Twenty-one feline plasma proteins, albumin, α₁-acid glycoprotein, α₁-antitrypsin, anti-thrombin III, ceruloplasmin, haptoglobin, α₂-macroglobulin, hemopexin, transferrin, apolipoprotein (apo) A-I, apo A-II, apo B, apo C-I, apo C-III, immunoglobulin (Ig) G, IgA, IgM, complement (C)3, C4, plasminogen, and fibrinogen, were purified and identified in immunoelectrophoretogram and polyacrylamide gel electrophoretogram. Methods used were polyacrylamide gel electrophoresis (PAGE), SDS-PAGE, agarose electrophoresis, immunoelectrophoresis (IEP), crossed immunoelectrophoresis (CIE), and electro transfer blotting (ETB) for the identification. The positions of 21 feline plasma proteins on immunoelectrophoretogram or polyacrylamide gel electrophoretogram were determined.

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The general structure of the spleen of the Afghan pika (Ochotona rufescens rufescens), recently noticed as a new laboratoly animal of Lagomorpha, was studied by light and electron microscopes. Scanning electron microscopy of micro corrosion casts was also used to visualize microvascular pathways in the pika spleen. The basic cellar compartments of the pika spleen and intrasplenic micro circulations were similar to those of rabbits, but active extramedullary myelopoiesis was noted in the splenic cords of adult pikas. It was also notewarthy that the marginal sinus had perforated wall as well as the splenic sinus wall in the pika spleen. Scanning electron microscopy of vascular casts suggested that mostly open routes existed in the red pulp, and both open and closed circulation did in the marginfal zone, though a possibility as a functionally closed circularation may remain. The marginal sinus was initially suggested to function as a filter against foreign particles or antigenic materials, because masses of resin and carbon particles injected intravenously leaked out firstly from the marginal sinus wall and further, the marginal sinus had perforated wall.

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The bronchoalveolar cell response was studied by pulmonary lavage after intratracheal inoculation of Rhodococcus (Corynebacterium) equi in mice. The phagocytic cell response in the lungs depended on the size of the inoculum. A few cells were recruited to the lungs following deposition of small inocula (10³ and 10⁵ CFU). However, a large number of neutrophils were recruited to the lungs following deposition of a large inoculum (10⁷ CFU). The effect of nitrogen mustard on early pulmonary clearance was investigated with this infection model in mice. Nitrogen mustard induced leukopenia, resulting in a slight influx of neutrophils into the lungs and permitted the multiplication of the bacteria in the lungs. After accumulation and dispersion of neutrophils, which caused rapid reduction of the number of inoculated bacteria, a number of macrophage infiltrated into the lungs. The effect of carrageenan on pulmonary clearance of R. equi became more prominent after 3 days post infection. These results indicated that resident alveolar macrophages were adequate to clear small numbers of R. equi, but neutrophils were required to clear larger numbers of R. equi in the antibacterial pulmonary defense of mice.

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Weanling mice were infected with avianized H virus of low pathogenicity, and various virological and immunological parameters were examined. In comparative study with other strains, mice infected with F and Str. viruses showed very high virus multiplication in the brain and all the infected mice died. In mice infected with H virus, the virus multiplied and, then, disappeared rapidly in the brain, and the majority of mice survived. CF antigen could be demonstrated in the brain of mice infected with H virus, even after the disappearance of the virus. In mice infected with Fl. H virus, the virus multiplied at such a low level that it was difficult to detect a distinct CF antigen from these mice. Mice infected with H and Fl. H viruses produced high virus-neutralizing antibody and CF antibody in serum. When a higher dose of H virus in the inoculum was employed, the virus multiplied earlier and disappeared earlier in the mouse brain compared with those infected with lower doses. In addition, in these mice, neutralizing antibody were produced earlier in serum. Among mice inoculated with a low dose of H virus, there was a decrease in the incidence of infection, and the onset of illness was delayed. However, their death rate was higher than that of those infected with high doses. No virus was demonstrated in the brain of the dead mice. In the early stage of H virus infection, no inhibitors or incomplete virus particles were found in the brain. When various types of antigen harvested from the brains of mice infected with H virus were examined, the protective ability or ability to produce neutralizing antibody in serum was correlated with the virus titers, and the ability to produce CF antibody in serum was correlated with the CF antigen titer in the brain. The disappearance of the virus from mouse brains infected with H virus in early stage, recovery from clinical illness and the rapid development of virus neutralizing antibody were discussed.

