Survey of rotavirus was conducted in 40 fecal samples which were divided into 4 groups according to diarrheal conditions to clarify the relationship between rotavirus and foals with diarrhea. Particles characteristic of rotavirus were directly detected by electron microscopy from 5 of 14 fecal samples collected from foals with acute diarrhea. Moreover, rotaviruses were isolated from 4 of 5 fecal samples with MA-104 cells and with trypsin treatment, and were capable of being successively passaged in these cells. On the other hand, no rotavirus was detected in 26 fecal samples collected from foals with various non-acute diarrhea. In the cross serum neutralization test, 4 isolated strains were antigenically recognized as the same serotype, which was slightly differed from BI strain of equine rotavirus and clearly separated from Lincoln strain of bovine rotavirus antigenically. From these results, it was revealed that rotavirus was closely related only to acute diarrhea in foals. Moreover, it was confirmed that equine rotavirus in field cases could be isolated in MA-104 cells with trypsin treatment.

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On the thrombus artificially formed in the saphenous vein of beagle dogs, the thrombolytic process was observed by scanning electron microscopy (SEM) and angiography. Thrombolysis occurred spontaneously in most dogs 24 hr after thrombus formation. The coagulative and fibrinolytic indices were determined in the thrombolytic process. The levels of fibrin degradation products (FDP) increased significantly from 12 to 24 hr after thrombus formation. The activated partial thromboplastin time (APTT) and the plasma plasminogen (Pg) level slightly increased 9 hr after confirmation of the thrombus. All these factors, however, were in the physiological range within a week. These results indicate that experimental thrombosis in beagle dogs was acute and reversible with little influence on systemic blood coagulation and fibrinolysis.

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Various kinds of treated cells were prepared from variants derived from serotype 1 strains of Haemophilus paragallinarum by enzymatic or physicochemic treatments. Hemagglutinating properties of the treated cells were investigated against freshly collected and glutaraldehyde-fixed chicken, human being (type O), or rabbit erythrocytes (RBC). At least 3 types of hemagglutinin showing different hemagglutination patterns, designated as hemagglutinins HA-L, HA-HL, and HA-HS, were defined from the results of their biologic properties. Heat-labile, trypsin-sensitive HA-L hemagglutinin agglutinated all RBC used, whereas trypsin-resistant heat-labile, HA-HL or heat-stable, HA-HS antigens agglutinated only fresh chicken or rabbit RBC except those from day-old chicks. Because of the presence of capsule surrounding bacterial cells, nontreated cells prepared from certain variant lacked the ability to agglutinate any RBC used; hyaluronidase-treatment rendered them agglutinate, and the hemagglutinin appeared by this treatment was HA-L hemagglutinin. After trypsin-or heat-treatment of HA-L hemagglutinin, hemagglutinins HA-HL or HA-HS appeared. Electron microscopic studies showed that the outer membrane of the treated cells adhered directly to the surface of RBC. The result suggests that the location of the 3 types of hemagglutinin HA-L, HA-HL, and HA-HS are in the outer membrane of bacterial cells, and indicates the necessity in use of proper hemagglutinin and RBC for experiment.

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Seventeen very young calves were made hypomagnesemia by feeding them a magnesium-deficient milk diet. The plasma magnesium concentration gradually fell during the feeding period, but it was not accompanied by hypocalcemia and there was no significant change in the plasma parathyroid hormone concentration. After the development of severe hypomagnesemia (plasma magnesium less than 0.8 mg/100 ml, six of the calves were anesthetized, and the vein of the parathyroid gland was cannulated to determine the parathyroid hormone-secretion rate. The induction of hypocalcemia was accompanied with pronounced increase in the parathyroid hormone secretion rate in all calves. Afterwards, a solution of magnesium chloride was infused into the jugular vein of these hypocalcemic, hypomagnesemic animals. The intravenous infusion of magnesium into these calves did not result in an increased parathyroid hormone-secretion rate. Compared with the calves fed on a magnesium adequate diet, the parathyroid hormone secretion rate in magnesium-deficient calves was greater during normocalcemia and mild hypocalcemia (plasma calcium between 8 and 10 mg/100 ml) but was the same during severer hypocalcemia (plasma calcium less than 8 mg/100 ml). The greater secretion rate than normal in the normocalcemic state was probably due to reduced inhibition of magnesium to the parathyroids. These results demonstrate that the severe hypomagnesemia in these calves does not interfere with the secretory response of parathyroid hormone to a hypocalcemic stimulus.

