http://www.cnr.it/ontology/cnr/individuo/prodotto/ID272072
Different activity levels of Dehalococcoides mccartyi revealed by FISH and CARD-FISH under non-steady and pseudo-steady state conditions (Articolo in rivista)
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- Different activity levels of Dehalococcoides mccartyi revealed by FISH and CARD-FISH under non-steady and pseudo-steady state conditions (Articolo in rivista) (literal)
- Anno
- 2013-01-01T00:00:00+01:00 (literal)
- Alternative label
Matturro B., Tandoi V. , Rossetti S. (2013)
Different activity levels of Dehalococcoides mccartyi revealed by FISH and CARD-FISH under non-steady and pseudo-steady state conditions
in New biotechnology (Online)
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- Matturro B., Tandoi V. , Rossetti S. (literal)
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- Water Research Institute, CNR, Via Salaria km 29.300, Monterotondo (RM) 00015, Italy (literal)
- Titolo
- Different activity levels of Dehalococcoides mccartyi revealed by FISH and CARD-FISH under non-steady and pseudo-steady state conditions (literal)
- Abstract
- A mixed culture capable of dechlorinating perchloroethylene (PCE) to ethene was analyzed under non
steady and pseudo-steady state conditions. Dehalococcoides mccartyi, considered to be the primary
dechlorinating bacterium able to completely degrade chlorinated hydrocarbons to non toxic ethene,
could be detected by CAtalyzed Reporter Deposition Fluorescence In Situ Hybridization (CARD-FISH)
since the beginning of culture operation but highlighted by conventional FISH only during active PCE
dechlorination to ethene and vinyl chloride (VC). Data generated from FISH and CARD-FISH analyses
were compared to those generated from applying PCR-based techniques directed at defining cell
abundances (Real Time PCR, qPCR) and assessing cell activities (Reverse Transcription qPCR, RT-qPCR)
of Dehalococcoides strains involved in the PCE reductive dechlorination (RD) process. qPCR targeting
reductive dehalogenase genes coding for enzymes involved in the individual steps of the RD process,
showed that Dehalococcoides strains carrying the tceA gene dominated the community. This observation
was consistent with PCE conversion products detected under pseudo-steady state (ethene and VC
production), since this tceA gene is known to be associated to strains capable of reducing chlorinated
solvents beyond 1,2-cis-dichloroethene (cis-DCE). In line with the FISH data, Dehalococcoides 16S rRNA
and tceA genes were expressed only during pseudo-steady state conditions when PCE was converted
completely to the final metabolic product ethene. Furthermore, Dehalococcoides cell abundances
estimated by CARD-FISH correlated positively with their 16SrRNA gene copy numbers quantified by
qPCR. This is consistent with the ability of both these methods to estimate total Dehalococcoides cell
numbers including those with low metabolic activities. Thus, this study shows that application potential
of FISH analysis to quantify rapidly and efficiently only active dechlorinators in complex communities (literal)
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