PCE Dechlorination Research Articles

Tetrachloroethylene dechlorination from sulfide-containing aquifers by biochar depends on hydrogeochemical conditions in two aspects: Adsorption and catalysis

Biochar can facilitate sulfide to reduce organic contaminants, such as tetrachloroethylene (PCE). However, the biochar interface catalytic performance in complex water matrices is rarely investigated, and the combined roles of sulfide and hydrogeochemical component in the dechlorination of PCE remains unclear. This study explore the influence of environmental parameters on PCE dechlorination by a wood-based biochar (WW800). The enhanced reductive dechlorination by WW800 was a two-site heterogeneous catalytic process in two stages: (1) adsorption and (2) reduction. In the absence of ions, the removal of PCE included both stages. Ca2+ cation bridge was important in enhancing the reducibility of –COOH on WW800 bound sulfide. The co-existence ionic conditions of six redox zones were simulated, HCO3– inhibited the binding of sulfide to WW800, and SO42− inhibited both adsorption and reduction stages. These inhibitory effects were prior to the promotion of Ca2+. The combination of multiple ions with WW800 consumed the active groups (pyridine N, C=O and O-C=O), which limited the transformation of sulfides to strong nucleophilicity. Among the single influencing factors, Ca2+, HCO3–, initial concentration of PCE and temperature were positively correlated with the removal kinetics of PCE (P<0.05). The low concentration of Fe2+ and Mn2+ in the simulated zones did not significantly promote the removal of PCE. According to the kinetic changes of dechlorination reaction, the inhibition of PCE conversion by humic acid was due to the change of sulfide species caused by pH value change. The aquifer environment with low temperature (10–15 °C) and DO (0–6.6 mg/L) had little effect on PCE dechlorination. Our results show that the hydrochemical components in aquifer enhance PCE dechlorination by biochar with different influence mechanisms, which provides theoretical guidance for application of this method in the aquifer.

Metagenomic 16S rRNA analysis and predictive functional profiling revealed intrinsic organohalides respiration and bioremediation potential in mangrove sediment

BackgroundMangrove sediment microbes are increasingly attracting scientific attention due to their demonstrated capacity for diverse bioremediation activities, encompassing a wide range of environmental contaminants.Materials and methodsThe microbial communities of five Avicennia marina mangrove sediment samples collected from Al Rayyis White Head, Red Sea (KSA), were characterized using Illumina amplicon sequencing of the 16S rRNA genes.ResultsOur study investigated the microbial composition and potential for organohalide bioremediation in five mangrove sediments from the Red Sea. While Proteobacteria dominated four microbiomes, Bacteroidetes dominated the fifth. Given the environmental concerns surrounding organohalides, their bioremediation is crucial. Encouragingly, we identified phylogenetically diverse organohalide-respiring bacteria (OHRB) across all samples, including Dehalogenimonas, Dehalococcoides, Anaeromyxobacter, Desulfuromonas, Geobacter, Desulfomonile, Desulfovibrio, Shewanella and Desulfitobacterium. These bacteria are known for their ability to dechlorinate organohalides through reductive dehalogenation. PICRUSt analysis further supported this potential, predicting the presence of functional biomarkers for organohalide respiration (OHR), including reductive dehalogenases targeting tetrachloroethene (PCE) and 3-chloro-4-hydroxyphenylacetate in most sediments. Enrichment cultures studies confirmed this prediction, demonstrating PCE dechlorination by the resident microbial community. PICRUSt also revealed a dominance of anaerobic metabolic processes, suggesting the microbiome’s adaptation to the oxygen-limited environment of the sediments.ConclusionThis study provided insights into the bacterial community composition of five mangrove sediments from the Red Sea. Notably, diverse OHRB were detected across all samples, which possess the metabolic potential for organohalide bioremediation through reductive dehalogenation pathways. Furthermore, PICRUSt analysis predicted the presence of functional biomarkers for OHR in most sediments, suggesting potential intrinsic OHR activity by the enclosed microbial community.

Interspecies Mobility of Organohalide Respiration Gene Clusters Enables Genetic Bioaugmentation.

