Aerobic metabolic trichloroethene biodegradation under field-relevant conditions.

Chloroethenes belong to the most widely distributed groundwater contaminants. Since 2014, it has been known that trichloroethene (TCE) can be degraded aerobically and metabolically as growth substrate by a mixed bacterial enrichment culture (named SF culture). In this study, the degradation capabilities under a range of field-relevant conditions were investigated in fixed-bed reactors as well as in batch experiments. Aerobic metabolic TCE degradation was stable over the long term, with degradation optima at 22 °C and pH 7. Degradation of up to 400 µM TCE was observed. The longest starvation period after which degradation of TCE was regained was 112 days. The possible co-contaminants perchloroethene, trans-1,2-dichloroethene, and cis-1,2-dichloroethene did not inhibit TCE degradation, even though they were not degraded themselves. The presence of equimolar amounts of 1,1-dichloroethene and vinyl chloride inhibited TCE degradation. Experiments with groundwater from different chloroethene-contaminated field sites proved the potential of the SF culture for bioaugmentation. Thus, aerobic metabolic TCE degradation should be considered as a promising method for the bioremediation of field sites with TCE as the main contaminant.

The ecotoxic potential of a new zero-valent iron nanomaterial, designed for the elimination of halogenated pollutants, and its effect on reductive dechlorinating microbial communities.

The purpose of this study was to assess the ecotoxic potential of a new zero-valent iron nanomaterial produced for the elimination of chlorinated pollutants at contaminated sites. Abiotic dechlorination through the newly developed nanoscale zero-valent iron material and its effects on dechlorinating bacteria were investigated in anaerobic batch and column experiments. The aged, i.e. oxidized, iron material was characterization with dynamic light scattering, transmission electron microscopy and energy dispersive x-ray analysis, x-ray diffractometry and cell-free reactive oxygen measurements. Furthermore, it was evaluated in aerobic ecotoxicological test systems with algae, crustacean, and fish, and also applied in a mechanism specific test for mutagenicity. The anaerobic column experiments showed co-occurrence of abiotic and biological dechlorination of the common groundwater contaminant perchloroethene. No prolonged toxicity of the nanomaterial (measured for up to 300 days) towards the investigated dechlorinating microorganism was observed. The nanomaterial has a flake like appearance and an inhomogeneous size distribution. The toxicity to crustacean and fish was calculated and the obtained EC50 values were 163 mg/L and 458 mg/L, respectively. The nanomaterial showed no mutagenicity. It physically interacted with algae, which had implications for further testing and the evaluation of the results. Thus, the newly developed iron nanomaterial was slightly toxic in its reduced state but no prolonged toxicity was recorded. The aquatic tests revealed a low toxicity with EC50 values ≥ 163 mg/L. These concentrations are unlikely to be reached in the aquatic environment. Hence, this nanomaterial is probably of no environmental concern not prohibiting its application for groundwater remediation.

Aerobic biodegradation of trichloroethene without auxiliary substrates.

Trichloroethene (TCE) represents a priority pollutant and is among the most frequently detected contaminants in groundwater. The current bioremediation measures have certain drawbacks like e.g. the need for auxiliary substrates. Here, the aerobic biodegradation of TCE as the sole growth substrate is demonstrated. This new process of metabolic TCE degradation was first detected in groundwater samples. TCE degradation was stable in an enriched mixed bacterial culture in mineral salts medium for over five years and repeated transfers of the culture resulting in a 10(10) times dilution of the original groundwater. Aerobic TCE degradation resulted in stoichiometric chloride formation. Stable carbon isotope fractionation was observed providing a reliable analytical tool to assess this new biodegradation process at field sites. The results suggest that aerobic biodegradation of TCE without auxiliary substrate could be considered as an option for natural attenuation or engineered bioremediation of contaminated sites.

Effect of chloroethene concentrations and granular activated carbon on reductive dechlorination rates and growth of Dehalococcoides spp.

