The author has already informed in the previous report, that the rabbit poisoned by the inhalation of carbon disulphide gas dies without fail by the paralysis of the respiratory centre. This report is based on the results obtained from the research with regard to the determination of the minimal lethal dose in the case of very acute intoxication by the inhalation of evaporated carbon disulphide gas. The experimental method is as follows:
10c.c. that is about 12.7g. of carbon disulphide is put in a small gas washing bottle, which has a capacity of about 100c.c. and is weighing about 80g. The bottle is weighed immediately after the two openings of it are corked closely by means of gum hat. The expiratory opening of the bottle is then connected with the inspiratory opening of the tracheal tube, which is inserted into the trachea of rabbit. Thus the evaporated gas of carbon disulphide in the bottle, together with the air passing through this liquid, is allowed to enter the lung of the animal, which will consequently be poisoned at once. When the respiration of the poisoned animal is ceased, the bottle is quickly taken off from the connection, thereupon the two openings being corked with the gum hat, and is weighed. The difference between the two weights is the total dose, evaporated in the bottle and mixed with the inspired air.
Four of the same gas washing bottle, each of them containing 30c.c. of 10% caustic kali alcoholic solution, are connected with each other by means of gum tube. The inspiratory opening of these bottles on one end is connected with the expiratory opening of the tracheal tube, inserted into the trachea of the animal, and the expiratory opening on the other end is also connected witd a large bottle containing about five litres of water, which flows out in the constant pressure. In this way a negative pressure arises in this large bottle, and it induces the expired air of animal into the latter through the caustic kali alcoholic solution contained in the four gas washing bottles.
The carbon disulphide, while passing through the caustic kali alcoholic solution together with the expired air, is changed into kali xanthogenate and is dissolved in this solution, thereby only the air gets off from it and comes into the large water bottle. The quantity of the kali xanthogenate to be found in the four gas washing bottles is calculated by means of 1/20 normal cupper sulphate solution. The calculated quantity is then substituted from the total quantity consumed as above mentioned, and the remaining corresponds to the absorbed quantity in she blood of animal, that means the minimal lethal dose.
As the water in the large bottle is kept at about the same temperature as that of the experimental chamber, and flows out in the constant pressure so as to make a negative pressure in this bottle as well as in the four connected gas washing bottles, thus inducing the expired air containing carbon disulphide into the bottles, so the volume of the water flowing out from the large bottle corresponds on the whole to the volume of the air (except the carbon disulphide gas contained in it.) inspired by the animal. It has been found by calculation that the quantity of poisonous gas in the inspired air averages about 17vol.%.
From the experiments on nineteen cases the author came to the following results:
The total quantity of the carbon disulphide evaporated in the bottle and mixed in the inspired air of the animal amounts to about 1g. on an average, of which about the half (0.5g.), that is 0.3g. for 1 kilogram of bodily weight, is absorbed into the blood of the animal. This is the real minimal lethal dose pro kilogram of bodily weight in the case of very acute intoxication in the rabbit by the inhalation of carbon disulphide gas. And the animal, inspiring the air which contains about 17vol.% of carbon disulphide dies in only 4 minutes on an average owing to intoxication.

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1. Artificial insemination was performed with two series of female rabbits.
2. Females of the one series, which were inseminated without any other treatment, showed only 8.3 percent fertility.
3. Females of the other series, which were inseminated after copulation with a sterile male, showed 62.5 percent fertility.
4. Duration of pregnancy was one day longer in the latter case than in the former.
5. Thus the proof is fully established that in the rabbit in the majority of cases ovulation does not take place spontaneously, but needs the stimulus of copulation.
6. Successful results with artificial insemination in the rabbit, therefore, can be obtained with a greater degree of certainty if preceded by copulation with a sterile male.