Anthropogenic organohalide pollutants pose a severe threat to public health and ecosystems. In situ bioremediation using organohalide respiring bacteria (OHRB) offers an environmentally friendly and cost-efficient strategy for decontaminating organohalide-polluted sites. The genomic structures of many OHRB suggest that dehalogenation traits can be horizontally transferred among microbial populations, but their occurrence among anaerobic OHRB has not yet been demonstrated experimentally. This study isolates and characterizes a novel tetrachloroethene (PCE)-dechlorinating Sulfurospirillum sp. strain SP, distinguishing itself among anaerobic OHRB by showcasing a mechanism essential for horizontal dissemination of reductive dehalogenation capabilities within microbial populations. Its genetic characterization identifies a unique plasmid (pSULSP), harboring reductive dehalogenase and de novo corrinoid biosynthesis operons, functions critical to organohalide respiration, flanked by mobile elements. The active mobility of these elements was demonstrated through genetic analyses of spontaneously emerging nondehalogenating variants of strain SP. More importantly, bioaugmentation of nondehalogenating microcosms with pSULSP DNA triggered anaerobic PCE dechlorination in taxonomically diverse bacterial populations. Our results directly support the hypothesis that exposure to anthropogenic organohalide pollutants can drive the emergence of dehalogenating microbial populations via horizontal gene transfer and demonstrate a mechanism by which genetic bioaugmentation for remediation of organohalide pollutants could be achieved in anaerobic environments.

Characterization of Reductive Dechlorination of Chlorinated Ethylenes by Anaerobic Consortium

Tetrachloroethylene （PCE） and trichloroethylene （TCE） are typical volatile halogenated organic compounds in groundwater that pose serious threats to the ecological environment and human health. To obtain an anaerobic microbial consortium capable of efficiently dechlorinating PCE and TCE to a non-toxic end product and to explore its potential in treating contaminated groundwater， an anaerobic microbial consortium W-1 that completely dechlorinated PCE and TCE to ethylene was obtained by repeatedly feeding PCE or TCE into the contaminated groundwater collected from an industrial site. The dechlorination rates of PCE and TCE were （120.1 ±4.9） μmol·（L·d）-1 and （172.4 ±21.8） μmol·（L·d）-1 in W-1， respectively. 16S rRNA gene amplicon sequencing and quantitative PCR （qPCR） showed that the relative abundance of Dehalobacter increased from 1.9% to 57.1%， with the gene copy number increasing by 1.7×107 copies per 1 μmol Cl- released when 98.3 μmol of PCE was dechlorinated to cis-1，2-dichloroethylene （cis-1，2-DCE）. The relative abundance of Dehalococcoides increased from 1.1% to 53.8% when cis-1，2-DCE was reductively dechlorinated to ethylene. The growth yield of Dehalococcoides gene copy number increased by 1.7×108 copies per 1 μmol Cl- released for the complete reductive dechlorination of PCE to ethylene. The results indicated that Dehalobacter and Dehalococcoides cooperated to completely detoxify PCE. When TCE was used as the only electron acceptor， the relative abundance of Dehalococcoides increased from （29.1 ±2.4）% to （7.7 ±0.2）%， and gene copy number increased by （1.9 ±0.4）×108 copies per 1 μmol Cl- released， after dechlorinating 222.8 μmol of TCE to ethylene. The 16S rRNA gene sequence of Dehalococcoides LWT1， the main functional dehalogenating bacterium in enrichment culture W-1， was obtained using PCR and Sanger sequencing， and it showed 100% similarity with the 16S rRNA gene sequence of D. mccartyi strain 195. The anaerobic microbial consortium W-1 was also bioaugmented into the groundwater contaminated by TCE at a concentration of 418.7 μmol·L-1. The results showed that （69.2 ±9.8）% of TCE could be completely detoxified to ethylene within 28 days with a dechlorination rate of （10.3 ±1.5） μmol·（L·d）-1. This study can provide the microbial resource and theoretical guidance for the anaerobic microbial remediation in PCE or TCE-contaminated groundwater.

