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The epidemiological and bacteriological investigations on four foodborne outbreaks caused by a new type of enterotoxinproducing Clostridium perfringens are described. C. perfringens isolated from patients of these outbreaks did not produce any known enterotoxin and did not carry the C. perfringens enterotoxin gene. However, the culture filtrates of these isolates induced the accumulation of fluid in rabbit ileal loop tests. The molecular weight of the new enterotoxin may be between 50,000 and 100,000, although the known C. perfringens enterotoxin is ca. 35,000. This new enterotoxin was heat labile, and its biological activities were inactivated by heating for 5 min at 60°C. The new enterotoxin was sensitive to pH values higher than 11.0 and protease treatment but was resistant to trypsin treatment. These results suggest that the new enterotoxin may be a protein. Although C. perfringens enterotoxin induced morphological changes in Vero cells, the changes induced by the new enterotoxin differed from those by the known C. perfringens enterotoxin. The new enterotoxin also induced morphological changes in L929 cells, whereas the known C. perfringens enterotoxin did not, because L929 cells lacked an appropriate enterotoxin receptor. Although C. perfringens enterotoxin is recognized as the only diarrheagenic toxin responsible for C. perfringens foodborne outbreaks, the results of the present study indicate that C. perfringens isolated from these four outbreaks produced a new type of enterotoxin.

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The detection of VT produced by Escherichia coli is very important for the identification of verotoxin-producing Escherichia coli (VTEC). The latex agglutination reagents (Denka Seiken Co. Ltd, Tokyo) which was developed to detect VT was compared with the vero cell bioassay or polymerase chain reaction method. A total 147 VT-positive strains (109 serotype O157:H 7/-and 38 non-O157 serotype) and 31 VT-negative strains which were isolated from human were investigated. In addition, a total of 79 VT-positive strains (14 serotype O157:H7i and 65 non-O157 serotype) and 79 VT-nagative strains which were isolated from animals were also examined. The latex agglutination assay for the human isolates showed the 100% sensitivity , specificity and agreement. The assay for the animal isolates showed 94 .9% sensitivity, 100% specificity and 97.5% agreement. Although 4 of 8 strains isolated from swine which produce VT2 variant toxin (VT2e) failed in detecting verotoxin by latex agglutination assay , VT2e was not related to human infections. We conclude that this latex agglutination reagent is highly sensitive and specific for detecting and characterizing VT of E. coli. The method is reliable, easy to perform at any laboratories.

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