater. Due to their bioaccumulation and carcinogenesis, DNAPLs in groundwater are a human health hazard and pose environmental risks. Therefore, decontamination of these contaminants is necessary. This study presents the in-situ remediation of trichloroethylene (TCE), perchloroethene (PCE), and 1,2-dichloroethene (1,2-DCE) DNAPLs through the direct injection of polyethylenimine (PEI)-coated ZVIN (PEI-ZVIN composite materials) to facilitate the reduction of contaminants in low-permeability media. A field test was conducted at the premises of a petrochemical company, situated in the Miaoli County of Northern Taiwan that discharged significant amounts of DNAPLs. After in-situ injection and one-day of reaction with groundwater contaminants, ZVIN was further characterized to examine its efficacy in the reduction of pollutants. After the direct injection of PEI-ZVIN, a notable reduction in the concentration of DNAPLs was recorded with conversion from toxic to non-toxic substances. Use of resistivity image profiling (RIP) technique suggested similar conductivity data for the PEI-coated ZVIN suspension and groundwater samples. X-ray absorption near edge structure (XANES) and X-ray absorption fine structure (EXAFS) studies depicted that the oxidation of ZVIN and PEI-ZVIN was occurring after the reductive reaction with contaminated groundwater. The reacted samples had bond distance values of 1.98, 2.00, 1.96, and 1.94 Å. Combining floating surface-coated ZVIN and RIP technique seems promising and environmentally attractive., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

While bioremediation technologies for trichloroethene (TCE), a suspected carcinogen, have been successfully demonstrated in neutral pH aquifers, these technologies are often ineffective for remediating TCE contamination in acidic aquifers (i.e., pH < 5.5). Acidophilic methanotrophs have been detected in several low pH environments, but their presence and potential role in TCE degradation in acidic aquifers is unknown. This study applied a stable isotope probing-based technique to identify active methanotrophs that are capable of degrading TCE in microcosms prepared from two low pH aquifers. A total of thirty-five clones of methanotrophs were derived from low pH microcosms in which methane and TCE degradation had been observed, with 29 clustered in γ-Proteobacteria and 6 clustered in α-Proteobacteria. None of the clones has a high similarity to known acidophilic methanotrophs from other environments. The presence and diversity of particulate MMO and soluble MMO were also investigated. The pmoA gene was detected predominantly at one site, and the presence of a specific form of mmoX in numerous samples suggested that Methylocella spp. may be common in acidic aquifers. Finally, a methane-grown culture at pH 4 was enriched from an acidic aquifer and its ability to biodegrade various chlorinated ethenes was tested. Interestingly, the mixed culture rapidly degraded TCE and vinyl chloride, but not cis-dichloroethene after growth on methane. The data suggest that aerobic biodegradation of TCE and other chlorinated solvents in low pH groundwater may be facilitated by methanotrophic bacteria, and that there are potentially a wide variety of different strains that inhabit acidic aquifers.

The degradation of trichloroethylene (TCE) and tetrachloroethylene (PCE), in incubations where ammonium was oxidized while iron was being reduced indicates that these compounds can be degraded during the Feammox process by Acidimicrobiaceae sp. A6 (ATCC, PTA-122488). None of these compounds were degraded in incubations to which no ammonium was added, indicating that they were degraded during the oxidation of ammonium. Degradation of TCE and PCE (ranging between 32% and 55%) was observed in incubations with a pure Acidimicrobiaceae sp. A6 culture as well as an Acidimicrobiaceae sp. A6 enrichment culture over a 2-week period. In addition to these batch studies, a column study, with a 5-h hydraulic residence time, was conducted contrasting the degradation of TCE in iron-rich soil columns that were either seeded with a pure or an enrichment culture of Acidimicrobiaceae sp. A6 to achieve ammonium oxidation under iron reduction, and a control column that was initially not seeded and later seeded with Geobacter metallireducens. While there was ∼22% TCE removal in the columns seeded with Acidimicrobiaceae sp. A6, there was no removal in the unseeded column or the column seeded with G. metallireducens which was being operated under iron reducing conditions. Feammox is an anoxic process that requires acidic conditions. Hence, these results indicate that this process might be harnessed where other bioremediation strategies are difficult, since many require neutral or alkaline conditions, and supplying ammonium to an anoxic aquifer is relatively easy, since there are not many processes that will oxidize ammonium in the absence of dissolved oxygen., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Chloroethenes are common groundwater and soil contaminants due to extensive historic utilization and inappropriate discharge. The tendency for chloroethenes to become sequestered as dense non-aqueous phase liquids (DNAPL)-a point source to groundwater contamination and causing high concentrations of chloroethenes in proximal aquifers poses a great challenge for remediation of chloroethene contaminated sites. In this study, we report isolation and characterization of a Dehalococcoides mccartyi strain 11G which couples growth with reductive dechlorination of trichloroethenes (TCE), dichloroethene (DCE) isomers and vinyl chloride (VC) to ethene at a growth yield ranging from 2.47 ± 0.23 × 108 to 5.64 ± 0.43 × 108 cells/µmoles Cl- released and co-metabolically dechlorinates tetrachloroethene (PCE) in the presence of TCE. Compared with previous D. mccartyi strains showing dechlorination of TCE at up to 2.0 mM, strain 11G is distinguished by its capacity to dechlorinate chloroethenes at initial concentrations of DCE isomers as high as 4 mM and TCE as high as 3.5 mM to ethene. Bioaugmentation of a contaminated microcosm with strain 11G resulted in complete detoxification of a mixture of 5 mM chloroethenes (2.5 mM of each TCE and cis-DCE) after 40 days. Strain 11G is a promising candidate for in situ bioremediation of high-concentration-chloroethene contaminated sites.