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The effect of onion ingestion on red cell anti-oxidizing agents such as superoxide dismutase (SOD), catalase, glucose-6-phosphate dehydrogenase (G-6-PD), glutathione peroxidase (GPx), NADH-methemoglobin reductase and reduced glutathione (GSH) was studied in relation to Heinz body hemolytic anemia in dogs. To dogs was given the onion soup equivalent to 30g of raw onion/kg body weight once a day for three days. A few days after the initiation of feeding, there was a transient decrease of SOD, catalase and G-6-PD activities to 60%, and GSH value to 70% of their predosing values with a sudden and marked Heinz body formation. These values then began to increase when Heinz bodies were observed in almost every red cells. On day 10, when red cells carrying Heinz bodies were almost cleared from blood circulation, catalase and G-6-PD reached peak values of nearly 250% of their predosing levels. Changes of GPx and NADH-methemoglobin reductase were not correlated with Heinz body counts. Methemoglobin concentration was significantly elevated and maintained in the level of 2-3% during the presence of Heinz bodies in the red cells.

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Specific-pathogen-free neonatal pigs were intranasally inoculated with serotype D strains of Pasteurella multocida or with strain L3 of Bordetella bronchiseptica, and changes in the nasal mucosae were examined with a scanning electron microscope (SEM) or a light microscope. P. multocida caused slight chronic catarrhal inflammatory changes in the infected nasal mucosa, but most of the epithelial cells appeared intact. A small number of lymphocytes and neutrophils infiltrated the lamina propria and submucosa, and a small amount of a mucopurulent substance accumulated on the mucosa. P. multocida was not readily observed on the nasal mucosa by SEM, though a low number of the organisms were constantly recovered from the nasal cavities of the infected pigs throughout the experimental period. In contrast, marked inflammatory changes were observed after the infection by B. bronchiseptica. Degeneration and desquamation of the nasal epithelial cells were noted throughout the infected nasal mucosa. Cilia were lost from most of the ciliated epithelial cells. Marked hyperplasia of the epithelial cells was also observed in many parts of the epithelium. B. bronchiseptica was frequently noted on the remaining cilia or on the microvillous cells by SEM, and numerous organisms were constantly recovered from the nasal cavities of the infected pigs during the period. Clinical atrophic rhinitis was induced only in the pigs given B. bronchiseptica.

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Alterations of the heart rate with and without autonomic blockade were traced through the growing period in rats and pigs. The heart rates following the atropine and/or propranolol administration were measured to estimate the autonomic nervous tone. The heart rate per body weight was calculated to make a comparison between rats and piglets. The heart rate in rats under autonomic blockade (a combined injection of atropine and propranolol) increased progressively during 2 to 4 weeks of age and thereafter decreased gradually with advancing age. On the other hand, the heart rate in piglets under the same treatment decreased gradually with age tracing a similar changing pattern as of the control heart rate. The parasympathetic tone of rats was low at 2 weeks of age and became higher during 2 to 4 weeks of age, whereas that of piglets at 2 weeks of age was at almost the same high level as other weeks of age. The sympathetic tone of piglets as well as rats was highest at 2 weeks of age. In piglets, the net autonomic tone, expressed as a sum of the sympathetic and the parasympathetic tones, was maintained at the sympathetic side through the experimental period. Although the net autonomic tone of rats was also in the sympathetic side at 2 weeks of age, it inverted to the parasympathetic side later on.

