As one of the means for the serological diagnosis of neurotropic viral infections, the complement-fixation test proved to be of definite value and was widely applied at present. Complement-fixation test was performed on 226 Japanese encephalitis cases who had been admitted into the Ebara Hospital during the epidemic of 1948 in Tokyo. The results obtained from the sera withdrawn at their early period after onset, have been already reported, There are some reports described by other workers on the same problem. It seems, however, there is no report made on the tests performed covering a long period of time after convalescence.
For this reason, 14 out of above-mentioned 226 cases were bled continuously after discharge from the hospital. Their bleeding was continued for about two and half years till March 1951. Complement-fixation test was performed on each serum sample and collected neutralization test too was performed as much as possible. There are several papers on the neutralizing antibody examined for a long period after convalescence. An attempt was made to re-examine the neutralizing antibody curve in Japanese encephalitis cases to know whether the curve of neutralizing antibody has any relation to that of complement-fixing antibody.
Besides, complement-fixation test was performed on the sera obtained from the patient suffering from non-encephalitic diseases, who had been admitted to the Ebara Hospital and had no anamnesis of Japanese encephalitis, at several months intervals from 1948 to 1951. The results of complement-fixation test made on their sera were compared with those from convalescent cases.

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A new anti-tubercular antibiotic was obtained from the broth filtrate of a new species of streptomyces, Streptomyces cinnamonensis n.sp.. When this substance was obtained in a crystalline state, the authors learned a report of Sobin on a thiazolidone antibiotic. At the same time another research group of Takeda's laboratories gave out a new streptomyces antibiotic, acidomycin. Comparing the chemical and physical characteristics of these substances, the authors came to the conclusion that they are identical. Though it is strongly tuberculostatic in vitro, it is ineffective in vivo. It suggests the existence of a certain antagonist in animal tissues. The structural formula of thiazolidone antibiotic proposed by McLamore and others indicates its close resemblance to biotin and especially to desthiobiotin. Moreover, imidazolidone caproic acid, which also resembles these substances, was shown to be an antagonist to biotin and desthiobiotin by Dittmer and du Vigneaud, and Pope and Smith. The authors' preliminary experiment indicated the antagonistic effect of this antibiotic to biotin. In the present paper the competitive nature of antagonistic relationship between thiazolidone antibiotic and either biotin or desthiobiotin was recorded.

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In the previous report, we compared several technics of the complementfixation test in Q fever. In those experiments, antigens and antisera of Q fever used were all given from other investigators or laboratories, because it was required to use materials of which titer had been already decided. We could not, however, get sufficient quantity of them to estimate preliminarily the titer of every antigen and to decide the dilution in which . a certain unit of antigen was included by all methods used, and so we used them in the dilution described on labels. Therefore, we were not quite sure to decide there was what sort of relationship between antigen and antiserum of Q fever in the complement-fixation test.
This time, we compared several methods of preparation or purification of antigen from the emulsion of yolk sac infected with Coxiella burnetii, which had been cultivated in our laboratory. And we examined the shape of reaction in the complement-fixation test between antigen and antibody. Moreover, we carried out the agglutination test with the antigens proved to be possibly used by the complement-fixation test, comparing a several technics of the former, and examined the relationship between complement-fixation test and agglutination test in Q fever.
As for antigen of Q fever for both tests, Bengtson tried, in 1941, to purify the materials, such as the emulsion of mouse spleen or of yolk sac, and got the antigen able to be used. In her case, the purification was done by only centrifugation with controlling pH and temperature. Later, since Craigie succeeded, for the purpose of preparing typhus vaccine, to purify the rickettsiae cultivated in embryonated yolk sacs, by dealing the yolk sac emulsion with ethyl-ether, this method was used for the preparation of rickettsial antigens by many other investigators, including Plotz, Reynolds and Pollard, Damon and Johnson, Bengtson and so on, having been applied to the preparation of Q fever antigen by Robbins, Rustigian, Snyder and Smadel and the Commission on Acute Respiratory Diseases, and was proved to be an excellant method.
Whatever method mentioned above is used, the last stage of the antigen preparing process is to collect and concentrate rickettsial bodies by centrifugation. It matters whether or not the supernatant of the centrifugation keeps any antigenicity as well as the sediment. Topping and Shear, Craigie, Watson, Clark and Malcomson and Shepard found many rickettsial bodies micro scopically in the sediment of centrif ugalized emulsion of embryonated yolk sac infected with rickettsiae of typhus group, after dealing it with ethyl-ether according to Craigie's method, at the same time they proved the presence of high antigenicity in the supernatant. They undertook many experiments using this so called soluble antigen as the antigen for the complement-fixation test. Shepard and Wyckoff succeeded in taking the electronmicroscopic photograph of the soluble antigen of typhus group rickettsia to clarify its serological signi-ficance.
As for the soluble antigen of Q fever rickettsia, Bengtson carried out the precipitation test, using the filtrate of centrifugal supernatant of the emulsion of animal organs as antigen, and got the results which could be thought in fact negative. There seemed not to be any other researcher who discussed on the significance of the soluble antigen especially as antigen in the complementfixation test.
Rickettsial body antigen for the complement-fixation test, as well in Q fever as in other groups of rickettsia, can be used also in the agglutination test.

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