Trichloroethylene (TCE) is one of the most common groundwater contaminants in the United States; however clean-up efforts are a challenge due to its physical and chemical properties. TCE and several of its degradation products were detected in the groundwater of the Beaver Dam Road Landfill site (Beltsville, MD) at concentrations above accepted maximum contaminant levels. A permeable reactive barrier (i.e., biowall) was installed to remediate the groundwater. Microbial infiltration and colonization of the biowall with native site bacteria was expected to occur. An array of molecular biological tools was applied to survey the microbial community for presence of organohalide-respiring microorganisms at the site. Microorganisms belonging to methanogens, acetogens, sulfate-reducing bacteria, and chlorinated aliphatic hydrocarbon-metabolizing bacteria were identified, thus making way for the application of the microbial populations in the biowall bioaugmentation efforts. In concomitant laboratory studies, molecular approaches were used to monitor continuously-fed column reactors containing saturated biowall material spiked with a commercially-available, Dehalococcoides-containing culture (SDC-9), with or without zero-valent iron (ZVI) shavings. The column without ZVI had the highest abundance of Dehalococcoides spp. (2.7 × 10 6  cells g -1 material, S.D. = 3.8 × 10 5  cells g -1 material), while the addition of ZVI did not affect the overall population. Although the addition of ZVI and biostimulation did change ratios of the Dehalococcoides strains, the results suggests that if ZVI would be applied as a biowall material amendment, biostimulation would not be required to maintain a Dehalococcoides population. These experimental results will be utilized in future remediation and/or biowall expansion plans to utilize the natural resources most effectively at the biowall site., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The glutathione (GSH) conjugates, S-(1,2-dichlorovinyl)-glutathione (DCVG) and S-(1,2-dichlorovinyl)-L-cysteine (DCVC), have been implicated in kidney toxicity and kidney cancer from trichloroethylene (TCE) exposure. Considerable differences in blood and tissue levels of DCVG and DCVC have been reported, depending on whether HPLC/UV (High Performance Liquid Chromatography-Ultraviolet) or HPLC/MS (HPLC-Mass Spectrometry) was used. A side-by-side comparison of analytical results with HPLC/UV and HPLC/MS/MS (High Performance Liquid Chromatography-Tandem Mass Spectrometry) detection was undertaken to quantitatively compare estimates for DCVG and DCVG using rat and human tissues. For the HPLC method, DCVG and DCVC were initially derivatized with fluorodinitrobenzene (DNP). The results from the HPLC/UV method showed that derivatized-DCVC eluted at the solvent front and could not be quantified. Derivatized-DCVG, however, was quantified but significant interference was observed in all four control tissues (rat blood, liver, kidney; and human blood), resulting in average spike recoveries of 222-22,990%. In contrast, direct analysis of spiked tissues by HPLC/MS/MS resulted in recoveries of 82-127% and 89-117% for DCVG and DCVC, respectively. These differences in analytical results were further confirmed in tissues from TCE-treated rats, e.g., DCVG levels in rat liver were 18,000 times higher by HPLC/UV as compared to HPLC/MS/MS. Fraction collection of the derivatized-DCVG peak (obtained with the HPLC-UV method), followed by peak identification via an HPLC/UV/Q-TOF/MS/MS method, identified DNP-derivatized endogenous glutamate as the primary interfering substance that contributed to and exaggerated recoveries of DCVG. Thus, estimates of DCVG based on the HPLC/UV methods are not reliable; they will over-estimate the formation of the GSH conjugates of TCE and will artifactually exaggerate the potential cancer risk in humans from TCE exposure. Therefore, it is recommended that any characterization of cancer risks from TCE exposure attributable to the GSH conjugates of TCE rely on results obtained with the more specific and reliable HPLC/MS/MS method., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The dechlorinating Dehalococcoides mccartyi species requires acetate as carbon source, but little is known on its growth under acetate limiting conditions. In this study, we observed growth and dechlorination of a D. mccartyi-containing mixed consortium in a fixed-carbon-free medium with trichloroethene in the aqueous phase and H 2 /CO 2 in the headspace. Around 4 mM formate was produced by day 40, while acetate was constantly below 0.05 mM. Microbial community analysis of the consortium revealed dominance by D. mccartyi and Desulfovibrio sp. (57 and 22% 16S rRNA gene copies, respectively). From this consortium, Desulfovibrio sp. strain F1 was isolated and found to produce formate and acetate (1.2 mM and 48 µM, respectively, by day 24) when cultivated alone in the above mentioned medium without trichloroethene. An established co-culture of strain F1 and D. mccartyi strain 195 demonstrated that strain 195 could grow and dechlorinate using acetate produced by strain F1; and that acetate was constantly below 25 µM in the co-culture. To verify that such low level of acetate is utilizable by D. mccartyi, we cultivated strain 195 alone under acetate-limiting conditions and found that strain 195 consumed acetate to below detection (5 µM). Based on the acetate consumption and cell yield of D. mccartyi, we estimated that on average 1.2 × 10 8 acetate molecules are needed to supply carbon for one D. mccartyi cell. Our study suggests that Desulfovibrio may supply a steady but low amount of fixed carbon to dechlorinating bacteria, exhibiting important implications for natural bio-attenuation when fixed carbon is limited.

Trichloroethylene (TCE) and tetrachloroethylene (PCE) are structurally similar olefins that can cause liver and kidney toxicity. Adverse effects of these chemicals are associated with metabolism to oxidative and glutathione conjugation moieties. It is thought that CYP2E1 is crucial to the oxidative metabolism of TCE and PCE, and may also play a role in formation of nephrotoxic metabolites; however, inter-species and inter-individual differences in contribution of CYP2E1 to metabolism and toxicity are not well understood. Therefore, the role of CYP2E1 in metabolism and toxic effects of TCE and PCE was investigated using male and female wild-type [129S1/SvlmJ], Cyp2e1(-/-), and humanized Cyp2e1 [hCYP2E1] mice. To fill in existing gaps in our knowledge, we conducted a toxicokinetic study of TCE (600 mg/kg, single dose, i.g.) and a subacute study of PCE (500 mg/kg/day, 5 days, i.g.) in 3 strains. Liver and kidney tissues were subject to profiling of oxidative and glutathione conjugation metabolites of TCE and PCE, as well as toxicity endpoints. The amounts of trichloroacetic acid formed in the liver was hCYP2E1≈ 129S1/SvlmJ > Cyp2e1(-/-) for both TCE and PCE; levels in males were about 2-fold higher than in females. Interestingly, 2- to 3-fold higher levels of conjugation metabolites were observed in TCE-treated Cyp2e1(-/-) mice. PCE induced lipid accumulation only in liver of 129S1/SvlmJ mice. In the kidney, PCE exposure resulted in acute proximal tubule injury in both sexes in all strains (hCYP2E1 ≈ 129S1/SvlmJ > Cyp2e1(-/-)). In conclusion, our results demonstrate that CYP2E1 is an important, but not exclusive actor in the oxidative metabolism and toxicity of TCE and PCE.

