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Development of a heat-pasteurization process is described for picked meat of Dungeness crabs (Cancer magister) contained in oxygen-impermeable flexible pouches, For each time-temperature treatment, 30 samples, each inoculated with an equal mixture of three strains of C. botulinum nonproteolytic type B, for a total of 107 spores, provided the basis for calculation of the thermal resistance (a 7D process). Following heat processing, the crabmeat was removed from the pouches and transferred to enrichment medium where it was incubated anaerobically for 150 days. Endpoints at which spores survived were determined by the presence of toxin in the enrichment medium. Process times ranged from 90 min at 88.9°C to 20.3 min at 94.4°C. D values (the time at each temperature required to reduce the inoculum by 1 log) ranged from 12.9 for the 88.9°C process to 2.9 for the 94.4°C process. The relative sterilization value, F0 was .054 and the pasteurization value, , was 240. This pasteurization process safely extends refri...

This study demonstrates the feasibility of a heat-pasteurization process for certain vacuum-packaged hot-smoked fishery products for inactivation of the spores of the nonproteolytic Group II Clostridium botulinum types B, E, and F. This process permits the use of lower concentrations of salt and other inhibitors without jeopardizing safety and quality of the products during prolonged refrigerated storage. The pasteurization treatment was developed based upon the inactivation of nonproteolytic types B or E in hot-process (smoked) salmon. Smoke was not applied to the samples inoculated with types B and E because of its possible inhibitory effects. After processing in the smokehouse, each sample was cooled to 34°F (1.1°C), injected with 106 spores, vacuum-packaged, and then heat-pasteurized in a water bath held at a constant temperature. A total of 85, 65, and 55 min in the 185°F (85°C), 192°F (88.9°C), and 198°F (92.2°C) baths, respectively, prevented toxin production by type E during 21 d of incubation at 25°C. Longer times, 175, 85, and 65 min, respectively, were required to prevent toxin production by nonproteolytic type B. Toxin production by type E during 120 d of storage at 10°C was prevented by a 45-minute treatment in the 198°F (92.2°C) bath. When heat-pasteurized samples were transferred into TPGY broth and incubated anaerobically for 150 d at 25°C, outgrowth and toxin production by type E was prevented by a 55-minute process at 198°F (92.2°C) and type B was prevented by a 65-minute process. This process does not, however, inactivate the more heat-resistant proteolytic strains of C. botulinum Group I or other spore-formers. The packages and master cartons of these pasteurized products therefore should follow the existing recommendations for smoked fishery products and be labeled "Keep refrigerated - Store below 38°F (3.3°C)."

Whitefish steaks were brined in NaCl, KCl, or equimolar NaCl:KCl to contain similar chloride ion concentration and inoculated intramuscularly with 10 or 100 spores of Clostridium botulinum type E per gram of fish. Steaks were then heated in a simulated (i.e., without smoke) hot-smoke process to internal temperatures of 62.8° to 76.7°C (145°-170°F) for the final 30 min of a 2- to 3-h process, packaged under vacuum in oxygen-impermeable film, and stored at abuse temperature of 25°C. During 7 d of storage, toxin production was inhibited in steaks containing more than 0.66 ionic strength NaCl, 0.64 KCl, or 0.71 equimolar NaCl:KCl. The results indicate that it is feasible to substitute KCl for NaCl in hot-process smoked fish for inhibition of outgrowth and toxin production by Clostridium botulinum type E.

Ready-to-eat picked meat of Dungeness crab (Cancer magister) packaged under vacuum in hermetically sealed (No. 10, five pound) cans, has become a public health concern because the product does not receive a process to inactivate Clostridium botulinum and refrigeration is the only barrier to prevent this bacterium from growing and producing neurotoxins. However, crab meat packaged in this manner has been distributed on ice to retail stores and restaurants for over 50 years without any reported illnesses from C. botulinum. Studies were therefore made to determine conditions that may have contributed to the safety of this product. C. botulinum type E was shown to be present in live Dungeness crab entering the processing facility, but was not detectable after the crabs were cooked in boiling water for ten minutes, nor in picked crab meat. Crab meat samples were inoculated with 10, 100, 1000 and 10,000 type E or type A spores/50 g, packaged in oxygen-permeable or vacuum packaged in oxygen-impermeable ...

