It was stated in the first report that, the experimental animals (mouse, white rat, guinea-pig, rabbit) die within a short time by the inhalation of air containing 1.5-3 volume % of carbon disulphide gas. As these experiments have been performed by confining the animals in a vessel containing a calculated quantity of carbon disulphide gas, only the volume % of gas necessary to kill the animals could be asserted; but the quantity which must have been absorbed in the blood during inhalation can not be known.
As described in the second report, the experiment was carried out in order to determine the quantity of gas absorbed in the blood as follows. A certain quantity of evaporated carbon disulphide gas contained in a small gas washing bottle and mixed with the air passing through this bottle, was allowed to enter the lungs of an experimental animal, and the carbon disulphide gas to be presented in the expired air was caught by means of a 10% alcoholic solution. of caustic potassium The quantity of gas absorbed into the animal's blood could thus be asserted by subtracting the quantity of gas found in the solution from that of the evaporated. The results from 19 experiments showed that, the total evaporated quantity was about 1g., and the half which (0.5g.) was absorbed in the blood, this being 0.3g. for 1kg. of body weight. In these cases, the inspired air contained, as already mentioned, about 17 volme % of carbon disulphide gas, and the poisoned animals died quickly in an average of 4 minutes. Comparing this 17 volume % of carbon disulphide gas with the 1.5-3 volume % of the gas mentioned in the first report, it can be easily seen that the difference between them is too great. For this reason, some changes have been made in the arrangements used in the experiments described in the second report, so as to diminish the volume % of the carbon disulphide gas in the air to be inspired. These changes are as follows:-
(1) The ball-form end of the inspiratory tube of the gas washing bottle was changed into a small pointed canal, so that even the minutest particle of air passing through. the carbon disulphide liquid could be clearly visible in the form of small bubbles.
(2) Between the bottle of carbon disulphide and the tracheal tube inserted into the trachea of rabbit, there was placed a T-shaped tube, through which the fresh air could come into the air containing carbon disulphide gas, which had passed through the tube during the inspiration of the animal. Thus the concentration of carbon disulphide gas in the inspiratory air was materially diluted.
(3) In order to absorb the carbon disulphide gas in the expi- red air, instead of using 4 gas washing bottles, each containing 30c.c. of the 10% alcoholic solution of caustic potassium, as in the former experiment, 3 bottles were employed, the first containing 50c.c. of the same solution, the second 40c.c., the third 30c.c. Should the inspiratory tube of the first bottle be clogged by the crystal of potossium xanthogenate, this bottle should be immediately taken off, and then the second and third bottles continuously are used. In 12 experiments there was never a case, in which the second bottle was clogged by the crystal.
(4) In this experiment, a very large water bottle of 20 litres capacity was used, because firstly the concentration of carbon disulphide gas in the inspiratory air should by far be less than that mentioned in the second report, secondly the time of intoxication should consequently be much longer, and thirdly the quantity of the flowing water which will necessary to suck the expired air into the 10% caustic kali alcoholic solution contained in 3 gas washing bottles as well as into the last water bottle must accordingly be very great.
(5) To prevent the contrary current of inspiratory as well as expiratory air, a remarkable improvement was made in the construction of the valves situated at the correspondiug places.

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I have informed in my previous paper (No. 1, Vol. IV, 1925, of this journal) that a number of cases of contagious pleuro-pneumonia occurred among cattle imported from China in September 1924.
8 months later viz. in May 1925, the disease again broke out in the city of Osaka and has rapidly propagated to the five neighbouring prefectures. On this occasion 124 cows were affected and 656 were treated as exposed to infection
Taking advantage of this opportunity I have made some further experiments upon the disease, the results obtained being briefly summarized as follows:
1. The virus of contagious pleuro-pneumonia grew well in 4-8 days in stab culture in beef-bouillon-agar or Martin's agar, both containing 9% bovine serum The virus generally formed a white network consisting of many small colonies along the stab line.
2. In the shake cultures in bouillon agar or Martin's bouillon agar, colonies developed semi-anaerobically, that is, principally in a definite stratum, about 1/2cm. below the surface of the culture medium in the course of 3-7 days.
3. The organism grew well in oxygen-free atmosphere quite anaerobically in the course of 7-8 days.
4. In a medium prepared from the transparent fluid of the anterior chamber of cattle eye, the organism grew in 7-8 days, showing a fine opalescent turbidity.
5. By inoculating a pure culture (0.2-0.6c.c.) into the anterior chamber of rabbit eye, I have recognized that the organism can survive there for two weeks. An acute iritis was observed in the inoculated eye.
6. 4-5c.c. of the pure culture introduced into the rabbit testicle were apparently absorbed in 1-2 days, but a few days later the testicle began to swell up and after 2 or 3 weeks it recovered into normal size or even smaller than normal. No symptom of suppuration was observed in the swollen testicle.
7. The serum from a rabbit immunized by inoculation of pure culture into the anterior chamber of the eye or into the testicle, showed a precipitine reaction to the serum from affected cattle.