Methanotrophs not only oxidize CH 4 , but also can oxidize a relatively broad range of other substrates, including trichloroethylene, alkanes, alkenes, and aromatic compounds. In this study, Methylosinus sporium was used as a model organism to characterize toluene metabolism by methanotrophs. Reverse transcription quantitative PCR analysis showed that toluene enhanced the mmoX expression of M. sporium. When the toluene concentration was below 2000 mg m -3 , the kinetics of toluene metabolism by M. sporium conformed to the Michaelis-Menten equation (V max  = 0.238 g g dry weight -1  h -1 , K m  = 545.2 mg m -3 ). The use of a solid-phase extraction technique followed by a gas chromatography-mass spectrometry analysis and molecular docking calculation showed that toluene was likely to primarily bind the di-iron center structural region of soluble methane monooxygenase (sMMO) hydroxylase and then be oxidized to o-cresol. Although M. sporium oxidized toluene, it did not incorporate toluene into its biomass. The coexistence of toluene and CH 4 could influence CH 4 oxidation, the growth of methanotrophs, and the distribution of CH 4 -derived carbon, which were related to the ratio of the toluene concentration to biomass. These results would be helpful to understand the metabolism of CH 4 and non-methane volatile organic compounds in the environment.

Two anaerobic bacterial consortia, each harboring a distinct Sulfurospirillum population, were derived from a 10 year old consortium, SL2, previously characterized for the stepwise dechlorination of tetrachloroethene (PCE) to cis-dichloroethene (cis-DCE) via accumulation of trichloroethene (TCE). Population SL2-1 dechlorinated PCE to TCE exclusively, while SL2-2 produced cis-DCE from PCE without substantial TCE accumulation. The reasons explaining the long-term coexistence of the populations were investigated. Genome sequencing revealed a novel Sulfurospirillum species, designated 'Candidatus Sulfurospirillum diekertiae', whose genome differed significantly from other Sulfurospirillum spp. (78%-83% ANI). Genome-wise, SL2-1 and SL2-2 populations are almost identical, but differences in their tetrachloroethene reductive dehalogenase sequences explain the distinct dechlorination patterns. An extended series of batch cultures were performed at PCE concentrations of 2-200 μM. A model was developed to determine their dechlorination kinetic parameters. The affinity constant and maximal growth rate differ between the populations: the affinity is 6- to 8-fold higher and the growth rate 5-fold lower for SL2-1 than SL2-2. Mixed cultivation of the enriched populations at 6 and 30 μM PCE showed that a low PCE concentration could be the driving force for both functional diversity of reductive dehalogenases and niche specialization of organohalide-respiring bacteria with overlapping substrate ranges.

Dehalococcoides spp. were extensively studied and applied to in-situ trichloroethylene (TCE) remediation since it is the only genus that can reduce TCE to harmless ethene completely. However, this technology was hindered because of the requirement of electron donor (i.e. hydrogen or fermentable organic substrate). Considering the spontaneous electric field and in-situ hydrogen production capacity of tourmaline, this mineral was used as an environmental-friendly bio-promoter for dechlorinating mixed culture containing Dehalococcoides spp. in this work. Research results showed that biodegradation of TCE and the intermediates were both significantly improved by tourmaline. The first-order TCE degradation rate coefficient increased from 0.0125 h -1 for the tourmaline-free system to 0.0306 h -1 for the system combined with 5 g L -1 tourmaline, and ethene production increased by 36%. The outstanding TCE-degrading ability in the tourmaline-bacteria system without the addition of fermentative electron donor (i.e. methanol) indicated that tourmaline could also produce electron donor to support dechlorinating bacteria. Tourmaline could have direct electric biostimulatory effect and indirect enhanced effect associated with water-derived H 2 production in the electric field of tourmaline. Meanwhile, PCR-DGGE analysis exhibited that tourmaline could accelerate the succession of a bacterial, dechlorinating community. The distinctive effects of tourmaline on bacteria were related to its stable electric properties. Therefore, tourmaline could be continuously used in the bioremediation. The present study provided a safe, convenient and persistent alternative to the commonly used enhancement approaches for anaerobic reductive dechlorination process., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Trichloroethylene (TCE) is a ubiquitous environmental toxicant that is a liver and kidney carcinogen. Conjugation of TCE with glutathione (GSH) leads to formation of nepthrotoxic and mutagenic metabolites postulated to be critical for kidney cancerdevelopment; however, relatively little is known regarding their tissue levels as previous analytical methods for their detection lacked sensitivity. Here, an LC-MS/MS-based method for simultaneous detection of S-(1,2-dichlorovinyl)-glutathione (DCVG), S-(1,2-dichlorovinyl)-L-cysteine (DCVC), and N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine (NAcDCVC) in multiple mouse tissues was developed. This analytical method is rapid, sensitive (limits of detection (LOD) 3-30 fmol across metabolites and tissues), and robust to quantify all three metabolites in liver, kidneys, and serum. The method was used to characterize inter-tissue and inter-strain variability in formation of conjugative metabolites of TCE. Single oral dose of TCE (24, 240 or 800 mg/kg) was administered to male mice from 20 inbred strains of Collaborative Cross. Inter-strain variability in the levels of DCVG, DCVC, and NAcDCVC (GSD = 1.6-2.9) was observed. Whereas NAcDCVC was distributed equally among analyzed tissues, highest levels of DCVG were detected in liver and DCVC in kidneys. Evidence indicated that inter-strain variability in conjugative metabolite formation of TCE might affect susceptibility to adverse health effects and that this method might aid in filling data gaps in human health assessment of TCE.

