The optic nerve fiber layer (NFL) of the chicken retina was studied quantitatively and morphologically at 17 positions along seven radially arranged bands from the dorsal tip of the pecten oculi using electron microscopy. The number of nerve fibers was counted in areas 6 μm in width × the full thickness of the NFL. Myelinated nerve fibers in the NFL were also identified immunohistochemically using anti-myelin basic protein serum. The dorsal area (dorsal, dorso-temporal and dorso-nasal bands) in the retina had thin NFL and contained the largest number of nerve fibers, which were mainly thin and unmyelinated. The ventral area (ventral and ventro-temporal bands) had a thick NFL and contained a relatively small number of nerve fibers, many of which were myelinated. The nasal band had the thickest NFL and contained as many nerve fibers as the dorsal area, with the temporal band showing a high ratio of myelinated fibers. The band had a thick NFL and contained many nerve fibers with a relatively low ratio of myelinated fibers. The relationship between the number and composition of nerve fibers in the NFL to the chicken visual characteristics was discussed. Although the myelin in the chicken retina was loose type, the myelin-forming cells were similar in appearance to dense oligodendrocytes.

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In the present study, we investigated the sexual dimorphisms, effects of castration and sex hormone treatment, and strain differences in the morphology of mouse kidney by measuring the percentage of renal corpuscles with cuboidal cells in the parietal layer of the glomerular capsule, the diameter of renal corpuscles and the number of nuclei of epithelial cells in the proximal convoluted tubule. In ICR mice, the percentage of renal corpuscles with cuboidal cells and the diameter of renal corpuscles were larger in males than in females, and the numbers of nuclei were lower in males than in females. All of these parameters became similar to those of intact females by orchiectomy, and restored to levels similar to those in intact males after testosterone treatment to orchiectomized males. On the other hand, ovariectomy and estradiol treatment gave no effects to any of these parameters. Similar sexual dimorphisms were observed in BALB/c, C57BL/6, C3H/He and DBA/2 mice as in ICR mice, although the diameter of renal corpuscles was not significantly different between males and females in BALB/c mice. Strain differences were observed in each of these three parameters among the 5 strains. In conclusion, the present study demonstrated sexual dimorphisms, effects of castration and sex hormone treatment, and strain differences in the morphology of mouse kidney by morphometrical analysis.

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Since we have clarified the manipulation mechanism using the radial sesamoid (RS) in the giant panda (Ailuropoda melanoleuca), our aim in this study is to examine the position, shape and function of the RS morphologically, and to observe the attachment to the RS of the M. abductor pollicis longus and the M. opponens pollicis in the other Ursidae species. So, we focused on the carpus and manus of the polar bear (Ursus maritimus) and the brown bear (Ursus arctos) in this study. The RS was tightly articulated to the radial carpal, and could not adduct-abduct independently of the radial carpal. The M. abductor pollicis longus tendon and the M. opponens pollicis belly were attached to the RS, independently. In the polar bear, the deep concave and the flat surface were confirmed in attachment area for these two muscles. The morphological relationship between the RS and the M. abductor pollicis longus and the M. opponens pollicis in the two species of bears were essentially consistent with that in the giant panda. It also demonstrated that the manipulation mechanism of the giant panda has been completely based on the functional relationship between the small RS, and the M. abductor pollicis longus and the M. opponens pollicis in Ursidae species.

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Uterine natural killer (uNK) cells belong to the large granular lymphocytes in the murine pregnant uterus and play essential roles in pregnancy success. We defined whether uNK cells can produce epidermal growth factor (EGF) important for implantation and embryo growth. The uNK cells were immunohistochemically positive for anti-EGF antibody especially during days 6 to 9 and at day 15 of pregnancy. Immunoreaction for EGF receptor was observed on the stromal cells in the metrial gland and trophoblasts in the placental labyrinth. EGF secretion (72.1 ± 2.25 ng/10⁴ cells) was noted in cultured uNK cells isolated from the metrial gland at day 15 of pregnancy. Treatment of anti-asialo-GM1 antibody raised the level of EGF (129 ± 21.5 ng/10⁴ cells). These results suggest uNK cell can produce and release EGF for placental development.

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We compared the receptor specificity of Newcastle disease viruses from a variety of avian species, including chickens and wild waterfowl, using hemagglutination tests with erythrocytes from different animal species. All isolates from wild waterfowl agglutinated horse erythrocytes, while the chicken isolates did not. The results showed that the receptor specificity of Newcastle disease viruses is different, depending on the avian species from which the viruses are isolated.

