The properties of the non-bacterial reduction in the muscles of domestic animals were examined with the following results:
(1) This reduction system is not affected by any condition in the regular sausage-manufacturing process.
(2) It differs from the bacterial reduction system. Therefore, it has no effect on the decomposition of the nitrate and nitrite upon which the color fixation of cured meat depends specifically.
(3) The optimum temperature for this reduction system is 35-40°C and the optimum pH falls in the neutral zone.

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In this experiment on the formation of nitric-oxide haem pigment in the muscles of domestic animals by the non-bacterial reduction, we have made optical observations and examinations, using ascorbic acid, for the purpose of modifying the properties of the reduction determined by the experiment described in report VIII of the series of our work, so that they may be applied to any practical process of meat industry.
As a result, it is proved that this reduction takes part in the formation of a NO derivative of haem pigment under the presence of NO2 and that the formation is promoted by addition of 1-ascorbic acid.
It is also shown that ascorbic acid modifies the properties of this reduction, which is inactive at a low temperature (5°C) and a low pH value(5.6-6.0) so much that it makes this reduction active even at such a low temperature and low pH value as required in a practical process.

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The effect of wing-cutting on the chemical composition of chicken meat was studied. Ninety-six Single-comb White Leghorn cockerels were divided into 8 groups, including one for untreated controls and the others composed of experimental birds whose wings were cut at different periods of time after hached. On the 71st day, 16 birds, 2 from each group, having almost the same weight, were sacrificed and the meat production in breast and legs was measured by group. Chemical analysis was carried out on the meat in each group to determine water, crude protein and crude fat contents.
1) There was no distinct difference between the average of the treated groups and the control group in respect to the body weight increase. However, 2 treated groups in which cutting was performed 32-and 38-hours after hatching showed a little more remarkable increase in body weight than the control group.
2) The comparison of the treated groups with the control group in respect to the chemical composition of meat in the breast and legs showed that water content of the breast meat showed no distinct difference between the average of the treated groups and the control group. Leg meat had much more water in the control group than in the treated groups. The water content of both kinds of meat of the 32- and 38-hour treated groups was larger than that of the equivalent of the control group. The crude protein content of breast meat, on a fresh matter basis, showed no difference between the average of the treated groups and the control. More crude protein was contained in leg meat of the treated groups than that of the control. The 32-, 38-, 50- and 56-hour treated groups had more crude protein in breast meat than the control group. The crude fat content, on a fresh matter basis, did not differ so much between the average of the treated groups. The 32-, 38- and 68-hour treated groups, however, had much more crude fat than the control group.
3) In the Single-comb White Leghorn cockerels the wing-cutting will cause an increase in meat production ratio, especially when the cutting was performed 32 or 38 hours after hatching. The 32-hour treated group gave 15% (as fresh matter), or 31% (as dry matter), and the 38-hour treated group 9% (as fresh matter), or 21% (as dry matter), more meat (breast and legs) production than the control group.
This tendency was distinctly recognized in the crude protein and the crude fat production. The 32-hour and 38-hour treated groups showed 30% and 23% more crude protein production, respectively, than the control group. The crude fat production in the same treated groups was increased by 89% and 36% more respectively than the control group.
4) The wing-cutting of cockerels performed at an adequate period of time may bring an increased production and better chemical composition of meat.

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Tsushima is an island in Nagasaki Prefecture which lies between Japan and Korea. Thanks to its location the island has played an important role to introduce the continental civilization into Japan and gives a dependable clue in elucidating the origin of the Japanese native horse. About one hundred of horses pure or nearly pure which can be called Taishu pony were kept in this island in 1952. The height at withers is 114-129cm; 122.6cm on the average in the male and 122.0cm in the female. The Taishu pony is classified in the same category of height with the island type horse approximating the Tokara pony(114.5cm) and the inland type horse approximating the Kiso(133.07cm) and Misaki(130.885cm). horses As the average height at withers is concerned, the Taishu pony is between these two types.
From its size, the prehistoric horse in the Iki Island located near the Tsushima Island, is classified between the small and the medium type horses of the prehistoric Japan, and it is very interesting to note that the Taishu pony is between the island type horse similar in size to the small type horse, and the inland type horse similer in size to the medium type horse of the Japanese prehistoric horse.
Unless our theory of dualism in respect to the origin of the Japanese native horse is completely overthrown, we believe that the Taishu pony must have originated in both the small and medium type horses in the Tsushima Island in the prehistoric age of Japan or after.