In a field study, aqueous cyclodextrin (CD) was investigated for its ability to extract chlorinated volatile organic compounds (cVOC), such as trichloroethylene (TCE), 1,1,1-trichloroethane (TCA), and dichloroethene (DCE) through in-situ flushing of a sandy aquifer. After cessation of aquifer flushing, a plume of CD was left. Changes in CD, cVOC, and inorganic terminal electron acceptors (TEAs) (DO, nitrate, sulfate, iron) were monitored in four rounds of wellwater sampling (20, 210, 342, and 425days after cessation of active pumping). Post-CD flushing VOC levels rebounded (850% for TCE, 190% for TCA, and 53% for DCE) between the first two sampling rounds, apparently due to rate-limited desorption from aquifer media and dissolution from remaining NAPL. However, substantial reduction in the mass of TCE (6.3 to 0.11mol: 98%) and TCA (2.8 to 0.73mol: 74%) in groundwater was observed between 210 and 425days. DCE should primarily be produced from the degradation of TCE and is expected to subsequently degrade to chloroethene. Since DCE levels decreased only slightly (0.23 to 0.17mol: 26%), its degradation rate should be similar to that produced from the decaying TCE. Cyclodextrin was monitored starting from day 210. The mass of residual CD (as measured by Total Organic Carbon) decreased from 150mol (day 210) to 66 (day 425) (56% decrease). The naturally anaerobic zone within the aquifer where residual CD mass decreased coincided with a loss of other major potential TEAs: nitrate (97% loss), sulfate (31%) and iron (31%). In other studies, TCE and 1,1,1-TCA have been found to be more energetically favorable TEAs than sulfate and iron and their degradation via reductive dechlorination has been found to be enhanced by the fermentation of carbohydrates. Such processes can explain these observations, but more investigation is needed to evaluate whether residual levels of CD can facilitate the anaerobic degradation of chlorinated VOCs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Cobamides such as vitamin B 12 are structurally conserved, cobalt-containing tetrapyrrole biomolecules that have essential biochemical functions in all domains of life. In organohalide respiration, a vital biological process for the global cycling of natural and anthropogenic organohalogens, cobamides are the requisite prosthetic groups for carbon-halogen bond-cleaving reductive dehalogenases. This study reports the biosynthesis of a new cobamide with unsubstituted purine as the lower base and assigns unsubstituted purine a biological function by demonstrating that Coα-purinyl-cobamide (purinyl-Cba) is the native prosthetic group in catalytically active tetrachloroethene reductive dehalogenases of Desulfitobacterium hafniense. Cobamides featuring different lower bases are not functionally equivalent, and purinyl-Cba elicits different physiological responses in corrinoid-auxotrophic, organohalide-respiring bacteria. Given that cobamide-dependent enzymes catalyze key steps in essential metabolic pathways, the discovery of a novel cobamide structure and the realization that lower bases can effectively modulate enzyme activities generate opportunities to manipulate functionalities of microbiomes.

Organohalide-respiring bacteria have key roles in the natural chlorine cycle; however, most of the current knowledge is based on cultures from contaminated environments. We demonstrate that grape pomace compost without prior exposure to chlorinated solvents harbors a Dehalogenimonas (Dhgm) species capable of using chlorinated ethenes, including the human carcinogen and common groundwater pollutant vinyl chloride (VC) as electron acceptors. Grape pomace microcosms and derived solid-free enrichment cultures were able to dechlorinate trichloroethene (TCE) to less chlorinated daughter products including ethene. 16S rRNA gene amplicon and qPCR analyses revealed a predominance of Dhgm sequences, but Dehalococcoides mccartyi (Dhc) biomarker genes were not detected. The enumeration of Dhgm 16S rRNA genes demonstrated VC-dependent growth, and 6.55±0.64 × 10 8 cells were measured per μmole of chloride released. Metagenome sequencing enabled the assembly of a Dhgm draft genome, and 52 putative reductive dehalogenase (RDase) genes were identified. Proteomic workflows identified a putative VC RDase with 49 and 56.1% amino acid similarity to the known VC RDases VcrA and BvcA, respectively. A survey of 1,173 groundwater samples collected from 111 chlorinated solvent-contaminated sites in the United States and Australia revealed that Dhgm 16S rRNA genes were frequently detected and outnumbered Dhc in 65% of the samples. Dhgm are likely greater contributors to reductive dechlorination of chlorinated solvents in contaminated aquifers than is currently recognized, and non-polluted environments represent sources of organohalide-respiring bacteria with novel RDase genes.

The ex situ treatment of N-nitrosodimethylamine (NDMA) and N-nitrodimethylamine (NTDMA) in groundwater was evaluated in a field-scale fluidized bed bioreactor (FBR). Both of these compounds, which originally entered groundwater at the test site from the use of liquid rocket propellant, are suspected human carcinogens. The objective of this research was to examine the application of a novel field-scale propane-fed fluidized bed bioreactor as an alternative to ultraviolet irradiation (UV) for treating NDMA and NTDMA to low part-per-trillion (ng/L) concentrations. Previous laboratory studies have shown that the bacterium Rhodococcus ruber ENV425 can biodegrade NDMA and NTDMA during growth on propane as a primary substrate and that the strain can effectively reduce NDMA concentrations in propane-fed bench-scale bioreactors of different design. R. ruber ENV425 was used as a seed culture for the FBR, which operated at a fluidization flow of ∼19 L-per-min (LPM) and received propane, oxygen, and inorganic nutrients in the feed. The reactor effectively treated ∼1 μg/L of influent NDMA to effluent concentrations of less than 10 ng/L at a hydraulic residence time (HRT) of only 10 min. At a 20 min HRT, the FBR reduced NDMA to <4.2 ng/L in the effluent, which was the discharge limit at the test site where the study was conducted. Similarly, NTDMA was consistently treated in the FBR from ∼0.5 μg/L to <10 ng/L at an HRT of 10 min or longer. Based on these removal rates, the average NDMA and NTDMA elimination capacities achieved were 2.1 mg NDMA treated/m 3 of expanded bed/hr of operation and 1.1 mg NTDMA treated/m 3 of expanded bed/hr of operation, respectively. The FBR system was highly resilient to upsets including power outages. Treatment of NDMA, but not NTDMA, was marginally affected when trace co-contaminants including trichloroethene (TCE) and trichlorofluoromethane (Freon 11) were initially added to feed groundwater, but performance recovered over a few weeks in the continued presence of these compounds. Strain ENV425 appeared to be replaced by native propanotrophs over time based on qPCR analysis, but contaminant treatment was not diminished. The results suggest that a FBR can be a viable alternative to UV treatment for removing NDMA from groundwater., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

