Stratification of chlorinated ethenes natural attenuation in an alluvial aquifer assessed by hydrochemical and biomolecular tools.

Biomolecular and hydrochemical tools were used to evaluate natural attenuation of chlorinated ethenes in a Quaternary alluvial aquifer located close to a historical source of large-scale tetrachloroethylene (PCE) contamination. Distinct stratification of redox zones was observed, despite the aquifer's small thickness (2.8 m). The uppermost zone of the target aquifer was characterised by oxygen- and nitrate-reducing conditions, with mixed iron- to sulphate-reducing conditions dominant in the lower zone, along with indications of methanogenesis. Natural attenuation of PCE was strongly influenced by redox heterogeneity, while higher levels of PCE degradation coincided with iron- to sulphate reducing conditions. Next generation sequencing of the middle and/or lower zones identified anaerobic bacteria (Firmicutes, Chloroflexi, Actinobacteria and Bacteroidetes) associated with reductive dechlorination. The relative abundance of dechlorinators (Dehalococcoides mccartyi, Dehalobacter sp.) identified by real-time PCR in soil from the lower levels supports the hypothesis that there is a significant potential for reductive dechlorination of PCE. Local conditions were insufficiently reducing for rapid complete dechlorination of PCE to harmless ethene. For reliable assessment of natural attenuation, or when designing monitoring or remedial systems, vertical stratification of key biological and hydrochemical markers should be analysed as standard, even in shallow aquifers.

Microbial degradation of chloroethenes: a review.

Contamination by chloroethenes has a severe negative effect on both the environment and human health. This has prompted intensive remediation activity in recent years, along with research into the efficacy of natural microbial communities for degrading toxic chloroethenes into less harmful compounds. Microbial degradation of chloroethenes can take place either through anaerobic organohalide respiration, where chloroethenes serve as electron acceptors; anaerobic and aerobic metabolic degradation, where chloroethenes are used as electron donors; or anaerobic and aerobic co-metabolic degradation, with chloroethene degradation occurring as a by-product during microbial metabolism of other growth substrates, without energy or carbon benefit. Recent research has focused on optimising these natural processes to serve as effective bioremediation technologies, with particular emphasis on (a) the diversity and role of bacterial groups involved in dechlorination microbial processes, and (b) detection of bacterial enzymes and genes connected with dehalogenation activity. In this review, we summarise the different mechanisms of chloroethene bacterial degradation suitable for bioremediation and provide a list of dechlorinating bacteria. We also provide an up-to-date summary of primers available for detecting functional genes in anaerobic and aerobic bacteria degrading chloroethenes metabolically or co-metabolically.

Occurrence of perchloroethylene in surface water and fish in a river ecosystem affected by groundwater contamination.

Long-term monitoring of the content of perchloroethylene (PCE) in a river ecosystem affected by groundwater contamination was performed at a site in the Czech Republic. The quality of surface water was monitored quarterly between 1994 and 2013, and fish were collected from the affected ecosystem to analyse the content of PCE in their tissue in 1998, 2011 and 2012. Concentrations of PCE (9-140 µg/kg) in the tissue of fish collected from the contaminated part of the river were elevated compared to the part of the river unaffected by the contamination (ND to 5 µg/kg PCE). The quality of surface water has improved as a result of groundwater remediation during the evaluated period. Before the remedial action, PCE concentrations ranged from 30 to 95 µg/L (1994-1997). Following commencement of remedial activities in September 1997, a decrease in the content of PCE in the surface water to 7.3 µg/L (1998) and further to 1 µg/L (2011) and 1.1 µg/L (2012) led to a progressive decrease in the average concentration of PCE in the fish muscle tissue from 79 µg/kg (1998) to 24 (2011) and 30 µg/kg (2012), respectively. It was determined that the bioconcentration of PCE does not have a linear dependence because the decrease in contamination in the fish muscle tissue is not directly proportional to the decrease in contamination in the river water. The observed average bioconcentration factors were 24 and 28 for the lower concentrations of PCE and 11 for the higher concentrations of PCE in the river. In terms of age, length and weight of the collected fish, weight had the greatest significance for bioconcentration, followed by the length, with age being evaluated as a less significant factor.

