Analysis of Clostridium botulinum Toxigenic Types A, B, and E for Fatty and Carbohydrate Content 1

Abstract

Lyophilized, 48-hr log-phase vegetative cells were extracted with chloroformmethanol (2:1, v/v) and ethanol-ether (3:1, v/v) and then saponified with methanolic KOH. Gas-liquid chromatography of the methyl esters of extractable fatty acids revealed distinctive "pattern profiles" of Clostridium botulinum toxigenic types "A," "B," and "E." C. perfringens type "A" and Escherichia coli strain "B" were also studied in a similar manner and were found to give pattern profiles which were distinct even from those obtained for the C. botulinum microorganisms. Amino sugar content of the five microorganisms was determined by using a Beckman amino acid analyzer. The molar ratio of glucosamine to that of galactosamine was found to be of further assistance in distinguishing the individual microorganisms.

Applications of cellular fatty acid analysis.

“…However, in comparison with headspace GLC, the Microbial Identification System method was thought to be too complicated. Toxigenic types A, B, and E of C. botulinum were studied by Fugate et al (52). In contrast to the results of CFA analysis with toxigenic types of C. perfringens (112), those investigators suggested that quantitative analysis of the CFAs could serve to distinguish types of C. botulinum.…”

Demonstration and Characterization of the Cell Wall Carbohydrate and Protein Antigens from Clostridium Botulinum Type E Saroma

“…Solomon et al (13) showed that C. botulinum type E shared common somatic antigens among the nonproteolytic strains of the organism. While non-peptidoglycan carbohydrate components of the cell wall of these strains were hexoses (glucose and galactose (3)) and hexosamine (glucosamine and galactosamine (5)), none of the antigens of these strains have yet been isolated, nor have their composition and antigenic specificity been elucidated.…”