In order to see the effectiveness of our stored whole blood agar media which were prepared for the inoculation of 0.1cc. of specimens either treated with 4% NaOH or neutralized, the authors made the following experiments.
Several serial ten-fold dilutions of the sputa were made according to their Gaffky scale, and then neutralized. Three dilutions of these were inoculated on the culture media to obtain 10 to 200 colonies by which the average number of colonies of the tested cases was calculated. By the average number of colonies and the statistical analysis, the efficacy of the stored whole blood agar media was evaluated and compared with various conventional media. Furthermore, the effect of the storage of the media upon their efficacy was investigated. And the following results were obtained.1. The medium for the inoculation of 4% NaOH treated specimen was examined with sputa from 158 cases. In regard to the average number of colonies, there was no difference in efficacy between our BIT medium and Sugita's stored whole whole blood agar medium, but more colonies were noted on ours tham Uno's stored whole blood agar medium, our modifica tion III Kirchner's agar medium containing serum, 3% Ogawa's egg medium, and 3% Ogawa's egg medium containing pyruvic acid. Secondly, However, statistically there was no significant difference between curs and others with the exception of Sugita's medium.
2. The medium for the neutralized specimens was tested with sputa from 213 cases. As to the average number of colonies, our BI medium showed more colonies than any other media. By the statistical analysis, there was no signif i cant difference among ours, Sugita's and Kamesaki's agar media containing stored whole blood, and Kirchner's agar medium containing serum, while a significant difference was seen between ours and other media, such as, Tarshis' and Uno's agar media containing stored whole blood, 1% Ogawa's egg medium and 1% Ogawa's egg medium containing pyruvic acid.
3. The effect of the storage upon the efficacy of our media was tested with sputa from 6 cases. Their efficacy was unaltered after 5 weeks' storage in a refrigerator.

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A total of 487 cases with pulmonary tuberculosis who were admitted to the Attatched Sanatorium of Research Institute during the period from 1954 to 1961 undergone original treatmet by chemotherapy and observed for longer than 6 months, were subjected to the study. Strains were defined as resistant, if there is any growth of tubercle bacilli on culture media containing 10γASM, 1γ IγNH or 1γA PAS. The results obtained were the following:
1. At the beginning of treatment, 281 cases (57.7%) were bacilli positive, and among them, 270 were positive by culture. Primay drug resist ance was found in 56 cases (20.7%).
2. Positive conversion of tubercle bacilli was found in 52 cases (10.7%) after continuing negative finding for longer than 4 months. Among them, 25 (48%) were smear positive and culture negative, and remaining 27 (52%) were culture positive. Among the latter, 20 cases (74%) were resistant to at least one of SM, INH and PAS.
3. Comparing the drug resistance at the beginning of treatment and at the time of positive conversion, multiple drug resistance arid INH resistance were found more frequently among the latter. (cf. Table 9).
4. The rate of drug resistant cases among positive converted cases showed no correlation with the quantity of bacilli discharge and the presence of cavity.
5. The clinical course of positive converted cases was different by the drug susceptibility and the quantity of bacilli discharge. Nearly all of the drug susceptible cases and most of the drug resistant cases with a small quantity of bacilli discharge converted to negative again under the same regimen of chemotherapy, while most of the drug resistant cases with a large quantity of bacilli discharge converted to negative by the surgical treatment.
6. Among 243 cases without cavity through out the whole observation period, 6 (2.5%) were resistant at the time of positive conversion.
7. The above mentioned results showed that most of the cases converted to positive after continuing negative finding for certain period were drug resistant, but their clinical course, especially that of the cases with a small quan tity of bacilli discharge, was different with that of the cases showing continuous bacilli discharge.

