Acid tolerance in Salmonella typhimurium induced by culturing in the presence of organic acids at different growth temperatures

The influence of growth temperature and acidification of the culture medium up to pH 4.25 with acetic, citric, lactic and hydrochloric acids on the growth and subsequent acid resistance at pH 3.0 of Salmonella typhimurium CECT 443 was studied. The minimum pH value which allowed for S. typhimurium growth within the temperature range of 25–37 °C was 4.5 when the pH was reduced using citric and hydrochloric acids, and 5.4 and 6.4 when lactic acid and acetic acid were used, respectively. At high (45 °C) or low (10 °C) temperatures, the growth pH boundary was increased about 1 pH unit. The growth temperature markedly modified the acid resistance of the resulting cells. In all cases, D -values were lower for cells grown at 10 °C and significantly increased with increasing growth temperature up to 37 °C, at which D -values obtained were up to 10 times higher. Cells grown at 45 °C showed D -values similar to those found for cells grown at 25 °C. The growth of cells in acidified media, regardless of the pH value, caused an increase in their acid resistance at the four incubation temperatures, although the magnitude of the Acid Tolerance Response (ATR) observed depended on the growth temperature. Acid adapted cultures at 10 °C showed D -values ranging from 5.75 to 6.91 min, which turned out to be about 2 times higher than those corresponding to non-acid adapted cultures, while higher temperatures induced an increase in D -values of at least 3.5 times. Another finding was that, while at 10 and 45 °C no significant differences among the effect of the different acids tested in inducing an ATR were observed, when cells were grown at 25 and 37 °C citric acid generally turned out to be the acid which induced the strongest ATR. Results obtained in this study show that growth temperature is an important factor affecting S. typhimurium acid resistance and could contribute to find new strategies based on intelligent combinations of hurdles, which could prevent the development or survival of Salmonella spp. in foods. The fact that moderately low temperatures (10 °C) markedly decrease the acid resistance and increase the growth pH boundary of S. typhimurium suggests the convenience to control the temperature during food processing as a strategy to prevent the growth and survival of this pathogenic microorganism.