Recovery in culture of viable but nonculturable Vibrio parahaemolyticus: regrowth or resuscitation?

Type Article
Date 2007-06
Language English
Author(s) Coutard Francois1, Crassous Philippe2, Droguet Michael3, Gobin Eric4, Colwell Rita5, Pommepuy Monique1, Hervio Heath Dominique1
Affiliation(s) 1 : IFREMER, Dept Environm Microbiol & Phycotoxines, Microbiol Lab, F-29280 Plouzane, France.
2 : IFREMER, Dept Etude Ecosyst Profonds, Lab Environm Profond, F-29280 Plouzane, France.
3 : Site CHU Morvan, Unite Physiol Comparee & Integrat, Grp Culture Cellulaire EA 3879, Brest, France.
4 : CHU Brest, Serv Anat Pathol, F-29285 Brest, France.
5 : Univ Maryland, Ctr Bioinformat & Computat Biol, College Pk, MD 20742 USA.
Source The ISME Journal (1751-7362) (Nature), 2007-06 , Vol. 1 , N. 2 , P. 111-120
DOI 10.1038/ismej.2007.1
WOS© Times Cited 48
Keyword(s) Transmission electron microscopy, Scanning electron microscopy, Resuscitation, Regrowth, Vibrio parahaemolyticus, VBNC state
Abstract The objective of this study was to explore the recovery of culturability of viable but nonculturable ( VBNC) Vibrio parahaemolyticus after temperature upshift and to determine whether regrowth or resuscitation occurred. A clinical strain of V. parahaemolyticus Vp5 was rendered VBNC by exposure to artificial seawater ( ASW) at 4 degrees C. Aliquots of the ASW suspension of cells ( 0.1, 1 and 10 ml) were subjected to increased temperatures of 20 degrees C and 37 degrees C. Culturability of the cells in the aliquots was monitored for colony formation on a rich medium and changes in morphology were measured by scanning ( SEM) and transmission ( TEM) electron microscopy. Samples of VBNC cells were fixed and examined by SEM, revealing a heterogeneous population comprising small cells and larger, flattened cells. Forty-eight hours after temperature upshift to 20 degrees C or 37 degrees C, both elongation and division by binary fission of the cells were observed, employing SEM and TEM, but only in the 10-ml aliquots. The results suggest that a portion of VBNC cells is able to undergo cell division. It is concluded that a portion of VBNC cells of V. parahaemolyticus subjected to cold temperatures remain viable. After temperature upshift, regrowth of those cells, rather than resuscitation of all bacteria of the initial inoculum, appears to be responsible for recovery of culturability of VBNC cells of V. parahaemolyticus. Nutrient in filtrates of VBNC cells is hypothesized to allow growth of the temperature-responsive cells, with cell division occurring via binary fission, but also including an atypical, asymmetric cell division.
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Coutard Francois, Crassous Philippe, Droguet Michael, Gobin Eric, Colwell Rita, Pommepuy Monique, Hervio Heath Dominique (2007). Recovery in culture of viable but nonculturable Vibrio parahaemolyticus: regrowth or resuscitation? The ISME Journal, 1(2), 111-120. Publisher's official version : , Open Access version :