The activity of rat liver glutathione-insulin transhydrogenase (GIT) was measured by HPLC. The degradation of fluorescein isothiocyanate-I (FITC-I)-labeled insulin is separated into several peaks, which are bound different amount of FITC-I. We selected mono-fluorescein-thiocarbamylated insulin to estimate the decrease of insulin content and it became possible to assay GIT activity. This novel method was time-saving and simple, and this system could utilize instead of previous method.

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Diagnostic significance of electrophoretic findings of serum protein in cows with traumatic pericarditis was evaluated. Affected cows were classified into 3 groups according to autoptical findings: fibrinous, sero-fibrinous, and purulent types. Slight hypoprotenemia, moderate hypo-albuminemia, slight hyper-α globulinemia and tendency of hyper-β globulinemia were commonly observed in the affected cows. The level of γ globulin tended to be lower in the cows with fibrinous or sero-fibrinous, and higher in purulent pericarditis than the level in healthy cows. In the serum protein electropherograms of the cows with fibrinous or sero-fibrinous pericarditis, there was pathognostic pattern composed of slender albumin, acute shape of α globulin with a broad rising accompanied by double peaks and with main peak migrating toward the albumin side, tendency of rising β globulin fraction, and large indentation between β and γ fractions. These findings except for the slender albumin fraction, however, was not or poorly observed in purulent pericarditis. Electrophoretic findings were subacute inflammatory pattern with non-selective serum protein losing in fibrinous or sero-fibrinous, and chronic inflammatory pattern in purulent pericarditis.

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Pamaquine and primaquine, the well known antimalarial 8-aminoquinolines, have not been reported for their anticoccidial activity. A series of battery experiments was conducted to investigate their activity against a laboratory strain of Eimeria tenella, E. necatrix, E. acervulina, E. maxima, or E. brunetti and revealed that both drugs were effective against E. tenella and E. necatrix, but not against the other three species. Pamaquine suppressed the symptoms of E. tenella induced coccidiosis at concentrations above 125 ppm in feed and primaquine controlled the clinical signs as well at levels above 31.2 ppm. The activity against E. necatrix was observed with pamaquine at 250 ppm and with primaquine at levels above 125 ppm. Pamaquine showed a tendency apparently to reduce body weight gain at 125-500 ppm, whereas primaquine showed the same tendency at 500 ppm. In a concomitantly conducted experiment, this adverse effect of pamaquine was averted in its molecular complexes with benzophenone, nitropyrazole, dinitrobenzoic acids and quinoline, and in its salts of sulfate or zinc chloride, and yet these compounds retained the same anticoccidial activity as of pamaquine. This suggests that these compounds had the broadened safety margin. Judging from their susceptibility to these compounds, E. tenella and E. necatrix will have similar metabolic functions to those of blood cell parasitizing protozoa like plasmodia and prioplasma, which are easily suppressed by this class of compound.

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Autoantibodies and erythrocyte membrane proteins were analyzed in a case of a dog with autoimmune hemolytic anemia (AIHA). In crisis phase, antiglobulin test was positive. The eluate from the erythrocytes of the dog with AIHA gave agglutination against autologus erythrocytes. Immunoglobulin subclasses in the eluate were revealed to be IgG and IgA by the double diffusion test. Comparing the SDS polyacrylamide gel electrophoretic patterns of erythrocyte membrane proteins between the crisis and remission phases, there was a change in the protein on protein 4.1 region. However, there were no changes in blood group typings in two phases.

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Yersinia enterocolitica O3, O4, O5A, O5B, O6, O6, 30, O9 and O13 and Yersinia intermedia were examined to clarify their survival in natural soil, river water and well water. The O3 strain disappeared most rapidly from soil at both temperature of 4°C and 20°C and from river water at 20°C. Although the O5B and O9 strains disappeared before the O3 strain did from river water at 4°C, the O9 strain survived as long as non-virulent strains did in soil at 4°C. The O3, O5B and O9 strains survived longer at 4°C than at 20°C in soil and river water. Non-virulent strains of O4, O5A and O6 survived longer in well water than in soil and river water. Although the O3 and O5A strains disappeared from supernatant filtered with 100μm and 5μm pore size filters, they maintained their viable cell numbers in supernatant of soil filtered with 0.22μm pore size filter and in the autoclaved supernatant.

