Tubercle bacillus and its adjuvant-active component, wax D, are known to produce a typical epithelioid granuloma and an adjuvant arthritis in animals. Two different derivatives of wax D were prepared: AD 6 and a polysaccharide portion of wax D. AD 6 was prepared by acetylating wax D and subseueqntly fractionating it by silicic acid column chromatography. AD 6 was found to be active as adjuvant. However, it possesses neither antigenicity nor toxicity nor anti-complementary activity nor adjuvant arthritis-inducing capacity. The polysaccharide portion was prepared by saponifying wax D. This was found to be antigenic but avoid of an adjuvant effect. None of these substances could produce the adjuvant arthritis or epithelioid granuloma, when injected alone. However, when they were injected together, the adjuvant arthritis and epithelioid granuloma could be induced. These results suggest that both the adjuvant effect and antigenicity may be required for the development of these changes in tissue. An alternative interpretation was also discussed.
In sensitization of mice with sheep red blood cells, the addition of adjuvant-active substances to the sensitizing antigen resulted in a highly significant increase in the size of plaques developed by spleen cells as measured by Jerne's plaque technique. The increase in plaque size was found to be a good measure for the adjuvant effect of substances as represented by mycobacterial adjuvants, because the phenomenon was specific for the adjuvant effect, highly reproducible, sensitive and can be performed with a limited number of mice.
The mode of action of immunological adjuvants was studied, using the ‘plaque size’ technique and purified mycobacterial adjuvants, wax D and AD 6. Data were obtained which suggest that wax D does not act directly on antibody-forming cells but on their precursors, causing the generation and/or proliferation of a special precursor cell designated as X'. Memory cells generated in the presence of wax D were found to ‘memorize’ its adjuvant effect, while normal memory cells generating by the stimulation of antigen alone were refractory to the adjuvant effect. Thus, wax D caused a generation of a special memory cells designated as Y'. It was thus strongly suggested that a special cell line X'-Y'-Z' is generated and/or proliferated in the animal bodies which received the mycobacterial adjuvants along with antigen.

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Authors reported the usefulness of STC for the earlier detection of mycobacterial growth. in previous papers (Kekkaku, Vol.46, p.335 and Vol.46, p.341). In these studies, 0.1ml of 0.5% STC solution (aqueous) was added onto the media after the inoculation. In the next paper authors reported the results of the application of STC-containing media in different concentration for earlier detection of mycobacterial growth and reported that 0.01% (100mcg/ml) was the appropriate concentration incorporating into the media (Kekkaku, Vol.47, p.269).
This paper presents the results of isolation culture using 0.01% STC-containing media from sputa.
The methods and results were summarized as follows:
Experiment 1;
Ogawa's media modified by Kudoh were used for this study. STC was added into the media before inspissation to make 100mcg/ml (0.01%) STC-containing media. Sputa were treated with 4% NaOH (volume ratio 1: 4) and inoculated 0.1ml for a pair of original and STC containing media. The reading of bacillary growth was performed at 2, 3, 4, 6 and 8 weeks, after the inoculation.
The results were shown in Table 1. As seen in this table, the difference in the culture positive rate between original and STC-containing media was not observed. Comparing the time of first growth between both media however 7.6% of positive culture on STC-containingmedia showed erlier growth than original media.
Experiment 2;
3% Ogawa's media were used for this study. The pretreatment of sputa, the method of inoculation and the time of reading were the same as experiment 1.
The results were shown in Table 2. The positive culture on original and STC-containing media was not different, however, the time of first growth was different on both media 14.6% ofpositive culture on STC-containing media showed earlier growth than on original media.
The result of contamination was shown in Table 3. The contamination rate was not different significantly between original and STC-containing media.
The niacin reaction using the stained colonies by STC was performed in small scale and reported in previous paper (Kekkaku, Vol.46, p.335). This time the niacin test using colonies isolated from each medium was performed and the results were shown in Table 4. As seen in this table, the difference of niacin reaction between colonies on original and on STC-containing media was not observed.
In conclusion, 0.01% STC-containing media was useful for earlier detection of mycobacterial growthfrom pathological specimens.

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Nagata-ku of Kobe City is a typical urban area with a population of about 210, 000 and an area of 11km². The southern part of the area is noted for a center of producing chemical shoes, and the majority of factories are small in size. Moreover, many smaller enterprises are related to these factories.
Kobe City showed the highest tuberculosis mortality figures in Japan, and the mortality rate from tuberculosis in Nagata-ku has been the highest or next to the highest in Kobe City. Nagata Health Center has been conducting the 3 years' special tuberculosis control programme including the mass survey for adult population and the supervison of detected cases since 1969.
The results obtained are as follows:
1) The rate of response to the mass survey was 52%, 65% and over 55%, respectively, in 1969, 1970 and 1971. The mass survey for smaller enterprises was done in 1971, and approximately 8, 000 were examined.
2) The detection rate of tuberculosis patients was 0.89% in 1969, and it gradually reduced to 0.7% in 1970 and 0.49% in 1971. Among workers in smaller enterprises, the detection rate was 1.26%.
3) The over-all detection rate was 0.74%, and the rate increased with age especially in the age groups over 50. The rate of male was 1.50%, and it increased markedly in the age groups over 40, while the rate of female was 0.49%.
4) The proportion of active infectious type to the whole detected cases was 33 to 39% in the age group over 50, and it was higher in male than in female in each year.
5) Among patients detected by the mass survey for general inhabitants, 34 to 52% of male patients and 20 to 31% of female patients were registered previously; in the case of workers in smaller enterprises, 12% of male case and 68% of female case were registered previously.
6) Among active infectious cases detected by the mass survey for general inhabitants, 64 to 78% were not registered previously, and among those found by the survey for smaller enterprises, the proportion was 79%.
7) Fifty eight to 67% of newly detected patients started treatment within 3 months after detection.
8) Seventy five per cent of cases found by the survey for smaller enterprises were residents of Nagata-ku.

