A relationship between luteal hypoplasia and ovulation was analyzed in cows. Adult cows of the luteal phase were injected with gonadotropin releasing hormone analogue (GnRH-A) 16 hr (group I of 4 head), 32 hr (group II of 4 head), or 48 hr (group III of 3 head) after injection with prostaglandin F_2α-analogue (PGF_2α-A), and then examined for the occurrence of luteal hypoplasia. Ovulation took place at 32-46 hr (37.5 hr on the average) in group I and 9-33 hr (20.0 hr on the average) in group II after injection with GnRH-A. Unsatisfactory luteinization and substantial shortening in estrous cycle were observed in all the cows of groups I and II. In group III, ovulation took place 12-32 hr (24.7 hr on the average) after injection with GnRH-A, and luteinization was satisfactory in one, but the corpora lutea formed were small in the other two cows. All the cows, except one in each of groups II and III were examined for changes in the plasma progesterone level after the induced ovulation. Low progesterone levels were observed in groups I and II, while in group III, the levels took almost the same course of changes as the normal estrous cycle, indicating the presence of a satisfactory luteinization. However, temporary increase was observed in another cow in this group. It is concluded that luteinization became unsatisfactory when ovulation was forced to take place in cows while the ovarian follicles were still developing.

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Usability of repeated liver biopsies was examined for the estimation of drug residual level in the calf liver. The liver biopsy was carried out under ultrasonic observation. Twenty four calves were given 20, 000 units of benzylpenicillin (PC-G) per kg body weight. Twelve calves were biopsied and sacrificed within 2 hrs after the biopsy at 3 and 7 days after the administration, and 7 calves were sacrificed at 3, 7, 14 and 21 days after the administration to take the liver samples (the serial sacrifice group). Five calves were biopsied repeatedly 15 times during 21 days following the administration to obtain the liver parenchyma (the repeated biopsy group). There was no significant difference (p=0.05) in PC-G concentrations between the biopsy samples and the samples obtained by sacrifice immediately after the biopsy, nor between the samples of "the repeated biopsy group" at 3 and 7 days after the administration and those of "the serial sacrifice group" at the corresponding points. Pharmacokinetic parameters and the decay lines calculated from the mean concentrations in both groups were almost identical. These results suggest that the estimation of drug residual level in the calf liver taken by repeated biopsies in place of the serial sacrifice is useful to determine the withdrawal time of drugs remaining predominantly in the liver.

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Gamma globulin fraction was obtained from serum or colostrum whey of cows vaccinated with bovine rotavirus type 1 and bovine coronavirus and reconstituted artificial colostrum (AC). A field study was conducted in 2 beef herds, where the prevalence of both viruses had been confirmed, in early spring when spontaneous diarrhea was prevalent. At the time of birth calves were divided equally into 2 groups. One group was fed an AC diet additionally 3 times at 2 day-intervals soon after birth. The second group was used as a control. The supply of AC did not entirely eliminate any clinical disease. There were some differences between the AC and control groups in the age in days of onset, duration, and severity of diarrhea. No differences were fround in the average level of circulating immunoglobulin or antibody in the calf serum between the 2 groups. In a twin the protective effect of the AC diet was directly observed on calf serum immunoglobulin level and on symptoms. The additional feeding of the AC diet would be useful for preventing clinical diarrhea in newborn calves.

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Superoxide dismutase (SOD) activity and the percentage of hemolysis were measured with bovine erythrocytes incubated for 24 hrs at 37°C in 0.1 and 0.5mM copper sulfate solutions. The SOD activity decreased rapidly, as only 50% and 15% of the initial erythrocyte content (2.8×10³units/g Hb) were found to be intact after 12 hrs incubation in 0.1 and 0.5mM copper sulfate solutions, respectively. Prior to the development of hemolysis and during the hemolytic phase, we detected a decrease in SOD activity. The percentage of hemolysis reached 30% and 80% after incubation for 24 hrs with 0.1 and 0.5mM copper sulfate solutions, respectively.