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An outbreak of fowl cholera occurred in a green pheasant (Phasianus colchicus) flock at a pheasantry in Saitama prefecture of Japan in February of 1984. Twenty-eight (19%) of 150 birds in the flock died of an acute disease. Pasteurella multocida was isolated in pure culture from hearts, lungs, livers, kidneys and spleens of three representative dead birds. Serovars of the isolates were identified as Carter's capsular type A, Heddleston's type 3, and Namioka's type 8, 9:A. Macroscopically, multiple necrotic foci were observed in livers and spleens, and hemorrhages in small intestines. Microscopically, multiple areas of necrosis, bacterial aggregates and moderate heterophilic infiltration were apparent in livers and spleens. Bacterial aggregates were also apparent in hearts and lungs. Hyperemia was apparent in the lungs, kidneys and spleens. Congestion and bacterial emboli were observed in small intestines. These etiological and pathological findings indicated that the infection was typical fowl cholera. This is the first report on the occurrence of fowl cholera in the green pheasant, which is an indigenous bird in Japan.

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Subcapsular cell hyperplasia in the adrenal glands of ICR mice, aged from 2 to 25 months, were investigated for incidence and morphological characteristics. The lesions were first observed in the 4-month-old mice of both sexes and increased gradually in incidence until reaching a plateau at 19 months of age when the incidence of males was 59% and of females was 91%. Females showed higher incidences than males at any ages examined. Subcapsular cells in the earliest lesion were of fusiform type (Type A cells) and formed scattered foci of small cellular aggregates just beneath the capsule. There were no morphological changes in the zona glomerulosa cells. The subcapsular cells then proliferated to expand their area vertically as well as horizontally and tended to form wedge-shaped hyperplastic foci. Large hyperplastic foci were composed of fusiform cells and polygonal cells (Type B cells) containing abundant cytoplasm. Fat staining showed lipid droplets in both Type A and B cells. Electron microscopic examination on the subcapsular cells revealed basement membranes, desmosomes and dense bodies which developed into intracytoplasmic vacuoles. Some subcapsular cells gathered together to form nests similar to the glands of zona glomerulosa.

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In order to examine the effect of particle size on the inhalation toxicity of liquid aerosols, male rats were exposed to mists of chlorfenvinphos (CVP), a liquid organophosphorus insecticide, in a newly developed size-selective inhalation chamber for 4 hr. Mists with two different particle size distributions were used: small mist consisting of particles smaller than 1μn, and large mist larger than 1μm in aerodynamic diameter. Lethality of the large mist determined by median lethal concentration (LC50) was about 5 times higher than that of the small mist. Almost all deaths occurred during the exposure. During the exposure the large mist caused two times higher blood CVP concentration in the same exposure concentrations. The area under the blood CVP concentration versus time curve for the 2-hr exposure and subsequent 5 hr was 4 times higher with the large mist compared with the small mist, suggesting a higher absorption rate of the large mist. The blood CVP concentration during the exposure of each LC50 was, however, significantly lower with the large mist compared with the small mist. Cerebral acetylcholinesterase (AChE) was inhibited during the exposure in relation to the blood CVP concentration. There was no difference in the relation of AChE inhibition and the blood CVP concentration between exposures of the large mist and the small mist. From these results, the higher blood CVP concentration during the exposure may only partly relate to the higher toxicity of the large mist.

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Immunologic relationships among Haemophilus pleuropneumoniae strains of serovar 1 to 5 were studied in guinea pigs by the complement-fixation (CF) and the protection tests. Most of the guinea pigs, which had been injected once or twice with each monovalent vaccine, had the homologous CF titers of 1:16 or higher, and survived the intraperitoneal challenge inoculation with the homologous strain, whereas none of the animals having titers of 1:8 or below survived, irrespective of the combinations of vaccines and chalenge strains. Most of the injected guinea pigs produced the CF antibodies whose titers were 1:16 or higher against the homologous strain, whereas below 1:8 against the strains of heterologous serovar. Thus, the production of CF antibodies and protection against the challenge strains were closely related each other and showed serovar specificity in the injected guinea pigs. On the other hand, especially in the guinea pigs injected twice, varying extents of cross-protection (maximum 37.5%) was observed among the strains of serovar 1 to 5. The cross-protection pattern was apparently associated with the cross-reaction pattern in the CF test. These results suggest that existence of common CF antigen(s) among the H. Pleuropneumoniae strains correlates with the cross-protection in guinea pigs.

