The effects of alteration of metabolic substrates and supplementation of amino acids on in vitro development of bovine embryos were examined. One-cell embryos derived from in vitro fertilization were cultured using a semi-chemically defined medium, Brinster's Medium, containing 3 mg/ml bovine serum albumin (BMOC-3) as a basal medium. BMOC-3 did not support embryonic development to the morula stage, but blastocysts were obtained by omitting glucose and lowering the lactate level from 20.12 to 3.0 mM and the pyruvate level from 0.5 to 0.3 mM (mBMOC-3). Supplementation of amino acids (glutamine plus the other 12 essential amino acids, as well as further addition of 7 non-essential amino acids) improved the embryonic developmental rate to the blastocyst stage with an increase in cell number equal to that for co-cultured embryos with bovine oviductal cells in TCM 199 with 10% fetal calf serum. These results clarified that the one-cell bovine embryo can develop to the blastocyst stage in a semi-chemically defined medium, and that amino acids support embryonic development in vitro.

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Unilateral degenerative atrophy of the optic nerve (ON) occurred in 6 of 80 male and 4 of 80 female Slc: Wistar rats. Two of these cases completely lost the intracranial portion of the unilateral ON and the remainder had the small ON. The optic disc and ON were located histologically in the posterior pole of the eyeball of 2 rats with no intracranial ON. ON lesions in all cases were characterized by a reduced number of axons with a small number of myelinated axons and marked astrogliosis. There were also swelling, fragmentation and spheroid formation of axons, as well as thickening of the connective tissue sheaths and vessel walls in the ON. One side of the optic chiasma and optic tract contralateral to the affected ON reduced in volume, became degenerated and were accompanied by gliosis. Focal of diffuse degeneration of the retina was observed in the eyeballs with affected ON. Retinal ganglion cells decreased in the number showing chromatolysis. These retinae became thin and developed degeneration of both inner and outer portions with sclerotic changes in the retinal vessels. The ophthalmic and ciliary arteries in the eyeballs with affected ON often developed broliferative or occlusive. endoarteritis, suggesting that retinal lesions may have resulted not only from axonal degeneration in the ON but also from ischemia. Histologic lesions suggestive of transneuronal degeneration were found in the contralateral lateral geniculate body and rostral colliculus. Based on the data presented, it was presumed that a primary lesion may have been induceds in the ON by a circulatory disturbance and followed by retrograde and anterograde degeneration in the visual pathways.

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Mechanisms of postinfectious pruritus and peracute death in mice by pseudorabies virus (PRV) were investigated by inoculating the Yamagata-S81 strain of PRV peripherally or intracerebrally into 4-week-old ICR and BALB/c mice. Clinical signs developed most rapidly in mice inoculated intracerebrally, with intermediate speed in mice inoculated intraocularly, and slowly in mice inoculated subcutaneously. Since intraocularly inoculated mice showed an acute reaction and this is considered a peripheral route, the distribution of viral antigens in the nervous system of intraocularly inoculated mice was examined immunohistologically. Viral antigens were mainly detected along the trigeminal and the oculomotor nerves, but neither necrosis nor an inflammatory response was observed in these areas. The infectious virus was efficiently recovered from the viral antigen-positive tissues. In the pruritic skin lesions, viral antigens were not observed. These findings indicate that the main route of viral spread in intraocularly inoculated mice is the trigeminal and oculomotor nerves and that the virus in the trigeminal nerve may trigger pruritus.

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By means of the enzyme-linked immunosorbent assay (ELISA), antibodies to Borrelia burgdorferi were examined in 970 cattle from southern Hokkaido, Japan. The seropositivity rate in the cattle less than 2 years old differed significantly from that in those 3 years old or more. A seasonal variation was also observed in the rate of seropositivity to B. burgdorferi. High levels of anti-B. burgdorferi antibody were detected even in sera from a healthy group of cattle, indicating that there are likely to be many inapparent cases of B. burgdorferi infection. This article discusses the syndromes associated with Lyme borreliosis and addresses the issues of diagnosis and treatment in animals.