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An experiment was carried out with 4 non-lactating ewes to determine the effects of replacing cold (C period: about 5.4°C) drinking water with warm (W period: about 26.8°C) on water, Mg or Ca metabolisms in winter. By changing from C to W period, the water intake and urine volume increased in 60-102% (P<0.05) and in 65-118% (P<0.05), respectively. Moreover, there was a significant positive correlation between the water intake and urine volume throughout the experimental period (P<0.001). Serum Mg concentration rose significantly in all ewes, and urinary Mg level decreased in three ewes. In the remaining ewe, water intake, urinary Mg and Ca excretions increased remarkably. The serum Ca concentration declined significantly in 2 ewes (inclusive of No. 3). There was a significant negative correlation between serum Mg and urinary Ca concentrations (P<0.01) throughout the experimental period. These results suggest that the water intake in sheep decreases if cold drinking water is supplied in winter, resulting in a decrease in the absorption of Mg and a decline in the serum Mg concentration. Renal function may be important for regulation of water, Mg and Ca metabolisms in cold environment. The absorption mechanism of Mg (mainly by diffusion) might differ from that of Ca.

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A cat which was fed muscles of a goat shed Toxoplasma-like oocysts (G-8 isolate) 43 days after feeding. The oocysts were subspherical to ovoid, measuring 12.7×11.5μm (unsporulated oocysts) and 13.0×11.4μm (sporulated oocysts). When mice were inoculated orally with 10⁵ oocysts, tachyzoites were observed in the kidney, lung, mesenteric lymph nodes, small intestine and spleen of the mice 1 to 15 days after infection. Cysts were seen in the brain and various muscles. Cysts measured 25-224×2-80μm in the skeletal muscle and 35×35μm in the brain 181 days after infection. Bradyzoites in mice were not infectious to other mice. Sometimes antibody to Toxoplasma was observed in the mouse sera but the titer was low. Cats fed bradyzoites began to discharge oocysts from 7 to 8 days after ingestion. But cats inoculated with 10⁶ oocysts of G-8 isolate shed no oocysts within 60 days after inoculation, and cats fed the muscles of these oocyst-inoculated cats shed no oocysts. The life cycle of G-8 isolate was obligatorily heteroxenous, and cysts in the muscles of the infected mice morpholoigcally resembled those of Hammondia hammondi (Frenkel and Dubey 1975). G-8 isolate was identified to be H. hammondi.

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To evaluate the systems of enzyme-linked immunosorbent assay (ELISA) for titration of monoclonal antibodies, a monoclonal antibody, E13, against Clostridiurn botulinum type E derivative toxin was examined for the binding property. E13 neutralizing the toxin did not bind the toxin but bound the toxoid fixed on the surface of the wells of the polystyrene assay plate. Preincubation of peroxidase-labeled E13 in a toxin solution suppressed the binding of E13 to the fixed toxoid. Radioimmunoassay demonstrated the direct binding of E13 to the toxin. When the surface of the wells was coated with polyclonal antibody against the toxin and the antigens were fixed on solid phase (sandwich ELISA), E13 reacted to both toxin and toxoid. These results indicate that the site of the toxin molecule binding E13 is masked by fixing onto the surface of polystyrene material, absorption of the toxin and toxoid to the surface may be attained in different modes, and random binding may result from formalinization of the toxin. Moreover, these findings may warn that the conventional ELISA in which the solid phase is coated directly with the antigen sometimes fails in finding the positive antibody reaction.

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The characteristics of virus (strain H-83) isolated from apparently healthy masu salmon (Oncorhynchus masou) were investigated. The isolate replicated in line cells derived from salmonid fish but not in those from non-salmonid fish. Cell monolayers infected with the virus showed cytopathic effect characterized by syncytia with intranuclear inclusion bodies and many metachromatic granules in the cytoplasm. Enveloped virions were estimated to be approximately 250nm in diameter and possessed hexagonal nucleocapsides of 103 to 118nm. The physicochemical and morphological features indicated that the strain H-83 is a herpesvirus. Viral antigens were detected in the cytoplasm and membrane of virus-infected cells by immunofluorescent antibody test. Cross neutralization test among strain H-83 and two other herpesviruses, Herpesvirus salmonis and Oncorhynchus masou virus (OMV), indicated that these three viruses were serologically different from each other. Further, the restriction endonuclease cleavage analysis of viral DNA showed that the cleavage pattern of the strain H-83 DNA was similar to that of OMV, while apparently distinct from H. salmonis.

