Nippon Saikingaku Zasshi

Three strains of Gram-negative rods which were non-motile and attacked glucose by fermentation with production of gas were isolated from human feces and temporarily named Nos. 71, 74, and 75 respectively. These strains were agglutinated by several kinds of anti-Shigella sera of A, B, C, and D groups.
Anti-No.71 serum was able to agglutinate Shigella dysenteriae 3, 4, 5, 6 and 7, Shigella flexneri 1_a, 1_b, 2_a, 2_b, 4_a, 4_b and 5, Shigella boydii 6 and 10, and Shigella sonnei.
Anti-No.74 serum was able to agglutinate Shigella dysenteriae 3, Shigella flexneri 1_a, 1_b, 2_a, 2_b, 3_a, 3_b, 4_a, 4_b, 4_c, 5 and X, Shigella boydii 7 and 10, and Shigella sonnei II.
Anti-No.75 serum was able to agglutinate Shigella dysenteriae 6, Shigella flexneri 1_a, 1_b, 2_a, 2_b, 3_a, 3_b, 4_a, 4_b, 4_c, 5, X and Y, and Shigella boydii 5 and 7.
It was impossible to identify these strains by using the method of Cowan and Steel, Bergey, Kauffmann, Sakazaki, or Edwards and Ewing.

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By the aid of the mouse foot-pad passage technic as a means of evaluation for the multiplication of Mycobacterium leprae, observation was made on the survival of this organism at different temperatures. Strain B2409, isolated by Dr. Shepard and maintained at the author's laboratory, and strain Yamamura, isolated at the same laboratory, were employed as sources of M. leprae. The procedures used for inoculation, harvesting, and quantitative evaluation were essentially the same as those described by Shepard.
Hanks' balanced salt solution (BSS) was used for preparing a bacillary suspension and its dilutions. The mice used were of the CF # 1 strain.
Storage of bacillary suspension at 5°C: A suspension containing 1.6×10⁵ acid-fast bacilli (strain B 2409) per ml in BSS was stored in a refrigerator at 5°C over a period from 24 hours to 2 weeks. At certain intervals of time during storage, 0.03ml of bacillary suspension, containing 4.8×10³ organisms, was inoculated into each mouse at the left hind foot-pad. Harvesting was made on the 120th, 180th, 300th. and 365th day after inoculation. The number of acid-fast bacilli in the foot-pad was counted by the microspot method.
There was no significant change in viability of the organisms among the samples at 24, 48, 72, and 96 hours of storage at 5°C. A considerable reduction in their viability, however, was observed in the sample at 7 days of storage.
Storage of bacillary suspension at -25°C: A suspension of M. leprae (strain Yamamura) containing 4.8×10³ bacilli per 0.03ml was stored in a deep freezer at -25°C over a period from 24 hours to 12 weeks. Then it was inoculated into mice at the foot-pad. The counting of bacilli was performed in the same way as mentioned above. The viability of M. leprae was not significantly affected by freezing and storage at -25°C within 12 weeks.
Inactivation of the organism at 60°C: A suspension of M. leprae (strain B2409) containing 4.8×10³per 0.03ml was kept in a water bath at 60°C for 5 to 60 minutes. Then 0.03ml of this suspension was inoculated into mice at the foot-pad. As a result, the viability of M. leprae was reduced gradually by the prolongation of time for treatment at 60°C, and completely lost by heating for 30 minutes.

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