Effect of wine pomace extract on dechlorination of chloroethenes in soil suspension

Chloroethenes are widely used as solvent in the metal industry and the dry cleaning industry, but their spillage into soil and groundwater due to improper handling has negatively impacted human health. Bioremediation using microorganisms is one of the technologies to clean up soil and groundwater contaminated with chloroethenes. In this study, we examined the bioremediation of chloroethene-contaminated soil using wine pomace extract (WPE). WPE is a liquid containing seven major carboxylic acids and other substances extracted from grape pomace produced in winemaking. WPE clearly promoted the anaerobic bioremediation of chloroethenes. In the tetrachloroethene (PCE) degradation test that used fractions derived from WPE, the water-eluted fraction containing l-lactic acid, l-tartaric acid, and others promoted the dechlorination of PCE, whereas the methanol-eluted fraction containing mainly syringic acid did not. In another PCE degradation test that used l-lactic acid, l-tartaric acid, and syringic acid test solutions, l-lactic acid and l-tartaric acid enhanced the dechlorination of PCE, but syringic acid did not. The results suggest that l-lactic acid and l-tartaric acid in WPE function as hydrogen donors in the anaerobic microbial degradation of chloroethene. This technology realizes environmental remediation through the effective use of food by-products.Graphical

Salinity determines performance, functional populations, and microbial ecology in consortia attenuating organohalide pollutants.

Organohalide pollutants are prevalent in coastal regions due to extensive intervention by anthropogenic activities, threatening public health and ecosystems. Gradients in salinity are a natural feature of coasts, but their impacts on the environmental fate of organohalides and the underlying microbial communities remain poorly understood. Here we report the effects of salinity on microbial reductive dechlorination of tetrachloroethene (PCE) and polychlorinated biphenyls (PCBs) in consortia derived from distinct environments (freshwater and marine sediments). Marine-derived microcosms exhibited higher halotolerance during PCE and PCB dechlorination, and a halotolerant dechlorinating culture was enriched from these microcosms. The organohalide-respiring bacteria (OHRB) responsible for PCE and PCB dechlorination in marine microcosms shifted from Dehalococcoides to Dehalobium when salinity increased. Broadly, lower microbial diversity, simpler co-occurrence networks, and more deterministic microbial community assemblages were observed under higher salinity. Separately, we observed that inhibition of dechlorination by high salinity could be attributed to suppressed viability of Dehalococcoides rather than reduced provision of substrates by syntrophic microorganisms. Additionally, the high activity of PCE dechlorinating reductive dehalogenases (RDases) in in vitro tests under high salinity suggests that high salinity likely disrupted cellular components other than RDases in Dehalococcoides. Genomic analyses indicated that the capability of Dehalobium to perform dehalogenation under high salinity was likely owing to the presence of genes associated with halotolerance in its genomes. Collectively, these mechanistic and ecological insights contribute to understanding the fate and bioremediation of organohalide pollutants in environments with changing salinity.

Sulfurospirillum diekertiae sp. nov., a tetrachloroethene-respiring bacterium isolated from contaminated soil.

Two anaerobic, tetrachloroethene- (PCE-) respiring bacterial isolates, designated strain ACSDCE T and strain ACSTCE, were characterized using a polyphasic approach. Cells were Gram-stain-negative, motile, non-spore-forming and shared a vibrioid- to spirillum-shaped morphology. Optimum growth occurred at 30 °C and 0.1–0.4 % salinity. The pH range for growth was pH 5.5–7.5, with an optimum at pH 7.2. Hydrogen, formate, pyruvate and lactate as electron donors supported respiratory reductive dechlorination of PCE to cis-1,2-dichloroethene (cDCE) in strain ACSDCE T and of PCE to trichloroethene (TCE) in strain ACSTCE. Both strains were able to grow with pyruvate under microaerobic conditions. Nitrate, elemental sulphur, and thiosulphate were alternative electron acceptors. Autotrophic growth was not observed and acetate served as carbon source for both strains. The major cellular fatty acids were C16 : 1 ω7c, C16 : 0, C14 : 0 and C18 : 1 ω7c. Both genomes feature a circular plasmid. Strains ACSDCE T and ACSTCE were previously assigned to the candidate species 'Sulfurospirillum acididehalogenans'. Here, based on key genomic features and pairwise comparisons of whole-genome sequences, including average nucleotide identity, digital DNA–DNA hybridization and average amino acid identity, strains ACSDCE T and ACSTCE, 'Ca. Sulfurospirillum diekertiae' strains SL2-1 and SL2-2, and the unclassified Sulfurospirillum sp. strain SPD-1 are grouped into one distinct species separate from previously described Sulfurospirillum species. Compared to Sulfurospirillum multivorans and Sulfurospirillum halorespirans , which dechlorinate PCE to cDCE without substantial TCE accumulation, these five strains produce TCE or cDCE as the end product. In addition, some cellular fatty acids (e.g., C16 : 0 3OH, C17 : 0 iso 3OH, C17 : 0 2OH) were detected in strains ACSDCE T and ACSTCE but not in other Sulfurospirillum species. On the basis of phylogenetic, physiological and phenotypic characteristics, 'Ca. Sulfurospirillum acididehalogenans' and 'Ca. Sulfurospirillum diekertiae' are proposed to be merged into one novel species within the genus Sulfurospirillum , for which the name Sulfurospirillum diekertiae sp. nov. is proposed. The type strain is ACSDCE T (=JCM 33349T= KCTC 15819T=CGMCC 1.5292T).

