Utilization of propylene glycol, glucose, fructose and sucrose by mycobacteria were tested on the following medium: (NH₄)₂SO₄, 2.64g; KH₂PO₄, 0.5g; MgSO₄-7H₂O, 0.5g; Purified agar (Wako), 30.0g; and distilled water, 1000ml. Propylene glycol was added at concentration of 0.5M, and sugars at concentration of 0.05M. The pH was adjusted to 7.0 by addition of 10% KOH. The media were incubated with one loopful of the stack cultures and incubated at 37°C. Growth was observed after three weeks in comparison with growth on control medium containing no carbon source.
Among mycobacteria tested (Table 1), M. smegmatis, M. fortuitum and M. parafortuitum utilized propylene glycol as sole carbon source, but not all strains. M. phlei and other rapid growers did not utilize it.
All strains of M. fortuitum, M. parafortuitum, M. smegmatis, and M. phlei utilized both glucose and fructose as sole carbon sources. Sucrose was utilized by a few strains of scotochromogens, M. fortuitum, and M. smegmatis. M. kansasii utilized only glucose.
Among M. avium, nonphotochromogens and scotochromogens, M. avium did not utilize all sugars, twelve of nineteen nonphotochromogens utilized only glucose, and fonrteen of fifteen scotochromogens utilized glucose. Five strains of scotochromogens utilized also fructose or sucrose or both (Table 2).

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As reported in the previous paper tubercle bacilli are deprived of their acid-fast stainability completely by the UV irradiation for a certain time period. Obviously longer time period of UV irradiation is required for the complete loss of acid-fastness in the virulent type strains than in the a- or low-virulent ones. This fact seems to suggesst the difference in the cell wall structure between these two types of tubercle bacilli and is likely to enable us to attribute the virulence of tubercle bacilli primarily to the proper cell wall structure. Based on the results obtasned a very simple method to distinguish, even roughly, these two types of strain by the staining of smears was devised. Further, the relation between the virulence and the cell wall structure of tubercle bacilli was discussed.

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Hyperimmunized anti-α-toxin horse serum failed to protect mice from lethal infection by virulent staphylococcus, while it protected mice from the intoxication by α toxin. On the other hand, it was elucidated that α toxin was able to promote the invasiveness of avirulent staphylococcus in mice. Therefore, it was suggested that a different factor in addition to α toxin ought to play a roll in virulent staphylococcal infection.
Passive immunization by antibacterial horse serum and active immunization of mice by vaccine or toxoid also failed to protect them from lethal infection. Thus, it seems that humoral factor in immunity in staphylococcal infection is quite limited.

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On the course of examination of sucrose-fermentability of Vibrio parahaemolyticus isolated from the stool of food poisoned patients, it was recognized that the sucrose-negative strains gave the intermediate appearance between positive and negative, or positive appearance in some case, with No.15 liquid medium (confirmation medium) or Artificial Marine Water medium, though they gave clearly negative on TCBS agar plates (isolation medium) or with the Simmons medium. The causative agents to decrease the pH of the medium were metals such as Cu⁺⁺, Pb⁺⁺ and Sn⁺⁺ which were contaminated in the distilled water used. These metal ions catalyze the hydrolysis of sucrose to glucose and fructose during autoclaving at 120C for 20min. Acids which were produced from those reduced sugars by sucrose-negative strains decreased the pH of the weakly buffered AMW medium, but not of strongly buffered Simmons medium. However, the amounts of the acids produced by sucrose positive strains from sucrose was large enough to decrease the pH of both AMW and Simmons media.
This conclusion came from the following observation: 1) The distilled water which gave the paradoxical results contained 0.25μg/ml of cupper ions as expected. 2) The medium made with deionized water did not give such a funny results. 3) If CuSO₄ was added to the sucrose solution before autoclaving at a final concentration of 1.0μg/ml, 2∼3% of sucrose was converted to the reducing sugars. The false positive results on the sucrose-fermentability was always found by the above sucrose solution. 4) When the sucrose, sterilized by membrane filter, was used, the pH changing curve was the identical with that without sucrose. If glucose, instead of sucrose, was added into such media, the extents of the decrease in pH were proportional to the amount of glucose added and independent of the amounts of sucrose. 5) When the medium was autoclaved in the presence of CuSO₄, the extent of the decrease in pH was also proportional to the amounts of reduced sugars produced from sucrose by autoclaving. 6) The buffering ability of Simmons medium was much stronger than AMW medium.
Thus, the sucrose fermentability can be determined accurately if one used the pure water which does not contain metal ions and a strong buffered medium as Simmons.