An environmental isolate Comamonas testosteroni strain RF2, which has been found to cometabolize trichloroethene (TCE) in the presence of phenol and sodium lactate as growth substrates, was tested to investigate its capacity for degrading 1,2-cis-dichloroethene (cDCE), 1,2-trans-dichlorothene (tDCE), and 1,1-dichloroethene (1,1DCE). Degradation assays were performed for single DCEs, as well as for a mixture of DCEs with TCE, which resembled contaminated plume in groundwater. RF2 was capable of efficiently removing all three dichloroethenes (DCEs) at the initial aqueous concentrations of 6.01 mg L -1 for cDCE, 3.80 mg L -1 for tDCE and 0.65 mg L -1 for 1,1DCE, with a removal efficiency of 100% for cDCE, 65.8% for tDCE, and 46.8% for 1,1DCE. Furthermore, complete removal of TCE, cDCE and 1,1DCE (122.5 μg L -1 , 84.3 μg L -1 and 51.4 μg L -1 , respectively) was observed in a mixture sample that also contained 72.33 μg L -1 of tDCE, which was removed to the amount of 72.3%. Moreover, degradation of cDCE (6.01 mg L -1 ) led to a 93.8% release of inorganic chloride, and 2,2-dichloroacetaldehyde was determined as the first intermediate of cDCE transformation. The findings of this study suggest that the strain RF2 exhibits the potential to remediate groundwater contaminated with less chlorinated ethenes., (Copyright © 2017. Published by Elsevier Ltd.)

Trichloroethylene (TCE), an industrial chemical and environmental contaminant, is a human carcinogen. Reactive metabolites are implicated in renal carcinogenesis associated with TCE exposure, yet the toxicity mechanisms of these metabolites and their contribution to cancer and other adverse effects remain unclear. We employed an integrated functional genomics approach that combined functional profiling studies in yeast and avian DT40 cell models to provide new insights into the specific mechanisms contributing to toxicity associated with TCE metabolites. Genome-wide profiling studies in yeast identified the error-prone translesion synthesis (TLS) pathway as an import mechanism in response to TCE metabolites. The role of TLS DNA repair was further confirmed by functional profiling in DT40 avian cell lines, but also revealed that TLS and homologous recombination DNA repair likely play competing roles in cellular susceptibility to TCE metabolites in higher eukaryotes. These DNA repair pathways are highly conserved between yeast, DT40, and humans. We propose that in humans, mutagenic TLS is favored over homologous recombination repair in response to TCE metabolites. The results of these studies contribute to the body of evidence supporting a mutagenic mode of action for TCE-induced renal carcinogenesis mediated by reactive metabolites in humans. Our approach illustrates the potential for high-throughput in vitro functional profiling in yeast to elucidate toxicity pathways (molecular initiating events, key events) and candidate susceptibility genes for focused study., (© The Author 2017. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Carbon tetrachloride (CT) and chloroform (CF) were transformed in batch reactor experiments conducted with anaerobic dechlorinating cultures and supernatant (ADC + S) harvested from continuous flow reactors. The Evanite (EV) and Victoria/Stanford (VS) cultures, capable of respiring trichloroethene (TCE), 1,2-cis-dichloroethene (cDCE), and vinyl chloride (VC) to ethene (ETH), were grown in continuous flow reactors receiving an influent feed of saturated TCE (10 mM; 60 mEq) and formate (45 mM; 90 mEq) but no CT or CF. Cells and supernatant were harvested from the chemostats and inoculated into batch reactors at the onset of each experiment. CT transformation was complete following first order kinetics with CF, DCM and CS 2 as the measurable transformation products, representing 20-40% of the original mass of CT, with CO 2 likely the unknown transformation product. CF was transformed to DCM and likely CO 2 at an order of magnitude rate lower than CT, while DCM was not further transformed. An analytical first order model including multiple key reactions effectively simulated CT transformation, product formation and transformation, and provided reasonable estimates of transformation rate coefficients. Biotic and abiotic treatments indicated that CT was mainly transformed via abiotic processes. However, the presence of live cells was associated with the transformation of CF to DCM. In biotic tests both TCE and CT were simultaneously transformed, with TCE transformed to ETH and approximately 15-53% less CF formed via CT transformation. A 14-day exposure to CF (CF max  = 1.4 μM) reduced all rates of chlorinated ethene respiration by a factor of 10 or greater., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Trichloroethylene (TCE) is a widespread soil and groundwater pollutant and clean-up is often problematic and expensive. Phytoremediation may be a cost-effective solution at some sites. This study investigates TCE degradation by willows (S. viminalis) and willows inoculated with three strains of B. cepacia (301C, PR1-31 and VM1330-pTOM), using chloride formation as an indicator of dehalogenation. Willows were grown in non-sterile, hydroponic conditions for 3 weeks in chloride-free nutrient solution spiked with TCE. TCE was added weekly due to rapid loss by volatilization. Chloride and TCE in solution were measured every 2-3 days and chloride and metabolite concentrations in plants were measured at test termination. Based on transpiration, no tree toxicity of TCE exposure was observed. However, trees grown in chloride-free solution showed severely inhibited transpiration. No or very little chloride was formed during the test, and levels of chloride in TCE-exposed trees were not elevated. Chloride concentrations in chloride containing TCE-free nutrient solution doubled within 23 days, indicating active exclusion of chloride by root cell membranes. Only traces of TCE-metabolites were detected in plant tissue. We conclude that TCE is not, or to a limited extent (less than 3%), aerobically degraded by the willow trees. The three strains of B. cepacia did not enhance TCE mineralization. Future successful application of rhizo- and phytodegradation of TCE requires measures to be taken to improve the degradation rates.

This study investigated the removal of hydrophobic trichloroethylene (TCE) in the presence of methanol (co-metabolite) in a biotrickling filter, which was seeded with fungi at pH4. Starvation was chosen as the biomass control strategy. Two systems, Biofilter I (methanol:TCE 70:30) and Biofilter II (methanol:TCE 80:20) were run in parallel, each with varying composition ratios. The TCE loading rates for both biofilters ranged from 3.22 to 12.88g/m 3 /hr. Depending on the ratio, methanol concentrations varied from 4.08 to 27.95g/m 3 /hr. The performance of the systems was evaluated and compared by calculating removal kinetics, carbon mass balance, efficiencies and elimination capacities. Methanol was observed to enhance TCE removal during the initial loading rate. However, methanol later inhibited TCE degradation above 6.44g TCE/m 3 /hr (Biofilter I) and 3.22g TCE/m 3 /hr (Biofilter II). Conversely, TCE did not impede methanol removal because over 95% methanol elimination was consistently achieved. Overall, Biofilter I was able to outperform Biofilter II due to its greater resistance towards methanol competition., (Copyright © 2016. Published by Elsevier B.V.)

