Thermal Resistance of Clostridium difficile Spores in Peptone Water and Pork Meat

The thermal resistance of four strains of Clostridium difficile spores (three hypervirulent and one nonhypervirulent) in peptone water (PW) and pork meat was evaluated individually at 70, 75, 80, 85, and 90°C using two recovery methods (taurocholate and lysozyme). PW or meat was inoculated with C. difficile spores and mixed to obtain ca. 5.0 log CFU/ml or 4.0 log CFU/g, respectively. The D-values of C. difficile spores in PW ranged from 7.07 to 22.14 h, 1.42 to 3.82 h, 0.35 to 0.59 h, 4.93 to 5.95 min, and 1.16 to 1.76 min at 70, 75, 80, 85, and 90°C, respectively, for the four strains using the taurocholate method. The D-values of the respective C. difficile spores were greater (P ≤ 0.05) using the lysozyme method, especially at higher temperatures (85 and 90°C). Greater thermal resistance of C. difficile spores was observed in meat than in PW using the lysozyme method. Hypervirulence of the C. difficile strains was not associated with greater thermal resistance in meat. The z-values for C. difficile spores in meat were between 6.21 and 7.21°C, and they were 11.24 and 12.12°C using the taurocholate and the lysozyme recovery methods, respectively. The D- and z-values of C. difficile spores were greater in both PW and pork than the values reported in the literature. C. difficile spores can survive traditional cooking or thermal processing practices and potentially grow in cooked, ready-to-eat products. The use of effective methods to recover heat-injured spores is necessary to obtain accurate thermal destruction parameters for C. difficile spores.