It was reported in the previous report that an antimicrobial activity against a virulent strain of M. tuberculosis could be conferred to dd mice, when they were transferred with peritoneal cells derived from the homologous (dd) mice which had been immuni zed with BCG strain. This effect of passive transfer was already apparent when the recipients were challenged 4 days after the transfer. In the present report the effects of some pretreatments of cells to be transferred and streptomycin (SM) administration to recipient mice after transfer were investigated. The results were judged by counting viable units of tubercle bacilli in the spleen and lung at various intervals after the challenge with a virulent strain resistant to SM, the culture being performed on media with and without SM.
A similar effect of passive transfer as by intact cells was obtained by using the material which had been oscillated (10 kc) for 2.5 minutes, but not by cells heated at 48°C 30 minutes. Viable units in organs of mice transferred with the heated cells showed the numbers similar to those in control animals. The effect of passive transfer with intact cells was also almost nullified when SM was administered during 4 days from immediately after transfer until challenge.
It was assumed from previous experiments that viable cells of BCG contained in transferred cells of immunized mice could withstand the pretreatments of heating or oscillation mentioned above, but they may be vulnerable to SM administered immediately after transfer.

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This is a report of a strain of human tubercle bacilli which showed inconstant niacin test results. This strain was isolated from a patient one month prior to his death, who expired inspite of chemotherapy for 1 year at a certain hospital and combined chemotherapeutic and surgical treatment at Kitasato Institute Hospital for the following 3 years.
After having completed monthly routine sensitivity tests during about 2 years of his first admission to the hospital, niacin tests were performed on 25 strains on 10 occasions, which had been grown on the various sensitivity test media. Among these, 12 strains showed weakly positive results, 2 strains doubtful and 11 strains negative. However, all the tests repeated after successive cultures of these negative strains were strongly positive.
The niacin tests performed on 6 strains which were isolated during the terminal phase of his illness were all negative in the first generation of cultures, and even after each succes sive culture of these strains until the 4 th generation, all the strains showed doubtful or negative results except for a single strain with weekly positive result.
From the results obtained, unless the results of niacin tests are constant, it is desired that various other bacteriological examinations are added to the niacin test in order to differen tiate human tubercle bacilli from other acid fast bacilli.

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Not only in the isolation of tubercle bacilli from specimens but also in the direct drug sensitivity test, 3% Ogawa's whole egg medium which contains KH₂PO₄ in a concentration of 3% is most widely used in Japan. Because, it is generally recognized that this medium is superior to Löwenstein-Jensen medium both in the simplicity in construction and in the practical usability. However, concerning the uniformity of the efficiency of all media prepared in different laboratories no observation has been made yet.
On the occasion of the Tuberculosis Mass-survey carried out in 1963 in Japan each 10 slants of 3 % Ogawa's medium prepared in 47 different local Public Health laboratories were collected to the National Institute of Health laboratory to make the comparative studies on the efficiency in the isolation of tubercle bacilli. Number of slants tested was 149 in total including each 3 slants of 3 different batches of media prepared in NIH laboratory.
Two sputum specimens from tuberculosis patients were homogenized strongly with equal volume of 2% NaOH solution employing homogenizer and were diluted ten-fold with 2% NaOH.
Two dilutions of 10^-3 and 10^-4 of one sputum and a dilution of 10^-3 of the other were. inoculated onto 149 slants of 3% Ogawa's media by three technicians.
From the former sputum (a) the growth of tubercle bacilli appeared after 4 weeks incubation, but from the latter (b) it was after 6 weeks as shown in Tables 1 and 3. Square root of the average colony number developed on 149 slants was 5.60.
Table 2 shows the statistical analysis of the variance of the colony number listed in. Table 1. According to the colony number these different 50 batches of media samples were grouped into 3, ie, A, B and C. To the group A 39, to B 3 and to C 8 batches belonged respectively as shown in Fig. 1. Square root of the average colony number in each group was 5.73, 6.78 and 4.60 respectively.
In each group the difference was statistically insignificant between different batches. But, the differences between group A and B, and group A and C were highly significant.
That the average number of group B was larger than that of group A may be effected by the rather larger content of the condensed water in slants. Contrary to this, the smaller mean number of group C than group A may be due to some special conditions in the process of media preparation.
The latter sputum (b) gave the results that growth of tubercle bacilli was greatly affected by the type of stopper on the slants; namely, retarded growth or no growth was observed on the slants with air passable rubber stopper.
From these observation it was shown that 3% Ogawa's media prepared in different laboratory in Japan are rather uniform and not so much different in the efficiency of isolating tubercle bacilli from specimens.

