鼠癩菌の無細胞液体培地での培養

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A cell-free liquid medium, which was referred to as NC-5, was established for cultivation of Mycobacterium lepraemurium. It was composed of the basal medium and some additives. The formerwas the Kirchner medium enriched with glucose, sodium pyruvate, and calcium pantothenate, andthe latter were goat serum, α-ketoglutaric acid, cytochrome C, hemin, and l-cystein hydrochloride. Allthe additives had to be added aseptically to the basal medium. When bacilli were cultivated in the NC-5 medium at 30 C, bacterial cells began to be elongatedgradually at 4-7 days of cultivation and started to multiply at 2 weeks. Finally, the growth of bacillicould be recognized macroscopically as a turbid mass. A possibility that the growth obtained in NC-5 might be due to the multiplication of other acid-fast bacilli which might have contaminated thestarting material was excluded, as no growth of bacilli was observed in the Kirchner medium, whichis the most suitable for the growth of most acid-fast bacilli. The growth of Mycobacterium lepraemurium reached a maximum in the NC-5 medium at 8 weeksof cultivation. The increase rate was about 100 or 1, 000 times. It is of great interest to note thatsmaller the inoculum size, the high was the growth rate. The possible generation time calculated fromthe bacillary count might be approximately from 8 to 14 days. The growth was inhibited by additionof isoniazid, streptomycin, and mitomycin C, but not by penicillin. In a preliminary trial, no bacilli multiplied in the second generation when the bacilli grown inthe NC-5 medium had been inoculated into a freshly prepared NC-5 medium. An abundant multiplicationof bacilli was observed however, when bacilli were transferred from a smear on the slide tothe NC-5 medium at definite intervals. Finally, it was indicated in animal experiments that thebacilli cultivated in the NC-5 medium for 223 days kept their pathogenic activity. On the contrary, no pathogenicity was observed in the bacilli cultivated in the Kirchner medium under the same conditions.
日本細菌学会の論文

鼠癩菌の無細胞液体培地での培養

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