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Chuzan virus agglutinated erythrocytes of several species of animals including bovine. The hemagglutinating (HA) activity against bovine erythrocytes was dependent on NaCl molarity and was expressed best at 0.6 M, but it was independent of pH and temperature. Three strains of Chuzan virus isolated from 2 cows and a pool of culicoides midges had indistinguishable HA antigenicity. All cattle infected with the virus developed high titers of hemagglutination inhibiting (HI) antibody which changed in parallel with neutralizing (NT) antibody titers. Correlation between HI and NT antibodies was very high and the antibodies persisted for one year or more. Therefore it was concluded that the HI test is applicable for survey of Chuzan virus infection among cattle in place of the NT test.

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The disposition and elimination of sulfadimethoxine (SDMX) in the skin of broiler-chickens were investigated. The administration of SDMX, in drinking water, at a concentration of 1, 000 ppm for 5 days demonstrated that the SDMX was eliminated much more slowly from the skin than from the other tissues or plasma. These results were duplicated and confirmed in another experiment, in which a single dose of 200mg/kg BW of SDMX was administered via a stomach tube. No significant difference in the SDMX residue level was observed between the broiler-chickens that had their skin surface sealed versus the non-sealed animals and that had ingested SDMX in their drinking water. This illustrated the higher SDMX residue in the skin was not attributable to external SDMX contamination from the drinking water, feces or urine. In addition, there was no significant difference among the SDMX residue concentrations in the thoracic, dorsal and leg skin samples, following an intravenous injection of SDMX (30mg/kg BW). This indicated that the SDMX was distributed evenly throughout the entire skin area of the broiler-chickens.

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The percentages of morphologically transferable embryos obtained from frozen-thawed demi-embryos which were embedded with or without agar, and from those with or without zonae pellucidae were 26.3% (5/19), 36.4% (8/22), 39.5% (15/38) and 40.0% (22/55), respectively. No significant differences were observed between these groups. Development to calves of frozen-thawed demi-embryos with or without zonae pellucidae was 25.0% (3/12) and 26.7% (4/15), respectively. There was also no significant difference between them. On the trial for production of identical twin calves of different ages, the pregnancy rates of fresh and frozen demi-embryos after transfer were 69.2% (9/13) and 11.1% (1/9), respectively. Out of 13 fresh demi-embryos and 9 frozen demi-embryos transferred, only one pair of identical twin male calves of different ages were produced. This frozen-thawed demi-embryo was stored for 43 days in liquid nitrogen before thawing and transfer. These twin calves were confirmed to be identical by blood typing. Although these calves had different birth dates, their growth rates indicated similar developmental patterns. We suggest that it is possible to produce identical twin calves of different ages. This possibility would be useful for predicting the sex, milk producing ability and progeny test of a pair of demi-embryos before a decision to transfer the other half of a pair is made.

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To detect oncorhynchus masou virus (OMV) in salmonid fish, we prepared recombinant plasmids from two distinct BamHI digests of OMV DNA as a probe. Viral DNAs could be detected two weeks earlier before the virus was isolated from asymptomatic or dead fish. Using the probe, viral DNAs were detected from fish reared in fish hatcheries or fish captured in a river in Hokkaido. The sensitivity limit of detection was estimated to be 10 copies of viral DNA per cell.

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In order to detect the presence of aflatoxin B₁ (AFB₁), the use of the enzyme-linked immunosorbent assay (ELISA) and recovery test was evaluated. The detection limit of ELISA for AFB₁ was 1pg/assay and the recovery from maize spiked with AFB₁ exceeded 80%. AFB₁ was detected by ELISA in seven out of twelve samples of imported food products including peanut, almond, red pepper, cocoa bean, black pepper, buckwheat, walnut, adlay, soybean, popcorn, and pistachio nut, and by high performance liquid chromatography (HPLC) in four of the samples. However, the content of AFB₁ in these samples was less than 10ng/g of the minimum value authorized by the Japanese sanitation law. These results demonstrate that ELISA is more sensitive than HPLC and imported food products are broadly contaminated with AFB₁

