[in Japanese]

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LePlOsPiraaUtUm77.aliS( ?? ?? ?? ?? ?? ?? ) ?? 100 ?? dd ?? ?? ?? ?? ( ?? ?? 1215g) ?? ?? ?? ?? ?? ??, ?? ?? ?? 14 ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? ?? 15 ?? ?? ?? ?? 3 ?? ??, ?? ?? ?? ?? ?? ??, ?? ?? ?? ?? 1· ?? ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? ??, 24 ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? ?? 8 ?? ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ( ?? ?? 「 ?? 」 ?? ?? ) ?? ?? ?? ??, ?? ?? ?? ?? ?? ?? ?? 2 ?? ?? ?? ?? ?? ?? ?? . ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? .1) ?? ?? ?? ?? ?? ?? ?? 2 ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? ?? ?? ?? 「 ?? 」 ?? ?? ?? ?? 90%, ?? ?? ?? 100% ?? ?? ?? ?? . ?? ?? ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ( ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? 2σ ?? 10g10 ?? ?? ??, ?? ?? ?? ?? ) ??, ?? ?? ?? ?? 5±1.8, 3.8±3.3 ?? ?? ?? ?? .2) ?? ?? ?? ?? ?? ?? 500μg/g( ?? ??, ?? ?? ?? ?? ), ?? ?? ?? ?? 1· ?? ?? ?? ?? ?? ?? ?? 50μg/g, ?? I· ?? ?? ?? ?? ?? ?? 50μg/g2, ?? ?? ?? ?? 3 ?? ?? ?? ?? ?? ?? ?? ?? ??, ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? .3) ?? ?? ?? ?? ?? 800U/g ?? ?? j′ ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? 50μg/g( ?? ?? 8 ?? ?? ?? ?? 16.6μglg ?? ?? ) ?? ?? ?? ?? ?? ?? ??, 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? 20% ?? ?? ?? 40% ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? . ?? ?? ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? 0.2±0.8 ?? ?? ?? 1.1±2.9 ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? .4) ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ??, ?? ?? ?? ?? ?? ?? 50μg/g ?? ?? 70%(2.0±3.2, ?? ?? ?? 2.4±3.6), ?? 5μg/g ?? ?? 100%(4.6±1.3), ?? ?? ?? ?? ?? ?? ?? ?? ?? 5μg/g ?? ?? 100%(3.3±2.5), ?? ?? ?? ?? ?? ?? ?? ?? 5μg/g ?? ?? 60%(1.4±2.4), ?? ?? ?? ?? ?? 800U/g( ?? ?? 1 ?? ?? ?? ?? ?? ) ?? ?? 70%(1.9±3.0), ?? 100U/g ?? ?? 100%(3.6±2.0), ?? ?? ?? ?? ?? ?? ?? ?? ?? 0.2cc ?? ?? 89%(2.9±3.O) ??, ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? .5) ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ??, 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ?? ?? ??, ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? ?? 「 ?? 」 ?? ?? ?? ?? ?? ?? ?? .

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[in Japanese]

