Pathogen thermal resistance studies on low-water activity foods (LWAF) use a variety of methods to inoculate food, as well as strategies to reduce water activity, which can influence thermal resistance observations. This study investigated effects of culture preparation method and osmolyte-induced water activity on thermal resistance of two Shiga toxin-producing Escherichia coli (STEC; O121:H19, 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 water activity on STEC thermal resistance, cells were suspended in osmolyte solutions with varying water activity: high (TSB, a w 0.99), intermediate (61% glycerol or 26% NaCl, a w 0.75), and low (82% glycerol, a w 0.5). In most instances, STEC in high-water activity broth exhibited greater heat resistance compared to reduced-water activity solutions, except the glycerol intermediate-water activity solution (a w 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 only some differences between high-a w and reduced-a w 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 water activity influence thermal inactivation of STEC by varying magnitudes. These results emphasize the challenges between extrapolating results from lab inactivation kinetic experiments to determine the inactivation of low water activity foods, especially those considered dry in nature.
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