Mixtures of proteolytic and nonproteolytic strains of toxigenic Clostridium botulinum types A, B, and F; nonproteolytic types B, E, and F; Clostridium sporogenes; and nontoxic E-like organisms resembling nonproteolytic C. botulinum were tested against each other for the purpose of selecting a mixture of compatible C. botulinum strains for inoculated pack studies on the basis of their sensitivity to bacteriophages and bacteriocin-like agents. All of the proteolytic strains produced bacteriocin-like agents that were inhibitory to three or more of the other proteolytic types and C. sporogenes. When selected strains of proteolytic types A and B were grown together, type A cultures produced neurotoxin, but type B toxin production was inhibited. Nonproteolytic strains of C. botulinum also produced bacteriocin-like agents against each other. Of these, type E strain EF4 produced bacteriocin-like agents against both proteolytic and nonproteolytic types of C. botulinum and C. sporogenes. EF4, however, was not inhibitory to the nontoxigenic E-like strains. When EF4 was grown with type A strain 62A, it had an inhibitory effect on type A toxin production. Strain 62A inactivated the type E toxin of EF4 after 7 to 21 days at 30 degrees C. On the basis of the production of these bacteriocin-like agents by different strains of C. botulinum and their potential effect on neurotoxin production, it is very important that compatible strains are used in mixtures for inoculated pack studies to determine the safety of a food process or product.

Water phase sodium chloride (WPS) levels of 1.8 to 3.0% in combination with heat pasteurization for 15 min at temperatures of 75, 80, 85, and 90 degrees C were evaluated as methods for the inactivation or inhibition of nonproteolytic, psychrotrophic Clostridium botulinum types B and E in crab analogs (imitation crab legs) subsequently stored at 10 and 25 degrees C. Samples inoculated with 10(2) type B or E spores per g prior to pasteurization remained nontoxic for 120 days at 10 degrees C and for 15 days at 25 degrees C. With 10(4) type E spores per g and 80 degrees C pasteurization, > or = 2.4 and 2.7% WPS was required for inhibition at 10 and 25 degrees C storage, respectively. Pasteurization at 85 degrees C decreased the inhibitory level of WPS to 2.1% at 10 degrees C and to 2.4% at 25 degrees C. When the inoculum was 10(4) type B spores per g, samples with 2.7% WPS were toxic after 80 days of storage at 10 degrees C. Samples inoculated with 10(3) type B spores per g and processed at 85 degrees C remained nontoxic for 15 days at 25 degrees C with a WPS of > or = 2.4%. When pasteurization was carried out before inoculation and packaging, 1.8% WPS prevented toxin production by 10(2) and 10(4) type E spores per g for 30 days at 10 degrees C, and this time period increased as the WPS concentrations increased. Three percent WPS prevented toxin production by 10(4) type E spores per g in vacuum-packaged analogs stored 110 days at 10 degrees C. Pasteurization processes used in these experiments, however, do not inactivate the heat-resistant proteolytic types of Clostridium botulinum. Therefore, the most important factor controlling the growth of this bacterium is continuous refrigeration below 3.0 degrees C or frozen storage of the finished product.

Liquid smoke in combination with NaCl was an effective inhibitor of outgrowth and toxin production by Clostridium botulinum types A and E spores in hot-processed whitefish, chub and carp stored at an abuse temperature of 25°C for 7 or 14 d. Surface-inoculated type E produced toxin in control samples containing 3.7% water-phase NaCl, but not in liquid smoke-treated samples having less than 2.0% water-phase NaCl. Liquid smoke was less effective when type E spores were injected intramuscularly. Liquid smoke lowered the concentration of NaCl required to inhibit toxin production by surface-inoculated type A from 4.6 to 2.8% in samples stored 7 d. Liquid smoke enhanced the ability of NaCl to prevent toxin production, but should not be considered a substitute for NaCl or refrigerated storage (below 3.3°C).

When Clostridium botulinum type C is cured of its prophage it simultaneously ceases to produce toxin. This nontoxigenic culture can then be converted to another toxigenic bacterial species, Clostridium novyi type A or to toxigenic Clostridium botulinum types C or D, by specific bacteriophages. The toxigenicity and type of toxin produced by these cultures depends upon the continued presence of these bacteriophages.