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1. The hemolytic zone, which is produced by the hemolytc streptococcus on the blood agar plate, is influenced by the hydrogen-ion concentration of the medium as well as by use of the blood of the horse or rabbit. It is, however, larger as the sizes of pH increase, i. e. on the acid medium it would be smaller than on the alkaline medium. The optimal alkalinity of the medium as it considered on the prodnction of the hemolytic zone is pH 7.5-8.0.
2. The hemolytic zone of the βg and βm, both of the atypical strains, as it is produced on the horse blood as well as the rabbit blood agar, has an obscure margin irrespectful of the hydrogen ion concentration. It will, however, become more and more clear as the alkalinity is augumented. That of the typical strains, e. g. βt, has a clearly definable margin irrespectful of the reactions of the medium. It is clearer on the rabbit blood agar than on the horse blood agar.
3. The timely relation of the development of the hemolytic zones is: That the time required for the development of the hemolytic zone by βg differs as the horse blood or rabbit blood is employed. On the horse blood agar, having an acid reaction, the hemolytic zone will not cease to grow even after 72 hours of cultivation, but on the alkaline medium, or pH should be 7.5 or more, it will not grow any more after an elapse of 48 hours, the sizes of the hemolytic zone after 72 hours of cultivation will be practically equal to those of the 48 hour old culture. On the rabbit blood agar, Just irrespectful of the reactions as on the horse blood agar, the sizes of the hemolytic zones will grow even after 72 hours of cultivation, the sizes of the zone at the 72nd hour being just eqnal to those of the 48 hour old culture on the horse blood agar.
4. The hemolytic zones of the βm strain will not cease to grow even after an elapse of 72 hours irrespectful of the use of the horse or rabbit blood or of the reactions of the medium. The sizes of the zone on the horse blood agar or on the rabbit blood agar are practically the same.
5. The strain βt, which had been isolated from the normal horse and a strain which had been isolated from glanders produced each different sizes of the hemolytic zones on the horse blood and rabbit blood agar irrespectful of the reactions of the medium, both of them ceased to grow after 48 hours of cultivation. The sizes of the zone after 72 hours of cultivation on the rabbit blood agar will practically be the same as those of the 24 hour old culture on the horse hlood agar.
6. The hemolytic zones on the medium are larger on the horse blord agar than on the rabbit blood agar, which fact may be explained on the differences of the hemolytic power against the homogenous and heterogenous erythrocytes because all the strains I employed for my experiments were isolated from the horse.
7. Instrains the common broth, both typical, e. g. βt and atypical, e. g. βg and βm. proliferated just the same, irrespectful of the differences of the reactions. The longevity of the culture on the medium, however, differs as the reactions of the medium is different. On the medium having a larger pH than 7.5, the differences would become more manifest, because on such a medium βg and βm will live 43-47 days, while the βt strain will live only 3-5 days.
8. The pnoliferation and longevity of both typical and atypical strains on the 1.0-0.5% glucose-brototh agreed with each other. In the 0.1% glucose broth, however, there were met with certain differences concerning the proliferation and longevity of each different strains.
9. Both the typical anb the atypical strains of the hemolytic streptococci of the horse proliferate very good in the common and the glucose broth having the pH of 7.5 and 8.0. Although the pH indices would be a little higher, the same good proliferation would be the result,