Iron nitride nanoparticles for rapid dechlorination of mixed chlorinated ethene contamination.

Sulfidation and, more recently, nitriding have been recognized as promising modifications to enhance the selectivity of nanoscale zero-valent iron (nZVI) particles for trichloroethene (TCE). Herein, we investigated the performance of iron nitride (FexN) nanoparticles in the removal of a broader range of chlorinated ethenes (CEs), including tetrachloroethene (PCE), cis-1,2-dichloroethene (cis-DCE), and their mixture with TCE, and compared it to the performance of sulfidated nZVI (S-nZVI) prepared from the same precursor nZVI. Two distinct types of iron nitride (FexN) nanoparticles, containing γ'-Fe4N and ε-Fe2-3N phases, exhibited substantially higher PCE and cis-DCE dechlorination rates compared to S-nZVI. A similar effect was observed with a CE mixture, which was completely dechlorinated by both types of FexN nanoparticles within 10 days, whereas S-nZVI was able to remove only about half of the amount, most of which being TCE. Density functional theory calculations further revealed that the cleavage of the first C-Cl bond was the rate-limiting step for all CEs dechlorinated on the γ'-Fe4N(001) surface, with the reaction barriers of PCE and cis-DCE being 29.9, and 40.8kJmol-1, respectively. FexN nanoparticles proved to be highly effective in the remediation of PCE, cis-DCE, and mixed CE contamination.

Effects of Heavy Metal Ions on Microbial Reductive Dechlorination of 1, 2-Dichloroethane and Tetrachloroethene

Microbial reductive dechlorination has been considered an effective process for the clean-up of organohalide-contaminated sites. Heavy metal ions are commonly present as co-contaminants in various organohalide-contaminated sites. To understand the impacts of heavy metal ions on the environmental fate of organohalides, we investigated the effects of Zn2+, Cu2+ and Cd2+ on reductive dechlorination of tetrachloroethene (PCE) and 1,2-dichloroethane (1,2-DCA) in sediment microcosms and transferred enrichment cultures. PCE and 1,2-DCA-dechlorinating enrichment cultures could be consecutively transferred in the presence of up to 10 mg/L Cu2+ or 10 mg/L Zn2+; by comparison, up to 50 mg/L Cd2+ had minor impacts on the microbial reductive dechlorination of PCE and 1,2-DCA. The inhibitory effects of tested heavy metal ions on microbial reductive dechlorination ranked in descending order are Zn2+, Cu2+, and Cd2+. Community profiling and principal component analysis indicate that the concentration and type of contaminants (e.g., heavy metal ions, organohalides) shaped the microbial community structure, an observation similar to a prior report. The enrichment of certain organohalide-respring bacteria (e.g., Dehalococcoides, Dehalogenimonas) during continuous transfers exposed to heavy metal ions suggests that they are capable of tolerating high concentrations of heavy metal ions. Our findings provide insights into the impacts of heavy metal ions on microbial reductive dechlorination and may be helpful for in situ bioremediation at sites contaminated with organohalides and heavy metals.

Enhanced perchloroethene dechlorination by humic acids via increasing the dehalogenase activity of Dehalococcoides strains.

Perchloroethene (PCE) is a widely used chlorinated solvent. PCE is toxic to humans and has been identified as an environmental contaminant at thousands of sites worldwide. Several Dehalococcoides mccartyi strains can transform PCE to ethene, and thus contribute to bioremediation of contaminated sites. Humic acids (HA) are ubiquitous redox-active compounds of natural aquatic and soil systems and have been intensively studied because of their effect in electron transfer. In this study, we observed the dechlorination of PCE was accelerated by HA in mixed cultures containing Dehalococcoides strains. Anthraquinone-2,6-disulfonic acid (AQDS), a humic acid analogue, inhibited PCE dechlorination in our cultures and thus induced an opposite effect on PCE dehalogenation than HA. We observed the same effect on PCE dechlorination with the pure culture of Dehalococcoides mccartyi strain CBDB1. Not only in mixed cultures but also in pure cultures, growth of Dehalococcoides was not influenced by HA but inhibited by AQDS. Enzymatic activity tests confirmed the dehalogenating activity of strain CBDB1 was increased by HA, especially when using hydrogen as electron donor. We conclude that HA enhanced PCE dechlorination by increasing the reaction speed between hydrogen and the dehalogenase enzyme rather than acting as electron shuttle through its quinone moieties.

