The role of dermonecrotic toxin (DNT) of Bordetella bronchiseptica and Pasteurella multocida, purified by repeated chromatography in Sephacryl S-200 gel, in the pathogenesis of atrophic rhinitis (AR) of swine was studied bacteriologically, clinically and pathologically. Two-week-old specific pathogen-free (SPF) piglets were parenterally treated with 30μg of DNT 3 times at 2-day interval and 7-week-old piglets were treated with 15μg of DNT twice a week for 5 weeks. In 2- to 3-week-old piglets, both B. bronchiseptica DNT and P. multocida DNT produced nasal turbinate lesions with similar severity, characterized by damage of the cilia, epithelial metaplasia, intensive proliferation of osteoblasts, regressive changes, and diffuse osteocytic osteolysis. In 7- to 12-week-old piglets, treatment with B. bronchiseptica DNT failed to produce progressive changes in the nasal turbinates. Histopathological examination revealed osteogenic processes and osteoid synthesis besides the proliferation of osteoblasts and mild osteocytic osteolysis. Moreover, severe gross pathological lesions developed in the stomach, liver, kidneys, and lymphoid organs. The piglets' appetite and body weight gain gradually decreased during the DNT treatment and in the last week when the toxic signs appeared. Treatment of 7- to 12-week-old piglets with P. multocida DNT resulted in progressive AR. Histopathologically, diffuse osteocytic osteolysis was observed in the nasal turbinates. Neither clinical signs nor pathological lesions of the visceral organs developed in these piglets. The authors emphasize that the DNT of B. bronchiseptica basically differs from that of P. multocida in biological properties, though there are certain similarities between the DNTs.

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Chuzan virus at 2 to 3 passage levels in cell cultures after isolation was inoculated intravenously into 15 seronegative pregnant cows at 89 to 150 days of gestation. All of the cows developed viremia a few days after inoculation and antibodies 2 weeks after inoculation. No clinical signs, except leukopenia, were observed throughout the experimental period. These 15 cows delivered 15 calves after normal gestation. One of the calves which was born to a dam inoculated at 120 days of gestation, showed impairment of movement, and the remaining 14 were healthy. Postmortem examination revealed that this calf had hydranencephaly-cerebellar hypoplasia (HCH) syndrome and that the remaining calves were normal. Two of the 15 calves, including the one that had HCH syndrome, had antibody to Chuzan virus in their precolostral sera. These findings provide additional evidence that Chuzan virus is the etiological agent of an epizootic of congenital abnormalities with HCH syndrome of calves in Japan, 1985 to 1986. We propose to name the HCH syndrome caused by Chuzan virus infection Chuzan disease.

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To develop a live virus vaccine for the prevention of bovine respiratory syncytial (BRS) virus infection in calves, an attempt was made to produce an attenuated virus. The RS-52 strain of BRS virus, isolated from the nasal secretions of a naturally infected calf, was subjected to serial passages in adult hamster lung established (HAL) cells at 30°C and the attenuated rs-52 strain as a live virus vaccine was established. The rs-52 strain multiplied better at 30°C than at 34 or 37°C in HAL cells. The differences in the highest virus titers of this strain between the culture temperature of 30°C and that of 34 or 37°C were more than 2.25 log TCID₅₀. Colostrum-deprived newborn calves and 2-4 months old calves inoculated with the rs-52 strain manifested no abnormal clinical sings at all. However, all inoculated calves produced serum neutralization antibody. When the colostrum-deprived newborn calves immunized with the rs-52 strain were challenged with the virulent NMK7 strain of BRS virus, they exhibited no pyrexia or other abnormal clinical signs at all. An attempt was made to recover the virus from nasal secretions of these calves, but in vain. On the other hand, a nonimmunized control colostrum-deprived newborn calf developed slight fever, mild cough, and slight serous nasal discharge after challenge exposure. The virus was recovered from nasal secretions of this calf. From these results, it was considered that the rs-52 strain could be used as an attenuated live virus vaccine for prevention of BRS virus infection.

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For the investigation of the cause of white muscle disease (WMD), tocopherol (Toc) and selenium (Se) levels and blood glutathione peroxidase (GSH-Px) activities were examined using lambs with WMD and their ewes. Serum Se levels of 4 lambs with WMD were low under 30 ppb, lambs showing very low levels below 15 ppb. The serum Se level was correlated with the blood GSH-Px activity showing remarkably low activities in the lambs with WMD. Se contents in the organs of lambs with WMD were lower than those of control lambs, and particularly liver Se contents were deficient levels below 50 ppb. Serum Toc levels were normal, but α-Toc contents in organs showed very low levels, especially in the liver. The serum Toc and Se levels and blood GSH-Px activities of their ewes and other sheep kept in the same farm revealed similar results to those of lambs with WMD. Feedstuffs supplied on the farm showed the deficient level of the Se content below 50 ppb and a very low level of α-Toc. It was concluded that WMD of lambs in Hokkaido was nutritional muscular dystrophy resulted from deficiencies of Toc and Se to their ewes.