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Histochemical methods, including special methods for cholesterol, neutral fats and phospholipids, were used for the study of lipids of the adrenal cortex of pig. Sixty-three pigs, from 6 months to 3 years old, consisting of 4 males, 46 females and 13 castrated males, were divided into 3 groups, i. e., (1) the small group composed of those of 6 to 9 months of age weighing 75 to 110 kg, (2) the medium group of those of 10 to 12 months of age weighing 110 to 150 kg, and (3) the large group composed of the remaining older pigs weighing more than 150 kg.
The following 4 types of the adrenal cortex were odserved among these pigs. (Figs. 1 and 2)
Type A1: Negative in the cholesterol test. The zona reticularis(subsequently referred to as ZR) was composed of small diffusedly sudanophilic cells. The younger ones of the small group belonged to this type.
Type A2: Negative in the cholesterol test. The ZR was composed of large cells containing many large sudanophilic lipid droplets, which were identified as sudanophilic pigment granules, described by NICANDER ('52). In this investigation, however, neither lipo-fuchsin nor any other pigment was observed in the droplets. The older ones of the small group belonged to this type.
Type B: positive in the cholesterol test. A cholesterol band was formed in the ZR, very close to the adrenal medulla. The cells of this band were of remarkably big size and included very large sudanophilic lipid droplets containing both cholesterol and neutral fats. The medium group belonged to this type.
Type C: Positive in the cholesterol test. The cholesterol band was found at a different distance from the medullary border. Such difference was likely to relate mostly to the age. The cells of this band seemed to be equivalent to the lipid-laden cells in the ZR belonging to type B. The large group belonged to this type. The ZR was rather wide in the females, but very narrow in the males and the castrated. Therefore, it can be said that the latter two belong to the transitional type of types B and C.
Consequently, the type of the adrenal cortex was transferred from A1 to A2, B and C in due course with the growth of animal. But in the adult some sex difference seemed to exist between the females and the males including the castrated.
The following zones were always distinguishable in the adrenal cortex of all the pigs, after at least 2 prenatal months, with regard to the density and structural difference of connective itssue fibers; the zona glomerulosa(ZG), intermediary zone, outer zona fasciculata(ZF), inner ZF and zona reticularis(ZR). But the ZR became wider with the growth of animal, and its constituent cells changed their nature and size due to the differentiation into 4 adrenocortical types.
The zona glomerulosa consisted of cells forming masses of irregular shape and included no connective tissue fibers in it. Coarse connective-tissue strands and the cords. The cells of the zona glomerulosa had a very small amout of sudanophobic cytoplasm containing a few extremely fine lipid granules.
In most cases, a narrow, inconspicuous intermediary zone was observed between the zona glomerulosa and the zona fasciculata, which were composed of smaller cells than those of the neighboring zones and a very small amout of connective tissue fibers.
The outer fasciculata consisted of large cells arranged in radial columns and had not so many connective tissue fibers. These cells became larger generally in the inner part of the zone and contained large lipid granules. The cytoplasm showed rather strong, diffused sudanophilia. The shape of the lipid granule varied in the inner parts; initially it became a crescent with a bright vacuole in the concave side, then a hemisphere, and at last a perfect sphere, increase in size being accompanied. BAKER'S test for phospholipids revealed some rather large granules of the same shape, which seemed to be identical with the above-mentioned lipid granules.