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As regards the saccharifying power of the saliva of the horse, various opinions have hitherto been published, but it is now widely known that the power is not a strong one. This has also been proved by the author's investigation. It is generally believed, on the other hand, that the starch digestion which takes place in the stomach of the horse is chiefly due to the action of ptyalin. This contradiction was the motive by which the author began to set his hand to the study of saccharization of food in the stomach. Having made a series of experiments the author has found that the starch digestion in the stomach is by no means due to the action of ptyalin but to an enzyme contained in the bronchial mucus. This enzyme, according to the author's experiment, has a strong saccharifying power and gives rise to the saccharization of food in the stomach. The author called the enzyme "lung amylase" after its origin. The following is the brief description of the experiments.
1) 50c.c. of 1% starch solution, added with 0.25g. of mucus taken from the surface of the tracheal mucous membrane, has been completely saccharized at 37°C in one hour on an average (20 minutes to 3 hours in 13 cases)
2) A piece of lung tissue taken from a horse killed by bloodletting was cut into smaller pieces and extracted with twice as much water as the volume of the piece. 50c.c. of 1% starch solution, added with 5c.c. of the extract, has also been converted into sugar at 37°C in 42 minutes on an average (15 minutes to 2 hours in 9 cases), while it has taken 10 hours and 20 minutes to saccharify the same starch solution by an addition of 5c.c. of horse serum, that means, the saccharifying power of the serum is about 1/150 as weak as that of the extract of lung tissue. From this fact it will be seen that the saccharifying power of the extract is not due to the action of serum amylase.
3) A tracheotomy was performed in the upper and lower part of the trachea and the communication between these two parts was shut putting a tamponade in the middle part. It was noticed, as the result of this operation, that the saccharifying power of the mucous substance taken from the upper part of the trachea became very weak and after 4-24 hours almost disappeared. It has been shown further that even after several hours no increase in the saccharifying power was constated in the mucus taken from the upper part which was then communicated with the lower part by means of a gummi tube to allow the air to pass through it. As for the saccharifying power of the mucus from the lower part no change has been resulted by the operation.
It may be thought from the foregoing that the amylase which is found in the tracheal mucus must be secreted in the lung, because there cannot be found any difference between the histological structure of the upper part and that of the lower part of the trachea.
4) The saliva taken from the oesophagus of a horse, intratracheally injected with a certain quantity of carmin solution, then stimulated to an increased secretion of saliva was coloured with carmin and proved to have a strong saccharifying power. A starch ball which was then given to the horse and taken from the oesophagus was also coloured with carmin. It has been found, moreover, that in the starch chyme which was kept at 37°C for one hour a large quantity of sugar was present. When oat was given, instead of starch ball, it was noticed that an increased quantity of carmin came out of the oesophagus.
The experiments, in which no injection of carmin was performed, gave the similar result, that is, no particular change was observed as regards the saccharifying power of saliva and the sugar formation in the chyme.
5) In the case of a horse in which the trachea was stopped in its upper part so as to prevent the forthcoming of the mucus from the trachea into the pharynx,

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Auf Grund meiner Untersuchungen nehme ich an, dass die Anämie bei der infektiösen Anämie des Pferdes zu einer hämolytischen Form gehört und zwar hauptsächlich durch die Erythrophagie der Histiozyten bedingt wird, welche deutlich in der Milz und im Knochenmark und weniger in der Lymphdrüse vorkommt. Die aufgenommenen roten Blutkörperchen verändern sich zu Hämosiderin, welches dann nach und nach zerfallt. Ein Teil von diesem Hämosiderin in der Milz wird wahrscheinlich auf dem Weg des Portalsystems nach der Leber zugeführt und ein anderer Teil blcibt für längere Zeit im Körpergewebe abgelagert. Das Hämosiderin oder seine Derivate kommen nicht leicht zur Ausscheidung aus der Niere oder aus dem Darm.
Schliesslich sei nosh bemerkt, dass bei infektiöser Anämie des Pferdes während des Fieberanfalls eine Vermehrung der Erythrozyten anstatt einer Verminderung manchmal beobachtet werden kann. Diese Vermehrung ist als Folge der durch das Krankheitsgift hervorgerufenen Überfunktion des Knochenmarks aufzufassen. Andererseits kann das Knochenmark im Gegensatz zur Überfunktion einer Atonie resp. Hypofunktion anheimfallen, wenn das Gift zu stark auf das Organ wirkt.