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The distribution of specific receptors on target organs is a major factor in the host range restriction of influenza A viruses. To assess the correlation between host receptors and the receptor specificity of influenza A viruses from sea mammals, we examined the receptors for influenza A virus in seal and whale lungs. A binding assay using two sialyloligosaccharide (SAα2, 3Gal and SAα2, 6Gal)-specific lectins showed that SAα2, 3Gal, but not SAα2, 6Gal, was found in both seal and whale lungs. Correspondingly, seal and whale influenza viruses preferentially recognized SAα2, 3Gal, but not SAα2, 6Gal. These results indicate that sialyloligosaccharides present at the replication site of influenza A viruses correlate with the receptor recognition of the viruses isolated from sea mammals.

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This experiment was conducted to ascertain whether the growth of Salmonella Enteritidis (SE) or Salmonella Typhimurium (ST) would be suppressed in the presence of antibodies contained in egg yolks. Specific pathogen-free chickens (102 days of age) were subcutaneously immunized with oil-adjuvanted bacterin of SE or ST, twice within a four-week interval. During 160 to 170 days of age, eggs were collected, the yolks were removed and mixed with an equal volume of physiological buffered saline, inoculated with ten colony forming units (CFU) of SE or ST, and incubated at 37°C or 20°C for 23 hr. The growth of organisms in each yolk solution was examined. The egg yolk derived from non-immunized hens was examined in the same manner as the controls. There was no difference in the growth titer between the antibody-positive yolk and the negative yolk. The result suggests that the antibodies in the yolk do not influence the growth of each organism, even if the hens are highly immunized.

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Feline serum amyloid A (SAA) cDNA clone was isolated from a hepatic mRNA of a mixed-breed cat. The feline SAA cDNA clone contains 333 nucleotides and deduced amino acid sequence shows 87.4%, 73.9%, and 65.8% homology with dog, human and mouse SAA respectively. Relative to the human and mouse SAA proteins, an additional peptide of eight amino acids is specified in the feline cDNA clone. Recombinant feline SAA (rfSAA) was expressed at high levels using pGEX bacterial expression system. Cleavage from the fusion moiety, and purification using glutathione-sepharose yielded pure soluble form of rfSAA. Antibodies generated against rfSAA were specific for feline SAA and showed no cross-reactivity with human SAA. Furthermore, antibodies against human SAA did not react with feline SAA. These results indicate that antigenicity of feline SAA is totally different from human SAA.

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In order to obtain available biochemical parameters related to the marbling score (MS), by which the meat was graded from 1 to 12 in Japan, serum biochemical values were examined in 13 Japanese Black steers during the fattening stages. The steers were divided into high MS (8-11) and low MS (4-5) groups and the fattening stage was also divided into 4 stages according to their feed contents; Stage I (11-13 month-old; 0-4 months after the fattening (af)), II (5-9 months af), III (10-14 months af), and IV (15-20 months af). High MS group showed significantly higher levels in glucose and urea nitrogen (at Stage I), albumin/globulin ratio (at Stage II), and magnesium (at Stage I and III), and lower level in vitamin A (at Stage III) than low MS group. These biochemical parameters, indicating conditions of steers during the fattening stages, are considered to have a possibility related to MS in fattening steers.

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A 3 year-old female Japanese Black cattle was diagnosed as diabetes mellitus (DM). Hyperglycemia (295 mg/dl), increase of serum fructosamine (487 μmol/l), elevated glycosylated hemoglobin A₁ (GHbA₁; 10.9%), low concentration of serum insulin (< 1.0 μU/ml), increased serum glucagon (399 pg/ml), and glucose intolerance (glucose disappearance rate; k=0.53) were noted. On the histopathologic findings in pancreas, insulitis with infiltration of mononuclear cells was found. This case suggests that serum fructosamine and GHbA₁ are available parameters for understanding of pathophysiological conditions of bovine DM.

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Primer designed on the basis of the conserved transcription start point (tsp) region of human and bovine gene transcripts, encoding the Interleukin-4 (IL-4), and subsequently the gene specific primer and an adaptor primer pair, was successfully used, to generate the full-length complementary DNA (cDNA) sequence coding for canine IL-4 (cIL-4), from pokeweed mitogen stimulated canine peripheral blood lymphocytes. The full-length cIL-4 contains an open reading frame of 399 nucleotides (nt), with a 5' ends of 66 base pairs (bp) and 3' ends of 125 bp. The nucleotide sequence contains six possible Asn-X-Thr or Asn-X-Ser linked glycosylation sites. Five sequence motifs of TATT or ATTTA, responsible for the regulation of gene expression, are found in the 3' untranslated region.