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Adult male mice, two to three months of age, were subjected to X-irradiation as follows: local (to the scrotum) with single doses ranging from 5, 000-250 roentgens. The testes were removed at varying intervals after irradiation, and the degenerative and regenerative processes of mitochondria in the tubular epithelium were studied with vital smears under the phase-contrast microscope, and by histological methods.
The restilts obtained seem to indicate that X-rays have many pathological effects on the mitochondria of spermatogenic and interstitial cells with the increase of doses.
1. The spermatogonia are the most radiosensitive germinal cells with respect to quantitative changes of mitochondria. They decreased in number by high dose irradiation (500-1000r), disappeared after twenty-four hours, and then regeneration was observed.
Mitochondria were mostly granular in the spermatogonia and spermatocytes, though a few short rods were found in them.In degenerating spermatocytes mitochondria showed a fragmentation or globulation.
In the transformation stages of spermatids into spermia, the breaking up of filamentous mitochondria into granules was observed.
Giant cells present in degenerated epithelium had large rounded mitochondria with vacuolizated structure and in the formation of the chromosome bridge were observed mitochondria) bridges.
2. The interstitial cells are more resistant to Xray injury than the tubular epithelium, but for not many days after the highest dose Xirradiation (2, 000-5, 000r)mitochondria showed bistinct qualitative changes (granulation, fragmentation, globulation or vacuolization) and comparatively rare quantitative changes.

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The normal shape and its denaturation of the casein particle in milk was investigated with the 50kV. electron-microscope.
Principal results obtained are as follows:
1) The size of casein particles was about 20 to 200 mμ in diameter in cow's and goat's milk, but it was much smaller in human milk than in cow's milk.
2) Some absorbed substance was observed around casein particles in the normal milk and seemed to have some relationship with calcium in milk.
3) At the iso-electric point of casein in milk, same casein particles were agglutinated, forming bigger particles, and the absorbed substance observed around the original casein particles disappeared in the newly formed particles. At pH 2.4, the casein particles underwent transformation and looked like "swelling particles",
4) When the pH value was adjusted to normal by adding alkali, these acid-denaturated, casein particles did not restored their original shape. Accordingly, this type of transformation was irreversible.

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For the preparation of the HCl-pepsin solution generally used in artificial digestion assay of protein, it is advantageous to know the distribution of the secretory cells of three types in the gastric mucosa of swine. They are the zymogenic, parietal, and mucous neck cells secreting pepsinogen, hydrochloric acid, and mucus, respectively.
In the present paper, the distribution of these cells in the gastric mucosa, mainly in the fundus, is described.
1) Sixteen pieces of mucosa about one cm square each collected from the fundus of the stomach, were treated, fixed, and stained by a specific method described below. Two more specimens of the same size were harvested, one from the pyloric and the other from the cardiac gland region, and manipulated in the same way.
Each specimen was fixed for 4 days in a solution consisting of one part of 35% neutral formalin and three parts of saturated corrosive sublimate solution, followed by rinsing in running water for 24 hours. After the treatment with Lugol's solution and rinsing, it was dehydrated, imbedded in paraffin, and sectioned 5μthick.
The sections were stained with Schiff's reagent (PAS) and aurantia.
2) Nuclei of the cells were counted under the microscope of 400-time magnification by the aid of a cross micrometer with 25 one-square-mm meshes.
In the cranio-caudal observation at, the fundus, the zymogenic cells were distributed in the base, body, and neck of the gland in the decreasing order, but the parietal cells in the neck, body, and base, in the decreasing order. The mucous neck cells were contained almost equally in those areas.
In the transverse observation, the zymogenic cells were contained in a larger number at a portion of the fundic gland region close to the cardiac gland region. On the contrary, the mucous neck cells were found mostly at another portion of the fundic gland region near the pyloric gland region. The parietal cells were distributed almost equally.
In conclusion, the zymogenic cells were contained in the largest number in the base of the glands in a portion of the fundic gland region near the cardiac gland region. Such portion of the region may be called the pepsinogen zone.
3) In the microscopic observation, no zymogenic nor mucous neck cells were found in the pyloric and cardiac mucosa, while some portions of the cardiac mucosa were stained yellow, where a few cells similar to the parietal cells, were observed.
The cells of the cardiac and pyloric gland regions were stained intensely red, especially remarkable in those of the latter region, which was very similar to the mucous neck cells in morphology and staining features. Consequently, from these observations the cells of the pyloric gland region may be a sort of mucus-secreting cells similar to the mucous neck cells.