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Attempts have been made by many investigators for classifying mycobacteria by the use of mycobacteriophage. However, little attention was paid hitherto to the identification of the phage strains, which were isolated from soil of various places independently. To carry out phage typing of mycobacteria, it seems indispen sable to make classification or identification of the phage strains to be used. In the present paper, studies on the classification of 32 myco phage strains collected from various laboratories of the world are presented.
Anti-phage sera were prepared by the repeated intravenous injections of phage suspension into rabbit. According to the neutralization rates calculated, 32 phage strains examined were classified into following 7 serological types: Y7, Y10, PP, Bl, Li, D29 and X. Here, X type phage strains did not show any sign of neutralization by the 6 kinds of anti-sera used. Further, from the standpoint of the activity to human type tubercle bacilli, these phage strains were classified into 3 lytic types, i. e. a, b and c. Thus, 32 phage strains were clas sified into 10 different types.
Typing phages were selected from these different types and some other strains were additionally selected in consideration of the specificity in the sensitivity to saprophytes and the plaque morphology to make up 14 typing phages. These14 phage strains are thought to be adequate for the phage typing on mycobacteria at the present step of the mycophage studies. However, more effort should be paid, of course, for discovering much more number of adequate phage strains for the typing or adaptive phage like Vi-phage II in Salmonella.

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Approximately one third of ca. 5, 000 households in the town of Iwaizumi, which is located in the Kitakami Mountain District in Iwate Prefecture in Japan, have been raising cattle. The sanitary conditions of the district were very poor. For instance, the members of the family were living under the same roof with their cattle. Therefore, it could be assumed that tuberculosis might be transferred from the members of the family to their cattle and vice versa. From this standpoint, the relation between the tuberculosis of human beings and that of cattle was investigated.
1) The incidence of positive tuberculin reaction was 43.3 per cent of 584 cattle-keepers and their relatives, whereas, that of 393 non-cattlekeepers was significantly lower, showing 32.8 per cent.
2) Eighteen relatives (41.8 per cent) of 43 cattle-keepers whose cattle were sacrificed because of positive tuberculin reaction were found as patients of pulmonary tuberculosis. On the other hand, 26 (23. 8 per cent) of 109 cattle-keepers whose cattle reacted negatively to the tuberculin test were detected as patients of pulmonary tuberculosis.
3) Twenty-three strains of tubercle bacilli were cultivated from the 100 people, but were determined as a human type of tubercle bacilli by using the niacin test.
Thirteen cattle were sacrificed because of positive tuberculin reaction, but were found to be “non-visible-lesion reactors”. Fifty samples of various kinds of organs taken from 13 sacrificed cattle gave neither human type nor bovine type of tubercle bacilli upon cultivation, except for 4 acid-fast strains which were proved as saprophytes by the biochemical classification.
4) To investigate the cause of the non-visiblelesion reactor cattle, pupils of primary andsecondary schools were tested with purified tuberculin-π, which was prepared from the avian type (A-71), the scotochromogen (Ishiistrain), the nonphotochromogen (Gamo-strain) and the photochromogen (P-16), along with the human strain H₃₇Rv-πI as a control.
The incidence of positive tuberculin reaction was generally higher by using the tuberculinπ of the H₃₇Rv-strain than by using the other tuberculin-π, however, there were about 4 percent of the reactors who did react to the other tuberculin-π.
5) Gastric juice, throat swabs, or sputum were collected from the pupils whose tuberculin test reacted more strongly to the other tuberculin-π than to the H₃₇Rv-tuberculin-π, and they were cultivated and examined for acid-fast bacilli. One strain of the acid-fast bacilli was cultivated from 75 cases and was determined as photochromogen. Guinea pigs were sensitized by the injection of 10 types of mycobacteria. These were H₃₇Rv, BCG, M. 11755, No.8, No.6, Yamamoto strain, M. smeg matis, M. phlei and Soil III and were grown on modified Sauton's media, from which the purified tuberculin-π was prepared for each by the method of Toda and co-workers.
6) The guinea pigs infected with H₃₇Rv-strain reacted positively not only to the H₃₇Rv-tuber culin-π, but also to the BCG-π, M. 11755-π and No.8-π, but none reacted positively to theπ-tuberculin of the remaining strains. Likewise, the guinea pigs infected with one strain reacted not only to the π-tuberculin of this strain but also to the tuberculin-π of some other strains. From this standpoint, it can be assumed that some strains of mycobacteria might have a common antigenicity.
7) As above mentioned, from an epidemio logical standpoint, some relation was found between patients of pulmonary tuberculosis and tuberculous cattle, but we could not find any bovine type of tubercle bacilli in human beings, and neither a human type nor a bovine type of tubercle bacilli in the cattle of “non-visible lesion reactors. ” This study will be continued.