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Sera and liver extracts of cows with fatty liver were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis to demonstrate the specific changes in protein profile of fatty liver cows. In the sera from cows with fatty liver, a 35K protein was found to be increased as compared to that of sera from normal cows. On the other hand, the increase of 35K protein was not observed in sera from cows with other diseases we so far tested. The increase of 35K protein in sera from fatty liver cows was further comfirmed by gel filtration of the sera using Sephadex G-100. The increase of 35K protein was also observed in liver extracts from fatty liver cows. These results suggested that the increase of this protein was specifically associated with fatty liver of cows.

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By pretreating mice with sublethal dose of mouse liver homogenate infected with Tyzzer's organism, resistance was precociously induced to fatal challenge inoculation made days 1 to 4 posttreatment. Serum antibody became detectable on day 4 postchallenge in both pretreated and challenged mice and serum interferon remained undetectable. Such resistance could be induced by pretreating with formol vaccine or lysate of the bacteria, lipopolysaccharide or BCG but not with Freund complete adjuvant, poly-inosinic: poly-cytidylic acid or inactivated mouse hepatitis virus.

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Of the six cats which were orally inoculated with 70, 000 protoscoleces of the Hokkaido isolate of Echinococcus multilocularis, only four were infected when examined on day 27 postinoculation. The recovery rates of the cestodes from the intestines of the cats were 0.1 to 1.2%. Most of the worms were recovered from the posterior half of the small intestine. The growth rate of the cestode was retarded as indicated by the slow segmentation, with the majority of the worms having less than two proglottides, as compared to those recovered from a dog which served as a control. On the contrary, the recovery rate of the cestodes from the control dog was 40%, with most of the worms having three proglottides.

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The four main types of pancreatic endocrine cells in the developing chick embryo were studied by light microscopic immunocytochemistry to describe their time of appearance, localization, and changes in the distribution during development in connection with the formation of pancreatic lobes. A cells first appeared at day 3 of incubation in the dorsal pancreatic bud, and increased in number thereafter to form the alpha islets. The alpha islets became huge and complicatedly diverged like those in the adult at day 11, but remained restricted to the third and splenic lobes derived from the dorsal pancreatic bud. B cells first appeared at day 4 of incubation in distinct beta islets in the dorsal pancreatic bud, but only a few of them showed the intense immunoreaction to anti-insulin serum at this stage. At day 6, most of B cells constituting beta islets in the dorsal primary lobe became immunopositive to anti-insulin serum. At day 11, beta islets spread into the dorsal and ventral lobes derived from right and left ventral pancreatic buds. D cells appeared first in the dorsal pancreatic bud at day 4 and joined the alpha islets at day 5. At day 11, D cells joined the beta islets and appeared in the dorsal and ventral lobes as did B cells. APP cells first appeared at day 9 in the exocrine portion of four lobes and remained restricted to the exocrine pancreas independently from pancreatic islets.

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A histopathological study using enzyme immunoassay (EIA) was carried out in mice experimentally infected with Yersinia pseudotuberculosis IB to elucidate the role of V antigen in vivo. The presence of V antigen and bacterial cells in the lesions was revealed in various organs by EIA using anti-V antigen serum and anti-O antigen serum. There was a distinct difference in the degree of V antigen production among the organs examined. V antigen was massively produced in the liver, spleen and mesenteric lymph nodes associated with lesions and bacteria. In the intestine, however, V antigen was weakly produced in the lesions under the intestinal epithelium despite the massive presence of a bacterial colony. The detailed staining pattern suggested that V antigen might be formed surrounding the bacterial cells in the lesions of the liver, spleen and mesenteric lymph nodes. Yersinia pseudotuberculosis IB mutant lacking in both VW antigen and plasmid did not cause lesions in any organs. In addition, fecal excretion of the organisms was closely associated with the formation of lesions in the intestine.