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Twenty immature male mice of bc-strain were hypophysectomized at the age of twenty-three to thirty-nine days (twenty-eight to thirty days in most cases), and sacrificed at various periods after ope-ration. Eleven litter-mates were used as controls, including six for reference controls. Fixing and staining were carried out as mentioned in report I of this series of paper. The results obtained were summarized as follows.
1) Macroscopic observations: After hypophysectomy the testes and adrenals regressed significantly in size and weight, and the ventral prostates and seminal vesicles did not developed. The testes turned brownish yellow, while the color of the adrenals and accessory reproductive organs faded. The changes in weight of the organs were shown in table 1.
2) Histological observations:
Testis: The diameter of the seminiferous tubules decreased, and a part of the germinal epithelium failed into the lumen and degenerated after hypo-physectomy. Spermatocyte disappeared completely by the thirtieth day after operation. The inter-stitial cells atrophied and creased in number after operation and by the tenth day their nuclei showed the wheel-cell-like figures. Cytolyses were also observed in some interstitial cells, the other cells being stained black strongly with HEIDENHAIN's iron hematoxylin.
Adrenal: A slight hypertrophy in the cells of the zona glomerulosa and atrophy in those of the zona fasciculata were observed and lipid decreased in these cells. These changes became more prominent in the zona fasciculata as days went. In the zona fasciculata cytolysis and karyopyknotic figures were also observed after hypophysectomy. Although the cells of the X-zone atrophied to a great extent and fuchsinophilia demonstrated by ZINSSER and ZINSSER'S ('51) Ponceau-fuchsin metnods dis-appeared from them completely, the X-zone still remained in the cortex after hypophysectomy.
Accessory reproductive glands: Both the ventral prostates and seminal vesicles did not grow after hypophysectomy, showing the immature type. The slightly developed accessory reproductive glands also regressed to the immature type after hypophy-sectomy.
In conclusion, the changes in the testes.adrenals and accessory reproductive glands of immature mice after hypophysectomy seem to be essentially similar to those observed in adult mice (NISHIDA and MOCHIZUKI, '54).

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Fourteen white Leghorn cockerels, 3 to 4 months of age, were adrenalectomized and sacrificed at different times for 12 hours to 8 days after adrenalectomy. Of these 2 birds were injected with desoxycorticosterone-acetate (DOCA, 5mg per day) for 4 days and sacrificed 5 days after the operation Two unilaterally adrenalectomized, 8 cortisone-tre-ated and 6 untreated cockerels of the same age were used as the controls. For the purpose of cyto-logical studies, the pituitaries of these birds were fixed in formol-sublimate and stained by GOLDERG and CHAIKOFF'S method ('52), HETDENHAIN'S azan te-chnique, GOMORI'S A-F method, PAS method and metachromatic staining with many basic dyes.
The results obtained are as follows:
(1) No alteration was shown in the number and the cytological structure of the α cells in the caudal lobe following adrenalectomy or injection with corticosteroid. This suggests that the α cells have no relation with the secretion of adrenocorticatrophin (ACTH).
(2) The adrenalectomized birds showed such remarkable changes as disappearance of granulation, pyknosis of the nuclei, and reduction in number and size of the β ce1s and were stained quite feebly.Degenerative cells commonly refered to as CROOKE'e cells appeared abundantly in the caudal lobe of the adrenalectomized or cortisone-treated birds. The βcells of the adrenalectomized birds were not completely recovered from these degenerative changes by the injection of DOCA. Accordingly, the cy tological picture of the βcells did dot seem to correlate to an increase or decrease in demand for endogenous ACTH.
(3) The most remarkable change in the cytological picture of the pituitary following adrenalectomy was observed in the V cells of the cephalic lobe. They were increased distinctly in number, enlarged to their maximum size, and showed a chromophobe appearance with little granulation in cytoplasm. Their uclei were markedly enlarged containing two or three(or occasionally more) clear, spherical nucleoli each, which became larger in accordance with the growth of the cells. The injection of DOCA prevented any changes following adrenalectomy in the V cells of the pituitary.
These observations made on the cytological changes in the pituitary indicate that only the V cells were associated with the ACTH secretion following adrenalectomy or treatment with corticosteroid. The PAS-positive granule of the V cells is not a chemical component of ACTH at all, but it may be cement or a carrier substance of ACTH.

