I,1t,III報では,各種の病畜(j]j(・鳥類・牛・人)に由来したE,r.の血清学的分類とそれらのFl:T来した動物の病型との関係,海産魚類の体表粘液から分離されたいわゆるE.r.の血清学的な検討及びそれらの.豚に対する病原学的な意義について,また,一倍1≦健康な.豚の扁桃腺からマウス接種で分離されE.r.と同定されたものについても検討され,それらのなかに,血清学的には,急性敗血症型から分離されるGroupA又は主として賭麻型からのGroupBと異るが,生物学的にはE.r.と同定せざるを得ない数群のものが,かなり高率に見出される事実についても触れた.本報告では青森県十和田市市営屠場において, 1年間に屠殺された1,045頭の健康な豚の扁桃腺を対象にE.r.の分離を試み,それら分離菌について,沈降反応のほかにのせガラス法凝集反応を併用して,先に」゜IR論したGroupA-D以外の群を探ね,また,その結果と野外の豚丹毒発生の実態とを比較しながら,これら一見健康な豚の扁桃腺などに保有されるE.?・.の疫学的意義を検討するとともに,本病の病件の本質について考察したところを述べた.それらを要約すると次のごとくである.I.一見健康な豚の扁桃腺からマウス接種によって分離され,生物学的性状からE.r.と同定されろく)ののなかには,GroupA,B,C,D以外にGroupE及びFが独立して存在することが知られた.2.これら6群(A-F)の菌の抗血清に対しての,被検E.r.菌株の濃厚菌液を抗原とするのせガラス法凝集反応による反応校様は,沈降反応(・こ代る簡易な方法として,E,r.の血清学的分類に充分慣用され得.る.しかも,これら6群の菌が描く夫々の反応模様からE.r.6群間の抗原的相互関係が,次報で述べる抗原分析の結果を待たずに予測された.3.十和田地方で生産され,食肉用として).1j号・殺された-見健.東な啄の扁桃腺から分離されなE.r.,特にGroupBの分離率従って分布度は,明らかに春.に低く秋に高い.4.見掛け上健康な豚の扁桃腺に保有されるE.r3は,群別に単-・である傾向が強く,臨床症状の有無にかかわらず,そこである程度増殖しつつ,何らかの族原学的・免疫学的影響を,その動物に与えつつある徴候がある.5.十和田地方における豚丹毒の発生は3.に述べた一見健康な豚の扁桃腺からのE.r.の分離率の季節的変動と反対に,明らかに春に多く不火から冬にかけては少いか殆ど見られない.6,扁桃腺からのE.r.特にGroupBの分F{{[l.率(ひいては分布度)の季節的変動と野外での豚丹毒の発生頻度との間にみられる注目すべき矛盾は,次のごとく説明された.・

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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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体(;コニ12~15gのdd系マウスに,それぞれ100ケのLePlOsPiraicterohaemorThagiae(TR-1株),およびLePtosPiraau/umnalis(佐伯株)を皮下接1111Fし,定められた時期にそれぞれ2匹づつ順次殺処分し,体内のレブトスピラを定量した.その結果を要約すれば次の通りである.1。 レプトスピラを接種されたマウスは,殆んど無症状に経過した.しかし,体内で,レブトスピラの組織だった増殖が認められた.2.佐伯株を用いておこなった実験によれば,感染初期に,マウス体内でレブトスピラの増殖が最初に認められる部位は血液であった.その他の部位,すなわちエキカおよびソケイリンパ節,接種部洗}條液,および肝においては,血液よりもおくれで,しかも血液よりも少数のレプトスピラが認められたにすぎなかった.3.マウスの血液中で,TR-1株は感染後4~91]{1-1,またイ/1-ミ・(白株は3~10日目にかけて,6,7日目を頂点とする相称性の発育曲線を示した.頂点のレフ斗スピラ数は,血液1cc当105(TR-1株)お3よび104(佐伯株)であった.対数的増殖期におけるMcangcncrat10ntimeは,TR-1株が7.2~11時間,佐拍株が6~8時間であった.感染後11日[11以降は両株とも血液から全く検出されなかった.4,感染後8~10日目頃,すなわち抗体がマウスの血液中に出現しはじめた時期に,抗体陽性のマウスの血液から,イ/{ミ・[}(-1株はしばしば検出されたのに反し,TR-1株は殆んど検出されなかった.Scrotypc間のおなにような差が,別のびと組のレプトスピラ,三河島株(LePtosPiraiclerohaemorr/lagiae)および岡IITI株(Leptospiraautumna/is)の間にも認められた。5.マウスの肝におけるTR-1株とイ左仙株の増殖は,それぞれの株の血液における増殖と,時期的にも量的にも略々似ていた.肝ではしかし,血淑におけるよりも1日おくれてレブトスピラが検出されはじめた.6.腎における両株の発育は,同じく血液におけるよりも1日おくれて認められたが,しかしその増殖曲練は,血液や肝における場合と全く異っていた.すなわち感染後7日目または8日目順に最初の頂点(TR-1株では106,佐伯株では105)が,ついで9~11口目,すなわち抗体出現の頃に下阿が認められ,さらにそれ以後に増殖曲線は再び上,11-]′.し,すくなくとも91日目までそれほど下降を示すことなく続く独得の形を呈した.最初の対数的増夕jl′(期におけるMcangcncrationtimcはTR-1株が凡そ6.8時間,佐拍株が6~9.6時間であった.7.尿中へのレブトスピラ排泄は,感染後21日目から頻繁に言忍められるようになった.8.マウスの牌,肺,副腎,お ,あった.11. この実験に用いた2つの型のレプトスピラによるマウスの実験的レプトスピラ病は,その細菌学的な姿が,相互に似ていることが認められた.

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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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