Comparative effects of hexavalent (K₂Cr₂O₇: Cr(VI)) and trivalent chromium (Cr(NO₃)₃: Cr(III)) on the development of lipid peroxidation, and the relationship between the lipid peroxidation and damage to tissues were studied using male ddY strain mice. The animals were administered with either of two chemicals at a dose of 20mg Cr/kg by a single intraperitoneal injection. The results obtained were as follows: (1) Lipid peroxidation in the liver, as measured by the synthesis of thiobarbituric acid reactive substances (TBARS), showed a significant increase at 24 and 48 hr after Cr(VI) injection, while in the kidney it was observed only at 48 hr. In the mice administered with Cr(III), TBARS formation in the liver went down below the control levels, while no change was observed in the kidney. (2) Chromium contents in the liver and kidney showed a maximum level at 6 hr after injection of Cr(VI) and then those declined to the half of the maximum level at 48hr, respectively. Chromium contents in the liver and kidney of the mice injected with Cr(III) were lower than those injected with Cr(VI) during the experimental period. (3) Increases of TBARS formation in the liver, chromium content in the liver and kidney, and ornithine carbamyl transferase (OCT) activity indicative of the liver cell damage, and urea nitrogen content in the serum, indicative of the kidney damage, observed at 24 hr after injection of Cr(VI) were inhibited by simultaneous injection of 100mg/kg of L-ascorbic acid, as antichrome agent, respectively. These observations might suggest a possible causative role of lipid peroxidation in Cr(VI) toxicity. (4) Incremental formation of TBARS in the liver after Cr(VI) injection was inhibited by simultaneous injection of N, N-diphenyl-p-phenylenediamine (DPPD), an antioxidant, but DPPD did not suppress the increase of chromium content of the organs examined (the liver and kidney), serum OCT activity and urea nitrogen content observed after Cr(VI) injection. This finding contradicted with the conclusion described above. The results of this study confirmed Cr(VI) induces lipid peroxidatioin in the liver and kidney of the mice, while lipid peroxidation is not responsible for the tissue damage induced by Cr(VI)

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Three strains of Fusobacterium necrophorum biovar C were injected into mice intraperitoneally and intraportally. All the mice survived. In one mouse out of 15 mice injected intraperitoneally, a few focal abscesses were formed in the liver. The microorganisms were recovered from the liver abscess and the tissue of liver with abscess. No changes were observed in the organs of other 14 mice and no bacteria were recovered from them. In the 15 mice injected intraportally, no liver abscesses and no macroscopic changes in the organs were formed. However, the inoculated bacteria were recovered from the liver of four mice. The pathogenicity of F. necrophorum biovar C was weaker than that of other two biovars.

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Twenty nine recent isolates of bovine viral diarrhea-mucosal disease (BVD-MD) virus, 17 from persistently infected cattle and 12 from mucosal disease, were compared antigenically with the reference strains by a serum neutralization test. The reference viruses were divided into 2 groups, tentatively designated as N and K, based on the antigenic relationships in the cross-neutralization test. Antigenic properties of recent isolates were considerably different with the sources of virus isolation. Seventeen isolates recovered from persistently infected cattle were divided into 3 groups in the neutralization test using antisera to the reference strains; 12 and 2 were considered as the possible members of groups K and N, respectively, and the others belonged to neither group. On the other hand, 10 of 12 isolates recovered from mucosal disease were considered as the possible members of group N, and the others were classified into neither group. Interestingly, none of BVD-MD viruses isolated from cases of mucosal disease belonged to group K. The results of serologic survey on sera collected from 713 cattle at the Hokkaido provinces in 1974 to 1988 indicated that infections of cattle with BVD-MD viruses other than group K were prominent before 1981. Cattle infected with group K BVD-MD virus were first detected in 1982, and increased in number thereafter. The results obtained in this study suggested that BVD-MD viruses with various antigenic properties spread widely among cattle herds, and also a possibility that clinical manifestations in cattle infected with BVD-MD viruses may differ with their antigenic properties.