The diversity of Dehalococcoides mccartyi (Dhc) and/or other organohalide respiring or associated microorganisms in parallel, partial, or complete trichloroethene (TCE) dehalogenating systems has not been well described. The composition of Dhc populations and the associated bacterial community that developed over 7.5 years in the top layer (0-10 cm) of eight TCE-fed columns were examined using pyrosequencing. Columns biostimulated with one of three carbon sources, along with non-stimulated controls, developed into complete (ethene production, whey amended), partial (cis-dichloroethene (DCE) and VC, an emulsified oil with nonionic surfactant), limited (<5 % cis-DCE and 95 % TCE, an emulsified oil), and non- (controls) TCE dehalogenating systems. Bioaugmentation of one column of each treatment with Bachman Road enrichment culture did not change Dhc populations nor the eventual degree of TCE dehalogenation. Pyrosequencing revealed high diversity among Dhc strains. There were 13 OTUs that were represented by more than 1000 sequences each. Cornell group-related populations dominated in complete TCE dehalogenating columns, while Pinellas group related Dhc dominated in all other treatments. General microbial communities varied with biostimulation, and three distinct microbial communities were established: one each for whey, oils, and control treatments. Bacterial genera, including Dehalobacter, Desulfitobacterium, Sulfurospirillum, Desulfuromonas, and Geobacter, all capable of partial TCE dehalogenation, were abundant in the limited and partial TCE dehalogenating systems. Dhc strain diversity was wider than previously reported and their composition within the community varied significantly depending on the nature of the carbon source applied and/or changes in the Dhc associated partners that fostered different biogeochemical conditions across the columns.

Trichloroethylene (TCE) is a chlorinated aliphatic organic compound often detected as pollutant in soils and ground water. "Green technologies" based on phytoremediation were proven to be effective to reclaim organic pollutants (e.g. TCE) and heavy metals from different environmental matrices. In this work, we use Zea mays L. for the removal of high TCE concentrations from medium cultures. In particular, we investigated a sealed bioreactor where the growth medium was contaminated with an increasing amount of TCE, in the range 55-280 mg/L; the removal capability of the maize plants was assessed by means of GC-MS and LC-MS analyses. An accurate mass balance of the system revealed that the plants were able to remove and metabolise TCE with an efficiency up to 20 %, depending on the total amount of TCE delivered in the bioreactor. Morphometric data showed that the growth of Z. mays is not significantly affected by the presence of the pollutant up to a concentration of 280 mg/L, while plants show significant alterations at higher TCE concentrations until the growth is completely inhibited for [TCE] ≃ 2000 mg/L. Finally, the presence of several TCE metabolites, including dichloroacetic and trichloroacetic acids, was detected in the roots and in the aerial part of the plants, revealing that Z. mays follows the green liver metabolic model. These results encourage further studies for the employment of this plant species in phytoremediation processes of soils and waters contaminated by TCE and, potentially, by many other chlorinated solvents.

BTEX (benzene, toluene, ethylbenzene, ortho-, meta-, and para-xylenes), methyl tert-butyl ether (MTBE), cis-1,2-dichloroethylene (cis-DCE), and trichloroethylene (TCE) are among the major soil and groundwater contaminants frequently co-existing, as a result of their widespread uses. Pseudomonas plecoglossicida was immobilized on waste scrap tyre to remove these contaminants mixture from synthetic contaminated water. The microbial activity was enhanced in the immobilized system, shown by the higher colony forming units (CFUs) (40%), while BTEX were used as growth substrates. The adsorption capacity of tyres toward contaminants reached a maximum within one day, with BTEX (76.3%) and TCE (64.3%) showing the highest sorption removal capacities, followed by cis-DCE (30.0%) and MTBE (11.0%). The adsorption data fitted the Freundlich isotherm with a good linear correlation (0.989-0.999) for the initial contaminants concentration range applied (25-125mg/L). The monoaromatic hydrocarbons were almost completely removed in the immobilized system and the favourable removal efficiencies of 78% and 90% were obtained for cis-DCE and TCE, respectively. The hybrid (biological, immobilization/physical, sorption) system was further evaluated with the contaminants spiked intermittently for the stable performance. The addition of mineral salt medium further enhanced the bioremoval of contaminants by stimulating the microbial growth to some extent., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Dehalococcoides mccartyi exhibits versatile capabilities to respire halogenated compounds under anaerobic conditions. In this study, we report the assembly and annotation of the complete genome of a chloroethene dechlorinating D. mccartyi strain 11a5. Bearing a 1461 973 base-pair chromosome, strain 11a5 distinguishes itself from other D. mccartyi strains by possessing a 5940 base-pair circular extrachromosomal genetic element which contains a reductive dehalogenase homolog. The whole genome of strain 11a5 harbors 31 putative reductive dehalogenase genes. Through transcriptional and proteomic analyses, we identified a new tetrachloroethene (PCE) reductive dehalogenase, PteA (11a5&#95;1355), which catalyzes reductive dechlorination from PCE to trichloroethene (TCE) and shares only 38% similarity in amino acid sequence with its closest relative PceA in D. mccartyi strain 195. The acquisition of the genome of strain 11a5 enlarged the database of D. mccartyi and enriched our understanding of this unique species, among which, the identification of a new PCE reductive dehalogenase can assist in understanding PCE dechlorination process. In addition, the discovery of the circular extrachromosomal genetic element in strain 11a5 may provide insights to investigate how reductive dehalogenase homologous genes are transferred and carried by organohalide respiring bacteria.