Carbon and chlorine isotope fractionation during microbial degradation of tetra- and trichloroethene.

Two-dimensional compound-specific isotope analysis (2D-CSIA), combining stable carbon and chlorine isotopes, holds potential for monitoring of natural attenuation of chlorinated ethenes (CEs) in contaminated soil and groundwater. However, interpretation of 2D-CSIA data sets is challenged by a shortage of experimental Cl isotope enrichment factors. Here, isotope enrichments factors for C and Cl (i.e., εC and εCl) were determined for biodegradation of tetrachloroethene (PCE) and trichloroethene (TCE) using microbial enrichment cultures from a heavily CE-contaminated aquifer. The obtained values were εC = -5.6 ± 0.7‰ (95% CI) and εCl = -2.0 ± 0.5‰ for PCE degradation and εC = -8.8 ± 0.2‰ and εCl = -3.5 ± 0.5‰ for TCE degradation. Combining the values for both εC and εCl yielded mechanism-diagnostic εCl/εC ratios of 0.35 ± 0.11 and 0.37 ± 0.11 for the degradation of PCE and TCE, respectively. Application of the obtained εC and εCl values to a previously investigated field site gave similar estimates for the fraction of degraded contaminant as in the previous study, but with a reduced uncertainty in assessment of the natural attenuation. Furthermore, 16S rRNA gene clone library analyses were performed on three samples from the PCE degradation experiments. A species closely related to Desulfitobacterium aromaticivorans UKTL dominated the reductive dechlorination process. This study contributes to the development of 2D-CSIA as a tool for evaluating remediation strategies of CEs at contaminated sites.

Dual carbon-chlorine stable isotope investigation of sources and fate of chlorinated ethenes in contaminated groundwater.

Chlorinated ethenes (CEs) are ubiquitous groundwater contaminants, yet there remains a need for a method to efficiently monitor their in situ degradation. We report here the first field application of combined stable carbon and chlorine isotope analysis of tetrachloroethene (PCE) and trichloroethene (TCE) to investigate their biodegradation in a heavily contaminated aquifer. The two-dimensional Compound Specific Isotope Analysis (2D-CSIA) approach was facilitated by a recently developed gas chromatography-quadrupole mass spectrometry (GCqMS) method for δ(37)Cl determination. Both C and Cl isotopes showed evidence of ongoing PCE transformation. Applying published C isotope enrichment factors (ε(C)) enabled evaluation of the extent of in situ PCE degradation (11-78%). We interpreted C and Cl isotopes using a numerical reactive transport model along a 60-m flow path. It revealed that combined PCE and TCE mass load was dechlorinated by less than 10%, and that cis-dichloroethene was not further dechlorinated. Furthermore, the 2D-CSIA approach allowed estimation of Cl isotope enrichment factors ε(Cl) (-7.8 to -0.8‰) and characteristic ε(Cl)/ε(C) values (0.42-1.12) for reductive PCE dechlorination at this field site. This investigation demonstrates the benefit of 2D-CSIA to assess in situ degradation of CEs and the applicability of Cl isotope fractionation to evaluate PCE and TCE dechlorination.

Major factors affecting in situ biodegradation rates of jet-fuel during large-scale biosparging project in sedimentary bedrock.

Biodegradation of petroleum hydrocarbons (TPH), mainly jet fuel, had taken place at the former Soviet Army air base in the Czech Republic. The remediation of large-scale petroleum contamination of soil and groundwater has provided valuable information about biosparging efficiency in the sandstone sedimentary bedrock. In 1997 petroleum contamination was found to be present in soil and groundwater across an area of 28 hectares, divided for the clean-up purpose into smaller clean-up fields (several hectares). The total estimated quantity of TPH released to the environment was about 7,000 metric tons. Biosparging was applied as an innovative clean-up technology at the site and was operated over a 10-year period (1997-2008). Importance of a variety of factors that affect bacterial activity in unsaturated and saturated zones was widely studied on the site and influence of natural and technological factors on clean-up efficiency in heavily contaminates areas of clean-up fields (initial contaminant mass 111-452 metric ton/ha) was evaluated. Long-term monitoring of the groundwater temperature has shown seasonal rises and falls of temperature which have caused a fluctuation in biodegradation activity during clean-up. By contrast, an overall rise of average groundwater temperature was observed in the clean-up fields, most probably as a result of the biological activity during the clean-up process. The significant rise of biodegradation rates, observed after air sparging intensification, and strong linear correlation between the air injection rates and biodegradation activities have shown that the air injection rate is the principal factor in biodegradation efficiency in heavily contaminated areas. It has a far more important role for achieving a biodegradation activity than the contamination content which appeared to have had only a slight effect after the removal of about 75% of initial contamination.

