Leptospira antibodies were examined, by means of agglutination lysis test, on sera of animals. The serum samples were collected from 2, 724 cattle, 669 swine and 208 horses in western districts of Japan, including Kinki, Chugoku and Shikoku, from 1958 to 1959. Antigens used in the agglutination lysis test were following 6 serotype strains: L. icterohaemorrhagiae, L. canicola, L. autumnalis, L. hebdomadis, L. australis A and L. pomona. The results obtained were as follows.
1. The positive serum reactions (1: 300 or more) were obtained in 349 of cattle (13%), in 68 of swine (19%) and in 75 of horses (36%). It was recognised that the positive reacting animals were distributed in almost all parts of western districts of Japan.
2. Among these positive reactions obtained, the most frequent ones were as to L. icterohaemorrhagiae followed by L. autumnalis, L. hebdomadis and L. australis A. As to L. canicola and L. pomona, there were a few cases reacting positively.
3. The positive rates of the leptospira agglutination tests in cattle and horses tended to increase with age. The positive rate in swine, whose ages were limited in 1 or 2 years, was higher than those in cattle and horses at the same age.

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From 1953 to 1959, leptospirosis in native black cattle was studied serologically and bacteriologically. The results obtained are summarised as follows.
1. The main symptom of the bovine leptospirosis was “hemoglobinuria”. However, the results of serological investigation on the bovine leptospirosis revaled that the cases of subclinical infection were much more than those showing the symptom of hemoglobinuria.
2. The hemoglobinuria caused by leptospira infection in cattle has been observed in the limted areas in Japan-nothern parts of Hyogo-Pref. and Kyoto-Pref. Therefore we regarded the hemoglobinuria as an endemic disease in cattle in Japan.
3. This disease in cattle has occur not generally in autumn months, particularly in September and October. The morbidity of this disease was low in calves and increased gradually with age.
4. Author encountered some cases of abortion in the infected cattle, in which both the cases of the hemoglobinuria and those of the subclinical infection were included.
5. Isolations of the etiological agents from the urine of cattle showing the hemoglobinuria were performed, and 25 strains of leptospira organism were obtained successfully. On these 25 strains, we identified 22 strains as L. autumnalis, 2 as L. hebdomadis and 1 as L. australis A. The isolations of L. autumnalis and L. australis A from cattle were considered to be the first case in the world.

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Relationship of the intensity of infection to the CF antibody titer at the time of infection made with attenuated virus was investigated by the author availing himself of the following 3 occasions in order to facilitate the selection of the virus strain to be attenuated for the use of vaccination and the decision of the amount of serum for the simultaneous inoculation of live virus and immune serum.
1. Simultaneous inoculation made on the calves of the Japanese black cattle with lapinized and avianized rinderpest virus (La strain) with varying amounts of immune serum.
2. Calves of the Japatese black cattle inoculated with the LA strain and those inoculated with the avianized rinderpest virus of bovine strain (BA strain) which has a stronger pathogenicity.
3. Inoculation of the same attenuated virus strain on the cattle of different resistance against rinderpest virus, namely, the inoculation of LA strain on the calves of the Japanese black cattle and adult black cattle and the inoculation of BA strain on the calves. of the Japanese black cattle and the calves of Holstein-Friestian. The results obtained were as follows:
I. On comparison of the calves receiving the simultaneous inoculation of LA strain and immune serum and those receiving LA strain alone, the latter were found to show more frequently the inoculation reaction and a greater “fever volume” of Nakamura et a1 and the result of complement fixation test showed that they possessed a higher rate of CF positive reactors with higher means of highest FT value and longer duration of the CF positive period.
As for the animals of simultaneous inoculation, the rate of inoculation reaction and the difference in the amount of fever volume were not recognizable. The smaller the amount of immune serum used, however, the rate of positive reactors was higher with higher means of highest FT value and longer duration of the CF positive period.
Challenge made with highly virulent strain of bovine origin on a part of the calves, receiving the single inoculation of LA strain and those receiving the simultaneous inoculation of LA strain and immune serum showed that the former had complete resistance against the challenge while in the latter case there were some which showed inoculation reaction and the changes in FT value after challenge though they presented the outward appearance of complete resistance. This intensification of FT value after challenge was; stronger amcng the animals receiving larger amounts of immune serum.
2. Calves of the Japanese black cattle receiving the inoculation of BA strain, in comparison with those receiving the inoculation with LA strain, showed a higher rate of inoculation reaction, larger amount of fever volume, higher means of highest FT valueand longer duration of the positive period.
3. On comparison of the calves and adult Japanese black cattle receiving LA virus, the former showed larger fever volume, higher means of highest and longer duration of CF positive period.
As for the calves of the Japanese black cattle and those of Holstein both receiving the inoculation of BA strain alone, the former showed more frequently inoculation reaction, larger fever volume, higher means of FT value and longer duration of CF positive period.
As for the calves of the Japanese black cattle and those of Holstein both receiving of BA strain alone, the former showed more frequenty inoculation reaction, larger fevervolume, higher mean of FT value and longer duration of CF positive period. In view of the findings stated above, it is concluded that the intensity of infection runs in parallel with the CF antibody titer and the duration of CF positive period when. inoculation is made with attenuated rinderpest virus.