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Localization of bacteria and bacterial materials was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium by a immunohistological method using rabbit antiserum against the outer membrane of those organisms and by a scanning electron microscope. The intact organisms of both inoculated bacterial species were detected on the rumen wall and in the lamina propria of the forestomach, and S. ruminantium also in the lymph nodes associated with the rumen. The bacterial materials were observed inside of macrophage-like cells in the lamina propria of the forestomach and of lymphoid cells in the lymph nodes. Number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the orally inoculated bacteria may act as antigenical stimulant in the mucosa of the forestomach and associated lymph nodes of calves.

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Distribution of immunoglobulin(Ig)-containing cells was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium. Pathological changes and many Ig-containing cells were observed in calves which inoculated three times at 2, 3 and 26 days of age. Follicular germinal center was increased in number and size of the lymph nodes associated with the forestomach, suggesting activation of lymph apparatus. In the associated lymph nodes, IgG-containing cells were predominant and were located in both cortex and medulla, mainly in the medullary cord, B lymphocyte areas. Only a few IgA- and IgM-containing cells were observed in the lymph nodes. Accordingly, the inoculated bacteria may stimulate IgG-containing B lymphocyte populations. A few IgG-containing cells were detected in the mucosa of the forestomach. Ig-containing cells, predominantly IgG, were observed in the mucosa of the abomasum and intestine, and in the mesenteric lymph nodes. However, number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the intraorally inoculated bacteria may stimulate the maturation of IgG positive lymphocytes in the lymph nodes associated with the forestomach.

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Calcium-dependent and -independent Sepharose 4B-binding bovine serum proteins were isolated by affinity chromatography on unsubstituted Sepharose 4B using 2 mM ethylenediamine tetraacetic acid followed by 0.1 M galactose. They appeared in two different protein peaks by gel filtration on Sephacryl S-200. The earlier eluted protein was demonstrated to be immunologically IgM, whereas the retarded one IgG. From these findings, Sepharose 4B-binding bovine serum proteins are suggested to be calcium-dependent and -independent immunoglobulins IgM and IgG.

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The cytotoxic activities of feline spleen cells treated with Toxoplasma lysate antigen (TLA) were assayed against feline leukemia virus (FeLV)-producing lymphoma, FL74 cells, and xenogeneic target lymphoma, mouse YAC-1 cells. The TLA treatments were performed in vivo alone, in vitro alone, and in vivo plus in vitro, respectively. In vivo plus in vitro treatments with TLA induced a marked augmentation in cytotoxic activity of spleen cells to FL74 cells. The treatment with TLA in vivo alone showed an enhancement of cytotoxic activity but in vitro alone did not. The cytotoxic effects of TLA-treated spleen cells obtained from the cats which had been previously immunized with live FL74 cells were similar to those of spleen cells from non-immunized cats treated with TLA. However, no increase of cytotoxicity was shown in the response to mouse YAC-1 cells regardless of TLA treatments. These results indicated that the in vivo TLA treatment augmented the cytotoxicity of feline spleen cells against FeLV-producing lymphoma cell.

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Improvement of pulmonary arterial lesions after heartworm removal using flexible alligator forceps was investigated by measuring pulmonary arterial pressure (PAP), and by angiographic and histopathological examinations in 11 dogs. PAP obtained immediately after worm removal corresponded well with angiographic abnormalities. In 2 dogs, high PAP immediately after worm removal fell gradually by 12 weeks, and obstructions on angiogram were resolved at 4 to 12 weeks. In 4 dogs, slightly high PAP fell to the normal range at 4 weeks, and angiographic abnormalities were considerably reduced at 4 to 12 weeks. In 5 dogs, PAP returned to normal range immediately after worm removal, and angiographic changes almost disappeared at 4 to 8 weeks. On biopsy immediately after worm removal, samples of the main pulmonary arteries showed severe intimal proliferations with villous or papillary protrusion into the lumen. Autopsy at 12 to 20 weeks indicated that the intimal protrusions were remarkably reduced as compared with the biopsy samples in all cases. However, villous intimal protrusions were seen in the caudal lobar arteries in cases with remaining alive worms. New vessels seemed to develop into thromboemboli with time. From these findings, it is clear that the pulmonary arterial lesions improved after heartworm removal, and the clinical signs disappeared following the improvement in hemodynamics. Aspirin therapy (5 mg/kg/day) for 4 weeks after worm removal in 5 dogs did not improve the intimal lesions as compared to 3 control dogs.

