Structure analysis and performance of a microbial community from a contaminated aquifer involved in the complete reductive dechlorination of 1,1,2,2‐tetrachloroethane to ethene

Rossetti, Simona; Aulenta, Federico; Majone, Mauro; Crocetti, Gregory; Tandoi, Valter

doi:10.1002/bit.21776

Cited by 39 publications

(27 citation statements)

References 33 publications

(49 reference statements)

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“…WL rdhA1 is distinct from the putative 1,2-DCA rdh recently identified by Marzorati (33,38). Given the involvement of Dehalobacter in the transformation of a number of different chlorinated compounds, including chlorinated ethanes (19,41) and ethenes (22,52), polychlorinated biphenyls (37), ␤-hexachlorocyclohexane (49), trichlorobenzenes (50), and 1,2-dichloropropane (42), there are few rdh genes identified from this genus.…”

Section: Discussionmentioning

confidence: 90%

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene

Grostern¹,

Edwards

2009

Appl Environ Microbiol

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Dehalobacter and "Dehalococcoides" spp. were previously shown to be involved in the biotransformation of 1,1,2-trichloroethane (1,1,2-TCA) and 1,2-dichloroethane (1,2-DCA) to ethene in a mixed anaerobic enrichment culture. Here we report the further enrichment and characterization of a Dehalobacter sp. from this mixed culture in coculture with an Acetobacterium sp. Through a series of serial transfers and dilutions with acetate, H 2 , and 1,2-DCA, a stable coculture of Acetobacterium and Dehalobacter spp. was obtained, where Dehalobacter grew during dechlorination. The isolated Acetobacterium strain did not dechlorinate 1,2-DCA. Quantitative PCR with specific primers showed that Dehalobacter cells did not grow in the absence of a chlorinated electron acceptor and that the growth yield with 1,2-DCA was 6.9 (؎0.7) ؋ 10 7 16S rRNA gene copies/mol 1,2-DCA degraded. PCR with degenerate primers targeting reductive dehalogenase genes detected three distinct Dehalobacter/Desulfitobacterium-type sequences in the mixed-parent culture, but only one of these was present in the 1,2-DCA-H 2 coculture. Reverse transcriptase PCR revealed the transcription of this dehalogenase gene specifically during the dechlorination of 1,2-DCA. The 1,2-DCA-H 2 coculture could dechlorinate 1,2-DCA but not 1,1,2-TCA, nor could it dechlorinate chlorinated ethenes. As a collective, the genus Dehalobacter has been show to dechlorinate many diverse compounds, but individual species seem to each have a narrow substrate range.

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Section: Discussionmentioning

confidence: 90%

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene

Grostern¹,

Edwards

2009

Appl Environ Microbiol

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“…Prior to the start of the bioelectrochemical experiments, the mixed culture was analyzed by fluorescence in situ hybridization (FISH), using previously developed probes and protocols (Fazi et al, 2008;Rossetti et al, 2008). This analysis revealed that Desulfitobacterium sp.…”

Section: Microbial Cultures: Cultivation and Characterizationmentioning

confidence: 99%

“…Subsequently, the carbon paper samples were processed for electron microscopy and for molecular analyses following previously described procedures (Aulenta et al, 2006;Rossetti et al, 2008).…”

Section: Scanning Electron Microscopy and Molecular Analyses Of The Cmentioning

confidence: 99%

Microbial reductive dechlorination of trichloroethene to ethene with electrodes serving as electron donors without the external addition of redox mediators

Aulenta

Canosa

Reale

et al. 2009

Biotech & Bioengineering

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144

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In situ bioremediation of industrial chlorinated solvents, such as trichloroethene (TCE), is typically accomplished by providing an organic electron donor to naturally occurring dechlorinating populations. In the present study, we show that TCE dechlorinating bacteria can access the electrons required for TCE dechlorination directly from a negatively polarized (-450 mV vs. SHE) carbon paper electrode. In replicated batch experiments, a mixed dechlorinating culture, also containing Dehalococcoides spp., dechlorinated TCE to cis-dichloroethene (cis-DCE) and lower amounts of vinyl chloride (VC) and ethene using the polarized electrode as the sole electron donor. Conversely, neither VC nor ethene formation occurred when a pure culture of the electro-active microorganism Geobacter lovleyi was used, under identical experimental conditions. Cyclic voltammetry tests, carried out on the filter-sterilized supernatant of the mixed culture revealed the presence of a self-produced redox mediator, exhibiting a midpoint potential of around -400 mV (vs. SHE). This yet unidentified redox-active molecule appeared to be involved in the extracellular electron transfer from the electrode to the dechlorinating bacteria. The ability of dechlorinating bacteria to use electrodes as electron donors opens new perspectives for the development of clean, versatile, and efficient bioremediation systems based on a controlled subsurface delivery of electrons in support of biodegradative metabolisms and provides further evidence on the possibility of using conductive materials to manipulate and control a range of microbial bioprocesses.

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“…Fluorescence In Situ Hybridization (FISH) and its variant CAtalysed Reporter Deposition FISH (CARD-FISH) enabled the microscopic enumeration of whole cells of 'Dehalococcoides' spp. [22][23][24][25]. This methodology is independent of DNA extraction and it is characterized by detection limits lower than those commonly reported for qPCR [26].…”

Section: Introductionmentioning

confidence: 99%

Field distribution and activity of chlorinated solvents degrading bacteria by combining CARD-FISH and real time PCR

Matturro¹,

Aulenta²,

Majone³

et al. 2012

New Biotechnology

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Nowadays several advanced molecular techniques are applied for quantifying bacteria involved in contaminant degradation processes. However, despite the fact that significant efforts have been taken to make these tools more reliable and specific, their application for the analysis of field samples is hardly ever applied. In this study, a combination of three methods (CARD-FISH, qPCR and RT-qPCR) was successfully applied to evaluate the distribution and the activity of known chlorinated solvent dechlorinating bacteria in a contaminated site where no remedial actions have been undertaken. CAtalysed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH) specifically provided the cell densities of known dechlorinating bacteria and was found to be more sensitive than quantitative PCR (qPCR) for the quantification of 'Dehalococcoides' cell numbers in the aquifer. Among the screened dechlorinators, 'Dehalococcoides' spp. were mainly found and nearly homogenously distributed in the aquifers at concentrations ranging from 8.1×10(5)±1.2×10(5) to 2.5×10(7)±5.6×10(6)cells per liter of groundwater (with a relative abundance out of the total Bacteria of 0.7-15%). Further, the dechlorination potentialities of 'Dehalococcoides' species living in the aquifer were evaluated by analyzing the abundance and the expression of 16S rRNA genes and reductive dehalogenase (RDase) encoding functional genes by qPCR and Reverse Transcription qPCR (RT-qPCR). 'Dehalococcoides'tceA gene, known to be associated to strains capable of reducing chlorinated solvents beyond cis-DCE, was found and expressed in the field. Overall, this study proved the existence of a well-established dechlorinating microbial community able to use contaminants as substrates for their metabolic activity and indicated the occurrence of reductive dechlorination at the site.

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Structure analysis and performance of a microbial community from a contaminated aquifer involved in the complete reductive dechlorination of 1,1,2,2‐tetrachloroethane to ethene

Cited by 39 publications

References 33 publications

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene

Microbial reductive dechlorination of trichloroethene to ethene with electrodes serving as electron donors without the external addition of redox mediators

Field distribution and activity of chlorinated solvents degrading bacteria by combining CARD-FISH and real time PCR

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