Acid resistance of twelve strains of Enterobacter sakazakii, and the impact of habituating the cells to an acidic environment

The association of powdered infant formula with cases of severe Enterobacter sakazakii infections in immunocompromised and premature neonates jas led to a need to learn about the basic behavior of this emerging pathogen in food systems and the environment. The current study examines the microorganism's stationary-phase acid resistance using 12 strains that had been previously used to characterize its thermal resistance. Acid resistance was determined by initially culturing the isolates for 18 h in brain heart infusion broth (BHI) at 36 degrees C, transferring the cells to tryptic soy broth (TSB) adjusted to pH 3.0 and 3.5, and determining E. sakazakii survival over the course of 5 h incubation at 36 degrees C. At pH 3.5, 10 of the 12 strains showed less than a 1 log cycle decline over the 5-h incubation period, with the most acid sensitive strain showing an approximate 3.5 log cycle decline. At pH 3.0, the decline over the 5-h incubation period ranged from 4.9 to > 6.3 log cycles; however, substantial diversity was evident when the 1-h/pH 3.0 results were compared. The effect of habituating the cells to a moderately acidic environment was determined by growing the strains in TSB with 0% (nonacidogenic) and 1% glucose (acidogenic), transferring the cells to acidified (pH 3.0) BHI, and determining E. sakazakii survival over the course of 5 h of incubation at 36 degrees C. While there was diversity observed among the strains, in general the stationary-phase acid resistances of several of the strains were enhanced, at least transitorily, by growth in the acidogenic medium. No apparent correlation between the stationary-phase relative acid resistances of the strains based on the 1-h/pH 3.0 acid inactivation values and the previously reported thermal D-values was observed.