Biodegradation of Tetrachloroethene in Batch Experiment and PHREEQC Model; Kinetic Study

Introduction: Bioremediation and biodegradation are considered as environmental friendly techniques for contaminants’ removal in polluted environment. In this study, the removal and kinetics of Tetrachloroethene (PCE) and Trichloroethene (TCE) microbial degradation, their inhibitory effects and the rate of dehalogenation capacity at high concentration of PCE were investigated.Materials and Methods: Dechlorinating culture was provided by Bioclear B.V. from a PCE-contaminated site (Evenblij in Hoogeveen -The Netherlands). The batch apparatuses were placed in an orbital shaker at 150 rpm at room temperature. In all the 18 batches, 6 different concentrations of PCE were measured from 0.1 mM to 0.6 mM. The degradation rate of PCE, Trichloroethene (TCE), and cis-1,2-dichloroethene (cDCE) were determined by the PHREEQC model.Results: The results revealed that the final product was ethene and the rate of dechlorinating of PCE increased gradually. The degradation process started after 3 days in batch modes (0.1 mM). After 10 days, the dechlorination of PCE to TCE was obtained in a low concentration of PCE (0.1 mM). Also, the TCE concentration became close to zero after 10 days. However, the start point was longer than PCE and the rate of biodegradation of TCE was faster than PCE. PCE did not show any progress in the dechlorinating procedure at 13th and 14th batch series and none of the daughter products were observed.Conclusions: It should be concluded that there was no single organism that could dechlorinate PCE to ethene, directly. Therefore, the best consortium of microorganisms to dechlorinate PCE to ethene faster, with less production of VC as the most hazardous compound, should be studied.

Constraints in Anaerobic Microbial Dechlorination, Fermentation, and Sulfate-Reduction Induced by High Concentrations of Tetrachloroethylene

Anaerobic bioremediation of tetrachloroethylene (PCE) under high concentration conditions is difficult. Anaerobic dechlorination of PCE occurs with synergetic reactions, fermentation, and sulfate-reduction; however, the way in which high concentrations of PCE affects these reactions is still poorly understood. This study aims to elucidate how high concentrations of PCE affect fermentation and sulfate-reduction, as well as PCE dechlorination. Laboratory dechlorination tests were performed using a wide concentration range of PCE between 2 and 125 mg/L added to a microbial consortium that had been continuously cultivated in the laboratory and completely dechlorinated PCE for over 4 years. Fermentation of lactate, reduction of sulfate, and dechlorination of PCE were monitored in addition to microbial activities based on RNA. All three reactions, fermentation, sulfate-reduction, and PCE dechlorination were observed to be inhibited. The inhibition for fermentation, sulfate-reduction, and dechlorination occurred when PCE concentrations were higher than 125, 75, and 30 mg/L, respectively. The fermenter, Anaerotignum, and the sulfate-reducer, Desulfosporosinus, were active when the dechlorination was inhibited with 30 mg/L of PCE. These findings suggest that there is interference of PCE dechlorination, despite the occurrence of fermentation and sulfate reduction. Bioaugmentation with a PCE dechlorinator that is tolerant to high PCE concentrations can be a possible solution for bioremediation of PCE when its concentrations are greater than 30 mg/L.

Catalytic performance and mechanism of biochars for dechlorination of tetrachloroethylene in sulfide aqueous solution

Biochar (BC) has been investigated as a natural and economical activator to treat organic contamination. Compared with commercial carbon materials, BCs have a better prospect of large-scale application. For the first time, this study certified degradation of tetrachloroethene (PCE) in sulfide-containing aqueous solutions catalyzed by wormwood-based BCs pyrolyzed at 600 °C, 700 °C, and 800 °C (BC600, BC700, and BC800). Interestingly, BC800 could catalyze PCE dechlorination and form acetylene and chloride ion with over 99 % nontoxic transformation in neutral and alkaline pH conditions. Furthermore, materials surface properties, BC dosages and sulfide concentrations were considered as limiting factors for dechlorination, and the last one had the strongest influence. XPS analysis demonstrated that catalytic ability of BC was attributed to pyridine nitrogen (N6) on surface, because C and O adjacent to N6 strongly favor nucleophilic reactions. These results evaluated the applicability of degrading toxic chlorinated alkenes mediated by natural carbon materials in sulfide-containing environment.