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Acid phosphatase as a representative lysosomal enzyme was detected in chicken juxtaglomerular (JG) cell, and then the phagocytic ability of JG cells was compared to that of glomerular endothelial cells (EN) and mesangial cells (MS) by injecting colloidal carbon into the renal vascular system. Acid phosphatase activities demonstrated in the granules and the so-called GERL of JG cells were more intensive than those found in MS and EN. However, considerable variation in the reactivity was noted among the individual granules in JG cells, varying from no reaction to remarkable reaction products. During the 1st day to 4 weeks of carbon administration, the EN of glomerular capillaries, and MS ingested variable amounts of carbon particles. In the JG cells, however, no particles could be observed during these periods. Based on the results, it was suggested that JG cells do not possess a phagocytotic nature and are clearly distinguishable from the MS and EN of chicken kidneys by intravascular administration of carbon particles. The roles of acid phosphatase in JG cells were also discussed from a viewpoint of renin production.

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Macroscopic and microscopic observations were made on the healing process of cut ends in the medial patellar ligaments during a period from 7 to 201 days after desmotomy in 24 normal cattle. Lameness was not seen in any of the cattle after the operation. Adherence at the severed site was not observed in any of the samples. The connective tissue already had bound up the cut ends 7 days after the operation. The repair tissue was grossly distinguishable from the ligament and surrounding connective tissue in all samples. On the basis of gross observations, the authors divided the repair period into three stages. In the early stage, there was no significant increase in the distance between the ends of the cut ligament since the operation, in the intermediate stage the distance tended to increase, and in the last stage the distance decreased. The mean distances observed in each of the stages were 19mm±S. D. (range 17 to 23mm), 23mm±S. D. (range 13 to 33mm) and 14mm±S. D. (range 7 to 27mm), respectively. Microscopic observations, demonstrated loose connective tissues in the gap during the early stage were loose connective tissues. Thereafter, fibroblasts closely aligned in longitudinal direction in the area closest to the cut ends. This arrangement progressed gradually towards the center of the gap. In the sample taken 201 days after operation, however, the cell density in the gap was identical to that of the ligament tissue. These results suggested that connective tissues in the gap gradually changed to ligament tissues.

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The structure of intra-erythrocytic stages of Babesia ovata was studied by electron microscope. The merozoite had piriform shape or paired piriform shape and was surrounded by an outer membrane and a double inner layer of the pellicle. Anterior and posterior polar rings, micronemes, rhoptries, mitochondrion and endoplasmic reticulum were observed in the cytoplasm. A single large nucleus and a characteristic spherical body were also recognized. A continuous single membrane and light electron dense region bounded the spherical body, which contained an aggregation of granular material stained with varying intensity. The trophozoite was pleomorphic in shape and surrounded by a single membrane. No conspicuous structures, except a nucleus and some micronemes, were observed in the cytoplasm. The trophozoite developed to the multiplication stage through a transformation process and became a typcal paired piriform merozoites after association with twin daugher-cells. The ultrastructure of intra-erythrocytic stages of B. ovata resembled that of B. bigemina.

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The effect of an intravenous injection of glucose (1.25 mmoles/kg) on plasma insulin, glucagon and 11-hydroxycorticosteroids (11-OHCS) was compared in castrated Landrace piglets between the thrmoneutral (T.N., 22°C) and the cold (2°C) environment. Glucose-induced insulin secretion was accelerated by the cold exposure, differently from the previous results obtained from sheep. Plasma glucagon did not change after the glucose injection in T.N. environment, but declined temporarily 3 min after the glucose injection in the cold environment. Plasma 11-OHCS after the glucose injectjion declined slightly in T.N. environment, and more evidently in the cold environment. The magnitude of the reduction in plasma 11-OHCS was the largest on day 4 of the cold exposure.