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Proliferative activity of murine interleukin-2 (IL-2) dependent T cells (CTLL-2) was detected in the culture supernatant of canine peripheral blood lymphocytes (PBL) stimulated with phytohemagglutinin-P (PHA-P), and was defined as dog IL-2. The highest production of IL-2 was obtained under the conditions in which a PBL population of 2×10⁶ cells/ml was stimulated with PHA-P at a concentration of 10μg/ml at 38°C for 48 hr. Dog IL-2 activity was significantly inhibited by heating at 65°C, acidification under pH 4, alkalification over pH 10, and trypsin exposure. A peak of dog IL-2 activity was detected in the fraction with a molecular weight of approximately 31, 000 by gel filtration. Long-term culture of canine lymphocytes was successful over 10 passages by using dog IL-2 with PHA-P-stimulation every 3 passages. The cultured cells mostly consisted of small- and medium-sized lymphocytes. These cells reacted to anti-dog thymocyte rabbit serum and anti-dog Thy-1 mouse monoclonal antibody. These cells were therefore considered to originate in T-lineage lymphocytes. Cytostasis of PBL from intact dogs reacting to canine transmissible venereal sarcoma cells was increased significantly when PBL was cultured for more than 30 days with homologous IL-2.

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Levels of 4-androstenedione (A), 5α-dihydrotestosterone (DHT) and testosterone (T) in plasma collected from the heart or the jugular vein were measured in 61 male fetal dogs at between 38 and 60 days of gestation and 55 male neonatal dogs at between 0 and 50 days after birth. Testicular descent and histology were investigated after euthanasia of the fetuses and neonates. Although the testis still lay on the caudal ridge of the kidney in the 38- and 40-day-old fetuses, the gonad had slightly migrated from the kidney by 42 days of gestation. The testis approached the internal inguinal ring at Day 0 after birth and had passed through the inguinal canal at Day 5 after birth. The inguinal passage was the active phase of testicular descent. Both testes were drawn into the scrotum after 35 days of age. The number of gonocytes in the semini ferous cords had increased in the 58-day-old fetus, and Leydig cells in the stroma had become obvious because of morphological development and the formation of cell clumps. Plasma A levels in the male fetuses decreased temporarily after birth and plasma DHT increased provisionally at 54 and 58 days of gestation. Plasma T levels in the male neonates increased markedly after birth. These findings suggest that testicular descent in the dog fetus is accelerated by the DHT and T secretory function of the Leydig cells which increased after 54 days of gestation.

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Anterior pituitary glands were removed from 18 sows at different stages of their estrous cycle to immunohistologically study the relationship between the stage of the estrous cycle and state of the luteinizing hormone (LH)-immunoreactive cells. The LH-immunoreactive cells were grouped into three types according to the amount of LH-immunoreactive granules stored in the cytoplasm (types A, B, and C). The number of LH-immunoreactive cells fluctuated along with the stage of the estrous cycle. In the 1st and the 6th stages, numerous LH-immunoreactive cells were observed (398±39, 391±60 cells/10^-2mm², respectively). In the 2nd stage, only a few cells were observed (6 cells/10^-2mm²) and from the 3rd to the 5th stages, their number gradually increased.

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We cloned and sequenced feline interleukin-6 (IL-6) cDNA using the polymerase chain reaction (PCR) technique to obtain information on regulation of the IL-6 gene. The feline IL-6 cDNA cloned in this study was 752 bp long and included all the amino acid coding region. At the amino acid level, the overall identities of the feline IL-6 to the human and murine counterparts were 58% and 39%, respectively. After transfection with the expression vector containing the cDNA, CRFK cells produced biologically active proteins that showed hybridoma growth promoting activity.

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The pineal gland of the Japanese quail (Coturnix coturnix japonica) retained numerous follicular structures, each of which was composed mostly of pinealocytes and supporting cells radially arranged around the follicular lumen during the first 6 weeks of age. In the pinealocytes, paraboloids and regular stacks of lamellar disks, which were demonstrated at 1 day of age in a previous study [14], rapidly decreased in number after 1 week of age. From 1 to 6 weeks of age, the pinealocytes had more developed lamellar whorls in the follicular lumen with the increased numbers of mitochondria in their apical protrusions and synaptic ribbons in their basal processes. In addition to these photosensory elements, pigment-containing cells occasionally accumulated in the basal side of the follicular structure. Their pigment granules, which were considered as melanin, gradually became larger and more roundish in shape after 1 week of age. The histological elements associated with the secretory function of the quail pineal were dense-cored vesicles and lysosomes in the pinealocytes and dense bodies in the supporting cells. The synaptic ribbons and the dense-cored vesicles in the pinealocytes rapidly increased in number at 1 week of age, scarcely exhibiting distinct changes thereafter. Thus, it is clarified that the foregoing photosensory and secretory elements change during the post-hatching development. Furthermore, it is suggested that, in respect of these elements, the pineal gland of the quail assumes more active appearances than that of the chicken.

