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 1.6×10
5 acid-fast bacilli (strain B 2409) per m
l 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, 0.03m
l of bacillary suspension, containing 4.8×10
3 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 4.8×10
3 bacilli per 0.03m
l 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 4.8×10
3per 0.03m
l was kept in a water bath at 60°C for 5 to 60 minutes. Then 0.03m
l 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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