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Seroepizootiologic survey among dogs and cats in the South-Kyushu area and serological tests for diagnosis of rotavirus infections were investigated by using canine RS 15 and porcine S80 strains. Although no clinical sign was observed in the puppies orally inoculated with RS 15 strain, all of them mounted an immune response and seroconversion within a week or so was detected by plaque reduction neutralization test (PRNT), hemagglutination-inhibition (HI) and reverse passive HI (RPHI) tests. It took more than 2 weeks to detect complement-fixation (CF) antibody in their sera. HI test was applied for detection of type-specific antibody instead of PRNT because antibody titers obtainable by both tests were generally correlative. RPHI test was considered to be applied routinely for serological diagnosis of rotavirus infections of dogs and cats. CF, RPHI and HI antibodies against RS 15 strain were detected in 25.0%, 42.1% and 41.9% of dog serum samples, respectively. About 60% of antibody-positive sera detected in the dogs were considered to be derived from infections with the same serotype as RS 15 strain. Although rotavirus antibodies were detected in the cats as well, it was supposed that different serotype(s) might have been epizootic in the population. HI antibody against RS 15 strain was detected in 13.8% of human sera.

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In total, 200 slaughtered pigs were bacteriologically, serologically, and pathologically examined. Haemophilus pleuropneumoniae was isolated from the nasal cavities and the lungs of clinically normal pigs. A total of 140 H. pleuropneumoniae isolates (serovar 2) were obtained from the nasal cavities and/or lungs of 106 (53%) carrier pigs originated at 48 farms; 103 (51.5%) from the nasal cavities and 37 (18.5%) from the lung lesions of porcine Haemophilus pleuropneumonia (PHP). Thirty-four of 37 pigs with PHP lesions harbored the organism both in the lung lesions and in their nasal cavities. Antibodies were detected in 103 (51.5%) of 200 pigs by the complement-fixation (CF) test. The organism was isolated from the nasal cavities of 93 (90.3%) of 103 seropositive pigs, whereas only 10 (10.3%) from those of 97 seronegative ones. None of the organism was isolated from the lungs of seronegative pigs. Most of the pigs with PHP lesions showed high CF antibody titers (≥1:16). Most of carrier pigs, therefore, might be detectable by the CF test, irrespective of location of the organisms in the host.

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Naturally occurring anti-Ca and anti-Cb isoantibodies that can be used for judging red blood cell groups were detected in the plasma of cats. Anti-Ca antibodies were observed in 66.7% of the plasma of the Cb(+) type cat, and anti-Cb antibodies in 27.3% of the Ca(+) type cat plasma. The agglutinin titers of anti-Ca and anti-Cb plasma were 1:64 to :128 and 1:2 or lower, but hemolysis was not seen with anti-Cb plasma. There was no relationship between the change in these titers and the season. Anti-Ca was seen in the IgM fraction. The Ca(+) type was seen in 99.0%, the Ca(-) type in 1.0%, the Cb(+) type in 10.7%, and the Cb(-) type in 89.3% of the 299 cats examined. Combining the red blood cell types, the Ca(+)Cb(-) type was observed in 89.3%, Ca(-)Cb(+) type in 1.0%, Ca(+)Cb(+) type in 9.7% of the cats, but the Ca(-)Cb(-) type was not found. There was no relationship between sex, or the breed and the red blood cell groups. The percentage of cats with agglutinin in the plasma reacting to the Ca(+) type and red cells tended to be higher with age.

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Two cell clones producing monoclonal antibodies (MAb) to the RT strain of Tyzzer's organism ('Bacillus piliformis'), of rat origin, were established by fusion of immunized BALB/c mouse splenocytes and NSl1 cells. Western blotting and immunofluorescence (IF) revealed that one of the MAb recognized an bacterial antigen having a molecular weight (M.W.) of 54, 000 and combined with the bacterial flagella, while the other recognized another antigen with M.W. of 73, 000 and combined with the bacterial cell surface but not with flagella. The former reacted also with antigens of mouse derived bacterial strain MSK of B. piliformis the latter did not. Both MAb were protective for the challenge infection with B. piliformis, reducing the number of necrotic foci in the liver of infected mice as well as that of cytopathic plaques on mouse hepatocyte culture.