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(1) The develpment of Infusoria found nummerously in the rumen, was rexamined by AKASHI'S method(this journal, 23;9, 1951) using bovine rumen contents.
Rumen contents were obtained from an indigenous goat through a permanent rumen fistula adapted to it. The rumen contents were dried up for a different period of time at a different temperature immediately or after centrifuged, and used for culture. No motile organisms could be found in any of the samples.
(2) Rumen contents not dried up nor soaked in acid were cultivated with goat's rumen fluid as culture medium. But contrary to the results reported by AKASHI, no motile organisms were found in all the specimens.

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Experiments were carried out on chickens for 91 days to elucidate the effect of a feed mixed with fish meal, yeast and fish oil, and the other feed of vegetable origin containing penicillin, Vitamin B12 and liver meal on an increase in body weight and quality and yields of chicken meat.
(1) When fish meal was replaced by yeast as a source of protein in the basic feed, 10mg (1mg=1, 000IU) of penicillin and 20 γ of vitamin per Kg of the feed and 5 per cent liver meal were added to the feed, the increase in body weight was most remarkable with larger yields of meat and the testes were found the largest. The quality of meat (calories and contents of extracts and fat), however, was better when fish meal was used in the basic feed instead of yeast.
(2) The vegetable ration with the addition of penicillin, Vitamin B12 and liver meal exerted a remarkable effect on the increase of body weight. However, the quality of meat proved better when the ration containing fish meal, yeast and liver meal was used.
(3) The ration, to which fish oil was added, gave unfavorable results causing little increase in body weight and producing only a little deposit of fat in the muscles.

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The present experiments were undertaken to confirm the presence of serological antigens in the bovine blood cells and frequency of their appearance in several breeds of cattle. For this purpose hemolysis by immunoisohemolysins was used, and the following results were obtained.
There are five type-specific antigens (A, B, C, D and E) in the bovine blood cells. As shown in Table 3, 32 combinations are possible from the five different antigens.
Frequency of appearance of these antigens in several breeds of cattle is shown in Table 4. The A antigen showed a distinct difference in fre-quency of appearance in the breeds, while the other antigens appeared almost regularly in the different breeds. It was 34.8% in the Holstein-Friesian, 16.5% in the Japanese indigenous, and only 2.4% in the Jersey cattle. The Guernsey cattle were found not to have the A antigen in their blood cells.
As indicated in Table 4, the relationship between the frequency ratio of the A antigen to the B in the Holstein and that in the Japanese indigenous cattle may be compared to the' Rassen index' of the human blood groups. That is, the frequency of the A antigen is higher in the Holstein than in the Japanese indigenous cattle, but that of B antigen is lower in the Holstein.
The Jersey cattle imported into Japan showed a distinct difference in frequency ratios of the A antigen due to the places where they were bred.

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