Proposed mechanism for sensitization by hypochlorite treatment of Clostridium botulinum spores

Hypochlorite-treated Clostridium botulinum 12885A spores, but not buffer-treated spores, could be germinated with lysozyme, indicating that their coats are made permeable to lysozyme by hypochlorite treatment so that the cortex is accessible. Hypochlorite-treated spores and spores extracted with 8 M urea-2-mercaptoethanol (pH 3.0) were sensitive to certain components of recovery media, but spores sensitized to lysozyme by other treatments were not. These data indicate that hypochlorite does more than increase coat permeability to lysozyme. Scanning electron microscopy revealed a more open-appearing surface of hypochlorite-treated spores, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that a greater amount of protein was removed from hypochlorite-treated and other lysozyme-sensitized spores than from buffer-treated spores. The data suggest that spore coat proteins may be removed by hypochlorite treatment, and this may be responsible for the sensitivity of spores and for their observed ability to germinate in lysozyme.