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Effects of short-chain fatty acids on pancreatic exocrine secretion were studied under anesthesia in calves within 2 weeks of age (2-wks calves) given only whole milk and milk replacement and in which rumen fermentation has not begun yet, and in calves at 13 weeks of age (13-wks calves) weaned at 40 days of age and in which rumen fermentation has already begun. Basal rate of juice flow and protein concentration and amylase activity in pancreatic juice under basal condition were significantly lower in the 2-wks calves than those in the 13-wks calves. Intravenous administrations of acetate, propionate and butyrate stimulated pancreatic juice secretion and protein and amylase output in both groups of calves. Those responses were increased with increasing carbon number in the molecule of fatty acids. Although the response of amylase output (/kg of body weight) in the 2-wks calves was significantly less than that in the 13-wks calves, the response of juice flow and protein output (/kg of body weight) in the 2-wks calves were equivalent to or greater than those in the 13-wks calves. These results indicate that the characteristic of pancreas, being stimulated by short-chain fatty acids, in calves and probably in other ruminants is not generated on the process of postnatal development, but has been already acquired before rumen fermentation begins.

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Studies were made on the development of Babesia ovata in the midgut of the nymphal tick vector, Haemaphysalis longicornis. In 12 hr post-repletion, merozoites were observed outside of erythrocytes infected with B. ovata in the contents of the midgut of the tick. After that, these merozoites were transformed into ring-forms which were comparatively large ring 2-3μm in diameter. Within 48-72 hr post-repletion, ring-form protozoa developed into spherical form 4/5μm in diameter. Within 3-4 days post-repletion, fission-forms were transformed into fission-bodies 2-3μm in diameter. Within 4-6 days post-repletion, fission-bodies developed into bizarre-forms 6-7μm in diameter. At this time, elongated form protozoa which were considered as microgametes, 6-8μm in length, are also seen. Within 6-8 days post-repletion, round-formed protozoa which were considered as zygotes in 9-10μm in diameter were observed in the gut. About 10 days after repletion, those round-formed protozoa were transformed into vermicule-formed and round-formed protozoa, 13-15μm in length, appeared again in the gut epithelial cells.

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Immunoperoxidase technique was applied for Pathological study on naturally occurring pneumonic tissues of calves from which Pasteurella haemolytica was isolated. Multifocal necrosis occurred in the lungs of 25 out of 42 calves (59.5%) and P. haemolytica antigen was detected in 22 out of the 25 calves (88.0%). The calves were divided into 3 groups according to the number of P. haemolytica isolated. The positive rate of the bacterial antigen detected by the technique was 66.6% (28/42) on the average, reaching up to 85.7% (18/21) in the group from which the largest number of P. haemolytica was isolated.

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Bovine herpesvirus 1 (BHV-1) isolates from respiratory tract and from vagina of bovine in Japan were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and the DNA restriction endonuclease cleavage pattern, and compared with European BHV-1 strains. Both protein profile and DNA cleavaged pattern of BHV-1 isolates from respiratory tract were the same as those of European infectious bovine rhinotracheitis (IBR) virus, whereas the protein profile and DNA cleavage patterns of one isolate (M1) from vagina was the same as those of the European infectious pustular vulvovaginitis (IPV) virus. The facts indicate that IPV virus has existed in Japan.

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The Prevalence of dental disorders in dogs was studied by applying index systems for human with some modifications. A total of 251 mongrel dogs including 143 stray dogs kept in the Animal Protection Offices in Tokyo and Hokkaido and 108 pet dogs visiting veterinary clinicians in Chiba Perfecture and Hokkaido were used. Periodontitis was prevalent among these dogs regardless of their sources and its incidence was increased with age. The lesion was more severe and more frequent in the premolar and molar regions than in the maxillary and mandibular incisor regions. Missing of teeth was observed at a high and increasing incidence with age. The tooth most commonly lost was the first premolar, followed by the other premolars and molars, where severe periodontitis was frequently found. Calculus was seen on many teeth, and aging agravated its prevalence and severity. Dental caries was observed in stray dogs, but neither to a serious degree nor at a significant level. These findings emphasize the necessity of dental hygiene, proper dental care and continuous periodical survey for dogs.