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In previous reports it was demonstrated in a series of experiments in vivo, concerninginfection of E. tenella, that the pancreatic juice of the chicken is a necessary factor for in-fection by E. tenella, acting on the excystation of the oocysts ingested in the intestinal canal, and that it may be induced only by the action of the pancreatic juice on the oocysts with-out the influences of the upper alimentary canal, including the gizzard. At present, nosatisfactory experiments have been made to prove the excystation of oocysts in vitro, as faras the oocysts of E. tenella are concerned. Based upon the above findings, the present ex-periments were undertaken in order to obtain further evidence, in detail, concerning theprocess of excystation in vitro of the oocysts of E. tenella in mediums containing pancreaticenzymes, and to study an essential component of the pancreatic juice in order to bringabout the liberation of the sporozoites from the oocysts and to determine the main site ofits action on the oocysts.For the purpose of this study on excystation, the E. tenella oocysts used were collectedfrom the cecal contents of chickens infected with the pure line of E. tenella, and obser-vations on the excystation, in the present experiments, were mady by employing the follow-ing hanging drop preparations, that is, a drop of enzyme solution containg a very smallnumber of oocysts is simply attached to the under surface of a cover glass which is mountedover a depression in a slide and then sealed with paraffin along the cover glass. They wereobserved ttnder a microscope placed in an incubator at 39 -4OC continuously for a longtime, instead of using the smear method of detecting excystation of oocysts, by which methodit might not be possible to observe the changes naturally occurring in the same oocystcontinuously, owing to the inevitable technical errors occurring during the experimentalprocedure. The results obtained were as follows :( l ) The excystation of the oocysts of E. tenella was induced by the emzymatic actiono selxes through both micropyles, initially of the sporocysts and then of the oocysts.( 2 ) The behaviors of the sporozoites, in the course of the excystation, were observedas follows :At the beginning of incubation the active appearances of oocysts were usually seen, and then the sprozoites started moving within a sporocyst, at first very slowly and thenbecoming more active, and occasionally, corresponding with their active rotating movements, the whole of the oocyst also moved slightly. In the meantime, the moving sporozoites cameout through the micropyle of the sporocyst and after having left the sporocyst, the freedsporozoites swam about within the oocyst, actively, as if sheeking for an opening, and soonsqueezed themselves through the region of the rnicropyle located at the narrow end of theoocyst wall, shox?xirtg their peculiar behaviors in leaving the oocyst through this region inexactly the same manner as the micropyle of the sporocyst.( 3 ) As to the essential enzyme in the pancreatic juice which brings about excystation, experiments were urrdertaken by using the following enzyme solutions ; the commercialpancreatic preparations, such as trypsin and pancreatin with a high tryptic activity, orcrystalline trypsin, and the physiological saline extracts of pancreas (chicken), and tlne duo-denal jtuice activated by enterokinase. In all cases, positive results were obtained by theexposure of the oocysts to such enzyme solutions, in regard to the effects on excystation.On the contrary, in the case of any enzyme preparation, such as a pancreatie preparationwith a IOXV trypLic a, cLivity, the enzyrne other than trypsir= in the pancreatic juice, crystal-line chyrrnotrypsin, and the comrnercial papain, if substituted for trypsirz, no excystation couldbe detected in any of the experirnents. [the rest omitted]

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Pigs have been said to play some role in an influenza epidemic. The author examinedthe antibody against myxovirus in sv.zine sera, in order to evaluate the role of pigs in thepandemic of Asian irnfluenza in 1957. During the course of these investigatioros, it wasrealized that the rnethods of measuring antibody against myxovirus presented various diffi-culties. The conditions and methods for the measuring of antibody were, therefore, examinedand the followint facts were clarified.I) There was a non-specifie HA ir, .hibitor against influenza A/Asia/57 virus. This in-hibitor seemed to be somewhat different from the inhibitors l<nown heretofore and couldnot be removed by heating at 56C for 30 min or by RDE treatrment. It could be removedby periodate treatment and the HAT test with the virus could be earried out only after theperiodate treatment of tlae sera. Unless adequate attention to the inhibitor was paid, theresults of the HAT test with the Asian influenza virus were apt to lead to a false conclusiort.The optirrnum condition for periodate treatment was found to be the rnixing of one volurrneof serum vith two volumes of M/ 100 K[O4 solution and then to neutralize the excess ofthe perioclate with glycerol after keeping the mixture overnight in the refrigerator.2) There w?as no stubborrt non-specific I-IA inhibitor in swine sera against swine in-fluenza virus and HVJ. The FIAT test of these viruses coruld be carried out easily.3) CF test of swine sera agairnsL Asian influenza, swine influenza virus and HVJ couldbe carried out using the modified Kolt"ner method (two units of cornplement, four units ofancigen, and serial dilution of scrap. Fhe CF test could also be carried out using tlte corn-plement dilution n?ethod (serial dilution of complement with constant units of antigen., andsera), but no special difference in the serodiagnostic value was found between the twornethods.4) The flocculation test of swine sera against Asian influenza virus arrd HVJ could becarried out using purified corncentrated virus

