1) A antigenic polysaccharide was isolated from the trypaflavine lysate of B. anthracis through ethanol precipitation and zone electrophoresis. 2) The substance was shown to be a single component in serological and physically by agar diffusion test and ultracentrifugation. 3) An antigenic polysaccharide from a heat dried cell extract and an antigenic polysaccharide from a lysate showed the identical precipitin titer, identical composition in respect to component sugar and amino acids, and similar infra-red absorption spectrum. 4) The substance and CA (antigenic substance from the heat dried cell extract) reacted with anti- B. cereus and anti-B. subtilis serum in equal degree.
In the past three years three strains of unclassified acid-fast bacilli were isolated fromthe sputum of patients with pulmonary disease in Kanto Central Hospital (Tokyo). For the identification of these organisms, various biological and biochemical test and animal experiments (mice) were used in this study. The following tests were performed: Cord formation, Preis test, Drug resistance test, Niacin test, Neutral red test, Auramin test, Catalase test, Peroxidase test, Urease test. Animal experiments: Mice were inoculated intravenously with approximately 0.3mg. of Dubos liquid medium, and sacrificed 2, 5 and 10 weeks after infection. Bacteriological properties of three strains is shown in Table 1. Its pathogenicity was strong for mice see Table 2, 3. From the experimental results, it can be supposed that three strains is an unclassifid acid-fast bacilli. According to Runyon's classification, these strains are similar to the strain of Group 1 photo.(Kato strain) and Group III nonphoto.(Uchida and Kekke strains), but different from the strains reported previrusly by many investigators in respect of various properties.
A study has been done on the cold trichloroacetic acid-soluble polysaccharides of C. albicans under several culture conditions and on the antigenic substances, and the following results were obtained. (I) The amounts of cold TCA-soluble polysaccharide of whole cells have been estimated by the anthrone value, and with synthetic medium it showed the maximumamounts in 2 or 3 days, and thereafter it decreased. Adding various organic acids in glucose medium, it was observed that the anthrone value of cells generally increased, especially with citrate and malate. Acetate or formate has a little effect for growth, but onthe contrary shows markedly effect on the anthrone value. On the other hand, stimulation of TCA-soluble polysaccharide formation of whole cells was not observed at addition of Casamino acid in the culture medium. (II) Extracts of tricholoroacetic acid from cells (SI) and that of hot water from cell residues (SII) were composed of mannose, glucose, galactose and glucosamine as the component sugars of antigenic substances, and further more SII fraction contained xylose, but both fractions did not contain uronic acid. It is interesting to note that SI and SII fractions contain several glyco-proteins which show resemble electrophoretic pattern. (III) In precipitin reaction, it was shown that SII fraction gave more strong reaction than SI.
When spores were suspended in distilled water and meat extract agar was used as a medium for colony formation, sublethal condition for the spores ofB. subtiliswas heating at80°for 30 minutes. What kinds of amino acids were released from spores suspended indistilled water by heating at various temperatures for 30 minutes, especially at sublethalconditions, were examined by paper electrophoresis and paper chromatography. It was shown that glutamic acid was released faintly at 60°C and 70°C and clearly at 80°C. Glutamic acid, aspartic acid, arginine, lysine and probably ornithine were released at 90°C, 100°C and 120°C. In all cases peptide appeared. By hydrolysis of this peptide it was shown that peptide consisted of glutamic acid, aspartic acid, arginine, lysine and neutral amino acids. As glutamic acid was released from spores by heating at under sublethal and sublethal temperatures, effects of glutamic acid on the colony formation of the spores heated at sublethal conditions were examined in synthetic medium. It was clearlyshown that glutamic acid was necessary for the colony formation of the spores heated at sublethal conditions in addition to amino acid which stimulated spore germination. Arginine had also good effect on the colony formation of that spores except glutamic acid. L-alanine, even if acted as a stimulative amino acid on spore germination, had an inhibitory effect on the colony formation with glutamic acid. The selationship between the requirement of amino acids for colony formation of the heated spores and the condition of heating of spores was as follows: a) When spores are heated at relatively low temperature, glutamic acid is not necessary for the colony formation. b) When spores are heated at relatively high temperature, only glutamic acid (or arginine) supports colony formation with amino acid which stimulates spore germination. c) When spores are heated at higher temperature, colony formation is impossible.