1) While non-specific polysaccharide is composed of peptide and polysaccharide, almost all of the antigenicity to the antianthrax serum exists in polysaccharide. 2) In a short hydrolysis accompanied by a little isolation of galactose, the serological activity is greatly reduced. 3) Immunospecific protein is not hydrolysed in N-H2SO4, and, accordingly, no reduction of antigenicity was seen. However, in the hydrolysis in N/2-HCl, a drastic reduction of antigenicity was brought about in a very short time.
The cross resistance of intermediate resistant mutants to streptomycin (SM) and kanamycin (KM) was studied genetically by several investigators. The present author attempted to observe the cross resistance of one-step highly SM resistant mutants and SM dependent mutants against KM, and to analyze the genetics of the cross resistance using SM dependent mutants. Strains used were Escherichia coli, strain B/r and its mutants. Phage P 1 was used for transduction technique. One-step highly SM resistant mutants showed the slightly increased resistance to KM, but SM dependent mutants showed various resistant levels, increased or decreased, to KM. From the results of reciprocal transduction between SM dependent mutants and original strain, it was clearly shown that a resistance level to KM was controlled by SM dependent mutation itself or by a closely linked mutation not separable from SM dependent mutation through transduction. The analysis of non-dependent (suppressor) mutants derived from SM dependent mutants indicated that resistance level to KM was also controlled by the suppressor mutation.
The character of R (umeda), an episome mediated multiple drug resistance factor, originating from Shigella flexneri 3 a (Umeda strain) was investigated. From the analysis on the phenotypic expression of the streptomycin (SM) resistance of this R factor as well as the inhibitory effect of streptomycin to its transfer to recipient E. coli, following results were obtained 1) Colonies of the recipient cell to which R (umeda) was transferred were hard to be obtained when donor and recipient were mix-incubated on the selective agar plate containing SM in order to transfer this factor to E. coli. This phenomenon is considered to coincide with the “SM effect” which was reported by Mitsuhashi et al. 2) So far examined, the SM effect has been observed only when E. coli is employed as recipient, such as upon the transfer of this factor from Shigella to Escherichia, but not when the recipient is Shigella, such as upon the transfer from Shigella to Shigella or from Escherichia to Shigella. Inspite of belonging to the same species, when B strain of E. coli was employed as recipient the SM effect was not so remarkable as when K-12 or C5o was the recipient. 3) The expression of SM resistance charactor of R (umeda) varies with the kind of recipient cell in which it enters. The SM resistance level expressed by this factor in other Shigella strains is as much high as that of the original strain Sh. flexneri 3 a-U (R), but it is extremely lower in Escherichia strains. And as far as Escherichia, R (umeda) can express a little higher SM resistance level in strain B than in K-12 or C50 strain. 4) The SM effects are assumed to be more remarkable when the bacteria, to which R (umeda) can not render an enough high level of SM resistance, are employed as the recipient of this R factor. In other words a parallelism could be observed between the SM expression level of R (umeda) expectable in a certain recipient and the sensitivity of that recipient to the SM effect. 5) It is assumed from the results of the analysis on the time course of the transfer of this R factor, that there is a certain lag phase until the R (umeda) incorporated into the recipient (E. coli) cell has rendered the latter the active resistance to streptomycin and in this lag phase occurs a series of indispensable events, one or some of which are sensitive to the SM effect. 6) On the other hand, the depressed expression of SM resistance of R (umeda) in E. coli can be made considerably better if chloramphenicol (CM) is added to the SM selective medium. 7) The SM effect could also be cancelled by adding CM to the SM agar plate on which donor and recipient are mix-inoculated. On this mechanism some possible explanations were discussed.
To observe in detail the influence of Streptomycin on the growth of SM dependent type, the author chose B. subtilis (ATCC 558) and made experiments on its responses to Streptomycin, with special reference to the relation of the SM concentration with spore formation of its SM dependent type together with germination and outgrowth of the spores formed. The results are as follows. 1. Among 18×109 cells of wild type of B. subtilis (ATCC 558) about 3, 200 cells of SM resistant and one cell of SM dependent are found as its mutant. 2. B. substilis (ATCC 558) shows normal growth at SM concentration below 10γ/ml, but the growth declines rapidly between SM 10γ/ml-20γ/ml, and the growth stops entirely at SM concentration 20γ/ml. 3. SM resistant type grows well between SM 0.1γ/ml-SM 2, 000γ/ml, the growth declines rapidly at about SM 10, 000γ/ml. 4. SM dependent type shows normal growth between SM 10, 000γ/ml-20γ/ml, the growth get worth below SM 10γ/ml, and the cell shape elongates several times, at SM below 1γ/ml each cell becomes filamentous form and has several ten times length of normal cell, as if the cells have stopped to divide at low SM concentration. 5. The spore formation of SM dependent type begins at SM 40 γ/ml, and the spore formation rate goes up with increase of SM concentration, and reaches 70 %-80 % when SM concentration is over 6, 000γ/ml. 6. The germination and outgrowth of spores of the SM dependent type slow down with decline of SM concentration, but the germinated spores grow in normal forms until SM concentration reaches 7.8γ/ml, then the germination becomes much slower, and the cells grow in filamentous forms or irregular forms. At extremely low SM concentration both germination and outgrowth nearly stop.
It is considered that the cellular factor plays a great role in the immunity of experimental typhoid. In order to clearify this cellular factor, we have tried to examine relations between the metabolic changes of host cells and immune process. The activities of several kinds of enzymes of the mouse peritoneal cells. were quantified during 4 weeks after vaccination with such vaccine as attenuated living vaccines, chrome-treated. killed vaccines or endotoxin. The results obtained were following. (1) Acid phosphatase activity of the peritoneal cells was increased after injection of those vaccines. Especially, 11 Rx living vaccine, which was known to give good immunity to the hosts, could greatly increase the enzyme activity of those cells. (2) Although the lysozyme activity was increased after inoculation of both living vaccines, 11 Rx. living vaccine appeared to be more effective to increase the activity than Jena living vaccine which was known to give poor immunity to the hosts. The injection with chrome-treated killed vaccines or endotoxin, which were known not to give cellular immunity to the hosts, decreased the activity of lysozyme of the host cells. (3) Cathepsin activity was slightly increased after injections of those various kinds of vaccines. (4) After inoculation of 11 Rx living vaccine, β-gluculonidase activity and succinic dehydrogenase activity were examined and were found that β-gluculonidase activity was usualy low in comparison with that of the normal controls and succinic dehydrogenase activity was variable.
In these experiments, two animals of Macacus cyclopis weighing 5 kg and one animal of M. irus weighing 3, 5 kg were used. These animals were treated as follows. Before infections, sonicate of Shigella flexneri type 2 a or chloramphenicol was given into the stomach as pretreatment. Three animals were infected with 20cc (50×108/cc) living cultures of Shigella flexneri type 2a by its introduction directly into the stomach by stomach tube. The results were as follows: (1) In the pretreated monkies of 5kg weight infected with Shigella, clinical and pathological symptoms of dysentery were recognized. (2) In the pretreated monkey weighing 3, 5kg death due to Shigella infection was observed within 7 hours. Sh. flexneri was isolated from the heartblood. In the pathological observation, lesions of the lungs were seen at the anterior caudal part. Gastritis and colitis with the hemorrhage were recognized in the gastrointestinal tract.