Detection of cytotoxin-hemolysin mRNA in nonculturable populations of environmental and clinical Vibrio vulnificus strains in artificial seawater

Type Article
Date 2002-11
Language English
Author(s) Fischer Le Saux Marion, Hervio Heath Dominique, Loaec Solen, Colwell Rita, Pommepuy Monique
Affiliation(s) IFREMER, DEL MP, Microbiol Lab, F-29280 Plouzane, France.
Univ Maryland, Inst Biotechnol, Ctr Marine Biotechnol, Baltimore, MD USA.
Source Applied and environmental microbiology (0099-2240) (American society for microbiology), 2002-11 , Vol. 68 , N. 11 , P. 5641-5646
DOI 10.1128/AEM.68.11.5641-5646.2002
WOS© Times Cited 48
Keyword(s) Artificial seawater, Reverse transcription PCR, Cytotoxin Hemolysin, Molecular detection method, Vibrio vulnificus
Abstract The objective of this study was to develop a molecular detection method that better estimates the potential risk associated with the presence of Vibrio vulnificus. For that purpose, we applied seminested reverse transcription-PCR (RT-PCR) to viable but nonculturable (VBNC) populations of V. vulnificus and targeted the cytotoxin-hemolysin virulence gene vvhA. Three strains, two environmental, IF Vv10 and IF Vv18, and one clinical, C7184, were used in this study. Artificial seawater, inoculated with mid-log-phase cells, was maintained at 4degreesC. VBNC cells resulted after 3, 6, and 14 days for C7184, IF Vv18, and IF Vv10, respectively. Our data indicate that seminested RT-PCR is sensitive for the detection of vvhA mRNA in artificial seawater when exclusively nonculturable bacteria are present. This is the first report of the expression of a toxin gene in VBNC V. vulnificus. Moreover, vvhA transcripts were shown to persist in nonculturable populations over a 4.5-month period, with a progressive decline of the signal over time. This result indicates that special attention should be given to the presence of potentially pathogenic VBNC cells in environmental samples when assessing public health risk.
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Fischer Le Saux Marion, Hervio Heath Dominique, Loaec Solen, Colwell Rita, Pommepuy Monique (2002). Detection of cytotoxin-hemolysin mRNA in nonculturable populations of environmental and clinical Vibrio vulnificus strains in artificial seawater. Applied and environmental microbiology, 68(11), 5641-5646. Publisher's official version : https://doi.org/10.1128/AEM.68.11.5641-5646.2002 , Open Access version : https://archimer.ifremer.fr/doc/00000/1269/