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A flexible alligator forceps was developed as a new instrument for removal of heartworms in the pulmonary arteries without conducting thoracotomy. It was 570mm to 620mm in overall length, and 380mm to 430mm in length of the insertion part. The forceps measured 3mm in diameter, and was operated by flex-manipulation and grasping mechanisms. In 9 dogs, 81.1% (90/111 worms) to 100% (9/9 worms) of heartworms were removed from the pulmonary arteries after 4 to 33 manipulations. Injuries after treatment found at autopsy were only a few petechiae in the right atrium and/or tricuspid valve and scratches in the jugular vein.

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Hemolysin was prepared from the culture supernatant of Haemophilus pleuropneumoniae, and its cytocidal and anti-phagocytic effects were investigated with pulmonary macrophages (PMP) obtained from the pig, mouse, guinea pig, or cotton rat. The hemolysin demonstrated both cytocidal and anti-phagocytic activities with the cotton rat, guinea pig, and pig PMP cells, but not with the mouse PMP cells. The species specificity in the cytocidal and anti-phagocytic effects with the PMP cells may be important in analyzing for the pathogenesis of the organisms to various animal species.

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An unknown metabolite of sulfadimethoxine (SDM) in chicken excreta was identified as N¹-(2-methoxy-6-hydroxy-4-pyrimidinyl) sulfanilamide by mass spectrometry, infrared spectroscopy, and etc. It was found to be a principal metabolite in the excreta of chicken that received SDM and considered as the causal substance of the nonlinear pharmacokinetics of SDM in chicken. This is the first report, in which the compond is identified as the major metabolite of SDM.

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A real-time ultrasound scanner was used for pregnancy diagnosis in rabbits. The images of fluid-filled round uteri were recognized first on the echograph after day 6 of gestation. Accuracy of the diagnosis approaches 100% on day 9 of gestation and the high accuracy was maintained throughout pregnancy period. After day 18 the fetal vertebra and the beating heart could be observed, and the death of fetuses was able to diagnose. Fetal age could be estimated by measuring the uterine diameter. Litter size also could be estimated with the errors of ±2. A real-time ultrasound can be used for the diagnosis of rabbit pregnancy as an accurate, rapid and easy method.

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Renal cell carcinoma in a 14 years old cow was reported. Grossly, the lesions were yellowish or whitish milky in color of various sizes in the renal cortex. The neoplastic cells were oval or polymorphic in shape with foamy eosinophilic cytoplasm. The round or oval nucleus was tended to be located eccentrically. Neoplastic transformation of the renal epithelial cells was suggested morphologically. Metastatic lesions were also found in the renal and hepatic lymphnodes, spleen and omentum.

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Hemagglutinin (HAin) of canine rotavirus RS 15 strain was investigated. Polyacrylamide gel electrophoretic and immune-blotting analysis of the virion indicated 5 major rotaviral polypeptides whose molecular weight ranged from 38K to 119K. The HAin of RS 15 strain existed in the outer capsid of virus. Results of hemagglutination-inhibition (HI) test with other rotavirus strains indicated a possibility that the HI test might be of use for detection of type-specific antibody.

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A method is described for isoelectric focusing of murine nonspecific esterases on polyacrylamide gel using hematocrit capillary tubes. For an analysis, less than 50μg of mouse tissues and only 30μl of gel mixture were needed. And it required only about two hours for a complete analysis. Using this method to analyze nonspecific esterases extracted from mouse tissues, it was possible to identify a composite total of 31 bands of nonspecific esterases.

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The effects of 105K glycoprotein (105Kgp), purified from sera of MDCC-MSB1-41C (MSB1-41C)-transplanted chickens, on the tumor cell growth were examined. The tumor growth in the 105Kgp-injected and cell-transplanted chickens was significantly accelerated in comparison with that of cell-transplanted control chickens. The enhancement of the in vitro cell growth of MSB1-41C, on the other hand, was stimulated by the addition of 105Kgp. These facts suggest that the 105Kgp may act as a enhancing factor for the tumor cell growth.