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Pathogenicity of the lapinized Nakamura-III (L) strain of rinderpest virus (RPV) was examined in four strains of rabbits consisting of two inbred strains (NW-NIBS and DUY-NIBS) and two outbred strains maintained in closed colony (NW-NIBS and JW-NIBS) using a marmoset lymphoblastoid cell line, B95a cell-passaged virus and tissue homogenates of virus-infected rabbits. The cell culture virus was found to maintain virulence for rabbits of both closed colony and inbred NW-NIBS strain similar to the homogenate virus. Among the strains investigated, inbred NW-NIBS strain showed the highest susceptibility to RPV. Thus experimental model in an inbred rabbit using cell culture virus became useful.

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We previously reported that the effects of footed stimulation on the monosynaptic reflexes (MRs) in the cat's lumbar spinal cord depended on stimulated footpads. In this study, the effects of different types of stimulation (tonic pressure, phasic pressure, squeezing and radiant heating at 50°C) on the MRs were studied in 12 adult cats Spinalized at T11-12. The effects of tonic pressure were very weak or not detected. The pattern for the effects of phasic pressure was similar to that for squeezing, but the effects of squeezing was stronger than those of phasic pressure. These results indicate that effects of mechanical stimulation were mainly induced from phasic afferent inputs. Radiant heating was effective, and differential effects between mechanical stimulation and radiant heating were observed in some MRs. Our results suggest a difference in neuronal pathways from various types of receptors in the footpads.

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On day 13 of gestation in Mongolian gerbil fetuses, the mesonephric (Wolffian) duct was already visible. On day 14, the anlage of the paramesonephric (Mullerian) duct was first observed close to the mesonephric duct in both sexes at the cranial part of the mesonephros. Thereafter, the paramesonephric duct grew caudally in parallel with the mesonephric duct. On day 19, the duct reached the urogenital sinus. At this stage, the anogenital distance was significantly longer in male than in female. Also in male, the mesonephric duct was increased and the paramesonephric duct was decreased in diameter compared with those in female. These suggest that both testosterone and anti-Mullerian hormone are produced in fetal Mongolian gerbils at least before day 19 of gestation.

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The proteins, genomic RNA and disassembly conditions and pathogenesis in vivo for a virulent mouse rotavirus (MRV) and a less virulent bovine rotavirus (BRV) were compared. An obvious difference in the mobility of several genomic RNA segments were observed in one-dimensional gels. Reassortants obtained by replacement of gene 4 in BRV with MRV gene 4 indicated that the dose of the virus used and the clinical outcome in vivo was determined by gene segment 4. Under in vitro conditions, a comparison of the inner capsid proteins by partial proteolytic peptide mapping did not reveal any difference between corresponding proteins. However, this technique did identify differences between the two corresponding outer capsid proteins of these viruses. These differences, in turn, may account for the increased stability of MRV, as compared to BRV, when subjected to calcium-chelating and chaotropic agents and may be one of the mechanisms involved in conferring virulence on the virus. The observed variability between the 84K proteins (VP4) provided a basis for further investigations in order to locate a potential virulence determinant, since it has been observed by us and others that the gene coding for this protein plays a role in determining virulence. A comparison of the carboxypeptidase digests of the MRV and BRV VP4 revealed an area of variability between amino acids 307 and 407, which may represent the site of a virulence determinant.

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In rat liver injured by carbon tetrachloride, ED-1-positive monocytes and resident macrophages significantly increased in number in damaged perivenular zones on days 2 and 4 after single or double dosing, peaking on day 2 and being two-fold more after the 2nd dosing. ED-2-positive resident macrophages significantly increased in number only on day 4 after the single dosing and on days 2, 4 and 6 after the 2nd dosing with a peak on day 2. A significant increase in number of muscle actin-positive perisinusoidal cells was seen on day 4 and on days 2 and 4 after the 1st and 2nd dosings, respectively, reaching a peak on day 4 and being more remarkable after the 2nd dosing.

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The process of occurrence of bright blindness, progressive retinal degeneration (PRD), in sheep was observed using two Suffolk ram lambs fed on a diet containing bracken powder. The first sign of the bright blindness was detected 4 months after the start of experiment. Based on these preliminary results, the amount of bracken powder necessary to induce PRD was estimated (experiment I). In the following experiment, ptaquiloside (PT), a norsesquiterpene glucoside of the illudane type isolated from bracken, which is a bracken carcinogen and a causative principle of cattle bracken poisoning was administered to two Suffolk ram lambs. It was clearly demonstrated in this experiment (experiment II) that PT present in bracken is also a causative principle of PRD.