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1) The Shwartzman type active factor was noted in cultured filtrate of Serratia marcescens and polysaccharide fractions were extracted from the filtrate.
2) The Shartzman active substance has been believed to be nonpathological but it is now believed that it plays a role in motivating the pathogenicity of the Serratia marcescens.
3) By the Shwartzman histological reaction it was possible to produce necrosis of the rabbit pancreas. This illustrates that localized circulatory damage caused by the Shwartzman reaction plays an important role in the formation of acute pancreatic necrosis in man.
4) The manifesting ability of the Shwartzman reaction is stronger in the pigment forming strain as compared to the nonpigment forming strain. This is believed due to the quantitative difference in the Shwartzman activating substance.

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The following evidence has already been observed in our department:
When staphylococci are suspended in saline solution and shaking-incubated at 37°C water bath, they all die at exponential rate, though with some differences according to the bacterial strain.
This phenomenon is not only interesting in its biological nature but also seems to have some important significances under the consideration of that the saline suspension is usually employed in many experiments of staphylococcus, for instance, serological reactions bioassay of various bacteriocidal action etc.
The present studies were carried out to analyse the biological nature of this phenomenon chiefly using one of the staphylococcus strains, No.66Y, which shows the typical pattern of this phenomenon.
And the results of the experiments are as follows:
1) The typical death could be observed when the concentration of cocci in the saline suspension were less than the so-called “M concentration” of this organisms in their bouillon cultures.
2) Under the condition above mentioned, the rapidity of the death of cocci is in proportion to the bacterial concentration in the saline suspension.
The less the concentration of cocci is at the biginning of incubation (O time), the greater the degree of the death. In the case of the typical strain 66Y, when the starting concentration of cocci is in order of 10⁶ cells per ml, the surviving cells after 3 hours incubation will be about 10^-4 less of the bacterial concentration at O time.
3) If the initial concentration is higher than the limits of M concentration, neither death nor proliferation of the cocci can be observed at least during the observation within 3 hours.
4) The death was much remarkable and rapid when the bacterial cells suspended in the saline solution were taken from the younger culture (of logarithmic phase) than that when they were taken from the old culture (of stationary or declining phase).
5) The closer the temperature of the incubation of the bacterial suspension was to the optimal temperature for the cultivation of staphylococcus, the greater degree of the death could be observed.
6) When KCN was added to the bacterial suspension in final concentration of 1/200mol. to stop the respiration of the bacterial cell, the rate of the death was much decreased.
7) Chloramphenicol did not give any influence to the course of this phenomenon when it was directly added into the saline suspension. However, when the cocci were preincubated for 2 hours at 37°C in the bouillon containg this antibiotics (as the inhibitor of protein synthesis) before taken and suspended in the saline solution, the death of such pretreated cocci in the saline solution could be greatly suppressed.
8) From the above mentioned experimental results, some possible explanations for the nature of this phenomenon were considered and discussed, presuming two metabolic cycles (growth cycle and autolytic cycle) which turn round to the direction reverse to each others in the staphylococcal cell.

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It was elucidated that 5mg. or 100mg. of Habu Snake Venom (Trimeresurus flavoviridis) were available as first immunizing doses in horses when it was treated by Freund's incomplete adjuvant or 100mg. of Dihydrothioctic acid. Four weeks after, 1mg. of plain venom was injected subcutaveously into horses, and the dose was then increased duplicatedly in every week. After ten or fifteen times of booster injection, 0.1ml. of the sera showed to neutralize sufficiently the lethal effect and local lesion due to the venom of 300γ or more. The sera were mixed in vitro with the various amounts of venom and injected intramuscularly into the legs of mice. Thus the total amount of venom used for the immunization and the periods of immunization were reduced to one half in compared with the immunization method without adjuvant adopted until now.

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