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Hemocytes of a marine gastropod, Nerita albicilla, but not those of an estuarine gastropod, Clithon retropictus, were observed to migrate to live and heat-killed cells of Vibrio parahaemolyticus and Escherichia coli through Nucleopore membrane in Blind well chamber. The defective migration of C. retropictus hemocytes might reflect, at least in part, the survival of V. parahaemolyticus in the estuarine gastropod.

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The change in activities of 3 major antioxidative enzymes in equine erythrocytes, superoxide dismutase (SOD), glutathione peroxidase (GSHpx), and catalase, was investigated in order to evaluate the effect of exercise. Blood samples were obtained from 11 thoroughbred horses before and immediately after vigorous exercise which induced the increase of plasma lipid peroxide (Lpx) concentration from 1.16+0.40nmol/ml to 1.29±0.34nmol/ml. Following the exercise, the GSHpx activity in erythrocytes was significantly reduced from 69±10IU/gHb to 65±8IU/gHb, whereas SOD and catalase activities were not changed. Effects of an antioxidative compound, containing selenium and vitamin E(Se-E), on the response of antioxidative enzyme activities following the exercise were examined. Seven horses were injected intramuscularly with Se-E(Se:25mg, vitamin E:54.8mg) and received the same vigorous exercise. After Se-E treatment, plasma Lpx levels before the exercise were decreased from 1.24±0.09nmol/ml to 0.86±0.03nmol/ml, however, SOD, GSHpx, and catalase activities were not varied. The Se-E treatment slightly prevented the decrease in GSHpx activity and the increase in plasma Lpx level after the exercise, having no effect on SOD and catalase activities. These results suggested that the changes of GSHpx activity in erythrocytes might reflect the protective condition against exercise-induced lipid peroxidation.

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Mice inoculated intracerebrally with parent, large-plaque (LP) and small-plaque (SP) strains of Kanagawa strain of Getah virus showed clinically recumbency and paralysis. The LP strain caused recumbency more rapidly and killed mice more early after inoculation than the parent and SP strains. Microscopically, skeletal muscles of the whole body were involved showing degenerative or inflammatory changes. In mice inoculated with the parent or SP strains, there were degeneration and necrosis of the muscle fibers with inflammatory cell infiltration and regenerative reaction. The lesions were particularly conspicuous in muscles of the hind legs. In mice inoculated with the LP strain, most of the muscle fibers revealed degeneration and necrosis, but reactive changes were poor. In addition, the periosteum and muscular connective tissue were thickened with karyorrhexis. Electron microscopically, virus particles were recognized mainly in cisternae of sarcoplasmic reticulum in skeletal muscle fibers of mice inoculated with the LP strain, while they were rare in those of animals injected with the parent and SP strains. From these findings, it was suggested that Kanagawa strain of Getah virus has the virulence to skeletal muscles of mice.

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The present study was conducted to examine how the social cue emanating from rams, the ram-effect, would influence the onset of melatonin-induced reproductive activity in anestrous ewes. Twenty non-lactating ewes were randomly allocated into 4 groups as follows based on a combination of the melatonin treatment (MEL) and the ram-effect (RAM): ewes of Groups A (MEL+RAM) and B (MEL) were subcutaneously implanted with melatonin capsules on April 18 (Day 0), which increased plasma melatonin levels by about 200pg/ml for at least 5 months, while Groups C (RAM) and D (control) were untreated with melatonin. Rams were introduced to Groups A and C on Day 0, whereas Groups B and D were isolated from rams. Ovarian function of the ewe was assessed on June 9-21 (Days 52-64) by monitoring plasma progesterone (P) profiles. Luteal function (plasma P>1 ng/ml for a week or longer) was evident in all the melatonin-treated ewes but only one in those untreated: 5/5 in Group A, 5/5 in Group B, 1/5 in Group C and 0/5 in Group D. By ultrasonography on Day 105 all the Group A ewes were diagnosed pregnant but none in the Group C despite that both the two groups had been run with rams. These results indicate that chronic melatonin treatment is capable of advancing the reproductive recrudescence in seasonally anestrous ewes, and that progonadal effects of rams are subtle, if any, during the mid-anestrous period.

