Limits...
Survival, induction and resuscitation of Vibrio cholerae from the viable but non-culturable state in the Southern Caribbean Sea.

Fernández-Delgado M, García-Amado MA, Contreras M, Incani RN, Chirinos H, Rojas H, Suárez P - Rev. Inst. Med. Trop. Sao Paulo (2015 Jan-Feb)

Bottom Line: At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state.Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells.Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología de Organismos, Universidad Simón Bolívar, Caracas, Venezuela.

ABSTRACT
The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Show MeSH

Related in: MedlinePlus

Resuscitation of Vibrio cholerae cells from the VBNC stateby adding spent media (SM) at logarithmic (SM1) and stationary (SM2) phases insamples diluted 10–2. SM day 0 () and SM day 7 (). Data are mean ± SE of a minimum of 23 and a maximum of 36replicate values. The symbol * shows significant effect (p< 0.05) for paired data of SM addition on the resuscitation of VBNCcells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4325519&req=5

f03: Resuscitation of Vibrio cholerae cells from the VBNC stateby adding spent media (SM) at logarithmic (SM1) and stationary (SM2) phases insamples diluted 10–2. SM day 0 () and SM day 7 (). Data are mean ± SE of a minimum of 23 and a maximum of 36replicate values. The symbol * shows significant effect (p< 0.05) for paired data of SM addition on the resuscitation of VBNCcells.

Mentions: Results from resuscitation experiments initially showed regrowth ofV. cholerae non-culturable cells in the undiluted samples by theaddition of BHI and HP media. However, after serial dilutions no growth was evident withboth media. The addition of SM at logarithmic and stationary phases to those wellscontaining 10–2 diluted samples, previously supplemented with either HP orBHI, showed the recovery of non-culturable cells and was found to be significant(p < 0.05) (Fig. 3).


Survival, induction and resuscitation of Vibrio cholerae from the viable but non-culturable state in the Southern Caribbean Sea.

Fernández-Delgado M, García-Amado MA, Contreras M, Incani RN, Chirinos H, Rojas H, Suárez P - Rev. Inst. Med. Trop. Sao Paulo (2015 Jan-Feb)

Resuscitation of Vibrio cholerae cells from the VBNC stateby adding spent media (SM) at logarithmic (SM1) and stationary (SM2) phases insamples diluted 10–2. SM day 0 () and SM day 7 (). Data are mean ± SE of a minimum of 23 and a maximum of 36replicate values. The symbol * shows significant effect (p< 0.05) for paired data of SM addition on the resuscitation of VBNCcells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4325519&req=5

f03: Resuscitation of Vibrio cholerae cells from the VBNC stateby adding spent media (SM) at logarithmic (SM1) and stationary (SM2) phases insamples diluted 10–2. SM day 0 () and SM day 7 (). Data are mean ± SE of a minimum of 23 and a maximum of 36replicate values. The symbol * shows significant effect (p< 0.05) for paired data of SM addition on the resuscitation of VBNCcells.
Mentions: Results from resuscitation experiments initially showed regrowth ofV. cholerae non-culturable cells in the undiluted samples by theaddition of BHI and HP media. However, after serial dilutions no growth was evident withboth media. The addition of SM at logarithmic and stationary phases to those wellscontaining 10–2 diluted samples, previously supplemented with either HP orBHI, showed the recovery of non-culturable cells and was found to be significant(p < 0.05) (Fig. 3).

Bottom Line: At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state.Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells.Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología de Organismos, Universidad Simón Bolívar, Caracas, Venezuela.

ABSTRACT
The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Show MeSH
Related in: MedlinePlus