The conversion of heat-resistant spores of type A into germinated heat-sensitive cells in the peritoneal cavity of mice has been observed. This in-vivo germination of spores was depressed in mice passively immunised with type-A botulinal antitoxin. Studies, in which the release of free Ca in the urine of mice challenged with labelled spores provided an indicator of spore germination, showed that spore germination occurred normally at 8 hr . However, the rate of germination as determined from Ca excretion was substantially inhibited when antibody-treated spores were used as the challenge.

In in-vitro studies with guinea-pig leucocytes, spore germination was similarly assayed by monitoring the release of Ca from labelled spores. Spore germination in these leucocyte systems was also suppressed when the spores were first treated with type-A antitoxic serum, although phagocytic indices for antibody-treated spores and vegetative cells were increased. Release of type-A botulinal toxin was either completely prevented or substantially suppressed when spores and vegetative cells were treated with specific botulinal antibody and incubated with leucocytes.

These results demonstrate that combination of spores with type-specific antibody prompted phagocytic engulfment of antibody-coated spores, but strongly inhibited subsequent phagocytic digestion of engulfed spores.


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