Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock.

2.50
Hdl Handle:
http://hdl.handle.net/10033/15676
Title:
Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock.
Authors:
González-Escalona, Narjol; Fey, Axel; Höfle, Manfred G; Espejo, Romilio T; A Guzmán, Carlos
Abstract:
We performed a comparative analysis of the Vibrio cholerae strain El Tor 3083 entering the viable but non-culturable (VBNC) state and starvation after incubation in artificial seawater (ASW) at 4 and 15 degrees C respectively. To this end, we determined bacterial culturability and membrane integrity, as well as the cellular levels of 16S rRNA and mRNA for the tuf, rpoS and relA genes, which were assessed by real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). Bacterial cells entering the VBNC state showed a 154, 5.1 x 10(3), 24- and 23-fold reduction in the number of copies of 16S rRNA and mRNA for tuf, rpoS and relA, in comparison to exponentially growing cells. The differences were less striking between cells in the VBNC and starvation states. The mRNA for relA was selectively increased in VBNC cells (3.2-folds), whereas a 3.9-fold reduction was observed for 16S rRNA. The obtained results confirmed that key activities of the cellular metabolism (i.e. tuf representing protein synthesis, and relA or rpoS stress response) were still detected in bacteria entering the VBNC state and starvation. These data suggest that the new Q-RT-PCR methodology, based on the selected RNA targets, could be successfully exploited for the identification (rRNA) of V. cholerae and assessment of its metabolic activity (tuf, rpoS, relA mRNA) in environmental samples.
Affiliation:
Vaccine Research Group, Division of Microbiology, GBF-German Research Centre for Biotechnology, Braunschweig, Germany.
Citation:
Quantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock. 2006, 8 (4):658-66 Environ. Microbiol.
Journal:
Environmental microbiology
Issue Date:
Apr-2006
URI:
http://hdl.handle.net/10033/15676
DOI:
10.1111/j.1462-2920.2005.00943.x
PubMed ID:
16584477
Type:
Article
Language:
en
ISSN:
1462-2912
Appears in Collections:
publications of the research group vaccinology and applied microbiology (VAC)

Full metadata record

DC FieldValue Language
dc.contributor.authorGonzález-Escalona, Narjol-
dc.contributor.authorFey, Axel-
dc.contributor.authorHöfle, Manfred G-
dc.contributor.authorEspejo, Romilio T-
dc.contributor.authorA Guzmán, Carlos-
dc.date.accessioned2008-01-04T15:13:11Z-
dc.date.available2008-01-04T15:13:11Z-
dc.date.issued2006-04-
dc.identifier.citationQuantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock. 2006, 8 (4):658-66 Environ. Microbiol.en
dc.identifier.issn1462-2912-
dc.identifier.pmid16584477-
dc.identifier.doi10.1111/j.1462-2920.2005.00943.x-
dc.identifier.urihttp://hdl.handle.net/10033/15676-
dc.description.abstractWe performed a comparative analysis of the Vibrio cholerae strain El Tor 3083 entering the viable but non-culturable (VBNC) state and starvation after incubation in artificial seawater (ASW) at 4 and 15 degrees C respectively. To this end, we determined bacterial culturability and membrane integrity, as well as the cellular levels of 16S rRNA and mRNA for the tuf, rpoS and relA genes, which were assessed by real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR). Bacterial cells entering the VBNC state showed a 154, 5.1 x 10(3), 24- and 23-fold reduction in the number of copies of 16S rRNA and mRNA for tuf, rpoS and relA, in comparison to exponentially growing cells. The differences were less striking between cells in the VBNC and starvation states. The mRNA for relA was selectively increased in VBNC cells (3.2-folds), whereas a 3.9-fold reduction was observed for 16S rRNA. The obtained results confirmed that key activities of the cellular metabolism (i.e. tuf representing protein synthesis, and relA or rpoS stress response) were still detected in bacteria entering the VBNC state and starvation. These data suggest that the new Q-RT-PCR methodology, based on the selected RNA targets, could be successfully exploited for the identification (rRNA) of V. cholerae and assessment of its metabolic activity (tuf, rpoS, relA mRNA) in environmental samples.en
dc.language.isoenen
dc.subject.meshBacterial Proteinsen
dc.subject.meshColden
dc.subject.meshColony Count, Microbialen
dc.subject.meshCulture Mediaen
dc.subject.meshRNA, Bacterialen
dc.subject.meshRNA, Ribosomal, 16Sen
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen
dc.subject.meshSeawateren
dc.subject.meshVibrio choleraeen
dc.titleQuantitative reverse transcription polymerase chain reaction analysis of Vibrio cholerae cells entering the viable but non-culturable state and starvation in response to cold shock.en
dc.typeArticleen
dc.contributor.departmentVaccine Research Group, Division of Microbiology, GBF-German Research Centre for Biotechnology, Braunschweig, Germany.en
dc.identifier.journalEnvironmental microbiologyen

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