A Study of Antigens of the Aerobic Sporeforming Bacteria

Abstract

Spore, vegetative 0 and vegetative H agglutinogens were demonstrated as separate entities in Bacillus species. Spore precipitinogens were distinct from those of the vegetative stage, the latter being associated with the H antigen.Injection of viable spores into rabbits resulted in the formation of antibodies for the vegetative phase as well as spore antibodies, while injection of autoclaved spores gave rise to spore antibodies only.Spore agglutinogens varied in their strain specificity, B. cereus and B. megaterium especially showing much subspecies specificity, but the spore precipitinogens showed a high degree of species specificity and could be of considerable taxonomic value. The multiplicity of spore precipitinogens in certain species was demonstrated and interspecies cross reactions were studied.THE SPORE, somatic and flagellar antigens of Bacillus polymyxa were studied by Davies (1951) using the agglutination technique. The three antigens proved to be xerologically distinct and there was a species specific antigen present in the spores of all 39 strains studied. Lamanna (1942) and Doak & Lamanna (1948) showed that the spores and vegetative cells of several Bacillus species possessed precipitinogens and that the spore precipitinogens were of some taxonomic value. The aims of the present work were to extend Davies' observations to other members of the genus. to include a study of spore and vegetative cell precipitinogens and to try to assess the taxonomic value of the different types of antigen. MATERIALS AND METHODS Preparation of antigensWhether intended for injection into rabbits or for use in agglutination or precipitation tests, only freshly prepared antigens were used and all cultures were checked microscopically for purity. Vegptative cell antigensCultures were grown in nutrient broth (Oxoicl) in 4 oz McCartney bottles. These were inoculated with a loopful of an overnight broth culture and incubated a t 30" for 12 hr with continuous gentle shaking. The cultures were centrifuged and the cells washed 6 times in 25 ml amounts of sterile saline. The final deposit was resuspended in saline, shaken rapidly for 10 min and centrifuged lightly to throw down cell aggregates. The homogeneous supernatant suspension was pipetted off, diluted with

Ecology and genomics of Bacillus subtilis

“…Does B. subtilis actually grow in soil or is this a place where spores accumulate until they are once again presented with conditions propitious for their germination and proliferation? The use of fluorescent anti-bodies to distinguish vegetative and spore forms of B. subtilis in diverse soil samples 4 revealed that the organism was most often in its vegetative form when associated with decaying organic material 5 . Further support for the idea that B. subtilis can lead a saprophytic lifestyle comes from experiments in which spores were inoculated into artificial soil microcosms saturated with filter-sterilized soluble organic matter extracted from soil 6 .…”

Spore Antigens of Clostridium Sporogenes

“…The highest dilution showing visible agglutination was taken as the serum titre. Negative serum and saline controls were included (Noble, 1927;Lamanna, 1940Lamanna, , 1942Norris and Wolf, 1961 ;Walker, 1963).…”

The Classification of Bacillus thuringiensis

“…Ant e&& analy& H agglutinations were performed by a method based on that of Noble (1927) and developed from that of Norris & Wolf (1961).…”