In the previous paper, the death phenomenon of staphylococci in saline solution was described and the results of the studies on the biological nature of this phenomenon were reported. This phenomenon is not only interesting in its biological nature but also seems to have some important significances because saline suspensions of staphylococci are usually employed as the control sample in various kinds of biological reactions. In these reaction, the control sample of bacterial suspensions should be in the resting condition under which the bacterial cells would not be killed and at the same time not proliferate. From this point of view, experiments were carried out to study on the inhibitors for the above mentioned death of staphylococcus in saline solution and to devise the idealistic medium for the suspension of staphylococcus composing of minimum set of amino acids. Results are as follows: (1) The death rate of staphylococcus in saline suspension can be slightly decreased if glucose is added to the saline medium. (2) If the staphylococci are suspended in simple casamino acid medium at 37°C and shaking-incubated, there occurs neither death nor growth of cocci even after overnight incubation. (3) Of the eight kinds of amino acids which are known to be required as the essential nutrient for the propagation of the St. aureus, three amino acids namely 1-tryptophane, glutamic acid, and hystidine which have been reported to promote the propagation of the staphylococci but are not indispensable, are proven to be unable to inhibite the death of staphylococci even when all of them are added into the saline medium. (4) Subsequently four amino acids; 1-arginine, 1-cystine, 1-glycine and 1-proline, which are known to be indispensable for the propagation of the staphylococci, were examined for their ability to inhibit the death of staphylococci when they were added to saline medium. The former three were proven to give a considerable suppressing effect to this phenomenon, and among them, the effect of cystine was the strongest. But the latter one could not show any effect. (5) Although the above mentioned suppressing effects of these three amino acids are less than that of casamino acid, they could show a remarkable effect as much as equivalent to that of the casamino acid when all these three amino acids were added together. (6) Consequently the mixture of the above mentioned 3 amino acids and glucose could be recommended as the most suitable medium for staphylococcus suspension.
This paper outlines the natures of Habu snake and Erabu sea snake and properties and actions of their venoms. Moreover was described the effect of tannic acid on the venoms. Habu snake (Trimereserus flavoviridis) is venomous, landinhibiting on the Amami islands and about 150cm long. The victims of Habu snake bite was estimated at 250 to 300 each year. The death rate during recent 7 years was more than 1 per cent. The minimal lethal dosis for mice, weighing 15 to 17g. was about 150γ/0.1ml by intramusculare injection. It was considered that the venom was composed of haemorrhagic, angiotoxic and myolytic factors, which were completly inactivated by heating at 100C for 10 minutes, and heat-stable myolytic factor. Erabu sea snake (Laticauda semifasciata) lives on the coast of Amami Oshima, and has strong fatal venoms. Minimal lethal dosis, in experiments with mice weighing 15 between 17g., was about 6γ/0.1ml by the intramusculare injections. Erabu sea snake venom is considered to have chiefly neurotoxic component which was relatively stable in heating. It was recognized that the toxic activities of the venoms of these different species were inhibited by aqueous solution of tannic acid; a 8.5% solution inactivated lethal and local haemorrhagic activities of 500γ/0.1ml of Habu snake venom, and fatal toxicities of 25γ/0.1ml of Erabu sea snake venom. The above mentioned effect of tannic acid on the venoms may be due to coagulations of the venom and tissue proteins by tannic acid.
A lysogenized Mycobacterium, strain F 21 (LI), was successfully induced by mitomycin C. The induction rate was maximum at 10 minutes after the addition of mitomycin C at a concentration of 1μg/ml. However, the induction rate was relatively lower than that of E. coli K 12 (λ) reported by Otsuji et al. Burts size of induced F 21 (LI) was calculated as 14.5. Two strains of Mycobacteria, F 21 and Jucho, both belongs to Mycobacterium smegmatis from their phage patterns, were lysogenized by four strains of temperate phage, LI, Y 13L, Y 13S and A 6, respectively, and each of them was treated with mytomycin C. The results obtained, revealed that in Mycobacteria as well as in other bacterial species, induction might depend upon the phage strain rather than bacterial strain which was lysogenized by the phage. Mitomycin C resistant F 21, lysogenized by the phage LI, could not be induced by mytomycin C but it could be induced by ultraviolet irradiation. Nitromin, which is known as an inducing agent, could not induce the phage LI from the F 21 (LI) under the present experimental conditions.