Some serological diagnosis methods and examinations for detection of antibodies to sheep-associated malignant catarrhal fever (MCF) infection were investigated. The wildebeest-associated MCF virus strain WCll propagated on fetal bovine thyroid cell cultures was used as an antigen. Antibodies were detected by complement fixation (CF) tests in cattle pathologically diagnosed as having sheep-associated MCF, as well as in cattle experimentally infected with MCF virus strain WCll. However, immunodiffusion precipitation was only detected in cattle infected with MCF virus strain WCll. The results of serological investigation by CF tests indicated that 64.3% of sheep possessed antibodies to MCF virus in the Hokkaido district of Japan and all serum samples which contained CF antibody titers greater than 1:4 had antibody titers larger than 1:8 in indirect immunofluorescence tests. The CF test we demonstrated here is available to quantitatively detect MCF virus antibody titers in epidemiological surveys.

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Histological and ultrastructural development of the olfactory epithelium (OE) and vomeronasal organ (VNO) was investigated in the Japanese reddish frog, Rana japonica. Tadpoles, from hatching to the end of metamorphosis, and adult frogs were examined. In the adult, olfactory cells of the OE were equipped with olfactory vesicles with long cilia, but supporting cells with microvilli. The supporting cells of the OE contained secretory granules, PAS-positive by light microscopy, in their apical cytoplasm. On the contrary, sensory cells of the VNO were equipped with microvilli, and supporting cells of the VNO were equipped with cilia, but without secretory granules. Embryologically, the olfactory cells were indistinguishable from the supporting cells in the olfactory placode (primitive OE) lining the nasal pit, at hatch. The VNO appeared as a concave of the ventral part of the OE at 4 days after hatch. At this time, the olfactory and supporting cells of the OE became distinguishable from each other. Secretory granules were formed in the supporting cells of the OE at 36 days after hatch, and the OE was similar in fine structure to that in the adult. While, the VNO remained immature at 24-36 days after hatch, and did not complete its ultrastructural development at 60 days after hatch, the end of metamorphosis. These findings suggest that the OE may take part in the olfaction earlier than the VNO in ontogeny.

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The olfactory organs are equipped with their own associated glands, Bowman's glands (BG) of the olfactory epithelium (OE), and Jacobson's glands (JG) of the vomeronasal organ (VNO). Histology and ultrastructure of these glands in the adult were investigated in the Japanese reddish frog, Rana japonica, along with their development from hatching to the end of metamorphosis. In the adult, BG cells contained large, electron-opaque secretory granules, intensely PAS-positive by light microscopy, while JG cells contained middle-sized secretory granules with moderate electron density, only faintly PAS-positive. Embryologically, BG appeared within the OE at 44 days after hatch, increased in number and were situated in the lamina propria under the OE at 52 days after hatch. BG cells contained large, electron-opaque granules, and well-developed rER at this time. While, JG appeared much earlier than BG. The VNO appeared as a concave of the ventral part of the OE at 4 days after hatch, and JG appeared under the VNO at 10 days after hatch. JG cells contained well-developed rER at 12 days after hatch. Secretory granules appeared in a small number in JG at 24 days after hatch, and increased thereafter. These findings suggest that JG may take part in secretion earlier than BG in ontogeny.

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Conditions for cytopathic effect (CPE) inhibition assay of canine interferon (IFN) activity in Madin-Darby canine kidney (MDCK) cells and in canine tumor cell line A72 was investigated using the New Jersey strain of vesicular stomatitis virus (VSV). The culture supematant from canine splenocytes stimulated with ultraviolet-irradiated Newcastle disease viruses was used as reference IFN. MDCK cells were resistant for growth of VSV when the cells were confluent. Full CPE was observed only in a sparsely growing culture. Canine IFN activity could be assayed on less than 10⁴ MDCK cells/well of a 96-well microplate, and more than 10⁵ TCID₅₀/ml of VSV was required. In A72 cells, VSV growth was not as dependent on cell density as in MDCK cells, requiring 10³ TCID₅₀/ml of VSV. MDCK-VSV system showed a higher IFN sensitivity than A72-VSV, whereas reproducibility was higher for the latter than the former. Based on these findings, A72-VSV system for canine IFN assay is recommended for practical use due to its easy handling characteristics.

