FUNDAMENTAL STUDY ON THE VERTICAL DIFFUSION METHOD:III Report. Culture Media and Reliability of the Vertical Diffusion Method
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In the present report, studies were made on the various physical states of the media andtheir influence on the inhibition zone and further on the reliability of this method itself.<BR>1) We have compared the inhibition zone of INH, using two kinds of test tubes, 695 ordinary slant tubes and 608 crook-necked tubes, each with 3% Ogawa medium. The onlydifference of these two groups of medium was that the latter had the same thickeness allover.<BR>After 2 days' incubation of the media inoculated with BCG, INH was poured on them.<BR>The reading was made after 3 weeks' incubation thereafter. The inhibition zone was measuredfrom the bottom of the test tube.With regard to all of the doses tested, there was no difference between the length of theinhibition zone of two groups. But among the culture media of one INH dose, it was variedin wide range, although it was almost the same in average in two groups (Table 1, Fig. 1, 2).<BR>2) Among the 695 slant tubes above mentioned, there was no difference in the lengthand the distribution of the inhibition zone between the 160 tubes whose media were from 8. 2cm to 9. 9 cm long and the 157 tubes whose media were from 12 cm to 17. 2 cm (Fig. 3).<BR>3) Into the groups of 10 tubes, each containing 8, 6, 4 and 2ml of 3% Ogawa medium, 0.2A of INH was poured immediately after the inoculation of 0.05mg of BCG. The length ofthe inhibition zone was in average 3.9, 4.0, 4.6 and 5.1cm.<BR>The growth of bacilli was similar between the tubes containg 8 ml of media and 6 ml ofmedia, but revealed the decreasing trend in the order of tubes containing 4 and 2 ml of themedia (Fig. 4).<BR>In the next experiment, M. phlei was inoculated on two groups of 10 tubes of 1% Ogawamedium, one group containing 8 ml and the other 2ml. After 4 days, there was 9 times asmany bacilli propagation in 8 ml of medium as that in 2ml of medium when inoculated with 10-5mg, and 4 times more in the former when inoculated with 10-7mg (Table 2). We considerthat there is a difference of about 10 times between 8 ml and 2ml of media concerning thepropagation of bacilli, but the difference of 1cm of the inhibition zone seems to be too largeeven if this difference of the bacilli number is taken into consideration. However, the decreaseof the bacilli number will certainly be one of the causes of the elongation of inhibition zone. in 2 ml media.
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