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狂犬病ノ蔓延ガ雜犬ノ繁殖ニ由テ助長セラルルコトハ明白ナル事實ナリ本邦ニ於テハ古來仔犬ヲ屠殺セズシテ遺棄スル習慣アリ此故ニ野犬ハ益々其數ヲ増加シ野犬捕獲隊ノ努力モ遂ニ豫期ノ効果ヲ擧ゲ能ハザル状況ニ在リ
雜犬ノ増加ヲ防グ爲メニハ或ル程度迄牝犬ヲシテ不姙ナラシメザル可ラズ之レ本研究ノ動機ニシテ之レガ完成ニ依リ必ズヤ犬ノ繁殖ハ制限セラレ從テ狂犬病ノ傳播上ニ大ナル影響ヲ與フ可シ
手術ニ據リ犬ヲ不姙ナラシメントスル方法ハ從來卵巣摘出ノ形式ヲ以テ行ハレタリ1923年べるぐ氏ハ卵巣摘出ガ内分泌ノ方面ヨリ考ヘテ賞ス可ラザルモノナリトシ子宮摘出ヲ行ヒテ不姙ノ目的ヲ達セリ本法ニ據リ犬ヲ避姙セシメ得ルコトハ明ナレドモ子宮摘出法ハ術式稍繁雜ニシテ一般的ニ廣ク實施セラル可キモノニ非ラズ依テ余ハ全然他ノ術式ヲ考案シ從來ノ卵巣摘出法ト比較シテ之レガ實驗的手術ヲ行ヘリ手術名手術頭數及ビ平均手術時間次ノ如シ
1.卵巣摘出術 16頭 36分
2.子宮體截斷術 27頭 13分
3.輸卵管截斷術 12頭 26分
4.輸卵管結紮術 20頭 16分
合計75頭ノ比較研究ヨリ次ノ結論ヲ得タリ
1.輸卵管截斷術或ハ輸卵管結紮術ヲ以テ卵巣摘出術ニ代ユルコトヲ得而シテ兩者共ニ卵巣摘出術ヨリ其方法簡單ナリ
2.子宮體截斷術ハ術式簡單ナルモ術後子宮腫嚢ヲ發生スルコト及ビ姙娠中ノモノヲ手術シ能ハザル缺點アルガ故實用的ナラズ
3.輸卵管手術ハ結紮法ニ據ルヲ一層簡便ナリトス
4.輸卵管手術ヲ行フモ發情ヲ止ムルコトヲ得ズ避姙ノ目的ノミヲ達シ得ルモノナリ
5.子宮體截斷術ヲ除キ他ハ總テ從來人ノ試ミザリシ腹壁横截法ニ據ルヲ良シトス

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In 1923 the author observed the prevalence of an infectious diarrhea (commonly called by the name of "Kiifun" or "Ohri") among young chicks from Nagoya and its neighbourhood. He isolated the anaerogenic strain of Bact. Pullorum, one of the causal agent of Bacillary White Diarrhea of Young Chicks in Europe and America, from the internal organs of chicks which had fallen dead from the disease, and he was able to demonstrate that the isolated organism was the real cause of their death. (Chuojuikaizasshi, Vol. 37-10)
The author again went on searching for the aerogenic strain of Bact. Pullorum, and in 1924 he met with several outbreaks of the so-called "Ohri" among young chicks from Nagoya and Okayama, and was able to isolate 2 types of Bact. Pullorum from the dead chicks. From this finding, the author has arrived at the conclusion with firm conviction that what has been called by the name of "Ohri" prevalent in the middle part of Japan must have contained the same kind of disease as Bacillary White Diarrhea of Young Chicks in foreign countries.
The results of his investigation are summarized as follows:
1) Two kinds of organism belonging to the category of the typhoid-paratyphoid group were isolated from the internal organs of chicks which had fallen victim to the so-called "Ohri" prevailing in Nagoya and Okayama. One of the two was the anaerogenic type of Bact. Pullorum (termed B-type by the author) and the other, resembling the former, was that which formed gas in glucose agar (termed A-type). The isolation ratio between the A-and B-type during the examination of the dead chicks from 3 outbreaks averaged 6:11.
2) The difference between the cultural characteristics of A-and B-type lies in: (a) that the former forms gas in glucose agar, while the latter does not, (b) that the humidity being deficient in the colonies on agar which seem somewhat fragile, the former, even if emulsified in the physiological salt solution, will not make the homogeneous suspension, while the latter, the colonies of which being soft and moist, easily makes the homogeneous suspension in the physiological salt solution, (c) and that the former, in the broth culture, attaches minute granulae substance annularly along the glass wall on the surface of the medium and forms pellicle and granular sediment after several days' incubation, while the latter lacks such characters.
3) The two types intersectionally correspond to each other so that it is difficult to distinguish one from the other, so far as the agglutination reaction goes. But the former agglutinates to a high degree in the immune serum of Enteritidis group, while the latter is almost negative.
4) The isolated organisms have a powerful pathogenic character against young chicks, causing a fatal septicemia either by inoculation or by feeding culture.
5) Comparing the characteristics of the isolated organisms with those of Bact. Pullorum, it may be seen that the B-type entirely corresponds to the anaerogenic type while the A-type does not corresponds, to the aerogenic type on account of the characters of colonies on agar and of broth culture. It may, however, be considered that such a difference is due to the minuteness or roughness with which the observation is made, for the characteristics of broth culture above referred to cannot be observed within 4-5 days' incubation and also those of the colonies on agar may be apt to be neglected. Therefore it may be considered that the A-type should be alleged to correspond to the aerogenic type of Bact. Pullorum.
In the agglutination reaction the relation between the two types and the other members of the typhoid-paratyphoid group is quite the same as the relation between Bact. Pullorum and those of the typhoid-paratyphoid group.

