Since M. lepraemurium Hawii was isolated on 1% Ogawa yolk medium, 1)-9) Kozeki et al 10)11) and Mori 12) succeeded in the follow-up-experiments of Ogawa's method, but successive inoculation with smooth suspension was not succeeded, just as described at first by Ogawa.
It was still obscurre that some growth-negative tubes were always observed in cases of successive inoculation and primary isolation, in despite of bacterial mass inoculation. The difficult problem is to solved when the single cell culture of M. lepraemurium would be succeeded. As it was very difficult to get 100% success of the successive inoculation even in the bacterial mass inoculation, some conditions for getting the better efficiency were carefully studied by the method of bacterial mass inoculation.
1) Successive inocualation of M. lepraemurium was not affected on the 1% Ogawa yolk medium without malachite green.
2) Successive inoculation of M. lepraemurium was independent on any modified 1% Ogawa yolk media supplemented respectively with the following substrates: cystin, cys-tein special agar noble, sodium citrate, sodium α-ketoglutarate, glucose, protoporphyrin and water- or butanol-extract of 1% Ogawa yolk medium on which M. lepraemurium had grown.
3) Successive inoculation of M. lepraemurium was slightly inhibited on the modified 1% Ogawa yolk medium supplemented with sodium pyruvate.
4) Addition of hemin to 1% Ogawa yolk medium gave a good result for successive inoculation of M. lepraemurium.
5) In successive inoculation of M. lepraemurium, air-loose condition through a small hole in gum stopper gave the better result than air-tight condition without a small hole in it.
6) The organism from old culture over 2 months was not good for successive inoculation, and young rough organism from 30 to 40 days culture was most suitable More over, the organisms should not be scattered on the surface of medium. It was important to inoculate crowdly for getting good results.

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