The aim of this study is to obtain a systems-level understanding of the interactions between Dehalococcoides and corrinoid-supplying microorganisms by analyzing community structures and functional compositions, activities, and dynamics in trichloroethene (TCE)-dechlorinating enrichments. Metagenomes and metatranscriptomes of the dechlorinating enrichments with and without exogenous cobalamin were compared. Seven putative draft genomes were binned from the metagenomes. At an early stage (2 days), more transcripts of genes in the Veillonellaceae bin-genome were detected in the metatranscriptome of the enrichment without exogenous cobalamin than in the one with the addition of cobalamin. Among these genes, sporulation-related genes exhibited the highest differential expression when cobalamin was not added, suggesting a possible release route of corrinoids from corrinoid producers. Other differentially expressed genes include those involved in energy conservation and nutrient transport (including cobalt transport). The most highly expressed corrinoid de novo biosynthesis pathway was also assigned to the Veillonellaceae bin-genome. Targeted quantitative PCR (qPCR) analyses confirmed higher transcript abundances of those corrinoid biosynthesis genes in the enrichment without exogenous cobalamin than in the enrichment with cobalamin. Furthermore, the corrinoid salvaging and modification pathway of Dehalococcoides was upregulated in response to the cobalamin stress. This study provides important insights into the microbial interactions and roles played by members of dechlorinating communities under cobalamin-limited conditions. IMPORTANCE The key chloroethene-dechlorinating bacterium Dehalococcoides mccartyi is a cobalamin auxotroph, thus acquiring corrinoids from other community members. Therefore, it is important to investigate the microbe-microbe interactions between Dehalococcoides and the corrinoid-providing microorganisms in a community. This study provides systems-level information, i.e., taxonomic and functional compositions and dynamics of the supportive microorganisms in dechlorinating communities under different cobalamin conditions. The findings shed light on the important roles of Veillonellaceae species in the communities compared to other coexisting community members in producing and providing corrinoids for Dehalococcoides species under cobalamin-limited conditions., (Copyright © 2017 American Society for Microbiology.)

In order to elucidate interactions between sulfate reduction and dechlorination, we systematically evaluated the effects of different concentrations of sulfate and sulfide on reductive dechlorination by isolates, constructed consortia, and enrichments containing Dehalococcoides sp. Sulfate (up to 5 mM) did not inhibit the growth or metabolism of pure cultures of the dechlorinator Dehalococcoides mccartyi 195, the sulfate reducer Desulfovibrio vulgaris Hildenborough, or the syntroph Syntrophomonas wolfei In contrast, sulfide at 5 mM exhibited inhibitory effects on growth of the sulfate reducer and the syntroph, as well as on both dechlorination and growth rates of D. mccartyi Transcriptomic analysis of D. mccartyi 195 revealed that genes encoding ATP synthase, biosynthesis, and Hym hydrogenase were downregulated during sulfide inhibition, whereas genes encoding metal-containing enzymes involved in energy metabolism were upregulated even though the activity of those enzymes (hydrogenases) was inhibited. When the electron acceptor (trichloroethene) was limiting and an electron donor (lactate) was provided in excess to cocultures and enrichments, high sulfate concentrations (5 mM) inhibited reductive dechlorination due to the toxicity of generated sulfide. The initial cell ratio of sulfate reducers to D. mccartyi (1:3, 1:1, or 3:1) did not affect the dechlorination performance in the presence of sulfate (2 and 5 mM). In contrast, under electron donor limitation, dechlorination was not affected by sulfate amendments due to low sulfide production, demonstrating that D. mccartyi can function effectively in anaerobic microbial communities containing moderate sulfate concentrations (5 mM), likely due to its ability to outcompete other hydrogen-consuming bacteria and archaea. IMPORTANCE Sulfate is common in subsurface environments and has been reported as a cocontaminant with chlorinated solvents at various concentrations. Inconsistent results for the effects of sulfate inhibition on the performance of dechlorination enrichment cultures have been reported in the literature. These inconsistent findings make it difficult to understand potential mechanisms of sulfate inhibition and complicate the interpretation of bioremediation field data. In order to elucidate interactions between sulfate reduction and reductive dechlorination, this study systematically evaluated the effects of different concentrations of sulfate and sulfide on reductive dechlorination by isolates, constructed consortia, and enrichments containing Dehalococcoides sp. This study provides a more fundamental understanding of the competition mechanisms between reductive dechlorination by Dehalococcoides mccartyi and sulfate reduction during the bioremediation process. It also provides insights on the significance of sulfate concentrations on reductive dechlorination under electron donor/acceptor-limiting conditions during in situ bioremediation applications. For example, at a trichloroethene-contaminated site with a high sulfate concentration, proper slow-releasing electron donors can be selected to generate an electron donor-limiting environment that favors reductive dechlorination and minimizes the sulfide inhibition effect., (Copyright © 2017 American Society for Microbiology.)

Organohalide-respiring bacteria can be difficult to enrich and isolate, which can limit research on these important organisms. The goal of this research was to develop a method to rapidly (minutes to days) enrich these organisms from a mixed community. The method presented is based on the hypothesis that organohalide-respiring bacteria would be more hydrophobic than other bacteria as they dehalogenate hydrophobic compounds. The method developed tests this hypothesis by separating a portion of putative organohalide-respiring bacteria, those phylogenetically related to Dehalococcoides mccartyi , at the interface between a hydrophobic organic solvent and an aqueous medium. This novel partial separation technique was tested with a polychlorinated biphenyl-enriched sediment-free culture, a tetrachloroethene-enriched digester sludge culture, and uncontaminated lake sediment. Significantly higher fractions, up to 20.4 times higher, of putative organohalide-respiring bacteria were enriched at the interface between the medium and either hexadecane or trichloroethene. The selective partial separation of these putative organohalide-respiring bacteria occurred after 20 min, strongly suggesting that the separation was a result of physical-chemical interactions between the cell surface and hydrophobic solvent. Dechlorination activity postseparation was verified by the production of cis -dichloroethene when amended with tetrachloroethene. A longer incubation time of 6 days prior to separation with trichloroethene increased the total number of putative organohalide-respiring bacteria. This method provides a way to quickly separate some of the putative organohalide-respiring bacteria from other bacteria, thereby improving our ability to study multiple and different bacteria of potential interest and improving knowledge of these bacteria. IMPORTANCE Organohalide-respiring bacteria, bacteria capable of respiring chlorinated contaminants, can be difficult to enrich, which can limit their predictable use for the bioremediation of contaminated sites. This paper describes a method to quickly separate Dehalococcoides -like bacteria, a group of organisms containing organohalide-respiring bacteria, from other bacteria in a mixed community. From this work, Dehalococcoides -like bacteria appear to have a hydrophobic cell surface, facilitating a rapid (20 min) partial separation from a mixed culture at the surface of a hydrophobic liquid. This method was verified in a polychlorinated biphenyl-enriched sediment-free culture, an anaerobic digester sludge, and uncontaminated sediment. The method described can drastically reduce the amount of time required to partially separate Dehalococcoides -like bacteria from a complex mixed culture, improving researchers' ability to study these important bacteria., (Copyright © 2017 American Society for Microbiology.)