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A male dog orally inoculated with 3×10⁸ viable cells of Brucella canis showed curl tails of spermatozoa at the 16th week and head-to-head type auto-spermagglutinations from the 18th to the 27th week, when the dog was killed. Serum spermagglutinating activity was always detected after the 14th week with a maximum titer at the 24th week. Serum IgM anti-sperm antibodies were not clearly detected, whereas serum IgG and IgA anti-sperm antibodies began to appear from the 12th and 20th weeks onward, respectively. F(ab')₂ fragments of the IgG agglutinated motile spermatozoa with head-to-head type and bound on the acrosomes of spermatozoa and spermatids. IgA antibody was detectable on the acrosome of the auto-agglutirlated sperm in the ductus epididymides, semen or urine.

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Immunogenic properties of seven variants having different morphologic, serologic, and pathogenic properties, which were dissociated from serotype 1 (serotype HA-1) Haemophilus paragallinarum, were investigated in chicks. Protective immunity was induced by injection with any of the four variants possessed both heat-labile, trypsin-sensitive, hyaluronidase-resistant, serotype 1-specific L1 and HA-L1 antigens. The immuned chicks had antibodies specific to serotype 1, agglutinin against L1 antigen and hemagglutination-inhibition (HI) antibodies against HA-L1 antigen. In contrast, the other three variants lacking both the antigens did not induce protection. Hyaluronidase-treatment of the hi variant consisting of highly encapsulated organisms abolished its virulence without affecting the protective activity and antibody-producing ability against antigens L1 and HA-L1. These results support our hypothesis that the antigens L1 and HA-L1, are responsible for immunogenicity.

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An enzyme-linked immunosorbent assay (ELISA) using peroxidase conjugate was adapted to detect IgM and IgG antibodies of Japanese encephalitis (JE) in sera from two experimentally infected swine. Using anti-swine IgM or IgG conjugate, ELISA IgM and IgG antibody titers to JE Virus were compared with hemagglutination inhibition (HI) titers before and after 2-Mercaptoethano1 (2-ME) treatment of the sera. ELISA IgM antibody was detected immediately after the disapperence of viremia, and the titers reached maximum in the early stage of infection but decreased in the convalescent period. ELISA IgG antibody, however, appeared later than IgM antibody, and the IgG titers changed to become predominate over the IgM titers in the convalescent sera. The titers in ELISA corresponded well with HI titers. It was found that ELISA could detect IgM antibody even though HI test could not detect it. Anti-swine IgM and IgG conjugates for ELISA were specific enogh to differentiate IgM and IgG sub-class antibodies in the swine serum. Aceton-ether (AE) zonal antigen was found to produce less non-specific reaction in ELISA as compared with sucrose-aceton (SA) antigen. Thses results suggest that ELISA has advantages over the HI test for serodiagnosis of JE in swine.

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When live or killed Proteus morganii, strain 1510, was inoculated intravenously to normal mice, serum factors for opsonization of the inoculated bacteria increased daily from the 2nd day of postinoculation. After inoculation with killed cells, the serum factors did not increase in Sarcoma-180 tumor-bearing mice. When the immune serum from normal mice inoculated with P. morganii was pretreated with anti-mouse IgG or anti-mouse IgM, opsonic activity markedly decreased. Although no opsonic activity was noted in heat-inactivated immune serum, when tumor-bearing mouse serum as a complement was added to inactivated immune serum, opsonic value recovered. The immune serum from normal mice after 3 days postinoculation demonstrated significant protection against challenge with the same bacteria in tumor-bearing mice when passively transferred, but serum from tumor-bearing mice inocultated with P. morganii did not indicate protective effect. When the immune serum was treated with anti-mouse IgM, protection against challenge decreased significantly. However, there was no significant difference between immune serum treated with anti-mouse IgG and untreated immune serum. It was found that IgM antibody necessary for optimal opsonization and protection was markedly impaired in tumor-bearing mice.

