Culturing enterohemorrhagic Escherichia coli in the presence and absence of glucose as a simple means of evaluating the acid tolerance of stationary-phase cells.

Prior growth of seven enterohemorrhagic and one nonenterohemorrhagic strains of Escherichia coli in tryptic soy broth with (TSB+G) and without (TSB-G) 1% glucose was evaluated for its effect on acid tolerance. The final pHs of 18-h TSB+G and TSB-G cultures were 4.6 to 5.2 and 6.9 to 7.0, respectively. Cells were then transferred to brain heart infusion broth adjusted to pH 2.5 or 3.0 with HCl, incubated at 37 degrees C for up to 7 h, and assayed periodically for viable populations with brain heart infusion and MacConkey agars. All enterohemorrhagic strains were acid resistant (< 0.5 log decline after 7 h) when initially cultured in TSB+G, but substantial differences in acid tolerance were observed among strains cultured in TSB-G (log declines ranged from < 0.3 to > 3.8). The results indicated that prior growth in a medium with and without a fermentable carbohydrate is a convenient way to studying the induction of acid tolerance, that acid inactivation is preceded by a period of acid injury, and that pH-independent and pH-dependent stationary-phase acid tolerance phenotypes may exist among strains of enterohemorrhagic E. coli.