1) The antigen suspension, saline and formalin (0.4%) saline which were first adjusted to pH 7.4 became to pH 6.3 after two weeks stay in room temperature. These decline in pH, however, had no influence upon the agglutinability of
Clostridium perfringens antigen.
2) Neither the differences of NaCl concentration in saline nor the difference of the components (agar, yeast extract, peptone, meat extract, soluble starch etc.) of the media used for the cultivation of
Clostridium perfringens showed influences upon the agglutinability.
3) The differences of the intensity and the titer of agglutination reaction were observed between antigens cultured on the heart infusion agar plate and in its original broth.
4) The size of the inoculum cultures into the fluid medium for the preparation of antigen had an influence on the agglutinability. Less than 10% of the volume of medium should be inoculated for the purpose of obtaining the antigen of stable agglutinability.
5) As to the reading of tubes in the agglutination reaction, the shaking method showed more stable results than the non-shaking method.
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