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The ultrastructure of the endothelial cells of the deep circumflex iliac and caudal superficial epigastric arteries supplying the abdomino-inguinal mammary gland of female Wistar rats was studied throughout the reproductive cycle with an electron microscope and image analyzer and compared with that of mammary gland capillaries. The subendothelial space of the arteries was broader at the virgin and post-weaning stages but narrower in pregnancy and lactation. In some rats, the endothelial cells of the arteries radiated some blunt processes into the media through fenestrations in the internal elastic lamina. The density of pinocytotic vesicles (PV) in the arteries (number of PV per μm² of endothelial cytoplasm) significantly increased during pregnancy and reached its maximum value during lactation, then subsequently decreased during the post-weaning stage. The mammary gland capillaries showed the maximum changes of PV during pregnancy and lactation, with twofold and fourfold increases during the respective periods. The density of mitochondria increased significantly in the capillaries during pregnancy and lactation. The length of the marginal folds and microvillous processes increased significantly during lactation in both the arteries and capillaries, and especially, increased twofold in the mammary gland capillaries. It is assumed that the ultrastructural changes of the endothelial cells of the arteries and capillaries are closely associated during reproductive cycle with the functional demand of the mammary glands.

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Cerebral vascular mineralization was found in 12 (60%) of 20 3- to 10-year-old healthy horses collected at an abattoir. It was variable in degree and occurred mostly in the pallidal arteries showing two types of lesions; small globoid bodies along capillaries, and amorphous deposits in the wall of arterioles, small- or medium-sized arteries and veins. Both types were strongly positive for periodic acid-Schiff reaction, and weakly positive for von Kossa's and Berlin blue stains. Elemental analysis of the deposit revealed the presence of large amounts of aluminum, moderate amounts of phosphorus, zinc, calcium and iron, and a small amount of sodium.

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The longitudinal distribution of pulmonary vascular compliance was evaluated in isolated canine lung lobes using arterial-(AO), venous-(VO), and double-occlusion (DO) techniques. Total vascular compliance (C_T) was separated into pulmonary arterial (Ca) and venous compliance (C_v) in lumped model of pulmonary circulation. Under constant pulmonary venous pressure (P_v) at 5 mmHg, blood inflow to the lobe (Q) was gradually increased by changing pulmonary arterial pressure (Pa) from 10 to 22 mmHg at 4 mmHg ranges. Changes in vascular blood volume (ΔV) with each increment in Q were determined by decreased reservoir blood volume of perfusion system. DO was performed at each level of Q and allowing all vascular pressures to equilibrate at the same static pressure (Ps), which was equal to the compliance-weighted average pressure in the circulation. C_T was obtained from the slope of the relationship between Ps and ΔV. When Pa and Pv were 14 and 5 mmHg, AO, VO, and DO were performed to measure pressures at Ca (Pca) and Cv (Pcv) and Ps. The arterial-to-venous compliance ratio (Ca/Cv) was evaluated using Pca, Pcv, and Ps measurements. C_T was 0.113±0.012ml/kg/mmHg. Ca/Cv was 0.30. Ca and Cv were 0.026±0.013 and 0.087±0.007 ml/kg/mmHg, respectively. These data demonstrated the usefulness of AO, VO, and DO techniques in evaluating the longitudinal distribution of compliance in canine pulmonary vasculature.

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Bovine colostrum has growth factor activity for stimulating DNA synthesis in calf kidney epithelial cells (CKT-1), Madin-canine kidney epithelial cells (MDCK) and rat L6 myoblasts (L6), of which the DNA stimulation level and the activity change with the time elapsed after the birth of a calf varied with their respective cells. The growth factor activity of colostrum for CKT-1 was stable regardless of the collection time of colostrum, and it was purified about 3, 650-fold in an overall yield of 1.2% from colostrum obtained 30 min after the birth of a calf. The purified growth factor had a molecular weight (MW) of 5, 000 and an isoelectric point of pH 9.7, and the amino acid composition was: Asx₅, Thr₂, Ser₄, Glx₁₄, Pro₂, Gly₄, Ala₄, Val₂, Ile, Leu₂, Tyr, Phe, Lys, His and Arg. The stimulated DNA synthesis in CKT-1 and L6 by the addition of purified growth factor at a final concentration of 16 ng/ml was as the same extent as calf serum at a final concentration of 1.52 mg/ml, and the relative activity for CKT-1 was even greater than that for L6.

