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DatoValore
TitleQuantitative estimation of Dehalococcoides mccartyi at laboratory and field scale: Comparative study between CARD-FISH and Real Time PCR
AbstractDehalococcoides mccartyi detectability in the field is a fundamental tool to assess the efficiency of natural attenuation or engineered bioremediation in chlorinated solvent-contaminated sites. This study reports on the direct comparison of quantitative data obtained by Real Time PCR (qPCR) and CAtalyzed Reporter Deposition-Fluorescence In situ Hybridization (CARD-FISH) over a wide range of Dehalococcoides concentrations (10-10(8)cellsmL(-1)) both in three independent 10-fold serial dilutions of a laboratory dechlorinating enrichment and in 49 groundwater samples from 6 different contaminated sites. Dehalococcoides enumeration by CARD-FISH yielded a linear curve in the analyzed concentration range which was consistent with the expected concentrations and showed good reproducibility in triplicate assays. Alternatively, qPCR did not allow for the discrimination of 16S rRNA gene concentrations lower than 10(3)gene copiesmL(-1) either in the dechlorinating mixed culture or in field samples. Overall this study highlights the limits of qPCR quantification, especially in samples where low concentrations of this microorganism may be expected, and suggests the use of a confirmatory methodology under these particular conditions
SourceJournal of microbiological methods 93 (2), pp. 127–133
KeywordsDehalococcoides mccartyi; qPCR; CARD-FISH; Chlorinated solvents; Biomonitoring
JournalJournal of microbiological methods
EditorElsevier Science Ireland, Shannon, Paesi Bassi
Year2013
TypeArticolo in rivista
DOI10.1016/j.mimet.2013.02.011
Authors1) Matturro B., 2) Heavner G.L., 2) Richardson R.E., 1) Rossetti S.
Text209678 2013 10.1016/j.mimet.2013.02.011 Dehalococcoides mccartyi; qPCR; CARD FISH; Chlorinated solvents; Biomonitoring Quantitative estimation of Dehalococcoides mccartyi at laboratory and field scale Comparative study between CARD FISH and Real Time PCR 1 Matturro B., 2 Heavner G.L., 2 Richardson R.E., 1 Rossetti S. 1 Water Research Institute, IRSA CNR, Via Salaria km 29,300, 00015 Monterotondo RM , Italy 2 School of Civil and Environmental Engineering, Cornell University, 220 Hollister Hall, Ithaca, NY 14853, United States Dehalococcoides mccartyi detectability in the field is a fundamental tool to assess the efficiency of natural attenuation or engineered bioremediation in chlorinated solvent contaminated sites. This study reports on the direct comparison of quantitative data obtained by Real Time PCR qPCR and CAtalyzed Reporter Deposition Fluorescence In situ Hybridization CARD FISH over a wide range of Dehalococcoides concentrations 10 10 8 cellsmL 1 both in three independent 10 fold serial dilutions of a laboratory dechlorinating enrichment and in 49 groundwater samples from 6 different contaminated sites. Dehalococcoides enumeration by CARD FISH yielded a linear curve in the analyzed concentration range which was consistent with the expected concentrations and showed good reproducibility in triplicate assays. Alternatively, qPCR did not allow for the discrimination of 16S rRNA gene concentrations lower than 10 3 gene copiesmL 1 either in the dechlorinating mixed culture or in field samples. Overall this study highlights the limits of qPCR quantification, especially in samples where low concentrations of this microorganism may be expected, and suggests the use of a confirmatory methodology under these particular conditions 93 Articolo in rivista Elsevier Science Ireland 0167 7012 Journal of microbiological methods Journal of microbiological methods J. microbiol. methods Journal of microbiological methods. bruna.matturro MATTURRO BRUNA simona.rossetti ROSSETTI SIMONA TA.P07.017.001 Caratterizzazione di biomasse per l ottimizzazione dei processi biologici