Detection and Quantification of<i>Dehalococcoides</i>-Related Bacteria in a Chlorinated Ethene-Contaminated Aquifer Undergoing Natural Attenuation

Bürgmann, Helmut; Kleikemper, Jutta; Duc, Laurence; Bunge, Michael; Schroth, Martin H.; Zeyer, Josef

doi:10.1080/10889860802477218

Cited by 8 publications

(7 citation statements)

References 64 publications

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“…It should be noted that different strains of Dehalococcoides with the same 16S rRNA gene sequence can have different dehalogenating abilities, and therefore targeting strain-specific reductive dehalogenase functional genes (e.g., vcrA and/or bvcA) is necessary to provide information about actual dechlorination activity (Ritalahti et al 2006;Lookman et al 2007;Behrens et al 2008;Cupples 2008;Tas et al 2009). However, use of 16S rRNA for quantifying Dehalococcoides is an established technique (e.g., Cupples et al 2003;Lendvay et al 2003;Smits et al 2004;Buergmann et al 2008;Cupples 2008) and was deemed appropriate for the analysis presented here.…”

Section: Microbiological Analysismentioning

confidence: 99%

Effect of pore velocity on biodegradation of cis-dichloroethene (DCE) in column experiments

et al. 2009

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Column experiments were conducted to evaluate the effect of pore velocity on the extent of biodegradation of cis-dichloroethene (cis-DCE) during transport in porous media. Columns were filled with homogeneous glass beads and inoculated with a culture capable of complete dechlorination of tetrachloroethene to ethene. A constant concentration of cis-DCE was maintained in the columns' influent. Three different pore velocities were tested in duplicate, subjecting each column to a constant velocity. At high flow velocity, degradation of cis-DCE to ethene was nearly complete within the residence time of the columns. However, at medium and low flow velocities, incomplete dechlorination was observed. After 7 weeks, DNA was harvested from the columns to determine differences in the microbial populations. Results suggest that Dehalococcoides sp. were present in higher quantities in the high-velocity columns, consistent with the observed dechlorination. These results suggest that, at contaminated groundwater sites, heterogeneity of groundwater velocity may be one factor that contributes to heterogeneous distribution of biological activity.

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Section: Microbiological Analysismentioning

confidence: 99%

Effect of pore velocity on biodegradation of cis-dichloroethene (DCE) in column experiments

et al. 2009

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“…Despite the fact that in the past years in situ fluorescence hybridization technique has gained widespread acceptance in the scientific community as a powerful tool for the screening of microbial populations and their dynamics in environmental samples [27], only few applications on dechlorinating mixed cultures are reported and still fewer on groundwater contaminated by chlorinated solvents [17,[22][23][24][28][29][30][31]. Here we report the abundance, distribution and activity of chlorinated solvents degrading bacteria in groundwater samples ascertained by the simultaneous application of CARD-FISH, qPCR and RT-qPCR.…”

Section: Introductionmentioning

confidence: 99%

“…16S rRNA genes and to discriminate among strains with different metabolic peculiarities by targeting key-functional genes. In particular, quantitative PCR protocols were developed for tceA, vcrA and bvcA genes involved in the reductive dechlorination process and were successfully applied to quantify these genes and their expression level in pure cultures, lab-scale microbial enrichments [15,16] and, to a minor extent, at biostimulated or bioaugmented aquifers [17][18][19][20][21].…”

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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“…Interestingly, the members of M4 did not show affiliation with known cultivated obligate OHRB from the genera Dehalococcoides and Dehalogenimonas. The 16S rRNA gene sequences retrieved from M4 showed 93-96 percent sequence similarity to Dehalococcoides-related uncultured bacteria (DQ223088 and DQ223076) ascribed to the "Grenchen cluster" and reported to be from an aquifer contaminated with chlorinated ethenes (Burgmann et al, 2008). Two M4 sequences clustered separately and were related to uncultured bacteria in monochlorobenzene-contaminated groundwater (Alfreider et al, 2002).…”

Section: Active Members Of the Organohalide Respiring Bacterial Guildmentioning

confidence: 98%

Divergent PCB organohalide-respiring consortia enriched from the efflux channel of a former Delor manufacturer in Eastern Europe

Praveckova

Brennerova

Čvančarová

et al. 2015

Ecotoxicology and Environmental Safety

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Detection and Quantification ofDehalococcoides-Related Bacteria in a Chlorinated Ethene-Contaminated Aquifer Undergoing Natural Attenuation

Cited by 8 publications

References 64 publications

Effect of pore velocity on biodegradation of cis-dichloroethene (DCE) in column experiments

Effect of pore velocity on biodegradation of cis-dichloroethene (DCE) in column experiments

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

Divergent PCB organohalide-respiring consortia enriched from the efflux channel of a former Delor manufacturer in Eastern Europe

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