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We studied the resistance of the red blood cells of 12 healthy domestic animals (horses, cattle, swine. sheep and goats) respectively by the same method and under the same conditions. As examination method we took Hamburger's method in the essential points, modified slightly as conditions demanded. The blood was taken from the jugular vein in all the animals except swine, in which the blood was taken in a slaughter house directly from the aorta (it consists chiefly of arterial blood). We prepared the NaCl-solutions from 0.33 to 0.78% differing by 0.01% respectively with sodium chloride (Merck). The process of our examination is as follows: 5c.c. NaCl-solutions were put into each of the centrifugal tubes pointed at the bottom and two drops of the defibrinated blood which had been kept in an ice-box for one hour beforehand, were put into the NaCl-solution in the tubes with the same pipette and then well mixed and put in an ice-box for three hours. Then the tubes were taken out from the ice-box and all put in the centrifuge for five minutes and the results were read.
1. The results of the foregoing experiments are summarized in the following table:
2. The minimum resistance. The horses show the greatest resistance and resistance decreases in this order catte, swine, sheep and goats. Swine show the greatest individual differences and goats, sheep, cattle and horses less in order. In general the minimum resistance is proportional to the size of the erythroeytes and in inverse ratio to the erythrocyte number.
3. The maximum resistance. The cattle show the greatest resistance and resistance decreases in this order horses, swine, sheep and goats. Goats and sheep show the greatest individual differences and swine, horses and cattle less in order. Generally the maximum resistance is also proportional to the size of the red corpuscles and inversely proportional to the number of the erythrocytes.
4. The resistance-width. On the whole the swine show the greatest and goats and sheep the next and cattle and horses the smallest. On the average the goats show the largest and in the next come swine, sheep and cattle while horses the smallest.
5. In short, in the resistance of erythrocytes, cattle and horses rank first showing hardly any difference, swine are intermediate while sheep and goats show the least resistance.

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In Sapporo and its neighbourhood, especially in the Makomanai Livestock Breeding Farm there has been a piroplasmosis-like disease of cattle, so-called "Tick-fever", which chiefly attacks pastured cattle and ist damage is not small. As recently its damage has gradually become larger and larger, it is now a matter of importance scientifically and practically to investigate the cause and nature of the disease. We have engaged in the investigation of the disease for several years and have already examined about 90 diseased cattle in order to know the character of the illness. We obtained the following results from blood examination, postmortem results, inoculation experiments and trypanblue (Cassella) injection. But our investigation will continue since it has not come to a satisfacrory conclusion.
1. The so-called "Tick-fever" which is an epidemic disease among the cattle in Sapporo and its neighbourhood, occurring in Summer, is analogous in type to Piroplasmosis.
2. The main symptoms consist of fever, anaemia, icterus, haemoglobinuria and abnormal erythrocytes pathologically changed; but it is very seldom that a case has all five symptoms. Even in cases, in which both fever and anaemia are very slight and the other symptoms are absent, there are always abnormal erythrocytes in the blood. For this reason we think that the abnormal erythrocytes are the most important factors in the diagnosis of this disease.
3. It is not possible now to determine to what kind of Piroplasmosis this disease belongs; but it coincides with neither Texas-fever nor Coast-fever. The disease resembles the former in symptoms, but the Piroplasma in the blood of the diseased animals are quite different in form; and the disease differs from Coast-fever in symptoms, but the parasitic Piroplasma morphologically resembles each other very nearly.
4. This disease is analogous to Piroplasmosis, but the Piroplasma proved in the bloodcells can not be decide with certainty as the excitants of the disease. For the presence of Piroplasma in the blood does not always denote that the the animal is infected. We have often found that healthy cattle have rather more parasites in the blood than the diseased animals. As is proved by the inoculation experiments with the blood of infected cattle into healthy ones, the transmission of Piroplasma and invasion of the disease does not go hand in hand.
5. With respect to the therapeutic value of trypanblue (Cassella) we cannot express an opinion, for we used too few animals for this purpose. But according to our experiments the trypanblue is not good for this disease, being rather harmful to the infected animals.

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