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The bottle-nosed dolphin (Tursiops truncatus) interferon-γ (IFN-γ) cDNA was cloned from mitogen stimulated peripheral blood mononuclear cells (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The dolphin IFN-γ cDNA was 585 base pairs in length and contained an open reading frame of 498 nucleotides encoding an IFN-γ precursor of 166 amino acids, with cleavage of the putative signal peptide of 19 amino acids. Analysis of the mature amino acid sequences revealed two potential N-linked glycosylation sites. Comparison of the predicted amino acid sequence showed that dolphin IFN-γ shares 86, 85, 63 and 42% similarity with the bovine, ovine, human and mouse IFN-γ_s, respectively.

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We investigated the diet content and properties of blood and milk in 11 pairs of Japanese Black cows and their calves for the one month following delivery. Four pairs (group A) had been no cases of white diarrhea during the year prior to this study, and 7 pairs (group B) had been a high occurrence of white diarrhea in calves during the same period. Properties of dams&rsque; diet in groups A and B before the onset of white diarrhea in calves were as follows: TDN fullness rate 98 ± 2% vs. 110 ± 5%, DCP fullness rate 151 ± 2% vs. 200 ± 33%, and starch content 5 ± 2% vs. 14 ± 3%. Blood and milk samples were collected from cows weekly and at the onset of white diarrhea in calves. No calves in group A had white diarrhea, while 5 out of 7 calves in group B had the diarrhea in this study. All cows were fed mixed hay, but 3 out of 5 cows whose calves had white diarrhea had been additionally given 3.0-3.5 kg of corn-silage a day from 4-5 days before onset. In all cows in the white diarrhea development group, the fat content of milk increased by 2.94 ± 1.82% at the day of onset in comparison before. Serum TG and BUN, respectively, increased by 3.8 ± 1.3 mg/dl and 3.7 ± 2.0 mg/dl. Feeding cows a low-starch diet and additional corn-silage may induce a transient increase in blood TG, BUN and milk fat, which may contribute to the occurrence of white diarrhea in calves.

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Electron microscopic observation was performed on platelets activated by collagen stimulation in Japanese Black cattle with δ-storage pool deficiency (δ-SPD) to identify their morphological and functional abnormalities compared from normal bovine platelets. Platelets of normal Japanese Black cattle changed their shapes to spherical and were in the phase of release reactions 5 min after the collagen (90.9 μg/ml) stimulation, and most of platelets were aggregated. On the other hand, in δ-SPD cattle, most of a granules were still dispersed in activated platelets, although the spherical shape change of the platelets was observed. These results suggested that there are abnormalities in the release reactions in platelets of δ-SPD cattle.

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The effects of systemically administered fibroblast growth factor-2 (FGF-2) at doses of 0.1 and 0.3 mg/kg/day for 7 days were investigated 5-week-old male SAMP6 mice, a model of low turnover osteopenia. The bone histomorphometry in the distal epiphyseal growth plate of the femur showed that 0.3 mg/kg/day of FGF-2 decreased the longitudinal growth rate and cartilage cell production rate and increased the growth plate width. Growth plate chondrocytes showed the features of defective endochondral ossification at the same dosage level. In the distal one third of the femur, the marrow trabecular area, endocortical mineral apposition rate and/or bone formation rate were increased in both the SAMP6 mice given 0.1 and 0.3 mg of FGF-2/kg/day. In this region, the endocortical osteoblasts were hypertrophied with some layers of overlying proliferated fibroblastic mesenchymal cells. The presence of small foci of bone formation within the layers of these mesenchymal cells indicates their osteogenic potential. On the other hand, the periosteal bone formation rate in the mid-shaft of the femur was depressed in the 0.3 mg/kg/day group. These results suggest that systemically administered FGF-2 may have the possibility to increase the peak bone mass in SAMP6 by stimulating the osteoprogenitor cells to proliferate and differentiate into osteoblasts and enhancing endocortical bone modelling. The higher dose of FGF-2, however, inhibited both endochondral and periosteal bone formation.

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The effect of X-irradiation on the progression of the cell cycle in cell lines from LEC and WKAH rats was investigated by a flow cytometer. When the cells were exposed to 5 Gy of X-rays at S phase, the proportion of S-phase cells in both cell populations decreased with incubation time and that of G2/M-phase cells was approximately 80% at 6 hr post-irradiation. At 12 hr post-irradiation, approximately 45% of the WKAH rat cells appeared in the G1 phase. However, 80-90% of LEC rat cells remained in the G2/M phase and less than 5% in the G1 phase during 6-12 hr post-irradiation. Thus, the LEC rat cells irradiated at S phase remained in the G2/M phase for at least 6 hr longer than did the WKAH rat cells.