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Characteristics of bone crystallites at the insertion of the distal phalanx of the horse in the stage of fibrous bone formation were investigated using electron microscopy. In this tissue, most of the collagen fibrils are ordered in the axial direction and crystallized. The longitudinal section of the collagen fibril showed periodic bands approximately 640/70A apart and with interperiod bands A1, A2, A3, B, C1, C2, D, E and F. In general, these crystallites seemed to adhere to the surface of the collagen fibrils and in the fibrils on the bone surface, needle-or plaque-shaped crystallites were oriented in the same axis as collagen fibrils and arranged on the cross bands. In the bone, many needle-shaped crystallites lay between the parallel collagen fibrils, and the crystallites infiltrated into the collagen fibrils where the interperiod bands are dense. These dense interperiod bands extended approximately 380-400A between two periodic bands 640-670A apart. Infiltration of the crystallites seemed to start at the fibril edge and spread reticulately into the interior. Lucent spots diameters of 50-80A were observed within the collagen fibrils. The crystallites filled the space (20-30A in width) between the spots.

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After intravenous inoculation with a strain of mouse hepatitis virus, MHV-D, C3H mice survived with significant viral growth and interferon (IFN) production in the spleen at 12 to 24 hr postinfection. C57BL mice, however, died of severe hepatitis within 5 to 6 days with neither prominent viral growth nor significant IFN production in the spleen. In resistant C3H mice IFN originated from the spleen was considered to be playing an important role in inhibiting the viral spread at an early stage of infection followed by T-cell dependent immune response, resulting in lower mortality.

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The effects of various chelating agents on the cytotoxicity induced by hexavalent chromium, potassium dichromate, were examined. HeLa cells were incubated in Eagle's minimum essential medium (MEM) with or without the chromate alone, or both chromate and any one of DL-penicillamine, glutathione (reduced and oxidized form), L-cysteine, 2, 3-dimercapto-1-propanol (BAL) and ethylene-diaminetetra-acetic acid (2Na-EDTA and 2Na·Ca-EDTA). After a given period of incubation, cell growth and the amount of chromium in the cells were measured. The results obtained were as follows. 1) The growth of HeLa cells in MEM with 3.4 μM chromate at 3 days of incubation was approximately 15% of that of the control cells. The growth of HeLa cells in MEM containing 3.4 μM chromate and 670.2 μM DL-penicillamine was 96%. However, no remarkable antagonism against the chromate toxicity was induced by 0.3 to 325.4 μM reduced glutathione, 0.2 to 163.2 μM oxidized glutathione, 0.6 to 56.9 μM L-cysteine, 0.8 to 8.1 μM BAL, 0.3 to 268.6 μM 2Na-EDTA, 0.2 to 224.0 μM 2Na·Ca-EDTA. 2) DL-penicillamine could not restore the cell growth inhibited by a preincubation in MEM with 3.4 μM chromate for 6 or 24 hours. 3) The chromium content (as Cr) of HeLa cells in MEM with 3.4 μM chromate at 6 hours of incubation was 0.148 μg/10⁶ cells. When 670.2 μM DL-penicillamine was added to MEM containing 3.4 μM chromate, the chromium content decreased to 0.0141μg/10⁶ cells (70% reduction). Other chelating agents, 325.4 μM reduced glutathione, 569.3 μM L-cysteine and 805.2 μM BAL, also remarkably reduced the chromium content. However, 163.2 μM oxidized glutathione, 268.6 μM 2Na-EDTA and 224.0 μM 2Na·Ca-EDTA did not reduce the chromium content. These results suggest that DL-penicillamine might inhibit the chromium uptake by HeLa cells and thus prevent the chromium-induced cytotoxicity.