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We experimentally demonstrated the anti-asthmatic effects of M-711, the dry extract of a Chinese herb medicine called Moku-boi-to, using the rat model of allergic asthma. Allergic asthma was induced by the antigen-antibody reaction in the rats and they showed anaphylactic symptoms accompanied with hypotension and depression of respiratory function. More than 20 mg/kg body weight of M-711 was effective in relieving the asthmatic symptoms like methylephedrine. It could lessened the suppress of respiration and provided a good improvement. It also reduced a drop of the blood pressure and improved it quickly. Its ingredients alone were much less effective than M-711 as the total. Those data suggested that M-711 was effective for alleviating allergic asthma in the present study and that its action was provided by interaction of all ingredient raw herbs of M-711.

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Lactoferrin (LF) is known to have broad spectrum antimicrobial properties. In regards to its defense mechanism against parasitic infection, it has shown phagocytic activity in the destruction of amastigotes, an intracellular parasitic form of Trypanosoma cruzi in macrophages. The effect of bovine lactoferrin on the intracellular growth Toxoplasma gondii parasites was examined in murine macrophage and embryonal cells. Co-cultures of host cells with the parasites were supplemented with either lactoferrin, apo-lactoferrin, holo-lactoferrin or transferrin in the culture media for varying periods. The growth activity of intracellular parasites in the host cells was determined by the measurement of selective incorporation of ³H-uracil. Supplement of lactoferrin had no effect on the penetration activity of the parasites, while development of intracellular parasites was inhibited linearly in concentration of lactoferrin. Supplement of apo-lactoferrin and holo-lactoferrin, but not transferrin showed similar effects. These suggest that lactoferrin induces the inhibitory effects on the development of intracellular parasites. Pretreatment of lactoferrin to the macrophages, however, did not show any inhibitory effects. Whereas, mouse embryonal cells preincubated with lactoferrin suppressed the intracellular growth. Thus, the action of lactoferrin to macrophages would be different from that of mouse embryonal cells.

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Nine aborted fetuses and one newborn calf, diagnosed as Chlamydia psittaci (C.psittaci) infection, were pathologically examined. The characteristic lesions in the liver were focal necrosis in 9 aborted fetuses and granulomatous necrotic foci in the newborn calf. Moderate numbers of intranuclear inclusion bodies were found in necrotic foci of the liver, spleen, thymus, lymph nodes, adrenal gland, kidney, lung and forestomach. Immunohistologically, a small number of C.psittaci antigens was demonstrated in necrotic foci of the liver and correlated with distribution of elementary bodies. Moderate numbers of infectious bovine rhinotracheitis (IBR) virus antigens were also detected in degenerating and necrotizing parenchymal cells in various organs and correlated with distribution of intranuclear inclusion bodies. Thus, these aborted bovine fetuses and newborn calf were interpreted as being dually infected with C.psittaci and IBR virus during pregnancy.

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Recombinant Hantaan virus nucleocapsid protein expressed in silkworm larvae was applied as a serological diagnostic antigen in Western blots (WB) of human sera. The sensitivity of this method was similar to that of the IFA test. Hemorrhagic fever with renal syndrome (HFRS) and nephropathia epidemica diagnosed by their cross-reactivity in WB. The specificity of this method was higher than that of IFA test because the background was low. Sera that exhibited high background staining in the IFA test were readily diagnosed with this method. We recommended WB using recombinant Hantaan virus nucleocapsid antigen as a confirmatory procedure for the serodiagnosis of hantavirus.