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To clarify the clinicopathological features of canine epulides, 189 epulides were reviewed retrospectively. The incidence of the fibromatous, ossifying, acanthomatous and giant cell epulides were 56.6% (107/189), 23.3% (44/189), 18.0% (34/189) and 2.1% (4/189), respectively. The average ages of dogs with fibromatous, ossifying, acanthomatous and giant cell epulides were 8.8, 8.4, 7.8 and 8.7 years, respectively. The male/female ratio of dogs with the acanthomatous epulis (0.8) was lower than those of dogs with the fibromatous (1.9), ossifying (1.4) and giant cell epulis (3.0). There were slight breed differences among the types of epulides. The most noticeable result was that 38.2% of the acanthomatous epulis occurred in Shetland sheepdogs. 43.9% of the fibromatous epulis and 52% of the ossifying epulides arose around maxillary premolars, while 58.8% of the acanthomatous epulis arose around the mandibular canines. Dogs with the fibromatous and ossifying epulides had more severe dental plaque deposition than those with the acanthomatous epulides. Few of the fibromatous (6/104) or ossifying epulides (4/44) showed recurrence after excision, while the majority (21/23) of the acanthomatous epulides showed rapid and repeated recurrences after surgical excision. Epulides treated with hemimandibulectomy or bleomycin chemotherapy did not recur. Giant cell epulides showed no recurrence after surgical removal. These results indicate that the acanthomatous epulis differed from other types of epulides in biological and morphological features and poor prognosis.

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Five racehorses in apparently normal condition succumbed to sudden cardiac death (SCD) during or shortly after intensive training exercise. Cardiopathologic examination was performed. In 1 of the 5 horses, the use of an electrocardiogram (ECG) recording taken continuously for 440 sec enabled us to analyze some of the arrhythmias in the terminal event of SCD. The ECG tracing exhibited the R-on-T phenomenon following a pair of ventricular premature contractions (VPCs). The phenomenon rapidly degenerated into ventricular fibrillation, which led to cardiac arrest. In all 5 horses cardiopathologic examination revealed the following lesions: (i) foci of myocardial fibrosis in the right atrium located close to the sinoatrial (SA) node, (ii) fibrotic and/or fibroplastic changes in the upper portion of the interventricular septum, including the atrioventricular (AV) conduction system, and (iii) arterio- and arteriolosclerosis of the SA and AV node vessels. Pathogenetically, the process by which the focal lesions of myocardial ischemia secondary to vascular sclerosis progressed into fibrosis and/or fibroplasia could play a major role in the genesis of arrhythmias. Presumably the fibrotic and/or fibroplastic changes in the area of the AV bundle and its bundle branches are closely related to the onset of fatal ventricular arrhythmias such as VPCs, deteriorating into ventricular fibrillation. SCD in training and racing Thoroughbred horses appears to be due to arrhythmia.

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In a 3-year-old Holstein cow, a tumor mass replaced the left olfactory bulb. The tumor was highly or moderately cellular, and consisted of tumor cells showing pleomorphism and anaplasia, sometimes with intracytoplasmic granules. The tumor showed weak reactivity for neurofilaments (NF) in most cells with distinct staining in a minority, and it was extremely rare to see neoplastic cells with positivity for glial fibrillary acidic protein (GFAP). The neoplastic cells displayed some ultrastructural features reminiscent of ganglionic cells, and the cytoplasmic granularity was due to the presence of numerous lysosomes. This tumor expressing both NF and GFAP may be histogenetically related to brain tumors of pluripotential cell origin in calves.

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Thirteen primiparous and 41 multiparous Holstein-Friesian cattle were used to study the relationship between maternal plasma progesterone (P₄) and estrone sulfate (E₁S) concentrations and the prevalence of dystocia. The calvings in 4 heifers and 30 cows were normal (eutocia), while the calvings in 9 heifers and 11 cows were difficult (dystocia). Neither the concentrations of P₄ nor E₁S were different between the groups with eutocia and dystocia from days 90 to 270 of pregnancy. However, a few days prior to parturition, eutocial cows and heifers showed a sharp decline of plasma P₄, while dystocial cattle did not show such a remarkable decline of P₄ concentration. Plasma P₄ levels in dystocial cows a few days antepartum were significantly higher than in eutocial animals (P<0.05 or P<0.01). Prepartum E₁S concentrations were significantly lower (P<0.05) in dystocial than eutocial cattle during the prepartum period from days 6 to 1 in heifers and from days 3 to 1 in cows. These results suggest that insufficient production of E₁S and delayed regression of the corpora lutea are possible causes of dystocia in cattle.

