A Method for Evaluation of Antituberculous Drugs by Their Diffusion into Depth Culture of Tubercule Bacilli in Semi-Solid Agar Medium.

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The aim or our study is to investigate the properties of semi-solid agar medium for depth culture of tubercle bacilli as compared with those of solid medium for surface culture and with those of liquid medium for submerged culture. The results have been reported in part. The observations made by us disclosed that in cultures under oxygen environment or with INH solution superposed on the medium, the disc of colonies submerged below the surface of the medium, the distance from the surface of the medium being related to the concentration of INH, that is, the growth of tubercle bacilli was inhibited in the uppermost layer of the medium. Each of minimum inhibitory concentrati ons of SM, KM, VM, PAS and INH obtained with this culture method using semi-solid agar medium was very close to that obtained from the dilution method with liquid medium. The present method was also found to be useful for the detection of the antibacterial effect of sera of patients under treatment with various antituberculous agents. Following are the results obtained. 1) Serum level of KM after intramuscul ar administration The minimum inhibitory concentration of KM obtained with the present method was about 1 mcg/ml, and this method could be employed for measuring the concentration of KM ranging from 20 to 1 mcg/ml. Two-fold dilutions of the serum were used for the tests. Serum levels 4 hours after 1 gm intramuscular administration in 30 cases ranged from 80 mcg/ml to less than 10 mcg/ml. 2) Specific effect of seru m under combined drug therapy The depths of the inhibitory zones and appearances of discs varied in accordance with the various drugs combined as in the cases treated with INH or KM. Especially, serum 4 hours after SM-PASINH administration showed marked growth inhibitory effect in the tube cultures. Colonies in the discs were smaller and less in number than those in cases treated with INH or KM alone. The growth inhibitory effect of sera collected 12 hours after administration was less than 4 hour sera. At present, various antituberculous age nts are clinically available and usually given combined. Therefore, it is important to know the total effect of the combined drug therapy in vivo as well as the effect of individual drugs in vitro. The culture method described ca n be used for the evaluation of the total effect of the combined drug therapy with a single medium and sera of patients. Tubercle bacilli grow in 5 to 6 days w ith this culture method and the examination described above can be conducted in this short period.
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A Method for Evaluation of Antituberculous Drugs by Their Diffusion into Depth Culture of Tubercule Bacilli in Semi-Solid Agar Medium.

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