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The fine structure of tubulobulbar complex (TBC) of goat seminiferous epithelium was studied using testicular samples fixed by perfusion. During the maturation phase of the spermatid, the TBC appeared on the periphery of the head before spermiation, dorsal and ventral parts of the head, both sides of the acrosome and on the postnuclear sheath region. At an early stage of the development of the TBC, a partial loss of smooth endoplasmic reticulum in the ectoplasmic specialization of a neighboring Sertoli cell was observed; filament bundles disappeared and a small fragmented smooth endoplasmic reticulum was left. Subsequently, a spermatid vesicle (coated vesicle) expanded to be in contact with the Sertoli cytoplasm by a small channel. These tubular projections extended to form a bulbous balloon-like structure forming sometimes a bi-balloon appearance, until torn off from the spermatid into the Sertoli cytoplasm and probably digested by the Sertoli cell. The significance of this structural development was morphologically discussed.

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A low molecular weight protein was separated from urine samples obtained from a heifer with spontaneous renal disease and from cows with CaNa₂EDTA-induced renal dysfunction. The molecular weight and electrophoretic mobility of the separated protein were examined. The low molecular weight protein collected by gel filtration chromatography was further separated into two fractions by ion exchange chromatography using DEAE-cellulose. One of the two fractions, the lowest molecular weight protein showed a single band in SDS-PAGE, and its molecular weight was approximately 12, 000. An antiserum against this protein formed a single precipitin line with the urine from cows with experimentally induced renal dysfunction and a heifer with spontaneous renal disease by the double immunodiffusion technique. However, the antiserum did not form any precipitin line with the concentrated urine of healthy cow and human β₂-microglobulin. In cellulose acetate membrane electrophoresis, this protein migrated in the same position as that of serum γ-globulin from healthy cow.

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Characteristics of CPE produced by porcine enteroviruses (PEV) were examined by Immunoperoxidase (IP) staining method. Viral antigens were detected earlier than appearance of CPE. Distinctive characteristics of the three CPE types were clearly showed by the method. In cross reactions by IP staining, the titers of the staining were high (1:6, 400 to 1:25, 600), even though neutralizing titers of PEV with CPE II or III were low (1:200 to 1:400). The very close relationship was detected between PEV with the same CPE type, but the very low relationships were detected between PEV with the different CPE type. The relationships in CPE I group were various. The results by IP staining were more relative to CPE type than the serotype. Thus, IP staining is one method to classify PEV clearly.

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In order to better understand androgen secretion in the tom cat during the breeding season, observations of diurnal changes in peripheral blood testosterone (T) levels, were made and the relation between androgen levels (androstenedione (A), 5α-dihydrotestosterone (DHT), T) in testicular and peripheral vein blood was borne. Histologic examinations of the testis were also performed to investigate spermatogenic function. The 9 tom cats used in these studies were 2-3 years old, weighed 3.5-4.0kg, and were raised in a room under natural lighting. As a result, diurnal peripheral blood T levels in the tom cat fluctuated greatly according to each individual, but since no definite trend could be identified, secretion was determined to be episodic. Although fairly large differences in the 3 types of androgen were observed in testicular vein blood depending on the individual animal, levels on the left and right were almost equal. Moreover, a correlation was found between the testicular and peripheral vein blood concentrations of the 3 androgens (A: p<0.01; DHT: p<0.05; T: p<0.01). Therefore, secretion of testicular androgen can be inferred by measuring peripheral blood androgen levels. Testicular histological examination revealed active spermatogenesis in all of the cats, and no significant inter individual differences were detected in either seminiferous tubule diameters or any of the various germ cells.

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The caliber of the umbilical arteries was measured in fetal rats on day 14 to 21 of gestation. The fetuses were rapidly frozen in an acetone-dry ice mixture. The caudal half of each whole-body frozen fetus was gradually shaved with a knife from the ventral side to expose the umbilical arteries for measurement of their calibers. Observation reveals that, on day 15 of gestation, the umbilical arteries are asymmetrical in that the right artery is more dilated than the left. It is further seen that the left umbilical artery is closed on day 17 in contrast to the right which is increasingly more dilated toward term. When indomethacin, a synthetic compound having an action similar to that of glucocorticoids, was administered to pregnant rats 3 hr prior to sacrifice on day 21 of gestation, the right fetal umbilical artery was significantly constricted. The reason for this phenomenon is unknown, but is discussed in association with the manner of closure of the ductus arteriosus.

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Monoclonal anti-idiotype antibodies against monoclonal antibody (23C11) to intraerythrocytic merozoites of Theileria sergenti were prepared and examined by indirect immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA). Three anti-idiotype antibodies containing antibodies against interspecies cross-reactive idiotopes and/or an internal image of Theileria sergenti merozoite were found. The presence of antibody against Theileria sergenti was shown by ELISA in these anti-idiotype antibodies induced anti-anti-idiotype antibodies.

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