In the previous report, the virulence of 43 strains of tubercle bacilli to mice was presented.
The authors concluded that the virulence of tubercle bacilli newly isolated from the Japanesepatients was not of the same degree with each other, but was varied from high to relativelylow degree. Employing the same strains as used in the previously reported mice experimentsexcluding six out of eight Indian strains, the virulence to guinea pigs was examined. Thesuspensions of bacilli used in the experiments of guinea pigs were the same as used in theexperiments of mice.Three or six adult male guinea pigs were infected subcutaneously with 0.1mg of eachstrain. All the animals were sacrificed eight weeks after infection. The tuberculous changesof visceral organs and lymph glands were observed macroscopically.
The standard of the macroscopic tuberculous indices of guinea pigs used in this reportwas as follows: For the changes of lungs and spleens, no lesions…0, 1 to 9 tuberclescl, 10to 19 tuberclesc2, 20 to 29 tuberclesc3, more than 30 tuberclesc4, innumerable tuberclesc5.
If large conlfuent tubercles existed, or many necrosis were found, 0.5 or 1 was added to theprevious indices. For the liver, no lesionc0, few tuberclesc1, some tuberclesc2, many tuberdesc3, , innumerable tuberclesc4, innumerable tubercles with necrosisc5. For the lymphglands, tuberclous swelling of the gland 2 to 5mm in diameterc1, 5 to 10mm in diameter2, 10 to 20mm in diameterc3, more than 20mm in diameterc4.
Another three guinea pigs were infected intraperitoneally with 0.1mg of bacilli, and thedeath rate up to eight weeks after infection was observed. (Thirteen out of 43 strains were, omitted from the experiment of intraperitoneal infection.) The results were summarized as follows:
(1) The average macroscopic tuberculous indices of guinea pigs infected subcutaneously with 0.1 mg of bacilli were varied from 26.8 to 5.2. The average indices of visceral organsexcluding lymph glands varried from 10.9 in the strain of the maximum virulence to 0.9 inthe strain of the minimum virulence (Table and Fig. 1).
(2) The average weight of the spleens also showed the differences of virulence accordingto the strain from one strain to another. The maximum average weight was 8.9g and theminimum was 0.8g (Table 1).
(3) The root indices of virulence after Mitchison were calculated. In our experiments, infection was performed subcutaneously, infectious dosis being 0.1mg, and and all the guineapigs were sacrificed eight weeks after infection. Therefore, the methods of experiments werenot identical with that of Mitchison, et al.Thirty four strains out of 43 Japanese strains were more than 0.8 in indices. The indicesof 4 strains were 0.8 to 0.6. The indices of five strains were less than 0.6., of whichtwo wereresistant to INH (Tables 1 and 2).
(4) Death rate of guinea pigs infected intraperitoneally with 0.1mg of bacilli variedfrom 100% to 0% (Table 1). But, the death rate did not correlate with the macroscopic tuberculous indices.
(5) INH-resistant strains were less virulent to guinea pigs.

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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.
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.
After 2 days' incubation of the media inoculated with BCG, INH was poured on them.
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).
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).
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.
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).
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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Retreatment cases of pulmonary tuberculosis 17 in number, were treated with isonicotinicacid hydrazide methansulfonate (IHMS) and sulfamonomethoxine for 4 months, and the clinicaleffects and side effects were observed.
The appearance of INH-resistant tubercle bacilli was delayed by the combined use ofsulfamonomethoxine with IHMS as well as by other sulfa-drugs.
From these results it is considered that combined therapy of IHMS and sulfamonomethoxineis advisable, and the therapentic value of sulfamonomethoxine is equal to that of the othersulfa-drugs already reported.

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