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.
Cellular monosaccharide patterns of Neisseriaceae.
Sixty-four strains of Neisseria, Moraxella, and Acinetobacter were screened for cellular monosaccharides by gas-liquid chromatography and other chromatographic techniques. The four sugars ribose, glucose, glucosamine, and 2-keto-3-deoxyoctonate (KDO) were detected in all strains. Heptose was detected only in "true neisseriae" (Neisseria gonorrhoeae, N. meningitidis, N. sicca, N. cinerea, N. flavescens, and N. elongata) and in the tentaively named species Moraxella urethralis. Some marked interspecies dissimilarities within groups were revealed. Thus, N. ovis and M. atlantae were characterized by the presence of mannose. Intraspecies differences were also encountered. N. meningitidis strains of serogroups B and C were distinguished from strains of serogroup A by their sialic acid content. This sugar was also detected in two out of three examined strains of M. nonliquefaciens. In Acinetobacter, heterogeneity of monosaccharide patterns was rather pronounced. The results show the applicability of gas chromatographic "monosaccharide" profiles fo whole cells or extracted carbohydrate in bacterial classification and identification, including differentiation at the subspecies level. In addition, such profiles may be useful for monitoring during purification of cellular polysaccharides.
Some Serological Aspects of the S-R Change in Clostridium histolyticum
The variation in antigenic structure associated with the S-R change in aerobic pathogens seems to be fairly uniform. In general, there is a loss of the antigenic component characterizing the somatic surface of the bacteria in the smooth phase. This antigenic component is often, but not always, of the nature of a complex polysaccharide. The loss of the latter polysaccharide from the surface of the bacterial cells, whether present in the form of a capsule or not, renders the organism non-pathogenic and agglutinable not only by its own antiserum, but also by antiserum prepared against bacteria in the smooth phase, and sometimes by antisera prepared against rough forms of other species (Topley and Wilson, 1936). The variation in the antigenic structure of the spore-bearing anaerobes has not been studied as fully, but it appears that the problem may be more complicated in this group. Orr, Josephson, Baker, and Reed (1933), working with Clostridium welchii, found that an antiserum prepared by the injection of bacilli in the smooth phase would not agglutinate the rough variant of the same culture. Colef (1936) isolated two non-toxic variants of Clostridium oedematiens which formed smooth colonies. Antisera prepared against these variants agglutinated the original culture to titer. Conversely, the antiserum prepared against the original culture agglutinated these variants. This crossagglutination was probably not due to similar flagellar agglutinogens, for flagella could not be demonstrated on the variants, which were non-motile.