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Though the agglutination test has been generally applied for the diagnosis of Brucellosis, specific reactions may be sometimes inhibited, or non-specific reactions may occur. Among studies to remove these facts, the anti-globulin test by Coombs and others (1945), by which could be detected incomplete antibody of Rh-factor, has been investigated for this disease since 1951. For experiments of this study, mainly goat and sheep withBr. melitensis have been used, while few reports on cattles with Br. abortus have been seen.
The auther has examined on 808 Jersey cattles this test, which has been never reported in our country, comparing with the agglutination test and C. F. T., and consecutive observations were made on cattles, which indicated positiv reactions.
By bovine Brucellosis, also Coombs antibody appeared earlier than agglutinin, and a rise of its titer was abrupt, and attained to peak, while the titer of agglutinin was rising;but, it did'nt disappear, even after the disappearance of agglutinin.
C. F. antibody appeared following Coombs antibody, and its appearance has been recognized to be parallel to Coombs antibody.
Cattles, by which both Coombs antibody and C. antibody were provedeven if agglutination test was negativ, should be suspected to be infected by Brucella. It seems likely, that the combined examination of Coombs test and C. F. test could dissolve the problem of cattles, which indicate weak reactions, and contribute to differentiate nonspecific rections from infection.
Coombs antibody is highly stable, and no fall of potency has been recognized after refrigerating, dissolution or long-term storage in icebor.

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The sensitivity of Salmonella pullorum against P., CTC., OTC, TC., CP., FM., SM., B., and LM., were measured, using the tube method and the plate method. Among strains used in this study, two strains belonged to standard type, three to intermediate type, and three to variant type. At the same time, the disk method was examined, comparing with these two methods, and the results with these three methods have been recognized to be similar.
It has clear from the results, that the organisms were highly sensitive against KM., FM., CP., TC. OTC., moderately sensitive against PC. and resistant against SM., B., CM. Strains of variant type and long-term culture were more sensitive to all kinds of antibiotics, especially to PC.,
The combined actions between KM., FM., CP., OTC., and PC., were investigated, using the tube and plate method. Comparing with the results by these methods, the cup plate technique, deviced by the auther to study synergism, summation, indifference and antagonism among all kinds of antibioties was investigated.
It has been certified, that the cup plate technique could be reliable, as the similar results were obtained from these three methods.
The combinations, as P.-CP., P.-KM., OTC.-CP., CTC.-KM., FM.-KM., were effectiive, while those, as CP.-FM., CP.-KM., not effective.

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Two hundred fourty nine sera taken for Wassermann's reaction and 79 sera of patients of different diseases were examined on antihemolytic reaction to θ hemolysin of Cl. perfringens type A. The latter sera were also tested on antistreptolysin O titre. In this study, antihemolytic reaction of the sera in a dilution, above 1: 20 was regarded to be positive.
Among the former 249 sera 28 sera were positive. Clinical diagnoses of these positive cases were as follows: 23 cases of pregnancy, each 1 case of erythematodes, chronic hepatitis, hypertension, schizophrenia and an unidentified case.
Among the latter 79 sera, 25 cases were positive, of which 6 cases were clinically related to streptococcal infection: rheumatic fever or acute nephritis, showing relatively high antistreptolysin O titre. Such antihemolytic reaction to O hemolysin of cl. perfringens type A as these cases would be rather regarded as the neutralizing effect of antistreptolysinO. according to Todd's (1941) report that antistreptolysin O neutralizes θ hemolysin of cl. perfringens type A. Clinical diagnoses of other 17 cases of 25 positive sera were diseases of liver or bile ducts, showing jaundice. Clinical diagnoses of the remaining 2cases were high blood pressure. On the other hand, pregnancy, hypertension or obstructive jaundice usually demonstrate excess of serum cholesterol which neutralizes according to Todd (1941) θ hemolysin of cl. perfringens type A.
Concequently, antihemolytic reaction of 47 positive sera except 7 cases of streptococcal infection may be regarded as the neutralizing effect of cholesterol excess in blood. The cases were as follows: 23 cases of pregnancy, 3 cases of hypertension, 24 cases of liver and bile duct disease that caused obstructive jaundice (5 cases of liver cirrhosis, 11 cases of hepatitis, 1 case of cholecystitis and 7 cases of liver canser).

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