This study focused on the investigation of (i) the tetrachloroethene (PCE) toxicity threshold of a reductively dechlorinating mixed culture containing Dehalococcoides spp., (ii) the adsorption of PCE on different types of granular activated carbon (GAC), and (iii) the bioavailability and reductive dechlorination in the presence of GAC. The abundance of Dehalococcoides spp. detected by quantitative real-time polymerase chain reaction (qPCR) was found to increase by 2-4 orders of magnitude during degradation of PCE. No degradation occurred at dissolved concentrations beyond 420 µM (70 mg/L). Different adsorption isotherms were determined for thermally and chemically activated carbons. The addition of GAC to biological assays reduced the dissolved PCE concentration below the toxicity threshold. The combination of microbial reductive dechlorination with GAC adsorption proved to be a promising method for remediation of groundwater contaminated by high concentrations of chloroethenes.

Robustness of an aerobic metabolically vinyl chloride degrading bacterial enrichment culture.

Degradation of the lower chlorinated ethenes is crucial to the application of natural attenuation or in situ bioremediation on chlorinated ethene contaminated sites. Recently, within mixtures of several chloroethenes as they can occur in contaminated groundwater inhibiting effects on aerobic chloroethene degradation have been shown. The current study demonstrated that metabolic vinyl chloride (VC) degradation by an enrichment culture originating from groundwater was not affected by an equimolar concentration (50 µM) of cis-1,2-dichloroethene (cDCE). Only cDCE concentrations at a ratio of 2.4:1 (initial cDCE to VC concentration) caused minor inhibition of VC degradation. Furthermore, the degradation of VC was not affected by the presence of trans-1,2-dichloroethene (tDCE), 1,1-dichloroethene (1,1-DCE), trichloroethene (TCE), and tetrachloroethene (PCE) in equimolar concentrations (50 µM). Only cDCE and tDCE were cometabolically degraded in small amounts. The VC-degrading culture demonstrated a broad pH tolerance from 5 to 9 with an optimum between 6 and 7. Results also showed that the culture could degrade VC concentrations up to 1,800 µM (110 mg/L).

Sequential anaerobic/aerobic biodegradation of chloroethenes--aspects of field application.

Because of a range of different industrial activities, sites contaminated with chloroethenes are a world-wide problem. Chloroethenes can be biodegraded by reductive dechlorination under anaerobic conditions as well as by oxidation under aerobic conditions. The tendency of chloroethenes to undergo reductive dechlorination decreases with a decreasing number of chlorine substituents, whereas with less chlorine substituents chloroethenes more easily undergo oxidative degradation. There is currently a growing interest in aerobic metabolic degradation of chloroethenes, which demonstrates advantages compared to cometabolic degradation pathways. Sequential anaerobic/aerobic biodegradation can overcome the disadvantages of reductive dechlorination and leads to complete mineralization of the chlorinated pollutants. This approach shows promise for site remediation in natural settings and in engineered systems.

Aerobic biodegradation of cis-1,2-dichloroethene as sole carbon source: Stable carbon isotope fractionation and growth characteristics.

Cis-1,2-dichloroethene (cDCE) is a compound of concern at many chloroethene-contaminated sites, since it tends to accumulate during reductive dechlorination of the higher chlorinated ethenes. Stable carbon isotope fractionation during aerobic cDCE biodegradation was observed in groundwater microcosms under varying incubation conditions (room temperature/groundwater temperature; with/without inorganic nutrients), and resulted in an average stable carbon isotope enrichment factor of -15.2+/-0.5 per thousand. A new enrichment culture, obtained from groundwater microcosms, degraded cDCE concentrations up to 100mgL(-1), was active at temperatures between 4 and 23 degrees C, had a pH optimum of approximately 7, and could withstand prolonged periods (250d) of starvation. Microbial growth during degradation of cDCE as sole carbon and energy source was demonstrated by protein formation in mineral medium not containing any known auxiliary substrate. The obtained growth yield was 12.5+/-1.9g of proteinMol(-1) of cDCE, with a doubling time of 53+/-2h at 23 degrees C. Aerobic degradation of cDCE as sole carbon and energy source appears to be a promising biological process for site remediation.

