EFFECT OF HEAT-TREATMENT ON SURVIVAL OF, AND GROWTH FROM, SPORES OF NONPROTEOLYTIC CLOSTRIDIUM-BOTULINUM AT REFRIGERATION TEMPERATURES

Spores of five type B, five type E, and two type F strains of nonproteolytic Clostridium botulinum were inoculated into tubes of an anaerobic meat medium plus lysozyme to give approximately 10(6) spores per tube. Sets of tubes were then subjected to a heat treatment, cooled, and incubated at 6, 8, 10, 12, and 25 degrees C for up to 60 days. Treatments equivalent to heating at 65 degrees C for 364 min, 70 degrees C for 8 min, and 75 degrees C for 27 min had little effect on growth and toxin formation. After a treatment equivalent to heating at 80 degrees C for 23 min, growth occurred at 6 and 8 degrees C within 28 to 40 days, After a treatment equivalent to heating at 85 degrees C for 19 min, growth occurred in some tubes at 6, 8, 10, or 12 degrees C within 28 to 53 days and at 25 degrees C in all tubes within 15 days. Following a treatment equivalent to heating at 95 degrees C for 15 min, growth was detected in some tubes incubated at 25 degrees C for fewer than 60 days but not in tubes incubated at 6 to 12 degrees C. The results indicate that heat treatment of processed foods equivalent to maintenance at 85 degrees C for 19 min combined with storage below 12 degrees C and a shelf life of not more than 28 days would reduce the risk of growth from spores of nonproteolytic C. botulinum by a factor of 10(6). If foods intended for refrigerated distribution and storage are liable to be maintained at temperatures higher than 10 degrees C, growth and toxin production by proteolytic strains of C. botulinum must also be prevented.