Acetylene (C 2 H 2 ) can be generated in contaminated groundwater sites as a consequence of chemical degradation of trichloroethene (TCE) by in situ minerals, and C 2 H 2 is known to inhibit bacterial dechlorination. In this study, we show that while high C 2 H 2 (1.3 mM) concentrations reversibly inhibit reductive dechlorination of TCE by Dehalococcoides mccartyi isolates as well as enrichment cultures containing D. mccartyi sp., low C 2 H 2 (0.4 mM) concentrations do not inhibit growth or metabolism of D. mccartyi. Cocultures of Pelobacter SFB93, a C 2 H 2 -fermenting bacterium, with D. mccartyi strain 195 or with D. mccartyi strain BAV1 were actively sustained by providing acetylene as the electron donor and carbon source while TCE or cis-DCE served as the electron acceptor. Inhibition by acetylene of reductive dechlorination and methanogenesis in the enrichment culture ANAS was observed, and the inhibition was removed by adding Pelobacter SFB93 into the consortium. Transcriptomic analysis of D. mccartyi strain 195 showed genes encoding for reductive dehalogenases (e.g., tceA) were not affected during the C 2 H 2 -inhibition, while genes encoding for ATP synthase, biosynthesis, and Hym hydrogenase were down-regulated during C 2 H 2 inhibition, consistent with the physiological observation of lower cell yields and reduced dechlorination rates in strain 195. These results will help facilitate the optimization of TCE-bioremediation at contaminated sites containing both TCE and C 2 H 2 .

In anoxic groundwater aquifers, the long-term survival of Dehalococcoides mccartyi populations expressing the gene vcrA (or bvcA) encoding reductive vinyl chloride dehalogenases are important to achieve complete dechlorination of tetrachloroethene (PCE) and trichloroethene (TCE) to nonchlorinated ethene. The absence or inactivity of vcrA-containing Dehalococcoides results in the accumulation of the harmful chlorinated intermediates dichloroethene (DCE) and vinyl chloride (VC). Although vcrA-containing Dehalococcoides subpopulations depend on synergistic interaction with other organohalide-respiring populations generating their metabolic electron acceptors (DCE and VC), their survival requires successful competition for electron donor within the entire organohalide-respiring microbial community. To understand this dualism of synergy and competition under growth conditions relevant in contaminated aquifers, we investigated Dehalococcoides-level population structure when subjected to a change in the ratio of electron donor to chlorinated electron acceptor in continuously stirred tank reactors (CSTRs) operated over 7 years. When the electron donor formate was supplied in stoichiometric excess to TCE, both tceA-containing and vcrA-containing Dehalococcoides populations persisted, and near-complete dechlorination to ethene was stably maintained. When the electron donor formate was supplied at substoichiometric concentrations, the interactions between tceA-containing and vcrA-containing populations shifted toward direct competition for the same limiting catabolic electron donor substrate with subsequent niche exclusion of the vcrA-containing population. After more than 2000 days of operation under electron donor limitation, increasing the electron donor to TCE ratio facilitated a recovery of the vcrA-containing Dehalococoides population to its original frequency. We demonstrate that electron donor scarcity alone, in the absence of competing metabolic processes or inhibitory dechlorination intermediate products, is sufficient to alter the Dehalococcoides population structure. These results underscore the importance of electron donor and chloroethene stoichiometry in maintaining balanced functional performance within consortia composed of multiple D. mccartyi subpopulations, even when other competing electron acceptor processes are absent.

1,2,3-Trichloropropane (TCP) is an emerging groundwater pollutant and suspected human carcinogen. TCP, a recalcitrant contaminant, has been detected in the subsurface near TCP manufacture facilities and many superfund sites. Considering the toxicity and the occurence of TCP, there is a need to seek for cost-effective treatment technologies for TCP-contaminated sites. This paper investigated TCP biodegradation by propane-oxidizing bacteria (PrOB) which are known to express propane monooxygenase (PrMO). PrMO can cometabolically degrade many different contaminants. Four PrOB, Rhodococus jostii RHA1, Mycobacterium vaccae JOB5, Rhodococcus rubber ENV425 and one isolate Sphingopyxis sp. AX-A were examined for their ability to degrade TCP. All the four PrOB resting cells were able to degrade TCP. Strain JOB5 exhibited the best TCP degradation ability (v initial = 9.7 ± 0.7 μg TCP (mg protein) -1 h -1 ). No TCP was degraded in the presence of acetylene (an inhibitor for PrMO), suggesting that PrMO might be responsible for TCP degradation. Furthermore, competitive inhibition was observed between propane and TCP, and between trichloroethylene (TCE) and TCP., (Copyright © 2016. Published by Elsevier Ltd.)

This work evaluated the interaction of perchlorate and trichloroethene (TCE), two common co-contaminants in groundwater, during bioreduction in serum bottles containing synthetic mineral salts media and microbial consortia. TCE at concentrations up to 0.3mM did not significantly affect perchlorate reduction; however, perchlorate concentrations higher than 0.1mM made the reduction of TCE significantly slower. Perchlorate primarily inhibited the reduction of vinyl chloride (VC, a daughter product of TCE) to ethene. Mechanistic analysis showed that the inhibition was mainly because perchlorate reduction is thermodynamically more favorable than reduction of TCE and its daughter products and not because of toxicity due to accumulation of dissolved oxygen produced during perchlorate reduction. As the initial perchlorate concentration increased from 0 to 600mg/L in a set of serum bottles, the relative abundance of Rhodocyclaceae (a putatively perchlorate-reducing genus) increased from 6.3 to 80.6%, while the relative abundance of Dehalococcoides, the only known genus that is able to reduce TCE all the way to ethene, significantly decreased. Similarly, the relative abundance of Proteobacteria (a phylum to which most known perchlorate-reducing bacteria belong) increased from 22% to almost 80%., (Copyright © 2016 Elsevier B.V. All rights reserved.)