Inhibition of aerobic metabolic cis-1,2-di-chloroethene biodegradation by other chloroethenes.

The presence of other chloroethenes influences aerobic metabolic biodegradation of cis-1,2-dichloroethene (cDCE). A new metabolically cDCE degrading enrichment culture was identified as also being capable of degrading vinyl chloride (VC), but not 1,1-dichloroethene (1,1DCE), trans-1,2-dichloroethene (tDCE), trichloroethene (TCE), or tetrachloroethene (PCE). The fastest degradation of cDCE was observed in the absence of any other chloroethene. In the presence of a second chloroethene (40-90 microM), the rate of cDCE (60 microM) degradation decreased in the following order: cDCE (+PCE) > cDCE (+tDCE) > cDCE (+VC)>cDCE (+1,1DCE) approximately cDCE (+TCE). With increasing concentrations of VC, ranging from 10 to 110 microM, the rate of cDCE (60 microM) degradation decreased. This study demonstrates that the inhibiting effects of chloroethene mixtures have to be considered during laboratory studies and bioremediation approaches based on metabolic cDCE degradation.

Growth kinetics and stable carbon isotope fractionation during aerobic degradation of cis-1,2-dichloroethene and vinyl chloride.

Assessing changes in the isotopic signature of contaminants is a promising new tool to monitor microbial degradation processes. In this study, chloroethene degradation was proven by depletion of chloroethenes, formation of chloride, increase in protein content and stable carbon isotope fractionation. Aerobic degradation of vinyl chloride (VC) was found to proceed metabolically, with degradation rates of 0.48 and 0.29 d(-1); and growth yields of 9.7 and 6.4 g of protein/mol of VC at room and groundwater temperature, respectively. Cis-1,2-dichloroethene (cDCE) was degraded cometabolically under aerobic conditions when VC was provided as growth substrate. Aerobic degradation was associated with significant stable carbon isotope fractionation, with enrichment factors ranging from -5.4+/-0.4 per thousand for metabolic degradation of VC to -9.8+/-1.7 per thousand for cometabolic degradation of cDCE. Thus, it was demonstrated that stable carbon isotope fractionation is suitable for assessing aerobic chloroethene degradation, which can contribute significantly to site remediation.

Fungal inoculum properties and its effect on growth and enzyme activity of Trametes versicolor in soil.

The effect of fungal inoculum properties on colonization of nonsterile soil by three isolates of the white-rot fungus Trametes versicolor was investigated. Fungal inoculum properties were examined in separate experiments and were fungal inoculum composition, age of fungal inoculum, concentration of the inoculum and inoculation method. The fungal inoculum composition study compared pine versus poplar sawdust as the basic carrier with varying amounts of corn grit, corn meal and starch. The age of the fungal inoculum studied ranged from 3 to 21 days. The inoculum concentration gradually increased from 0 to 50% (v/v). The study assessing inoculation method compared mixing with layering techniques. The effect of moisture conditions of soil, sawdust and sand in combination with two inoculation methods (mixing versus point source inoculation) on colonization by T. versicolor was also determined. Colonization of soil was always assessed visually and enzymatically monitoring mycelial growth, biological potential (fluorescein diacetate assay) and laccase levels. Generally, the three different assessment methods correlated (P < 0.05) with each other. A fungal inoculum based on pine sawdust supported white-rot fungal growth in soil better than a poplar sawdust basis. Colonization of soil by T. versicolor was improved by increasing the corn content of the fungal inoculum. Younger (<7 days old) fungal inoculum resulted in better soil colonization than older (>10 days). A strong correlation (P < 0.001) was observed between the amount of fungal inoculum used in the soil augmentation and white-rot fungal colonization of soil. Inoculation of the fungal inoculum into soil by mixing was preferable over application in layers or point source inoculation. Moisture level did not influence biological potential measurements, but affected mycelial growth and laccase expression.