Biodegradation of tetrachloroethylene by a newly isolated aerobic Sphingopyxis ummariensis VR13

Chlorinated aliphatic solvents are major sources of groundwater and soil contamination. In this study, an aerobic bacterial strain, Sphingopyxis ummariensis VR13, which has been newly isolated from petrochemical wastewater sludge, was used for the dechlorination of PCE in relatively high concentrations. The addition of a co-substrate as glucose and yeast extract enhanced the dechlorination of PCE. An adaptation of the bacterial cells to PCE resulted in a significant increase in the PCE degradation yield (62.9–39.4%) at relatively high initial PCE concentrations (0.4–5 mM). The adapted cells achieved the highest biodegradation yield (64.8%) in 1.2mM. However, the maximum dechlorination percentage (41.6%) was measured in lower PCE concentration. The kinetic studies showed that PCE degradation was associated with the biomass growth because a higher removal of PCE (64.8%) occurred in a higher cell density. The degradation kinetics of PCE was properly fitted by Monod-like equation with the specific degradation rate of 7.2mmol PCE (g biomass)−1d−1, which was even faster than the reported anaerobic bacteria at this concentration. This strain can be used in the aerobic degradation of PCE.

Reductive dechlorination of 1,2-dichloroethane in the presence of chloroethenes and 1,2-dichloropropane as co-contaminants

1,2-Dichloroethane (1,2-DCA) is one of the most abundant manmade chlorinated organic contaminants in the world. Reductive dechlorination of 1,2-DCA by organohalide-respiring bacteria (OHRB) can be impacted by other chlorinated contaminants such as chloroethenes and chloropropanes that can co-exist with 1,2-DCA at contaminated sites. The aim of this study was to evaluate the effect of chloroethenes and 1,2-dichloropropane (1,2-DCP) on 1,2-DCA dechlorination using sediment cultures enriched with 1,2-DCA as the sole chlorinated compound (EA culture) or with 1,2-DCA and tetrachloroethene (PCE) (EB culture), and to model dechlorination kinetics. Both cultures contained Dehalococcoides as most predominated OHRB, and Dehalogenimonas and Geobacter as other known OHRB. In sediment-free enrichments obtained from the EA and EB cultures, dechlorination of 1,2-DCA was inhibited in the presence of the same concentrations of either PCE, vinyl chloride (VC), or 1,2-DCP; however, concurrent dechlorination of dual chlorinated compounds was achieved. In contrast, 1,2-DCA dechlorination completely ceased in the presence of cis-dichloroethene (cDCE) and only occurred after cDCE was fully dechlorinated. In turn, 1,2-DCA did not affect dechlorination of PCE, cDCE, VC, and 1,2-DCP. In sediment-free enrichments obtained from the EA culture, Dehalogenimonas 16S rRNA gene copy numbers decreased 1–3 orders of magnitude likely due to an inhibitory effect of chloroethenes. Dechlorination with and without competitive inhibition fit Michaelis-Menten kinetics and confirmed the inhibitory effect of chloroethenes and 1,2-DCP on 1,2-DCA dechlorination. This study reinforces that the type of chlorinated substrate drives the selection of specific OHRB, and indicates that removal of chloroethenes and in particular cDCE might be necessary before effective removal of 1,2-DCA at sites contaminated with mixed chlorinated solvents.

Functional Genes and Bacterial Communities During Organohalide Respiration of Chloroethenes in Microcosms of Multi-Contaminated Groundwater.