Pollutant plume delineation from tree core sampling using standardized ranks.

There are currently contradicting results in the literature about the way chloroethene (CE) concentrations from tree core sampling correlate with those from groundwater measurements. This paper addresses this issue by focusing on groundwater and tree core datasets in CE contaminated site, Czech Republic. Preliminary analyses revealed strongly and positively skewed distributions for the tree core dataset, with an intra-tree variability accounting for more than 80% of the total variability, while the spatial analyses based on variograms indicated no obvious spatial pattern for CE concentration. Using rank transformation, it is shown how the results were improved by revealing the initially hidden spatial structure for both variables when they are handled separately. However, bivariate analyses based on cross-covariance functions still failed to indicate a clear spatial correlation between groundwater and tree core measurements. Nonetheless, tree core sampling and analysis proved to be a quick and inexpensive semi-quantitative method and a useful tool.

Enhancement of the microbial community biomass and diversity during air sparging bioremediation of a soil highly contaminated with kerosene and BTEX.

In order to obtain insights in complexity shifts taking place in natural microbial communities under strong selective pressure, soils from a former air force base in the Czech Republic, highly contaminated with jet fuel and at different stages of a bioremediation air sparging treatment, were analyzed. By tracking phospholipid fatty acids and 16S rRNA genes, a detailed monitoring of the changes in quantities and composition of the microbial communities developed at different stages of the bioventing treatment progress was performed. Depending on the length of the air sparging treatment that led to a significant reduction in the contamination level, we observed a clear shift in the soil microbial community being dominated by Pseudomonads under the harsh conditions of high aromatic contamination to a status of low aromatic concentrations, increased biomass content, and a complex composition with diverse bacterial taxonomical branches.

Using tree core samples to monitor natural attenuation and plume distribution after a PCE spill.

The potential of using tree core samples to detect and monitor natural attenuation of perchloroethene (PCE) in groundwater was investigated at a PCE-contaminated site. In the area of the known plume with PCE concentrations between 0.004 and > 40 mg/L, cores were collected from tree trunks at a height of about 1 m above ground surface. Tree sampling of the site was completed in under six hours. Chlorinated ethenes were analyzed by headspace GC/MS. PCE (0.001 to 7 mg/ kg) and natural attenuation products, TCE (< 0.001 to 0.4 mg/ kg) and c-DCE (< 0.001 to 0.46 mg/kg), were detected in tree cores. 1,1-dichloroethene and vinyl chloride were not detected, corresponding to very low concentrations in the groundwater. The contaminant plume was mapped from the concentrations measured in trees, which delineated a probable hot spot area that had been undetected in decades of traditional groundwater monitoring. Natural attenuation products in tree cores increased with distance from the known source area. Concentrations of PCE and reductive dechlorination products in tree cores were correlated with the corresponding groundwater concentrations. Within a range of limitations, tree-core sampling provides a rapid, reliable and inexpensive method to investigate the extent of shallow contamination by chlorinated ethenes in soil and groundwater.

Capacity of the bioremediation technology for clean-up of soil and groundwater contaminated with petroleum hydrocarbons.

A column reactor was designed and used to simulate conditions affecting the bioremediations of petroleum hydrocarbons. The work illustratively describes the aerobic (model) clean-up of soil samples enabling to predict the efficiency of a technology installed in parallel on contaminated former airport. The data showing the performance of thus precharacterized technology are presented.