Diverse dechlorinators and dechlorination genes enriched through amendment of chlorinated natural organic matter fractions.

In uncontaminated environments, chlorinated natural organic matter (Cl-NOM) can act as an electron acceptor for organohalide-respiring bacteria. It is unknown, however, whether different types of Cl-NOM are preferentially dechlorinated or whether enrichment with Cl-NOM affects the ability of bacteria to dechlorinate contaminants. In this research NOM was extracted from sediment, fractionated based on hydrophobicity, and either amended to polychlorinated biphenyl-contaminated soil directly or chlorinated and then amended to soil. Amendments of the least hydrophobic Cl-NOM fraction were dechlorinated most rapidly, followed by the moderately hydrophobic Cl-NOM fraction. Soil that had been enriched on the moderately hydrophobic fraction of Cl-NOM was also capable of faster dechlorination of the contaminants trichloroethene and tetrachlorobenzene. Community analysis of the soil during enrichment showed that some known organohalide-respiring bacteria were present and may have played a role in dechlorination; nevertheless, many bacteria appeared to be enriched during both Cl-NOM and contaminant dechlorination. In addition, the quantities of two haloalkane dehalogenase genes increased during enrichment on Cl-NOM. These results show for the first time that Cl-NOM can prime contaminant dechlorination and also suggest that hydrolytic dechlorination processes were involved in both Cl-NOM and contaminant dechlorination.

Aqueous film forming foam and associated perfluoroalkyl substances inhibit methane production and Co-contaminant degradation in an anaerobic microbial community.

Aqueous film forming foams (AFFF) can contain gram per liter concentrations of per- and polyfluoroalkyl substances (PFAS) and are often released in large quantities directly to the environment as they are used to fight fires. AFFF composition is complex and contains many unknown PFAS in addition to ingredients such as hydrocarbons, solvents, and corrosion inhibitors. While biological effects of single PFAS have been studied, the effects of PFAS-containing mixtures, such as AFFF, are unknown. The effect of PFAS on microorganisms is also not well understood; nevertheless, we rely on microorganisms in locations containing elevated PFAS concentrations to perform certain functions, such as carbon cycling and co-contaminant degradation. This study focused on determining the functional consequences of AFFF and PFAS exposure in a microbial community in both the presence and the absence of a co-contaminant. AFFF, select PFAS, and a PFAS mixture were tested to determine the effect of AFFF on an anaerobic microbial community and the characteristics of the PFAS that drive toxicity in such mixtures. To study this, anaerobic digester communities were exposed to PFAS and a co-contaminant (2,4-dichlorophenol, DCP); methane production, as an indicator of toxicity and the community's ability to cycle carbon, and co-contaminant degradation were monitored. Results showed that PFAS and AFFF can alter the toxicity of DCP, inhibit DCP degradation, decrease the number of methanogens present, and change the microbial community structure. DCP was also able to decrease the toxicity of the PFAS perfluorooctane sulfonate (PFOS), possibly by changing the sorption of PFOS to the microorganisms present. Additionally, it was determined that while PFOS was responsible for AFFF toxicity, no single PFAS or simple PFAS mixture accurately accounted for the inhibition of DCP degradation caused by AFFF exposure.

Presence, Diversity, and Enrichment of Respiratory Reductive Dehalogenase and Non-respiratory Hydrolytic and Oxidative Dehalogenase Genes in Terrestrial Environments.

Organohalide-respiring bacteria have been linked to the cycling and possible respiration of chlorinated natural organic matter (Cl-NOM) in uncontaminated soils and sediments. The importance of non-respiratory hydrolytic/oxidative dechlorination processes in the cycling of Cl-NOM in terrestrial soil and sediment, however, is still not understood. This research analyzes the dechlorination potential of terrestrial systems through analysis of the metagenomes of urban lake sediments and cultures enriched with Cl-NOM. Even with the variability in sample type and enrichment conditions, the potential to dechlorinate was universal, with reductive dehalogenase genes and hydrolytic or oxidative dehalogenase genes found in all samples analyzed. The reductive dehalogenase genes detected grouped taxonomically with those from organohalide-respiring bacteria with broad metabolic capabilities, as opposed to those that obligately respire organohalides. Furthermore, reductive dehalogenase genes and two haloacid dehalogenase genes increased in abundance when sediment was enriched with high concentrations of Cl-NOM. Our data suggests that both respiratory and non-respiratory dechlorination processes are important for Cl-NOM cycling, and that non-obligate organohalide-respiring bacteria are most likely involved in these processes.

Rapid Enrichment of Dehalococcoides-Like Bacteria by Partial Hydrophobic Separation.

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.

Estrone degradation: does organic matter (quality), matter?

Understanding the parameters that drive E1 degradation is necessary to improve existing wastewater treatment systems and evaluate potential treatment options. Organic matter quality could be an important parameter. Microbial communities grown from activated sludge seeds using different dissolved organic matter sources were tested for E1 degradation rates. Synthetic wastewater was aged, filter-sterilized, and used as a carbon and energy source to determine if recalcitrant organic carbon enhances E1 degradation. Higher E1 degradation was observed by biomass grown on 8 d old synthetic wastewater compared to biomass grown on fresh synthetic wastewater (P = 0.033) despite much lower concentrations of bacteria. Minimal or no E1 degradation was observed in biomass grown on 2 d old synthetic wastewater. Organic carbon analyses suggest that products of cell lysis or microbial products released under starvation stress stimulate E1 degradation. Additional water sources were also tested: lake water, river water, and effluents from a municipal wastewater treatement plant and a treatment wetland. E1 degradation was only observed in biomass grown in treatment effluent. Nitrogen, dissolved organic carbon, and trace element concentrations were not causative factors for E1 degradation. In both experiments, spectrophotometric analyses reveal degradation of E1 is associated with microbially derived organic carbon but not general recalcitrance.