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The aim of the present study was to examine the LH response to exogenous estradiol in 4 heifers with ACTH-induced ovarian follicular cysts. During the control experiment, administration of estradiol 24 hr after PGF_2α in luteal phase heifers resulted in a LH response in all 4 heifers. The LH response was obtained between 16-20 hr after estradiol administration. The peak LH concentration (Mean ± SEM; 5.1 ± 0.8 ng/ml) during the control study was significantly different (P<0.05) from the concentration after cyst formation. None of the 4 heifers responded to estradiol after ovarian cyst formation. This result suggests that heifers with ACTH-induced ovarian follicular cysts may have a defective hypothalamio-pituitary response to exogenous estradiol similar to cows with spontaneous ovarian cysts.

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A sero-epizootiological survey was conducted for Japanese encephalitis virus (JEV) and Getah virus (GeV) at 10 to 20 regional horse race tracks from 1991 to 1997 in Japan. It was observed that geometrical mean (GM) antibody titer to JEV and GeV was 10 to 50 times higher than others at several race courses (RCs) almost every year. Of them, several race horses showing high antibody titer, which were suggested to be infected with the virus, were also observed in this survey. These data suggested that the viruses have spread among race horses almost every year in Japan, although, fortunately, no horse showing clinical illness due to these viruses was observed. The calendar years and the names of the race courses in which the spread of JEV was suggested were Sonoda and Nakatsu RCs in 1991, Nakatsu RC in 1992, Arao RC in 1993, Nagoya RC in 1994, Kaminoyama, Urawa, Saga and Arao RCs in 1995 and Ooi and Saga RCs in 1997. Spread of JEV was not observed in 1996. The calendar year and name of the race courses at which the spread of GeV was suggested were at Ooi, Sonoda and Nakatsu RCs in 1991, Arao RC in 1992, Nakatsu RC in 1994 and 1995, Funabashi RC in 1996, Saga RC in 1997. It was suggested that surveillance of JEV and GeV should be continued in the future in at least the southern to middle parts of Japan. It has also been suggested that it is necessary to promote the wider use of vaccines to persons related to horse racing.

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The type II feline infectious peritonitis virus (FIPV) epitopes for neutralizing and enhancing antibodies are present on large spike glycoprotein (S) protein. In this study, we established monoclonal antibody-resistant mutant viruses resistant to three different monoclonal antibodies with neutralizing activity in Felis catus whole fetus cells and enhancing activity in feline macrophages, recognizing distinct epitopes on type II FIPV S protein. By comparing the nucleotide sequences of these mutant viruses with that of wild-type virus, we attempted to identify the neutralizing epitopes. The mutations were localized in the region of amino acid residues from 480 to 649 from the N terminal of the S protein.

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To investigate the susceptibility of early bovine embryos to noncytopathogenic bovine viral diarrhea virus (NCP BVDV), 2- and 4-cell embryos produced in vitro from which zona pellucida had been removed by pronase treatment, and hatched blastocysts were exposed to 10⁶ TCID₅₀/ml of NCP BVDV No. 12 strain. The virus was detected in all embryo samples immediately prior to cultivation but not in the medium. After 24-hr culture, the virus was isolated from four media and two embryo samples in four experiments in the blastocyst group, and the viral antigen was demonstrated in the cytoplasm of the embryo cells by the immunofluorescent technique. By contrast, no virus was recovered from, or viral antigen detected in samples from the 2- and 4-cell embryo group in any of the experiments, even though they were exposed to the virus after removal of the zona pellucida. These findings suggest that 2- and 4-cell embryos are unlikely to be susceptible to NCP BVDV, but that blastocysts are capable of being infected with the virus.

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To establish the evolutionary association between the equine 1 H7 HA and M genes, phylogenetic analyses of the six internal gene segments of equine 1 influenza viruses (H7N7 subtype) were performed using partial nucleotide sequences. The results demonstrated that five internal genes (PB1, PB2, PA, NP and NS) of equine 1 viruses isolated after 1964 were replaced by those of equine 2 H3N8 viruses. However, the M gene was maintained during the evolution of these equine 1 viruses. These findings suggest a functional association between equine H7 HA and M gene products, most likely M2 protein.

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