Thermal Resistance of Single Strains of Shiga Toxin-Producing Escherichia coli O121:H19 and O157:H7 Based on Culture Preparation Method and Osmolyte-Reduced Water Activity.

Abstract: Bacterial exposure to stress, such as reduced water activity (aw), can increase thermal resistance. Pathogen thermal resistance studies on low-aw foods use a variety of methods to inoculate food, as well as strategies to reduce aw, which can influence observations. This study investigated effects of culture preparation method and osmolyte-induced aw on thermal resistance of two Shiga toxin-producing Escherichia coli (STEC) strains (O121:H19 and O157:H7) challenged with isothermal conditions, determining D- and z-values for each isolate (56, 59, and 62°C). Tryptic soy broth (TSB) and agar (lawn cultures) were compared. D-values of broth cultures were significantly and consistently larger than those of lawn cultures, and O121 was significantly more resistant than O157, but only at 56°C (P < 0.05). To compare potential effects of aw on STEC thermal resistance, cells were suspended in osmolyte solutions with varying aw: high (TSB, aw 0.99), intermediate (61% glycerol or 26% NaCl, aw 0.75), and low (82% glycerol, aw 0.5). In most instances, STEC strains in high-aw broth exhibited greater heat resistance compared to reduced-aw solutions, with the exception of the glycerol intermediate-aw solution (aw 0.75). Magnitudes varied with strain and temperature. The z-values of lawn cultures were significantly lower than those of broth cultures (P < 0.05), but there were few differences between high-aw and reduced-aw samples. There were no significant differences of z-values based on strain type. These results highlight that thermal resistance can be affected by culture preparation and that osmolyte-induced changes to aw influence thermal inactivation of STEC by varying magnitudes. These results emphasize the challenges of extrapolating results from laboratory inactivation kinetic experiments to determine the inactivation of low-aw foods, especially those considered dry in nature.