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A study has been made to obtain the indices which are useful to select the high prevalent areas of tuberculosis, on the basis of routinely available informations from the mass X-ray and tuberculin surveys of school pupils aged 6-14 years in Kawasaki City. Rate of markedly (redness over 10mm with induration and duplicate redness) and moderately (redness over 10mm with induration) positive reactors to Mantoux test (2.5 TU PPDs) among 1st grade pupils of primary school (6 years of age), and appearance rate of calcified pulmonary and/or hilar nodes primary foci on mass miniature radiography among all pupils of primary and/or junior high schools in the City, were drawn from the results of survey in 1971, and calculated for each city wards. Simultaneously, annual incidence rate of newly notified active tuberculosis (all forms) and infectious pulmonary tuberculosis, prevalence rate of registered tuberculosis (including inactive case, all forms), active tuberculosis (all forms) and infectious pulmonary tuberculosis at the end of the year, were drawn from the registry of tuberculosis of 1971 in Kawasaki City and calculated for each city wards.
Analyses were made to determine the relation between each rates of the former and those of the latter respectively. Rate of moderately positive reactors to Mantoux test among 1st grade pupils of primary school and appearance rate of calcified foci among all pupils of primary and junior high schools in each city ward directly correlated with prevalence rate of infectious pulmonary tuberculosis in each city ward respectively. Moreover, appearance rate of calcified foci among pupils of junior high school in each city ward directly correlated with annual incidence rate of infectious pulmouary tuberculosis in each city ward.
BCG vaccination during infantile and preschool periods has been applied in the City on a mass scale. Therefore, the rate of moderately positive reactors among 1st grade pupils of primary school was regarded to be of little value as an index of prevalence of tuberculosis.
In order to determine further relation between appearance rate of calcified foci among pupils and annual rates on registry of the City, records of school survey and registration data in 1962, 1965, 1968 and 1971 were analysed. Appearance rate of calcified foci among pupils of junior high school in every 3years directly correlated with annual incidence rate of newly notified active tuberculosis (all forms) and infectious pulmonary tuberculosis, and prevalence rate of registered tuberculosis (all forms) respectively, but not correlated with prevalence rate of infectious pulmonary tuberculosis. The discrepancy may be derived from the alteration of criteria on “infectious”.
It was suggested that the prevalence of calcified foci found in school pupils may be of value as an index of prevalence of tuberculosis in the area.

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Recently these authors have encountered a case of Mycobacterium marinum skin infection not related to exposure in a contaminated swimming pool. Since the skin disease due to the organism of this species is relatively rare in Japan, a rather detailed bacteriological study was made.
A 71-year-old woman fell on the cemented floor at a fish shop in Hiroshima and abraded her right bucca in October, 1971. When she was seen about ten days after the injury, there was reddened, swollen, well demarcated induration with a number of pustules on the bucca. By taking the various antimicrobial agents and steroid hormones, the lesions were slightly improved. A submandibular lymph node was swollen in November incised and drained on December 14. On February 29, 1972, she began taking kanamycin in a dose of 0.5g every other day by injection, which promptly improved lesions.
A chest roentgenogram showed no evidence of tuberculosis. A tuberculin test gave positive reaction.
Punch biopsy was done on November 17, 1971. Biopsy showed slight hyperkeratosis and parakeratosis. Deeper, there was a granulomatous inflammatory reaction with histiocyte-like cells, lymphocytes and giant cells. Ziehl-Neelsen stain revealed no acid-fast rods.
Pus specimens were inoculated directly onto nutrient agar and/or Sabouraud agar and incubated at 27°C. From the buccal lesions acid-fast bacilli were isolated on November 17 and 27 and December 14, 1971. From the submandibular lymph node similar acid-fast organisms were isolated on January 6, 1972. In all cases, the colonies appeared in from two to three weeks after inoculation, as white, moist, slightly elevated, smooth areas, having irregular margin. They have the characteristic of changing color from white to yellow following exposure to light.
Three of four strains isolated were submitted to the various biological and biochemical tests. At first transfers good growth occured at 30μ, 37μ and 22μ and the days required for visible growth were 5, 7, and 8 to 9 days, respectively. No growth was obtained at 40°C in two weeks. All strains were negative for niacin and nitrate reduction tests, while positive for Tween 80 hydrolysis, catalase, 68°C catalase, acid phosphatase, 70°C acid phosphatase and putrescine-oxydase. The amidase activity was positive for urease, nicotinamidase, pyrazina midase and allantoinase, whereas negative for acetamidase, benzamidase, isonicotinamidase, salicylamidase, succinamidase, malonamidase, propionamidase, acrylamidase and oxamidase.
Each of five female mice (#CF 1) was injected intravenously with 5.8×10⁷ viable units of the Irita I strain isolated on November 27, 1971. During the experiment nodular swellings covered with scales or crusts were seen on their tails. All of the animals died in from nine to thirteen days after infection. No grossly visible disease was noted on the visceral organs. A large number of organisms were recovered from the tail, and the moderate to numerous number of organisms from lungs, liver, kidneys and spleen.
In summary, the biological and biochemical characteristics as well as the mouse pathoge nicity of the organism reported in this paper are compatible with those of M. marinum strain.

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