Microcosm experiments with CE-contaminated groundwater from a former industrial site were set-up to evaluate the relationships between biological CE dissipation, dehalogenase genes abundance and bacterial genera diversity. Impact of high concentrations of PCE on organohalide respiration was also evaluated. Complete or partial dechlorination of PCE, TCE, cis-DCE and VC was observed independently of the addition of a reducing agent (Na2S) or an electron donor (acetate). The addition of either 10 or 100 μM PCE had no effect on organohalide respiration. qPCR analysis of reductive dehalogenases genes (pceA, tceA, vcrA, and bvcA) indicated that the version of pceA gene found in the genus Dehalococcoides [hereafter named pceA(Dhc)] and vcrA gene increased in abundance by one order of magnitude during the first 10 days of incubation. The version of the pceA gene found, among others, in the genus Dehalobacter, Sulfurospirillum, Desulfuromonas, and Geobacter [hereafter named pceA(Dhb)] and bvcA gene showed very low abundance. The tceA gene was not detected throughout the experiment. The proportion of pceA(Dhc) or vcrA genes relative to the universal 16S ribosomal RNA (16S rRNA) gene increased by up to 6-fold upon completion of cis-DCE dissipation. Sequencing of 16S rRNA amplicons indicated that the abundance of Operational Taxonomic Units (OTUs) affiliated to dehalogenating genera Dehalococcoides, Sulfurospirillum, and Geobacter represented more than 20% sequence abundance in the microcosms. Among organohalide respiration associated genera, only abundance of Dehalococcoides spp. increased up to fourfold upon complete dissipation of PCE and cis-DCE, suggesting a major implication of Dehalococcoides in CEs organohalide respiration. The relative abundance of pceA and vcrA genes correlated with the occurrence of Dehalococcoides and with dissipation extent of PCE, cis-DCE and CV. A new type of dehalogenating Dehalococcoides sp. phylotype affiliated to the Pinellas group, and suggested to contain both pceA(Dhc) and vcrA genes, may be involved in organohalide respiration of CEs in groundwater of the study site. Overall, the results demonstrate in situ dechlorination potential of CE in the plume, and suggest that taxonomic and functional biomarkers in laboratory microcosms of contaminated groundwater following pollutant exposure can help predict bioremediation potential at contaminated industrial sites.

Nitrous Oxide Is a Potent Inhibitor of Bacterial Reductive Dechlorination

Organohalide-respiring bacteria are key players for the turnover of organohalogens. At sites impacted with chlorinated ethenes, bioremediation promotes reductive dechlorination; however, stoichiometric conversion to environmentally benign ethene is not always achieved. We demonstrate that nitrous oxide (N2O), a compound commonly present in groundwater, inhibits organohalide respiration. N2O concentrations in the low micromolar range decreased dechlorination rates and resulted in incomplete dechlorination of tetrachloroethene (PCE) in Geobacter lovleyi strain SZ and of cis-1,2-dichloroethene ( cDCE) and vinyl chloride (VC) in Dehalococcoides mccartyi strain BAV1 axenic cultures. Presumably, N2O interferes with reductive dechlorination by reacting with super-reduced Co(I)-corrinoids of reductive dehalogenases, which is supported by the finding that N2O did not inhibit corrinoid-independent fumarate-to-succinate reduction in strain SZ. Kinetic analyses revealed a best fit to the noncompetitive Michaelis-Menten inhibition model and determined N2O inhibitory constants, KI, for PCE and cDCE dechlorination of 40.8 ± 3.8 and 21.2 ± 3.5 μM in strain SZ and strain BAV1, respectively. The lowest KI value of 9.6 ± 0.4 μM was determined for VC to ethene reductive dechlorination in strain BAV1, suggesting that this crucial dechlorination step for achieving detoxification is most susceptible to N2O inhibition. Groundwater N2O concentrations exceeding 100 μM are not uncommon, especially in watersheds impacted by nitrate runoff from agricultural sources. Thus, dissolved N2O measurements can inform about cDCE and VC stalls at sites impacted with chlorinated ethenes.

Strategy for the Rapid Dechlorination of Polychlorinated Biphenyls (PCBs) by Dehalococcoides mccartyi Strains.