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By biological and physical properties, mink enteritis virus (MEV) was compared with feline panleukopenia virus (FPLV) and canine parvovirus (CPV). MEV had about the same properties as FPLV in hemagglutination-inhibition tests, in the propagating ability of the viruses in MDCK cells and in the hemagglutinability, whereas the heat stability at 80°C, and the restriction enzyme digestion patterns of the viral replicating form DNAs were similar to those of CPV.

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Induction of the maternal behavior in sexually inexperienced male rats exposed continuously to young foster pups was tested. The vomeronasal organ ablation accelerated maternal behavior of males toward foster pups.

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A 24-month-old Holstein heifer was examined because of a loud heart murmur and exercise intolerance. Echocardiographically, a high ventricular septal defect with overriding aorta, right ventricular hypertrophy and pulmonary dilatation were detected. Besides, cardiac catheterization revealed slight elevation of the pulmonary arterial pressure, and Eisenmenger's complex was diagnosed.

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Twenty neurons in the subfornical organ (SFO) were antidromically activated by electrical stimulation of the hypothalamic supraoptic nucleus (SON) in the rat. Electrical stimulation of the lateral hypothalamic area (LHA) excited the activity of 12 identified units, but did not affect that of the remaining units. The excitatory responses to LHA stimulation were blocked by microiontophoretically applied saralasin (Sar), an angiotensin II (AII) antagonist. These results suggest that the LHA has an excitatory influence on the activity of SFO neurons projecting to the SON and that the influence may be mediated by AII receptors.

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Long term oral treatment with taurine effectively blocked spontaneous epileptic seizure in five cats. Without concomitant use of anticonvulsant drugs, taurine blocked or markedly decreased clinical symptoms of epilepsy including the characteristic spikes on electroencephalograms.

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The enzyme-linked immunosorbent assay (ELISA) method was used to detect antibodies to equid herpesvirus type 1 (EHV-1) in horses. The antigen was a preparation obtained by solubilizing virus-infected cells with Nonidet P-40 (NP-40). The ELISA values were almost constant in the antigen protein concentration range of 5-1μg/well in the reaction with EHV-1 positive serum. The average ELISA value of the negative serum in 11 cases was 0.236. Changes in ELISA values and neutralizing antibody titers were compared using sera collected at intervals of about 1 week after inoculation of EHV-1 inactivated vaccine. The ELISA values and neutralizing antibody titers showed similar changes, but the detection sensitivity of ELISA was excellent in sera collected 1-2 weeks after inoculation. From these results, it might be possible to apply ELISA for the detection of EHV-1 antibodies.

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T lymphocytes bearing receptors for IgG (T_γ cells) were isolated from bovine peripheral blood lymphocytes (PBL) by the two serial methods of sheep erythrocyte(E)-rosette sedimentation and bovine erythrocyte-antibody-rosette sedimentation. T_γ cells were cocultured with bovine non-E-rosette forming lymphocytes or PBL in the presence of pokeweed mitogen for 5 days at 37°C. Plaque forming cells (PFC) in these cultures were determined by the reverse hemolytic plaque assay. Bovine T_γcells were confirmed to provide the suppressive effect on the development of PFC response.

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Colchicine was subperineurially injected to the tibial nerve in chickens, and it caused a persistent partial paralysis of the leg on the injected side for 60 to 90 days. The injected chickens gradually recovered. Acetylcholinesterase accumulation in the nerve after 24 hr ligation was low until day 36, but high at day 128 of post-treatment. Weight of the gastrocnemial muscle on the injected side decreased to about a half of that of the other side at day 32, then recovered considerably at day 128. The chicken seemed to be a suitable experimental animal to study the mechanism of colchicine effect on the peripheral nerve.

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A 14-year-old female dog of Ainu breed was admitted to a veterinary hospital. It showed signs of congestive heart failure with accompanying chronic interstitial nephritis. In the pathological examination, a focal microinfarct was present in the myocardium and it was surrounded by an area of fibrosis in the left ventricular wall. Before death, there were no findings clearly suggesting the presence of this type of lesion obtained by electrocardiographic examination and serum analysis, but marked increase in CPK-MB/ total CPK activity was observed.

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