Affiliation:
1. Laboratoire de Microbiologie, Département Environnement Microbiologie et Phycotoxines, IFREMER, 29280 Plouzané, France
Abstract
ABSTRACT
A real-time reverse transcription-PCR method was developed to determine whether the recovery of culturability of viable but nonculturable (VBNC)
Vibrio parahaemolyticus
induced the expression of virulence genes coding for the thermostable direct hemolysin and for type III secretion system 2 (TTSS2). The culturability of clinical strain Vp5 of
V. parahaemolyticus
in artificial seawater at 4°C was monitored, and the VBNC state was obtained. One day after entry into the VBNC state, temperature upshifts to 20 and 37°C allowed the recovery of culturability. Standard curves for the relative quantification of expression of the housekeeping genes
rpoS
,
pvsA
,
fur
, and
pvuA
; the
tdh2
gene; and the TTSS2 genes (VPA1354, VPA1346, and VPA1342) were established. The levels of expression of the
pvsA
and
pvuA
genes were stable and were used to normalize the levels of expression of the other genes. No transcriptional induction of the selected virulence genes under the temperature conditions used to recover the culturability of the VBNC bacteria was observed. The study results demonstrate that the recovery of culturability of VBNC cells of pathogenic
V. parahaemolyticus
is restricted to regrowth, without correlation with the induction of virulence gene expression. Disease induction would depend mainly on host-pathogen interactions that allow the expression of the virulence genes. This is the first time that the use of mRNA to detect viable cells was evaluated by computing the half-lives of multiple mRNA species under conditions inducing the VBNC state.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
45 articles.
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