Summary: Fluorescent labelled specific antisera have been used to distinguish between types A, B and F, types C and D, and type E of Clostridium botulinum.
PLATE CITHB genus Chsrridium contains many species pathogenic to man and animals; their isolation and identification can be a ditlicult and timeansumhg procedure. A rapid method of differentiating closely related species in direct smears from pathological specimens would have many advantages. The use of fluorescent labelled antibodies (Coons, Creech and Jones, 1941) appears to offer such a method.Cl. septicurn and Cl, c h v o e i are two closely related species found in a variety of pathological conditions. Both are pleomorphic and exhibit forms of great similarity. The biochemical reactions of the two organisms are also similar and, although they usually differ in the fermentation of sucrose and salicin, anomalous strains do occur. The only certain method of discrimination has been by protection tests in animals, but even here an antiserum to Cl. septicum will sometimes give protection against challenge with Cl. Juucvoei.The results of Henderson (1934) suggested that in the 4 classical types of Robertson (1919-20), there were 3 distinct " 0 " antigens, and one common " 0 " antigen shared by strains in varying degrees. M o m (1959), however, divided 37 strains of Cl. septicurn, including representatives of Henderson's strains, into 2 groups on the basis of the " 0 " antigen, neither of which cross-reacted with Cl. chauvoei. All strains of Cl. chauvoei that Moussa studied possessed a common *' 0 " antigen and in this he coniirmed the earlier work of Roberts (1931) and Henderson (1932). Both organisms had common spore antigens and Moussa suggested that the presence or absence of spores in the antigen preparations used by various workers was responsible for much of the conflicting evidence for varying degrees of cross-reaction reported.Specific staining using fluorescent-labelled vegetative " 0 " antisera has been used to differentiate between these two organisms.MATBRLALS AND METHODS
Strains.The following strains were used:-CZ. septicwn: Wellcome Research Laboratories culture collection, English strains (CN 3204, 3205); Scottish strains (10553, 10568); New Zealand strains (CN 5115, 5116); Sinhalese strain (CN5021); Swiss strain (CN3232); South American strain (CN2779); Icelandic strain (CN368); origin unknown(CN1866); National Collection of Type Cultures, Robertson type I (NCTC 504, 547); Robertson type I1 (NCTC 281,282,549,550); Robertson type 111 (NmC 284, 501,551); Robertson type IV (NcTc286).
Summary. Ferritin labelled antibodies have been used to detect the presence of vegetative antigens in spores. The specificity of the labelled antibodies was demonstrated. Vegetative antigens were found along the core or cortical membrane in spore disintegrates, and after germination along the developing vegetative cell wall. The implications of these results are discussed in relation to morphological studies of ultra‐thin sections.
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