Immunological analysis of spores of Clostridium botulinum type E

Abstract

High spore yields (>90%) of C. botulinum type E were obtained by a pseudosynchronous growth in trypticase peptone glucose broth. Clean spore suspensions of eight type E strains were prepared by enzymatic digestion of the vegetative components followed by differential centrifugation. Agglutination tests of hyperimmune spore antiserum against type E 070 strain showed significant titers with spore suspensions of eight strains. Immunodiffusion tests with homologous antisera showed a multiplicity of antigens present in spore extracts of the Beluga and 070 strain. Some of the antigens appeared to be strain specific whereas others were shared by several strains. The precipitating antigens appeared to be type specific since precipitin lines did not form with spore extracts of C. botulinum types A and B. A line of identity was formed with spore extracts of seven toxigenic and one nontoxigenic strains, indicating the presence of a common spore antigen specific to type E strains.