Analysis of Clostridium botulinum toxigenic types A, B, and E for fatty and carbohydrate content.
Lyophilized, 48-hr log-phase vegetative cells were extracted with chloroform-methanol (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.
Gas chromatography of bacterial whole cell methanolysates; V. Fatty acid composition of Neisseriae and Moraxellae.
Twentyfive strains or substrains belonging to the bacterial species Moraxella nonliquefaciens, M. bovis, M. kingii, M. osloensis, M. phenylpyrouvica and M. urethralis (tentative designation) were examined by gas-liquid chromatography (GLC) of whole cell methanolysates. Each strain gave a reproducible chromatogram, usually with no difference between non-fimbriated and fimbriated variants. With a few exceptions, there was no significant difference between the elution profiles of strains belonging to the same species. The study revealed similarities as well as distinct dissimilarities in interspecies comparisons. When compared with M. bovis, the species M. nonliquefaciens and M. phenylpyrouvica showed most similarity, whereas M. kingii revealed the greatest chromatographic deviation. The remaining two species were intermediate in this respect. The results are discussed in relation to taxonomic data previously arrived at by other methods, in particular measurements of nucleic acid homologies and heterologies. Generally, the two groups of results were consistent, and it is concluded that GLC is a valuable tool for evaluation of relationship between species, adding significantly to the reliability of species allocation to genus. GLC appears also applicable for species identification of isolates of moraxellae and similar organisms. However, the sometimes great similarities between closely related species and the occasionally observed (minor) intraspecies variations in elution profile substantiate the need for supplementary methods.
The position of Clostridium tetani within the serological schema for the proteolytic clostridia.
In a serological study of strains of Clostridium sporogenes, Mandia and Brunershowed that identification of proteolytic clostridia was possible by means of antigenic formulas based on the reactions of the heat-labile antigens. Of the 2S Cl. sporogenes strains which they studied, 18 formed a homogenous serological group and were referred to as group I of the proteolytic clostridia. Three strains designated as Cl. sporogenes formed a second group with a strain of Clostridium parabotulinum. When a number of strains of Cl. parabotulinum and related organisms were included in the study, all reacted as members of the second group! This group was then referred to as serological group II of the proteolytic clostridia. A strain designated as Cl. sporogenes did not fall into either groups I or II, but showed, however, an agglutinative relationship to a strain of Clostridium histolyticum. When 13 strains of Cl. histolyticum were included in the study, they demonstrated none of the 12 previously determined antigenic factors possessed by organisms of groups I and II and they comprised a third homogenous serological group of the proteolytic clostridia, group lIP Serological identification of representatives of these three species was