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In order to clarify the relationship between three types of Indonesian native pigs (Bali, Batak of North Sumatra, and Toraja of South Sulawesi) and seven populations of Asian wild boars (Honshu, Kyushu, and Amami of Japan, Taiwan, Sumatra and Sulawesi of Indonesia, and Thai), the multivariate craniometric comparisons using 10 mandibular measurements were carried out in the present study. The results are summarized as follows: (1) Seven populations of Asian wild boars were separated into three groups by sizes: small, medium, and large, based on the discrimination by the first principal component acceptable as a size vector. The second principal component, acceptable as a shape vector, also contributed some towards the discrimination of the small-sized group from the other groups. (2) There is a complete overlapping of the principal component chart between the Batak native pigs and Toraja native pigs, both of which are short-eared. These two pigs occupy the intermediate situation between the small-sized wild boar and the medium-sized one, when they are projected into the axis of the first component. On the other hand, when they are projected into the axis of the second component, these two pigs fell in the medium-sized group of the wild boar. The Bali native pig, which is a Hainun type of the native pig, is probably to be separated both from the wild boars and from other native pigs. (3) The proportion of the ramus of the mandible to the body of the mandible seems to be an important discriminator in its shape, since the eigenvectors of measurements belonging to the ramus of the mandible show negative values, whereas those belonging to the body of the mandible show positive ones, in the second component.

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Expression of major histocompatibility antigens (MHAs) on six cell lines derived from Marek's disease (MD) lymphomas was investigated by the membrane immunofluorescence test. The results revealed varying expression of MHAs on the cell lines tested (MDCC-JP1, JP2, HP1, HP2, RP1 and MSB1). Antigenic diversity of MHAs was observed even among four sublines and 18 clones derived from the MSB1 cell line, indicating that this cell line consists of a mixture of cells expressing different MHAs. Furthermore, after more than six months' cultivation, a change and/or a reduction of the reactivity with antisera specific for antigens of the B complex were observed in some of these MD cell lines. These results suggested the possibility of changes in MHA expression on MD lymphoma line cells during long-term cultivation in vitro.

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Cleft palate was induced in all hamster fetuses by means of a single intramuscular injection of cortisone at a dose of 1.0 mg/g body weight on day 10+12 hr of gestation. The elevation of the palatine shelves in the cortisone-fetuses occurred without interruption but about one day later than in the control-fetuses. The opposing shelves occasionally came in contact with each other without the subsequent fusion.

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In 4 of 8 orally infected piglets with Aujeszky's disease virus, necrotizing vasculitis was observed in venules, arteioles and lymphatic vessels running around the submaxillary lymphnodes and tonsils. Intranuclear inclusions, specific immunofluorescence and herpetic virus particles were present in the degenerating vascular endothelial cells. The findings strongly suggested multiplication of the virus within endothelial cells providing a source of dissemination.

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Lymphoid tumor cells derived from a calf form case of bovine lymphosarcoma were successfully passaged in nude mice for over 25 passages. The lymphoid tumor cells from this transplantable cell line, BTL-T3, have been continuously cultured in vitro for over 6 months, and the descendent cells which had a T-cell marker and was designated as "BTL-C3", expressed a tumor-associated surface antigen (CBTL-ASA), which was not detected on the normal lymphocytes nor the other forms of bovine lymphosarcoma.

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Mycoplasmas were isolated from oral or nasal cavities of two lions, a Muscovy duck, a falcated teal, and three Indian white-backed vultures among zoo-animals comprising 19 mammals, six reptiles and 17 birds examined. The isolates from the lion were biochemically identical to those of M. felis but serologically different from them. No strains from the Indian white-backed vultures reacted with any antisera against the reference mycoplasma strains from birds.

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