Spore Antigens of Clostridium Sporogenes

Abstract

PLATE ISINCE Defalle (1902) first reported the antigenicity of the spores in Bacillus species, opinion has been divided regarding the value of spore antigens in the differentiation of serological types among spore-forming bacteria. Defalle (1902) and Mellon and Anderson (1919) were of the view that spores possessed specific spore antigens distinct from those of the vegetative forms of the same species. The later report of Starin and Dack (1923) that spores were not antigenic and of Krauskopf and McCoy (1937) that spores did not produce antibodies to themselves but gave rise to antibodies specific for the H and 0 antigens of the bacillary forms, perhaps discouraged further study, but Howie and Cruickshank (1940) were able to prove beyond reasonable doubt that specific spore antibodies could be produced that were distinct from H and 0 antibodies produced by vegetative cells.The work of Moussa (1956) established the potential of spore antigens in the taxonomy of the clostridia. Meisel and Rymkiewicz (1959) studied a limited number of strains in each of seven species of clostridia and found more than one type in most of these species on the basis of their spore agglutinogens. The present investigation covers the immunological reactivity of the spores of Clostridium sporogenes ; their relationship to the cultural and biochemical characters of this organism has already been reported (Princewill, 1978). MATERIALS AND METHODSCultures. The 84 cultures of C. sporogenes used in this investigation, and the sources from which they originated, have been described elsewhere (Princewill, 1978). Cultures were maintained in cooked-meat medium (CMM, Southern Group Laboratory, Hither Green Hospital, London, SE 13).Production of spores. The stock cultures were inoculated into fresh CMM and incubated at 37°C for 24 h. Subcultures were made in two transfers into reinforced clostridial medium (RCM, Hirsch and Grinsted, 1954), from which 2-ml inocula were made on RCM agar slopes in 1 14-ml (4-oz) screw-capped bottles. The cultures were incubated anaerobically at 37°C for 7 days and the growth from 9-10 bottles harvested by washing with chilled 30-40 nll of distilled water. The pooled washings were allowed to stand for 15-20 min. for coarse debris to settle out, and the supernatant was decanted and centrifuged at 400g for 10 min. The sedimented spores were resuspended and washed once more in chilled water. The turbidity of the final suspension was adjusted against Brown's opacity tube no. 10 (Burroughs Wellcome) and distributed in 12-ml amounts into 28-m1(1 oz) bottles and stood in ice ready for sonication in a 150-W ultrasonic disintegrator (MSE) with a 9.5-mm (# in.) titanium probe at an amplitude of 18 flpm (peak to peak) for 5 min. To prevent germination of spores

Immunofluorescent study of the spore antigens of proteolyticstrains ofClostridium botulinum

“…botulinum by the indirect fluorescent antibody test (FAT). These were similar to those used in previous reports (Princewill, 1978(Princewill, , 1979a, with the addition of the following:…”

“…sporogenes on the basis of physiological or biochemical characteristics (Lynt et al 1972). Princewill (1979a) divided Cl. sporogenes into types by means of the spore antigens.…”

Spore antigens in the classification of some clostridia

“…This confirms once more the taxonomic value of the spore precipitinogens (Norris and Wolf, 196 1;Princewill, 1979). <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 <20 …”

“…The seven antisera prepared against the three types of C. sporogenes (Princewill 1979) were also included in these tests. The results are shown in table I.…”

“…The method of producing the spore suspensions was similar to that described by Princewill (1979). The cultures were harvested and sonicated and the spores were freed from the vegetative debris by repeated washings.…”