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The morphology of blood cells in the carp was investigated by light and electron microscopy. Erythrocytes, thrombocytes, lymphocytes, granulocytes and monocytes were identified as the peripheral blood cells. Thrombocytes were round to long oval, each containing vesicular and microtubular structures and an oval nucleus with abundant heterochromatins. Lymphocytes were divided into three types in size, small, medium and large. Some of the small and medium lymphocytes were α-naphthyl-acetate esterase (ANAE) positive, while large lymphocytes were pyroninophilic. Granulocytes were distinguished into three types (type I, type II and type III) according to the morphology of the nucleus and granules. Type I granulocytes possessed lobulated nuclei and a large number of cytoplasmic granules, each of which was oval and contained electron-dense materials and a crystalloid. Type II granulocytes had small eccentric nuclei and were subdivided into IIa and IIb granulocytes by electron microscopic analysis. Granules of type IIa granulocytes were furnished with an electron-dense rim. Granules of type IIb granulocytes were larger than those of type IIa, containing randomly distributed electron-dense and electron-lucent materials. Type III granulocytes possessed round nuclei and a few large granules. The granules were filled with regularly arranged fibriform materials and some needle-like structures. Monocytes were morphologically similar to those of mammals.

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A series of experiments were undertaken to determine the most effective route of immunization with a mixture of killed Babesia rodhaini antigen (S antigen) and formalin-fixed Corynebacterium parvum (Propionibacterium acnes) bacterin (CPB) against challenge infection with B. rodhaini 3 weeks later. The mice pretreated with S antigen and CPB mixture intraperitoneally, but not intramuscularly, were significantly resistant to intraperitoneal (IP) or intravenous (IV) challenge with 10⁶ organisms. The survival rates were 70.0 (IP challenge) and 60.0% (IV challenge) respectively. Fairly protective activities were equally produced in mice intravenously pretreated with S antigen and CPB with survival rates of 60.0% against IV challenge, but 30% against IP. These results indicated that the IP injection of S antigen and CPB mixture is desirable route for immunization against subsequent IP or IV challenge with B. rodhaini. On the other hand, lower protective effect was reconfirmed in the mice treated with S antigen and Freund's Complete adjuvant, regardless of immunization routes in the additional experiment. The survival rates were 33.3, 14.3 and 11.8% in the intraperitoneally, intramuscularly and subcutaneouly-treated mice respectively against IP challenge with 10⁶ organisms.

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Gravid sclerosis of the uterine artery was histologically examined in multiparous cows. This sclerosis was characterized by a thickening of the intima and a lamination of the internal elastic lamina. It was clearly observed at non-pregnant and early-pregnant stages and in the late postpartum period, but it could not be detected at term-pregnant stages and in the early postpartum period. These results suggest that the gravid sclerosis disappears during pregnancy and re-establishes after parturition. The importance of local factors, e.g. hemodynamics, in the cyclic appearance of gravid sclerosis is discussed.

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Methemoglobin formation and reduction in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) were compared with those in normal canine cells (LK cells). Nitrite-induced methemoglobin formation in hemoglobin solutions indicated that the hemoglobin from HK cells was oxidized at essentially the same rate as that of LK cells. However, methemoglobin formation in HK cells was slower, due to the inhibition by high glutathione (GSH) concentration. Methemoglobin reduction was allowed to take place on nitrite-treated and washed erythrocytes in a glucose medium and was reduced more rapidly in HK cells than in LK cells. During the reduction, the amounts of lactate and pyruvate increased more rapidly in HK cells, indicating enhanced glycolysis in HK cells. It is thus evident that the hemoglobin of HK cells is more securely protected from nitrite-induced oxidation by the GSH presence in great excess and by the increase in glycolysis.