Anaerobic bacteria play critical roles in the environmental bioremediation of polychlorinated biphenyls (PCBs). However, in situ applications of PCB dechlorinating anaerobes have been largely impeded by difficulties in growing PCB dechlorinators to a high cell abundance in short time periods. Here, we report the accelerated onset of PCB dechlorination by pre-cultivating Dehalococcoides mccartyi strains on chloroethenes as alternative electron acceptors. The extensive dechlorination of Aroclor 1260 was achieved within 1 week by D. mccartyi CG4 pregrown on trichloroethene (TCE) or tetrachloroethene (PCE). Compared to control cultures fed solely with Aroclor 1260, the PCB dechlorination rate was up to 30 times greater in cultures pre-cultivated with chloroethenes. However, when CG4 was simultaneously exposed to multiple potential substrates (PCE, TCE, and PCBs), PCB dechlorination was not observed until PCE was completely depleted. The expression of the bifunctional PCE and PCBs reductive dehalogenase (RDase) gene pcbA4 was inhibited by the presence of both substrates. Furthermore, in the presence of >0.3 mM TCE (produced as an intermediate from PCE dechlorination), the PCB dechlorination rate was an order of magnitude lower than in cultures amended with Aroclor 1260 after the complete depletion of TCE. This reduced PCB dechlorination rate corresponded with a sharp decrease in pcbA4 transcripts in the presence of both TCE and PCBs. The utilization of chloroethenes and PCBs as substrates by CG4 was found to be largely sequential rather than concurrent, suggesting that PCE and TCE are preferred substrates for the RDase responsible for PCE, TCE, and PCB dechlorination.

Acceleration of perchloroethylene dechlorination by extracellular secretions from Microbacterium in a mixed culture containing Desulfitobacterium

The study was conducted to demonstrate the influence of extracellular secretions from Microbacterium on the reductive dechlorination of tetrachloroethene (PCE). A series of mixed cultures were established from a paddy soil sample. In the mixed cultures amended with extracellular secretions from Microbacterium, PCE was rapidly and completely converted into cis-1,2-dichloroethene (cis-DCE) and trans-1,2-dichloroethene (trans-DCE) within 40 days. The unamended mixed cultures showed weak signs of dechlorination after a pronounced lag phase, and trichloroethene (TCE) was accumulated as a major end product. This result means that amendment with extracellular secretions from Microbacterium shortened the lag phase, increased the dechlorination velocity and promoted the production of less-chlorinated chloroethene. The results were corroborated by defined subculture experiments, which proved that microorganisms from unamended mixed cultures could also be stimulated by extracellular secretions from Microbacterium. Desulfitobacterium was identified as the main dechlorinating population in all mixed cultures by direct PCR. Additionally, the 16S rRNA gene copies of Desulfitobacterium increased by one or two orders of magnitude with PCE dechlorination, which provided corroborative evidence for the identification result. The volatile fatty acids were monitored, and most interestingly, a close association between propionate oxidation and dechlorination was found, which has rarely been mentioned before. It was assumed that the oxidation of propionate provided hydrogen for dechlorination, while dechlorination facilitated the shift of the reaction toward propionate oxidation by reducing the partial pressure of hydrogen.

Multi-method assessment of the intrinsic biodegradation potential of an aquifer contaminated with chlorinated ethenes at an industrial area in Barcelona (Spain)

The bioremediation potential of an aquifer contaminated with tetrachloroethene (PCE) was assessed by combining hydrogeochemical data of the site, microcosm studies, metabolites concentrations, compound specific-stable carbon isotope analysis and the identification of selected reductive dechlorination biomarker genes. The characterization of the site through 10 monitoring wells evidenced that leaked PCE was transformed to TCE and cis-DCE via hydrogenolysis. Carbon isotopic mass balance of chlorinated ethenes pointed to two distinct sources of contamination and discarded relevant alternate degradation pathways in the aquifer. Application of specific-genus primers targeting Dehalococcoides mccartyi species and the vinyl chloride-to-ethene reductive dehalogenase vcrA indicated the presence of autochthonous bacteria capable of the complete dechlorination of PCE. The observed cis-DCE stall was consistent with the aquifer geochemistry (positive redox potentials; presence of dissolved oxygen, nitrate, and sulphate; absence of ferrous iron), which was thermodynamically favourable to dechlorinate highly chlorinated ethenes but required lower redox potentials to evolve beyond cis-DCE to the innocuous end product ethene. Accordingly, the addition of lactate or a mixture of ethanol plus methanol as electron donor sources in parallel field-derived anoxic microcosms accelerated dechlorination of PCE and passed cis-DCE up to ethene, unlike the controls (without amendments, representative of field natural attenuation). Lactate fermentation produced acetate at near-stoichiometric amounts. The array of techniques used in this study provided complementary lines of evidence to suggest that enhanced anaerobic bioremediation using lactate as electron donor source is a feasible strategy to successfully decontaminate this site.