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The sequence analysis of the genomic region of Marek's disease virus (MDV) serotype 2 (MDV2) which had homology with the gene encoding MDV serotype 1 (MDV1) homologue of glycoprotein C (gC) of herpes simplex virus revealed that there was a single open reading frame (ORF) capable of encoding 479 amino acids. The predicted protein from this ORF had strong hydrophobic regions in both the amino and carboxyl termini and several potential N-linked glycosylation sites, as seen in those from the ORFs of MDV1 and MDV serotype 3 (MDV3=herpesvirus of turkeys (HVT)) gC homologues. By northern-blot analysis using a DNA fragment located in the MDV2 gC homologue ORF as a probe, 3.6 and 1.9 kb RNA transcripts were identified in MDV2-infected cells. When the alignment of amino acid sequences of the gC homologous genes of three MDV serotypes were compared, the homology between MDV1 and MDV2 and that between MDV1 and HVT showed about 76% identity, whereas that between MDV2 and HVT was about 70% identity. These results suggested a possibility that the evolutionary distance between MDV1 and MDV2 and between MDV1 and HVT is shorter than that between MDV2 and HVT.

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In order to analyze the detoxication functions in rats with diabetes induced by streptozotocin, the authors administered to the diabetic animals two drugs, ethionine and benzo(a)pyrene, which affect mainly the liver and are metabolized through a glutathione conjugation process, and examined the changes in the content of glutathione and activities of related enzymes in the liver. In the liver of the rats with streptozotocin-induced diabetes, the total glutathione content, glutathione S-transferase activity and glutathione-insulin transhydrogenase activity were lower than those of normal rat livers, while the glutathione peroxidase activity showed high values. Although specific changes in the glutathione-related detoxication functions were observed in the rats to which ethionine or benzo(a)pyrene had been administered, these changes were not revealed under diabetic conditions. It is suggested that in diabetic rats responses to toxic stimuli are suppressed.

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To establish criteria of pulmonary edema by extravascular thermal volume (ETV), ETV and a roentgenogram of the thorax were compared in normal and experimental hydrostatic pulmonary edema dogs. ETV was measured by the double indicator dilution technique using heat as a diffusible indicator and sodium ions as a non-diffusible indicator (thermal-Na method). Mean ETV in 14 dogs under physiologic conditions was 8.20±1.44 ml/kg. Comparison of ETV with the roentgenogram indicated the density of thoracic the roentgenogram to increase when ETV exceeded 11 ml/kg. Pulmonary edema was recognized clearly in the thoracic roentgenogram at more than 15 ml/kg of ETV. Further increase in ETV did not occur with additional radiographic change. ETV was significantly (P<0.01) correlated with arterial oxygen tension (PaO₂) but not statistically correlated with colloid oncotic pressure (COP), pulmonary arterial wedge pressure (PAWP) or PAWP-COP. Measurement of ETV would thus appear to be a reliable method for evaluating the severity of pulmonary edema, which has been difficult to diagnose by roentgenography and should facilitate the early diagnosis and management of pulmonary edema.

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The therapeutic effect of dibutyryl cyclic AMP (DBcAMP) in endotoxic shock was evaluated, using 11 dogs with experimentally-induced endotoxic shock (5 in DBcAMP group and 6 in control group) under general anesthesia. The DBcAMP group was treated by single intravenous injection of DBcAMP (10 mg/kg) at 15 min before inoculation with endotoxin (3 mg/kg). After the inoculation of endotoxin, this group was given drip infusion of DBcAMP at a rate of 0.1 mg/kg/min over 180 min. Hemodynamic parameters and chemical mediators were measured until 360 min after endotoxin inoculation. The cardiac output and urinary volume, which were decreased in the control group, were significantly inhibited to decrease in the DBcAMP group (p<0.01). The increases in 6-keto-PGF₁α and thromboxane B₂, chemical mediators released in endotoxic shock, were significantly inhibited (P<0.05 and P<0.01, respectively). These results suggested that DBcAMP is useful for the treatment of endotoxic shock.

