Background: Nanoscale zero-valent iron (nZVI) is commonly used for remediation of groundwater contaminated by chlorinated ethenes (CEs); however, its long-term reactivity and subsurface transport are limited. A novel nZVI–AC material, consisting of colloidal activated carbon (AC) with embedded nZVI clusters, was developed with the aim of overcoming the limitations of nZVI alone. Results: Application of a limited amount of nZVI–AC to an oxic, nitrate-rich, highly permeable quaternary aquifer triggered time-limited transformation of CEs, with noticeable involvement of reductive dechlorination. Reductive dechlorination of CEs was dominantly abiotic, as an increase in the concentration of vinyl chloride (VC) and ethene did not coincide with an increase in the abundance of reductive biomarkers for complete dechlorination of CEs (Dehalococcoides, Dehalogenimonas, VC reductase genes vcrA and bvcA). Application of nZVI–AC under unfavourable hydrochemical conditions resulted in no dramatic change in the microbial community, the reducing effect resulting in temporal proliferation of nitrate and iron reducers only. At a later stage, generation of reduced iron induced an increase in iron-oxidizing bacteria. High concentrations and a continuous mass influx of competing electron acceptors (nitrate and dissolved oxygen) created unfavourable conditions for sulphate-reducers and organohalide-respiring bacteria, though it allowed the survival of aerobic microorganisms of the genera Pseudomonas, Polaromonas and Rhodoferax, known for their ability to assimilate VC or cis-1,2-dichloroethene. A potential for aerobic oxidative degradation of CE metabolites was also indicated by detection of the ethenotroph functional gene etnE. Conclusions: This pilot study, based on the application of nZVI–AC, failed to provide a sustainable effect on CE contamination; however, it provided valuable insights into induced hydrogeochemical and microbial processes that could help in designing full-scale applications.

A stall of cis-1,2-DCE and vinyl chloride (VC) is frequently observed during bioremediation of groundwater chloroethenes via reductive dechlorination. These chloroethenes may be oxidised by aerobic methanotrophs or ethenotrophs co-metabolically and/or metabolically. We assessed the potential for such oxidation at 12 sites (49 groundwater samples) using hydrochemical and molecular biological tools. Both ethenotroph (etnC and etnE) and methanotroph (mmoX and pmoA) functional genes were identified in 90% of samples, while reductive dehalogenase functional genes (vcrA and bvcA) were identified in 82%. All functional genes were simultaneously detected in 78% of samples, in actively biostimulated sites in 88% of samples. Correlation analysis revealed that cis-1,2-DCE concentration was positively correlated with vcrA, etnC and etnE, while VC concentration was correlated with etnC, etnE, vcrA and bvcA. However, feature selection based on random forest classification indicated a significant relationship for the vcrA in relation to cis-1,2-DCE, and vcrA, bvcA and etnE for VC and no prove of relationship between cis-1,2-DCE or VC and the methanotroph functional genes. Analysis of hydrochemical parameters indicated that aerobic oxidation of chloroethenes by ethenotrophs may take place under a range of redox conditions of aquifers and coincide with high ethene and VC concentrations.

In situ remediation is usually restricted by temperature, lack of substrate for reductive dechlorination (anaerobic respiration), the presence of dehalogenating microorganisms, and specific bedrock conditions. In this work, trichloroethene (TCE) degradation was studied by a number of methods, from physical–chemical analyses to molecular biological tools. The abundance changes in dechlorinating bacteria were monitored using real-time PCR. The functional genes vcrA and bvcA as well as the 16S rRNA specific for representatives of genera Dehalococcoides, Dehalobacter, and Desulfitobacterium were monitored. Furthermore, the sulfate-reducing bacteria and denitrifying bacteria were observed by amplifying the functional genes apsA and nirK. The elevated temperature and the substrate (whey) addition significantly affected TCE dechlorination. The chlorine index decreased after nine weeks from 2.5 to 0.1 at 22 °C, to 1.1 at 17 °C and 1.7 at 12 °C and complete dechlorination was achieved at 22 °C with whey addition. The achieved results of this work show the feasibility and effectiveness of biological dechlorination of TCE enhanced with elevated temperature and whey addition.