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Time-course observation of the acid phosphatase activity of the liver and the spleen wasa. conducted in mice after intravenous infection with a bovine strain of tubercle bacilli (Ravenel).
The mice were of dd-line commercially available in this country, weighing about 20g Nonspecific acid phosphatase activity of the tissue homogenates was measured using p-nitrophenyl phosphate as substrate at pH 5. 6 (0. 1 M acetate buffer) and specific glucose-6-phosphatase of the liver was measured using sodium salt of G-6-P as substrate at pH 6. 5 (0.1 M maleic acid buffer). Taking the swelling of the test organs into consideration, the activity was expressed as the amount of liberated p-nitrophenol or phosphate P by the whole organ in 60 minutes at 40°C for the former or in 15 minutes at 40° for the latter. Then, per cent increase of the activity from normal (uninfected) was calculated and presented graphically.
As shown in Fig. 1, 2, and 3, acid phosphatase activity of the liver and the spleen was elevated in the course of infection. In the spleen, the maximum elevation was observed in the later period than the peak of the bacilli-multiplication curve. Once increased activity showed a tendency to decrease thereafter, perhaps corresponding to the stage of host-parasite equilibrium in tuberculous infection. On the other hand, G-6-Pase activity of the liver did notincrease at all, but sometimes showed a lowered value.
G-6-Pase activity is the function of liver parenchymal cells, and acid phosphatase activity is that of Kupffer cells according to the cytochemical investigation of Thorbecke et al. Therefore, there is a good evidence to believe that the elevation of acid phosphatase activity of the spleen and the liver in tuberculous infection is an expression of the activated reticuloendothelial system. The swelling of these organs will be explained as the increase of cell numbers in the, system.

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Microbiologically active isoniazid (INH) concentration in the sensitivity test medium was assayed to the lower limit of 0.08μg/ml, by the thin-agar cylinder-plate method using INH highly sensitive rapid grower i. e., Mycobacterium H-7 as a test organism.
INH potency in the sensitivity test medium during storage or incubation was investigated as follows:
1) In the solid egg medium (Ogawa's 1 % KH₂PO₄), no remarkable decrease of INH activity was observed under the conditions of heating at 90°C for 60 minutes or storage at 4°C for 3 months. Whereas, deterioration of INH potency was observed after storage at 4°C for more than 3 months and within 2 months incubation at 37°C.
2) In Dubos' liquid medium, all of INH and its derivatives i. e., IHMS, INHG, IP, which proved to be microbiologically active, decreased in their potencies and were inactivated within 2 weeks incubation at 37°C.

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One thousand pulmonary resection for tuberculosis were performed in the past ten years from 1953 to June 1963. The mortality rate was 1.6% (16 deaths) and the incidence of postoperative complications was considerably lower than those reported previously, that is, 3.0% of bronchial Estula (30 cases), 0.2% of empyema thoracis (2 cases) and 0.3% of Hemothorax (3 cases).
Bronchial fistula has been consi dered to occur mainly in the case with resistant organisms or cavity formation. However, our study revealed that this complication is resulted mainly from faulty handling of bronchial stump except in the case of deteriorated general condition, accident or tuberculous change of bronchial stump.
Empyema thoracis has been considered to be due to contamination of intrapleural space caused by the rupture of cavity. However, this complication revealed to be preventable by sufficient intrapleural irrigation, postoperative drainage and chemotherapy.
The incidence of postoperative hemothorax can be kept minimum by paying adequate attention to hemorrhage during the operative procedure avoiding extrapleural dissection and to hemorrhagic tendency or fibrinolysis.

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