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Induction of arginase activity in mouse myeloid leukemic M1 cells by treatment with recombinant human tumor necrosis factor (rH-TNF) or TNF-elicited mouse serum (TNS) were examined in vitro. M1 cells differentiated into macrophage-like cells by addition of rH-TNF or TNS. The differentiated cells expressed phagocytic function and did not grow anymore. Cytolytic effect of rH-TNF or TNS was not observed. The differentiation of M1 cells into phagocytic cells by the TNS treatment was more rapidly than that by the rH-TNF treatment though TNS contained 25-time less amounts of TNF indicating species specificity of TNF action on myeloid leukemic cells or TNS containing other differentiation factor (s). ³H-ornithine formation from ³H-arginine is catalyzed by arginase (EC, 3.5.3.1). The enzyme product increased in the M1 cell culture medium by the treatment with rH-TNF or TNS. The arginase activity statistically correlated with the appearance percentage of differentiated cells with phagocytic function. These results suggest that in vitro differentiation of M1 cells is accompanied by induction of arginase activity.

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Cardiopulmonary values were determined in dogs with an artificial model of heartworm caval syndrome, which was produced by insertion of heartworm-like silicone tubes into the tricuspid valve orifice and right atrium. Fifteen to 25 tubes with some knots were inserted in 6 dogs (knot group), and 7 to 11 tubes (small-number group) or 29 to 37 tubes (large-number group) without a knot in 3 dogs, respectively. After tube insertion, angiographic contrast medium infused into the right ventricle regurgitated to the right atrium in all cases, and the regurgitation was the most severe in the large-number group. On electrocardiographic findings, the atrial and/or ventricular premature beat developed. The height of a- and v-wave of right atrial pressure curves elevated in all groups. The elevation in v-wave was obvious in the large-number group. The pulmonary arterial pressure tended to fall or to elevate slightly, and total pulmonary resistance increased in all groups. The right cardiac output decreased significantly in all cases. The right heart hemodynamics of the model might resemble those in spontaneous cases without disturbed pulmonary circulation.

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We established a new analytical condition to measure the canine glycated hemoglobin by high performance liquid chromatography (HPLC) using cation exchange column. The canine hemoglobin gave five peaks consisting of 2 major and 3 minor hemoglobin fractions such as HbA1a, HbA1b and HbA1c. Measurement was done in 38 clinically normal dogs and 10 diabetic dogs. Mean HbA1c values (% of total Hb) in normal and diabetic dogs were 2.60 and 6.41%, respectively. And mean HbA1 values were 3.58 and 7.41%, respectively. The mean values of the canine HbA1c and HbA1 in diabetic dogs was higher than those in normal dogs, significantly (p<0.01). Advantages of the HPLC method and applicability for monitoring effectiveness of insulin therapy in the canine diabetes mellitus are discussed.

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Vibrio parahaemolyticus strains D-3 and R-13 were found to be cleared within 7 days from a marine neritid gastropod mollusc, Heminerita japonica, maintained in artificial seawater with salinities of 15, 25 and 35 permil (‰) at 25°C. Escherichia coli strain YS-2 survived at a level of 10² colony forming units per gram in the mollusc maintained in 15‰ water for up to 14 days and fell to non-detectable level within 7 days in a 35‰ salinity group. The ability of H. japonica to clear these organisms seems to be less active than that of a marine species, Nerita albicilla, and more active than that of an estuarine species, Clithon retropictus.

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Protein composition was compared and epitopes were analyzed among the isolates of Anaplasma centrale and A. marginale by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting using bovine antisera and monoclonal antibodies, and enzyme-linked immunosorbent assay. Common and unique proteins were found among the isolates. All isolates tested had a major surface protein with an apparent molecular weight of 38 to 40 kilodalton which had slight molecular size variations between species. This protein was also a dominant immunogen to the host. At least two species-common epitopes, one of which might contain carbohydrate (s), were present on the major surface protein. One species-specific epitope was identified on the major surface protein of A. marginale isolates.

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The fractionation of serum bile acids was performed in the dog, cow, horse, and human by high performance liquid chromatography equipped with an immobilized 3α-hydroxysteroid dehydrogenase (3α-HSD) column. There were significant differences in the bile acid compositions, conjugation patterns and quantities of each bile acid among these animals. Cholic acid was the major primary bile acid in the dog and cow, which constituted 62.9% and 83.5%, respectively, whereas chenodeoxycholic acid was the major acid in the horse and human, which constituted 68.4% and 46.3%, respectively. Taurine conjugates were predominant in the dog and horse, which constituted 94.4% and 85.3%, respectively, whereas glycine conjugates were predominant in the cow and human, which constituted 31.0% and 49.4%, respectively. Although there were several unidentified peaks, it was confirmed that they had a hydroxy group at the C-3α position by chromatographing the samples without the 3α-HSD column.

