In Japan, vaccination is established for the prevention of influenza. Our influenza virus vaccines contain 100 CCA per mililiter of each strain of A1, A2 and B type.
According to author's experience, chief reactions to vaccines are redness, swelling and heat at the site of injection, which give discomfort to the recipients. While systemic reactions, such as the increase in body temperature are not common.
To eliminate these reactions, we tried the purification of the viruses by the chromatographic method with brushite form of calcium phosphate, which was reported by Taverne et al in 1958. We repeated many experiments with 3 types of the viruses and obtained average recovery of 65.3%, ranging from 39.0% to 96.4%, in 7 experiments with A1 type, 79.4%, ranging from 55.6% to 89.3%, in 6 experiments with A2 type, and 73.8%, ranging from 52.7% to 87.5%, in 4 experiments with type B.
By rechromatography, the recovery rate was as high as over 90%.
Purified monovalent vaccines of type A and B viruses, each containing 300 CCA units, were examined for their safety and potency comparing with the non-purified, which were treated only once with Sharples centrifuge and resuspended in buffered physiological saline. In safety test conducted by the guinea pig method, we could obtain far better results from the purified ones.
Potency test was conducted with mice, injecting 5 fold graded dilutions of vaccines intraperitoneally. They were bled after 2 weeks, blood collected, sera separated and used for neutralization test in embryonated eggs. The results showed that purified vaccines had potency not inferior to non-purified ones.
The same vaccines were examined for reactions and antigenicity to human bodies. Four groups of recipients, each consisting of 20, were inoculated with one of four vaccines, purified or non purified vaccines of type A2 and B.
Reactions (redness, swelling, heat at injection site and increase in body temperature etc.) were examined. Systemic reactions were weak in all vaccines. Redness began to appear 4 hours after inoculation and faded after 3 to 4 days. In the purified vaccine groups of both types, redness were 8.1mm at 24 hours after inoculation and 16.0mm at 48 hours by type B purified vaccine, wheras those of the non-purified of the same type were 60.2mm and 74.6mm. By A2 type purified vaccine, they were 7.2mm and 6.0mm, but non-purified control showed 23.5mm and 12.9mm.
Swelling and heat at injection site were far less in the case of purified vaccines of both types compared to the non-purified.
Antigenicity of vaccines was estimated by examining increase of HAI titer 4 weeks after vaccination. Purified vaccines of both types of A2 and B gave lower antigenicity to some extent. But, further large scale experiments may be necessary to confirm whether the purified vaccines are truly inferior or not.

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Sera obtained from normal guinea pigs, rabbits, chickens and monkeys were examined to characterize the non-specific hemaggultinin inhibitors contained in each. Although preliminary results on this p problem were previously described, reexamination was thought to be necessary after a discovery of the avid Asian virus. High sensitivity of this virus to various inhibitors, again made this problem as an unsettled one.
In the study, both heated Lee and Asian P viruses were used to see the different distribution of a and a inhibitors in these sera. Mumps, Sendai and Newcastle disease viruses were also used to characterize each inhibitor, because these hemolytic viruses were known to form irreversible union with red cells, like Asian P virus.
Obtained results will be summarized as follows:
1) a-inhibitor noticed in guinea pig sera by Shimizu and Ishida using FMI strain of influenza A virus as an indicator, was shown to be the same principle active against Asian set of P virus. This principle was different from a-inhibitor active against heated Lee.
2) An inhibitor specifically active against mumps virus was found in monkey sera of both rhesus and cynomologous. Furthermore, with serum specimens obtained from animals other than monkey, inhibitor titer against heated Lee virus.
3) Newcastle disease virus was found to be more sensitive to serum specimens received pretreatment of either RDE or KIO₄ than that without any treatment. This kind of change was particularly the case with monkey and guinea pig sera, and was assumed to be the same phenomenon as reported as the heat activation phenomenon.

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1. In the spring, 1954, there was an outbreak of so-called “mouse-influenza” at A institute in Tokyo, and in 1956, there was another outbreak of the same disease at B institute in Kanagawa prefecture. Four strains of Myxovirus para-influenzae 1 (HVJ) were isolated from the lung of four groups of the mice at B institute. Three-weeks-mice inoculated intranasally showed clear “mouseinfluenza” symptoms and all of them died between 2 to 7 days.
In the autumn, 1954, the same virus, once more, was isolated from the lung of two infant guinea-pigs produced in Gifu prefecture. Hemorrhagic lesion, edema and bronchial catarrh were observed in the lung.
2. During the period from April to October, 1954, the sera were collected from the market-mice in Chiba, Saitama, Tokyo and Shizuoka prefectures and the mice of in inbred-strain at six institutes in many districts of Japan. The positive high titers of H. I, C. F and neutralizing antibody for para-influenza 1 virus were observed in the some of the sera. It was supposed that the virus may be wildly prevalent among the mouse population in that year.
3. It was observed that the mice-passaged-virus shows the viscerotrope character, while the egg-passaged-virus shows the pneumotrope character in accordance with the results examined on the D D M strain.

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An epidemic of an infectious disease occurred among canaries, starting in Yokohama and Tokyo from February, 1958, and spreading almost throughout Japan. Affected birds showed swollen eyelids, lacrimation, ruffled feathers, and dyspnea and almost all the birds terminated fatally. Etiologic studies resulted in isolating a virus producing characteristic lesions on the chorioallantoic membranes of developing hen's eggs from naturally infected canaries. The isolated virus was found to be an member of the pox group on the bases of morphologic characteristics of cytoplasmic inclusions formed in affected skins of naturally or experimentally infected canaries and in infected chorioallantoic membranes and electron-microscopic properties of elementary bodies. The virus was found to have common antigens with fowl and pigeon pox viruses by complement fixation tests. Based on these findings together with its origin and pathogenicity for canaries it was concluded that the disease was caused by canary pox virus. This is the first report in which canary pox was definitely diagnosed in Japan.

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Canary pox virus was grown in cell culture of chick embryo.
1) The virus proliferated in the cultured cells with cytopathogenic effect. Serial passages were readily accomplished by inoculating infected culture fluid into fresh cultures.
2) The specific complement fixing antigen was found in infected culture fluid.
3) The characteristic cytoplasmic inclusions were formed in the infected cells.
4) A marked decrease of pH of the medium was noticed in the cell cultures showing marked the cytopathic changes.
5) The virus passaged in the cell culture produced the disease when inoculated to canaries.
6) The cell culture was successfully employed for primary isolation of the virus from naturally infected canaries.

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When cells of Escherichia coli whose synthetic capacity has been lost by heavy irradiation of ultraviolet light are infected with bacteriophage T₂ inactivated by the same irradiation, a considerable amount of protein is found to be synthesized. It is evident that this protein synthesis can be induced by infection with inactive bacteriophage. The protein synthesis occurs also in unirradiated normal cells infected with inactive bacteriophage. In view of various observations in these cases, it is most probable that these proteins are equally abnormal proteins produced by bacteriophage infection. None of them has phage antigenicity. The studies in order to get several informations upon these proteins are the principal subjects of this paper.

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