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Spongiform lesions were found in the gray matter of cerebrum, midbrain, pons and medulla oblongata, and the white matter of cerebrum in three scrapie sheep aged 4 to 5 years old. Ultrastructural evidence supports that the spongiform lesions were caused by vacuolation in the neuronal and glial perikarya, vacuolation or swelling of the neuronal and glial processes, dilatation of the periaxonal space, and splitting of the myelin sheath followed by the intramyelinic vacuolation. Association of the vacuoles in the neuronal perikarya with the endoplasmic reticulum was supported by enlargement of the rough endoplasmic reticulum and of cisterns of Golgi apparatus adjacent to the vacuoles. The vacuolation and swelling in the neurons or glias were regarded as primary pathologic changes.

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Eleven dogs from several veterinary hospitals which died of spontaneous parvoviral enteritis were studied histopathologically to determine pathognomonic lesions which may serve as diagnostic indicators in this disease. Giant epithelial cells with bizarre nuclei and intracellular microcysts suggesting dysplastic regeneration were often observed in the affected mucosa of the small intestine in all cases.

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The development of Theileria sergenti in the salivary glands of adult Haemaphysalis longicornis ticks was morphologically studied during 4 days of tick's feeding on rabbits. Theileria sergenti demonstrated three forms of development (primary, secondary and tertiary sporoblasts) in the process of budding into sporozoites in the glands for 1 to 4 days. It was clear that the sporozoites were formed in the acinar cells of those glands and that most of them were released from the hollow cytoplasmic area of these cells by the 4th day.

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A 12-month-old, grazing Holstein-Friesian cow with theileriasis and anaplasmosis suddenly fell down and showed nervous symptoms. She was slaughtered 29 days after manifesting nervous signs. The main macroscopical changes were marked anemia, generalized icterus and two necrotic foci in the white matter of the cerebellum. Histologically, granulomatous lesions with necrosis in the cerebellar medulla and subacute suppurative nephritis caused by invasion of zygomycotic organisms were observed in addition to hemolytic changes due to the protozoal infection.

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Fifteen neurons in the subfornical organ (SFO) were antidromically activated by electrical stimulation of the supraoptic nucleus (SON) of the hypothalamus in urethane-anesthetized rats. Microiontophoretically (MIPh) applied angiotensin II (AII) excited the activity of all identified SFO neurons and the effect of AII was blocked by MIPh-applied saralasin (Sar), an AII antagonist. Intravenously administered AII excited the activity of most (n=13) of these identified SFO neurons and this exciting effect by intravenous AII was also blocked by MIPh-applied Sar.

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Glucose-6-phosphate dehydrogenase (E. C. 1. 1. 1. 49) (G6PD) from pig red cell was purified by DEAE-sephadex A-50 chromatography and gel filtration on G-100 and G-25. Ammonium sulfate precipitation was used after the DEAE-Sephadex A-50 and Sephadex G-100 steps. The final enzyme preparation had a specific activity of 18 enzyme units/mg. The determination of apparent Km values for G6P and NADP displayed that the enzyme has greater affinity for NADP than for G6P. Based on high Ki value for NADPH, the pig red cell G6PD appears not to be significantly regulated by NADPH. On the other hand, the enzyme appears to be more susceptible to inorganic phosphate as an inhibitor than NADPH since the Ki value for inorganic phosphate is many-fold less than that for NADPH.