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A total of 132 embryos were recovered from 17 superovulated donor cows 7 d after estrus. Seventy-four embryos were selected and assigned to 2 treatment groups. The number of whole embryos that were directly transferred (Group A) and bisected (Group B) were 44 and 30 embryos, respectively. Sixty demi-embryos were produced from 30 morulae to blastocyst-stage embryos that were bisected. One hundred-three embryos, including whole and demi-embryos without zonae pellucidae, were nonsurgically transferred. Only one whole or demi-embryo was transferred to each recipients. The pregnancy rate for whole embryos (A) was 63.6% (28/44), while for demi-embryos (B) it was 74.6% (44/59). There was no significant difference between the pregnancy rates of whole embryos (A) and bisected embryos (B) transferred 7 d after estrus. Forty-three calves including the 14 sets of identical twins were obtained from 30 original embryos (143.3%) using the embryo bisection technique.

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Oviduct epithelia obtained from 32 cows were cultured. The oviducts were classified into follicular and luteal phases and divided into ampulla and isthmus regions. The epithelial cells were dissociated by enzyme digestion and cultured in plastic dishes with Dulbecco's modified Eagle's medium/Ham's F12 (1:1) containing 10% calf serum. After enzyme treatment, the epithelial suspension showed free ciliated and non-ciliated cells, and cell mass. The non-ciliated cells contained secretory granules in the cytoplasm. The cell mass was composed of ciliated and secretory cells. The cell mass adhered to the dish within 12-24 hr, while the free ciliated cells attached on Day 2 of the culture. The cells grew into confluent monolayers on Day 4. The cell monolayers contained ciliated and non-ciliated cells. The monolayered non-ciliated cells showed a few secretory granules. When the cells were further cultured without subculturing, ciliary activity diminished on Day 5 and was rarely detected on Day 9. When the cells were subcultured on Day 3, ciliary movement was detected on the monolayers for only 2 days. Cell mass that did not adhere to the dish and remained floating in the medium formed ball-like structures on Day 2. Active ciliary beating was observed on the cells that were cultured in the medium supplemented with 10^-5 and 10^-9 M estradiol-17β, however, the ciliary activity diminished on Day 5. No difference in the cell growth was observed between the follicular and luteal phases or between the ampulla and isthmus regions.

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The effects of vitamin A deficiency on the pituitary-gonadal function were examined by measurements of serum and pituitary level of pituitary hormones and serum testosterone concentration, and by investigations of histological changes in the testis and the pituitary gland in vitamin A-deficient (VAD) and supplemented (VAS) rats. The growth of VAD rats was retarded and their body weights were decreased after 9 weeks of experiments and attained about one half of the weight of control animals at 12 weeks. In the VAD rats, serum testosterone concentrations were decreased significantly compared with those in the VAS controls. Serum and pituitary concentrations of GH were significantly lower but those of LH were slightly lower in the VAD rats than those in the controls, while the serum FSH concentration was significantly higher than that in the control rats. The seminiferous tubules in the testes of VAD rats were comprised largely of Sertoli cells and a reduced number of spermatogonia and contained fibrous formation in their lumen. In the pituitary gland, GH cells were significantly reduced in number in the VAD rats, but gonadotropic (GTH) cells were increased remarkably in size and number, showing hypertrophy and vacuolation similar to those in castration cells. The cytological changes in the pituitary gland and the increased discharge of FSH represent a secondary and compensatory change similar to that seen following castration and vitamin E deficiency.

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The purpose of this study is to elucidate some morphological characteristics of the bar-structure in bovine erythrocytes infected with Theileria sergenti. The bar-structure, the veil or the both were observed in infected erythrocytes. Infected bovine erythrocytes were classified into four types according to the included structures. Infected cells containing bar-structures increased with the progress of parasitemia. In Giemsa-stained blood preparations, bar-structures appeared purplish-red and measured a mean of 1.6μm in length and up to 0.1μm in width. Bar-structures were usually straight, sometimes S-shaped, and located in the periphery of infected erythrocytes. In the direct fluorescent antibody test with T. sergenti-positive bovine serum both piroplasms and bar-structures exhibited fluorescence. However, in the indirect fluorescent antibody test with monoclonal antibodies against piroplasms only piroplasms showed a highly specific fluorescence. Electron microscopy revealed that bar-structures were vesicular in structure, surrounded by a double membrane connected to the host cell membrane.

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