Response of toxigenic Vibrio cholerae 01 to physico-chemical stresses in aquatic environments.

The survival and growth of toxigenic Vibrio cholerae 01 in water under various conditions of salinity, pH, temperature and cation composition and concentration were studied in an extensive series of laboratory experiments. Inter- and intra-strain variation in stress response (of 01 and non-01 strains) and the ability of V. cholerae to adapt to stressful environments were also studied. Toxigenic V. cholerae 01 were able to survive for at least 70 days at 25 degrees C in solutions of sea salt. The optimal salt concentration was 2.0% though all solutions in the range 0.25-3.0% gave good support. Substrains with enhanced capacity to persist at sub-optimal salinity (0.1%) were demonstrated. A great degree of inter-strain variation in stress response at low salinity (0.05%) was found among 59 strains, and this variation was unrelated to serogroup (01 or non-01), source (clinical or environmental) or country of origin (Tanzania or Bangladesh). At optimal salinity, inter-strain variation was less and 18 out of 20 strains remained viable at high concentrations for at least 40 months at 25 degrees C. V. cholerae 01 could not survive beyond 45 days at 4 degrees C and optimal salinity, either with or without nutrients. The optimal pH range for survival at 25 degrees C was 7.0-8.5 at optimal salinity, and 7.5-9.0 at low salinity. V. cholerae 01 require Na+ for survival in the absence of nutrients, and for enhanced growth in their presence. The presence of Ca2+ or Mg2+, in addition to Na+, further enhanced survival. These, and other results reported in this paper, suggest that toxigenic V. cholerae 01 are able to survive for extended periods in warm water containing no nutrients but having a salinity of 0.25-3.0% and a pH of around 8.0. With added nutrients and under the same conditions, rapid growth is possible. The implications of these findings for the identification of putative aquatic reservoirs of V. cholerae 01, and for the epidemiology of cholera, are considerable.