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Effects of vitamin D₃ (VD₃) injection on the activity of thyroid parafollicular cells (C cells) and calcium (Ca) metabolism were examined in rats of non-(NP), middle (MP) and late pregnancy (LP). At 3 days after injection, the average area of a C cell was significantly wider (P<0.01) in the VD₃ groups than that in their control groups in NP, MP and LP. On the other hand, the plasma Ca concentration in the VD₃ groups decreased significantly (P<0.05) in comparison with that in their control groups in NP and LP, and tended to decrease in MP. These results suggest that injection of VD₃ may accelerate the activity of C cells, which may result in the decrease of plasma Ca concentration in both non-pregnant and pregnant rats.

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Among the 23 Salmonella strains isolated from 21 diseased chickens and 2 embryonated eggs in 6 poultry farms near Lusaka City in Zambia, serovars identified were S.Gallinarum (11 strains), S.Agona (7 strains), S.Alamo (1 strain), S.Infantis (1 strain), S.Virginia (1 strain), S.Haifa (1 strain), and S.Dublin (1 strain). S.Gallinarum was detected at the highest incidence and from all the poultry farms. Fourteen serovars have been reported for the chickens in Zambia so far. Three serovars (S.Alamo, S.Haifa, S.Virginia) were newly identified in this study.

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Diaphragmatic muscles in two slaughtered Holstein-Friesian revealed slightly pale color, swelling, and stiffness on palpation. Histologically the muscle fibers showed internal nuclei, fiber-splitting, variation in diameter, central core-like structures, sarcoplasmic masses, and vacuolar degeneration. These lesions were the same as those in dystrophy of the diaphragmatic muscles in Holstein-Friesian cows. It was demonstrated that muscular dystrophy of the diaphragm in Holstein-Friesian cattle occured also in males, probably by inheriting an autosomal recessive trait.

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During the period from January to March 1995, the authors first isolated Bartonella henselae from the blood of three (9.1%) of 33 domestic cats in Japan. The three cats were a 1.5-year male pet cat-old with urinary retention, and 6-year-old female pound and age-unknown female pet cats with no abnormalities. The blood was taken in a lysis-centrifugation tube (Wampole Isolator tube) and cultured on 5% rabbit-blood heart infusion agar plates at 35°C in the 5% CO₂ atmosphere. Visible tiny rough colonies developed 14 days after incubation. The isolates showed Gram-negative and pleomorphic rods in microscopic observation. The DNA extracted from the isolates was amplified by PCR using two primers, which were specific for the rikettsial citrate synthase gene. The isolates were identified as B.henselae from the patterns of digestion with TaqI and HhaI of the amplified gene. It was confirmed that cats in Japan harbored B.henselae in their blood, and that cats play a significant role as the reservoir of the organism.

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A preparation of peptidoglycan (PG) of swine Bifidobacterium thermophilum was orally administered to SPF-BALB/C and ICR mice and its effect on phagocytosis splenetic neutrophils from PG administered mice was measured by chemiluminescent response (CL) and fluorometric analysis and the result was compared with that of non-treated mice. PG stimulated phagocytosis of neutrophils in a dose-dependent manner, whereas dosage exceeding the optimum concentration (500 μg) inhibited phagocytosis. The maximum effect on phagocytosis of neutrophils was observed at 3 days after administration of PG 500 μg. The result of fluorometric analysis was almost similar to that of CL. These results indicate that orally administered PG enhances the activity of the phagocytosis of splenetic neutrophils from mice.

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Five field strains of Erysipelothrix rhusiopathiae belonging to serotype 2 were compared for their growth ability, immunogenicity in mice, SDS-PAGE profile of cell surface proteins and their immunoblotting patterns. Strain Tama-96 showed the most stable growth in Feist medium and tryptose phosphate broth with Tween 80 (TPB), and its immunogenicity was highest in a mouse protection test using the inactivated vaccines prepared from 20-h TPB culture. The 50% mouse protective dose of the vaccine was only 12 μl. SDS-PAGE and immunoblotting patterns of the proteins were similar among the strains in general and indicated that 66 to 64 kDa protein antigens were dominant.

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