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It seems tlaat equine infectious anaemia is specific to the horse, but some investigatorsreported the infectivity of this virus to man. For a long time we have been employed inthe production of various immune sera, using horses, and have thought II11lC[l on this subject.In 1955, we started the studies on hog cholera and a few other viruses througlr thetissue culture method, and some interesting results were obtained. Therefore, similar in-vestigations were also performed on the equine infectious anaemia virus.At the beginning of this investigation, the author could not find any available reportsin this field and therefore, found it to be very important to complete the cultivating methodfor equine tissue, since the horse seemed to be the only animal susceptible to this virus.Various experiments were performed, and the following results were obtained. lissuecultures were performed by the plasma-clotting method and the trypsin-dispersing method.l. Synthetic medium 199 containing various percentages of cattle serum was used asthe culture medium, and it was found that the optimal ratio of medium 199 and cattleserum was 7 : 3 (Table l), and in subsequent experiments the same ratio of medium 199and cattle serum was employed.2. As a fundamental experiment to determine, indirectly, the most adequate organsfor the cultivation, in young or adult horses, the tissue cultures from equine embryos at thelate stage of pregnancy were tried first because of the possibility of aseptic processing. Theresults were shown in Table 2. These results were not always consistently obtained, but inmy experiments the most excellent growth of tissue was obtained in the spleen, and goodgrowth was also obtained in the kidnies and the Nymph nodes. In the testis, lungs, andbone marrow, good growth was sometimes obtained, but in the liver and heart the resultswere not definite.3. If fresh tissue was used without delay, excellent growth was obtained in young oradult horses, as weI[ as in the embryos.4. Through the monolayer tissue culture meth studies regarding preservation methods, and the passage method of tissue culture, were under-taken by many investigators. In my experiments, the attempts at delaying the maximumcell-growth by rneans of placing the culture in the cold room and thus inhibiting its meta-bolism, were performed. The results were as excellent, or even better than in the experi-ments in which the tissue was cultivated after a long storage in the cold room. The cellsof horse origin wzere more easily grown, more adaptable to the acidified culture medium, and could be maintained longer tlnan the cells of swine origin. A few cultures of tissuederived from young horses were maintained for more than 100 days, in rny laboratory.Frorru the results described above, the author considers that a renaarkable outgrowth ofcells is obtainable not orxly in the spleen, kidnies and Nymph nodes of horses, but also inthe heart, liver, testis, muscle and bone marrow, if fresh organs are used and adequate cut-ture methods are applied, and, also, that these tissue cultures will be most suitable for usein investigations on the equine infectious anaemia virus.

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The results of experimentation on the immune response of mice to the living swineerysipelas vaccine (acriflavine attenuated bacilli), are summarized as follows :1. From the results of correlation between the immune doses and the survival ratesobtained from the protection tests, the immune response was divided into two parts at thecritical point, corresponding to approximately one hundred bacilli. In the case of the testsdone with the doses below the critical, it was recognized th?tt the relation was regressive.On the other hand, when the tests XVCFC performed with tlte doses beyond the critical, theregressive relation was broken and in most cases, regardless of the size of the immune doses, showed a full percentage of sturvivals (Fig. l).In the range of small immune doses, below the critical point, values estimated fromthe regression line would widely vary because of error in bacterial counting and differencesin the susceptibility of mice. Accordingly, the inoculum at the upper portion of the line(order of 10) appears to be available for checking the antigenicity of the vaecine strain.2. On the bases of the above mentioned datum, the influence of the difference in sexand body weight of the test mice on the immunizabilit3r, was checked. From the resultobtained, it was found that the immunity developed in mice was influenced, significantly, bythe difference in body weight, but not by that of sex (Table 1).This finding appears to show that a selection should made based on the weight of themice to be ernployed, in order to prevent a wide variation in the potency values.3. The pattern of the dose response field, consisting of wide immune and challengedoses, show " ed that the size of the immue doses had a more marked influence on the sur-vival of mice than did the size of the challenge (Fig. 2). Namely>mice immunized withthe doses beyond the critical point were resistant against infection even with extremelysevere challenge doses, although there were some deaths. Tire fact that the size of thechallenge doses w

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