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Pooled colostral milk samples were collected from apparently healthy Holstein cows. Immunoglobulin (Ig) was partially purified from the colostral milk and concentrated by freeze drying. The Ig powders had neutralizing antibody titers of 1:6, 250 to 1:31, 250 against bovine rotavirus (RV), and reacted with four RV proteins of VP2, VP4, VP6, and VP7. No therapeutic effect of Ig powder showed on clinically serious diarrhea with RV in dairy and beef calves. However, the Ig powder had a preventive effect on the diarrhea in beef calves when it was given soon after calving.

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Reflex responses of the upper airway muscles during experimental nasal and tracheal occlusions were studied in 21 anesthetized dogs breathing through the nose or a tracheostomy (tracheal cannula). Out of them, five animals were examined for maintained negative and positive pressures applied to the isolated upper airway. When the nose was occluded at end-expiration during three consecutive breathings, the electromyographic (EMG) activity of alae nasi (AN) and posterior cricoarytenoid (PCA) muscle markedly increased. The upper airway was subjected to negative pressure, -2.41∼-3.04kPa (n=34) by these nasal occlusions. The activity of upper airway muscles was also increased by tracheal occlusions. However, such augmentation of PCA activity was much less in tracheal occlusions than nasal occlusions, whereas there was the smaller difference in AN activity between both occlusions. During nasal occlusions, a prolongation of the inspiratory time (T_I) was observed. Furthermore, maintained negative pressure applied to the isolated upper airway provoked an augmentation of PCA activity. Such an augmented EMG activity of PCA and the prolongation of T_I were largely diminished by sectioning the superior laryngeal nerves. These results proves that the upper airway muscles play an important role to protect the upper airway from collapsing by nasal occlusions. In addition, it is suggested that a lack of lung volume feedback is substantially concerned with the activation of AN, while negative pressure in the upper airway is essential to stimulate PCA.

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Calcium-dependent and -independent bovine IgG reactive with invertase were isolated by affinity chromatography on invertase-coupled Sepharose 4B and their specificities were determined by competitive binding assays. The binding of ¹²⁵I-labeled calcium-dependent and -independent bovine IgG reactive with invertase to invertase-coupled Sepharose 4B were most effectively inhibited by invertase and mannan. The binding was more weakly inhibited by mannose, glucose, and methyl α-D-mannopyranoside, whereas that was much more weakly inhibited by other monosaccharides and glycoproteins. These findings suggest that the combining sites of calcium-dependent and -independent bovine IgG reactive with invertase may be specific for invertase and/or mannan.

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In Japanese black cattle with large and long-existing hematomas, platelets was impaired in collagen aggregation function in vitro. There was no statistically significant difference from control animals in the tests of PT (prothrombin time) and PTT (partial thromboplastin time) for extrinsic and intrinsic blood coagulation system. Aside from impaired collagen aggregation function, platelets in the hematoma cattle showed the similar aggregation patterns as the normal cattle, when ADP, serotonin (5-HT), thrombin, arachidonic acid, epinephrine and ristocetin were used as agents for inducing aggregation. Decreased aggregation function as well as impaired collagen-induced release response in platelets suggested the hematoma cattle to be of storage pool disease (SPD). The impaired platelet was postulated to be a main cause of the large and long-existing hematomas. All of the hematoma cattle with impaired platelet functions had the eosinophils in peripheral blood of which granules were fewer and larger than normal ones. These large eosinophil granules were peroxidase positive and periodic acid Schiff (PAS) staining negative as typical eosinophil granules.

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A new type of Robertsonian translocation (1/21) was found in Holstein-Friesian breed, and transmitted through the three generations, namely from the bull to 6 out of 11 daughters and 4 out of 7 grand-daughters/sons. Although the father of the bull was not available for cytogenetical studies, it was ascertained that 5 half-sibs of the bull had a normal karyotype. By C-banded karyotypes, the 9 translocation carriers within the three generations had a dicentric centromere in the translocated chromosome. It seems that no morphological effects have been noted to be associated with the translocation in all cases.

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