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Die bekannte Erscheinung, dass das durch Erhitzen inaktivierte Rinderserum, wenn es mit frischem Pferdeserum vermischt ist, also Komplement zusetzt ist, in vitro rote Blutkörperchen zusammengeballt und dann zur Auflösung bringt, hat durch die Untersuchungen von Ehrlich und Bordet und deren Mitarbeiter festgestellt.
Für sich allein kann weder das aktive Pferdeserum noch das inaktivierte Rinderserum eine Zusammenballung der rote Blutkörperchen bewirken.
Ehrlich schreibt dem Rinderserum die Funktion des Ambozeptor zu, durch dessen Vermittelung das Komplement an die rote Blutkörperchen gebunden wird und diese konglutiniert werden. Nach Bordet Ansicht sollen die Blutkröperchen durch das konglutiniert werden, wenn die vorher durch den Ambozeptor des Pferdeserums gebunden haben. Neuerdungs schreibt Miesner vom Mechanismus der Konglutination, die durch 3 Komponente, Konglutinin im Rinderserum, Sensibilisin und Komplement im Pferdeserum hervorgeruft ist.
Diese Reaktion ist schon zur Serodiagnostik praktisch angewandt, obgleich manche Erklärungen auf den Mechanismus imstande sind, und erkannt man die genaue Resultate auf solchen an.
Ich habe zweifelhaft auf oben eräwhnten Beschreibungen, insbesondere vom Eingangsmechanismus nach verschiedenen Autoren, gehabt, so ist es mir wichtig, das festzustellen, welche Erklärung richtig zu sein.
Auf mancherlei Art habe ich meine Versuche ausgefuhrt und diese Resultate sind folgende:
1. Für sich allein kann aktives Pferdeserum eine Zusammenballung der rote Blutkörperchen aus Ziegen bewirken, aber Kaninchen und Meerschweinchenserum keine Wirkung.
In diesem Falle beide Sera ruften die Hämolyse hervor, aber Kaninchen-oder Meersciwehnchenserum konglutiniert rote Blutkörperchen aus Pferde, ohne auszulösen. Es scheint deshalb die Eigenschaft des Serums gegen Blutkörperchen nach der Arten des Tieres verschieden zu sein, wenn man die aus verschiedenen Tieres stammende Blutkörperchen auf denselben Versuche angewandt hat.
2. Beim Eintritt der Konglutination treten inaktiviertes Rinderoder aktives Pferdeserum die Kompensationsfähigkeit mit einander hervor, Kaninchen-und Meerschweinchenserum aber nicht.
Durch Inaktivierungsmanipulation des Serums mit Hitze, die man gewöhnlich braucht, wird Konglutinin zum Konglutinoid verändert, dann bewirkt sich, für sich allein rote Blutkörperchen nicht mehr zusammenzuballen.
3. Wenn man 3 Arten des Serums, inaktiviertes Rinder-und Pferdeserum, aktives Kaninchen-oder Meerschweinchenserum, zum Konglutinationsversuche, ruft heftige Hämolyse hervor, uud man kann keine Kompensationsfahigkeit auf derselben Reaktion erkennen.
Kaninchen-oder Meerschweinchenserum ist Ziegenblutkörperchen zuweilen zu konglutinieren, nach der Beschaffenheit der Individualität.
4. Als man Rinder-oder Pferdeserum zum Konglutinationsversuche braucht, wird die Konglutination positiv geworden, wenn man zum einen inaktiven Serum eine kleine Menge vom anderen aktiven Serum zusetzt. Wenn beide Sera aktiv bleiben, steigt ihre Kompensatsonfähigkeit ca. 10 fach.
5. Komplement, Konglutination zu beteiligen, ist das Endstück desselben Komplement. Obgleich das Enddtück gehaltene Menge des Pferdeserums nicht beständig oder sehr minimum ist, ruft diese Reaktion immer hervor, so kann man annehmen dadurch, dass die Mitarbeit des Kompements und einer im Albuminteil des Pferdeserums enthaltenden Substanz begründet ist.
6. Wenn man das inaktive Rinderserum durch Liefmannsche Mehode Albumin-und Globulinteil trennt, setzt die Fähigkeit des Konglutinoids herab.

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We detected 2 kinds of the spirochete on the ulcerated surface of the prepuce. beneath which was a large round-cell sarcoma of the penis. While I type (Fig. 1) is a flexible spiral 11, 0μ long with 3-7 coarse turns. II type (Fig. 2) reveals an inflexible body 8, 5μ long with 6-15 fine turns. having one minute terminal filament at each end. The spirochete of II type being very closely related to Spirochaeta pallida. the causative agent of syphilis. we on the one hand inoculated materials from the lesion into the testieles of rabbits, and on the other hand we tried the Wassermann test as well as the Sacks-Georgi reaction with the serum of this patient. but all with negative results.
As far as we know concerning spirochetes found by many investigators in a number of ulcers. it may be considered that the spirochetes in question were in all probability non-pathogenic and happened to subsist and multiply saprophytically in the prepuce ulcer produced by the growth of penis sarcoma.

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[in Japanese]

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