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BCG vaccination to tuberculin-positive individuals has hitherto been considered as a contraindication. One reason for this is that in the case of the present intracutaneous vaccination, the local reaction at the site of vaccination is stronger with these individuals than with tuberculinnegative ones. The other reason is that, because BCG vaccination is carried out in order to make positive the tuberculin reaction of the vaccinated, the vaccination to already positive individuals is not necessary.
However, from the point of furnishing a sufficient immunity, it is desirable to keep the tuberculin reaction induced by BCG vaccinationalways positive. With the booster vaccination system of BCG to the individuals only when their reactions have reduced to the negative level, as seen in the routine vaccination in Japan, there may be left a certain period of time during which the tuberculin reaction will remain as negative. Therefore it is desirable to vaccinate BCG, if possible, before the reaction converts into negative, which means the vaccination to tuberculin-positive individuals. Furthermore, if BCG can be inoculated to tuberculin positive-individuals, this will greatly facilitate the routine vaccination procedure, because it may be carried out without the preexamination of tuberculin reaction
But all these alterations may become possible only when repeated vaccinations with BCG to tuberculin-positive individuals, including naturally infected ones, cause no untoward reaction, either locally or systemically.

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Although a cavity in a living body which is under the dynamics of respiration undergoes physical and chemical changes constantly, only a few patho-physiological studies have been reported from this viewpoint. From our department, a series of such studies have already been published, and the present report is also a part of them.
Experimental lung cavities were produced in 120 rabbits by using heat killed BCG suspensions, according to Yamamura's method. The course of the production of the cavities were classified into the following 4 stages; the pre-cavitation stage (20 days after the introduction of secondary antigen into the lung); the early stage of cavitation (20-40 days); the completing stage of cavitation (40-60 days); the stage of secondary changes in the cavity wall (60 days and over). The course of cavitation was observed roentgenographically and macroscopically, the observation period covering 530 days at the longest.
At the same time changes of the optical and electron microscopic features of lung tissues in the main lesion (cavity wall), in the surrounding regions and in the normal region of opposite lung were compared. Chemical changes in the lung tissues of the regions were followed up with the following results:
1. Roentgenographical alterations in the cavity formation are not widely different from those reported hitherto, and are in good agreement with macroscopical findings.
2. From the completing stage of cavitation onward, experimental lung cavities produced with heat killed BCG suspensions partly begin to show a tendency toward roentgenographical and histological healings. Chemical changes in the tissues also reflect these facts to some extent. In the entire lung segments, diffuse productive lesions, smaller than millet grains, are sometimes formed centering around the alveolar septum. This is thought of as due to the passage of primary and secondary sensitizing antigens into the blood.
3. The followings are the electron microscopicchanges:
i) Early exudative changes begin with intraand extracellular edemata, and are due chiefly to the marked increase in pinocytosis.
ii) Epithelioid cells are considered as being formed when juvenile phagocytes develop into the mature type through the phagocytic type in a specific environment.
iii) Further studies will be required on the mechanism underlying lattice fiber formation or hyalinization.
iv) From 200 days onwards, a fairly great number of plasma cells are seen in fibrosed tissues together with lymphocytes and fibro cytes (fibroblasts).
4. From the early stage to the completing.

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