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Ultracytochemical localization of K⁺-dependent p-nitrophenyl-phosphatase (p-NPPase) activity using Mayahara's method was investigated on the choroid plexus of N-methylnitrosourea (MNU)-induced dysgenetic hydromicrocephalic rat offsprings aged 12 weeks. The reaction sites were characteristically observed in lysosomes, multivesicular bodies, coated vesicles and on basal infoldings in the epithelial cells, in microvesicles in the interstitial cells, and in an increasing number of pinocytotic vesicles and along junctional complex in the capillary endothelial cells. These changes suggest that leaking blood components in choroidal edema might pass the choroidal cells.

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Detection of myoglobin (Mb) in sera and urine of sixteen calves with nutritional myopathy was achieved by immunodiffusion method. Mb were detected in 12 serum samples out of 16 calves and in 9 urine samples out of 13 calves. Concentrations of Mb in serum and urine decreased markedly by treatment and were scarcely detected in any samples after one or two days. The minimal detectable concentration of Mb by immunodiffusion method was 5μg per ml.

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In Microtus species, diabetes was induced easily by feeding with low fiber and high concentrate diet. It was considered to be caused of their characteristic as herbivore that the renal threshold for glucose or glycolytic enzymes (GK, HK) activities were very low. In the diabetic voles, the glycolytic enzymes activities controlled by insulin changed greatly with the progress of the disease. So, Microtus species is considered to be useful as an animal model of the disorder of glucose metabolism in herbivore.

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Decreased plasma insulin levels in lactating rats were restored by the treatment with CB-154, but at the same time, lactation ceased. Low plasma insulin levels in lactating rats were also restored by the removal of pups. This rise due to pups-removal was not prevented by prolactin administration. Plasma insulin lowered again with resumption of nursing. Thus, milk production presumably serves to suppress insulin secretion from B-cells.

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Susceptibility of several cell cultures to the feline panleukopenia virus TU 1 strain and canine parvovirus Cp49 strain was investigated. All the feline cell cultures supported the replication of both strains, but the canine cells did only that of Cp49 strain. Contrary to expectation, the evidence for positive growth of TU 1 strain in some strains of Madin-Darby canine kidney (MDCK) cells was obtained when the inoculated cells were serially subcultured. However, TU 1 strain passaged 10 times in MDCK cells did not gain the ability to infect other canine cells.

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A chordoma-like tumor occurred in the coccygeal region of a 101-week-old female ICR mouse. The tumor cells were growing in sheets with a lobular pattern and showed cytoplasmic vacuolation in varing degrees. Markedly vacuolated cells closely resembled the physaliferous cells in human chordoma. Aggregates of cytoplasmic filaments or fibrillar materials in the large vacuoles were demonstrated by electron microscopy.

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In trypsin-treated paraffin sections of swine kidney samples fixed in 10% neutral buffered-formalin (NBF) for 6 to 11 months at room temperature and examined for IgG and C3 in the glomeruli by immunofluorescence, IgG was well preserved as in fresh frozen sections but C3 was remarkablly reduced. In samples fixed either in NBF or 10% formalin at 4°C or room temperature for one to 180 days, IgG was preserved better in NBF than formalin, while C3 fluorescence was apparently reduced during fixation, at room temperature.

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Insulin secretory responses to intravenous injections of glucose, ketone body or volatile fatty acids were investigated in herbivorous voles, Microtus arvalis Pallas. Glucose stimulated insulin secretion accompanying hyperglycemia. Propionate, butyrate, valerate or β-hydroxybutyrate stimulated insulin secretion to a similar extent as glucose did whithout changing blood glucose concentration, but acetate did not.

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A study was performed to assess use of tolazoline as an antagonist of xylazine in cattle. Intravenous injection of tolazoline markedly reduced the recovery period from sedation and analgesia when given 15 min after intravenous injection of xylazine in comparison with those that received xylazine+Ringer's solution. Rumen hypomotility and salivation evoked by xylazine were substantially reduced by tolazoline. These results indicate potential usefulness of tolazoline as an antagonist of xylazine in cattle.

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