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Cysts of the K strain of Toxoplasma gondii were administered orally to nine pregnant cats having no antibody to the organism at each of the three stages of gestation, namely early (14 days), middle (30 days), and late (40 and 51 days). Premature and weak or deformed fetuses were observed in the offsprings delivered by mother cats infected with Toxoplasma in the early or middle stage of gestation, while the cats infected at the late stage were delivered of apparently healthy fetuses. Mother cats developed parasitemia excreting Toxoplasma oocysts in their feces and formed the antibody to the organism after the oral administration. Toxoplasma was recovered from the organs or tissues of all fetuses delivered by the mother cats infected in the early stage of gestation. All kittens born healthy from the cats infected in the middle stage of gestation were free from Toxoplasma, but 80% of the premature kittens were found to be infected. Toxoplasma was isolated also from the organs of about half the offsprings born healthy from the mother cats infected in the late stage of gestation. These results indicate that congenital infection with Toxoplasma may occur commonly and cause damage to the fetuses when pregnant cats are infected orally with Toxoplasma cysts in the early or middle stage of gestation.

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Serum amyloid A protein (SAA) was isolated from equine acute-phase serum by repeating Sephadex G-75 gel filtration 3times. Quantitative measurement of equine SAA was performed by the single radial immunodiffusion technique with rabbit anti-equine SAA serum. In clinically normal horses, the SAA concentration remained relatively high from immediately after birth up to 1 week of age. After this the concentration showed periodic fluctiation in the range of approximately 13 to 30 μg/ml. The mean (±SD)concentration of SAA in foals (≤12 months old) and in adult horses (≤18 months old) was 19.37±9.41 and 21.53±9.81μg/ml, respectively. In mares during the perinatal period, the SAA concentration remained stable and within the normal range for 4 months before parturition. After foaling, it increased quickly and reached a peak value of 136.78±56.74μg/ml on day 3 postpartum, and then began to decrease at 2 weeks postpartum returning to within the normal range by 1 month postpartum. In horses with experimentally induced inflammation, the SAA concentration increased quickly, and reached the highest value, approximately 4 to 20 times higher than pre-treatment values, on day 2 after treatment. It then returned to the base line values within 10 days to 4 weeks, concurrent with the disappearance of local inflammatory signs. The SAA concentration was very high in most horses with clinical signs of inflammation. It was concluded from these data that equine SAA was a sensitive acute-phase reactive protein which increased in the early phase of various acute inflammations.

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Autonomic nervous function in Tsukuba high (THE) and low (TLE) emotional rats as well as Wistar rats (control) was studied by means of electrocardiographical examination in addition to the runway test. In the runway test, locomotive activity differed significantly among the three strains, the greatest activity in TLE and the smallest in THE. Electrocardiogram (ECG) and locomotive activity were successively recorded for 24 hr using a small size of telemeter system. Though the locomotive activity showed a clear nocturnal rhythm, no significant difference was observed among the three strains. The mean value of R-R intervals in THE tended to be smaller than those in other two strains; showing a significant difference between THE and TLE. The mean value of standard deviation (SD) of R-R intervals in TLE tended to be larger than those in other two strains; showing a significant difference between TLE and Wistar in the light and dark periods, and also between TLE and THE in the light period. As regards coefficients of variation of R-R intervals (CV), there was no significant difference among the three strains. Autonomic nervous tone in THE and TLE was evaluated by means of autonomic blockades using atropine and/or propranolol. The sympathetic nervous tone represented 21.6% in THE and 16.8% in TLE, while the parasympathetic nervous tone -9.0% and -12.3% in THE and TLE, respectively. There was a significant difference in the net autonomic nervous tone between THE (10.7%) and TLE (-6.6%). These results demonstrated that the sympathetic nervous activity was higher in THE than in TLE, while conversely, the parasympathetic nervous activity was higher in TLE than in THE.

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We examined the survival rate of C57BL/6 mice challenged with Babesia rodhaini 21 days post-killed Corynebacterium parvum (KCP) inoculation in comparison with ICR mice. All KCP-inoculated ICR mice survived the challenge infection, while all KCP-inoculated C57BL/6 mice died. To account in part for the difference in protective effects of KCP, stimulatory effects of KCP on splenic phagocyte chemiluminescence (CL) response were investigated in C57BL/6 and ICR mice. The KCP-inoculated ICR mice exhibited significantly higher CL values than ICR mice without KCP inoculation. No significant difference in CL value was observed between C57BL/6 mice with and without KCP inoculation. These results taken together, the lack of resistance of the KCP-inoculated C57BL/6 mice to B. rodhaini infection may be attributed in part to small stimulatory effect of KCP on splenic phagocytes.

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A direct colony hybridization method was used for the detection of Campylobacter jejuni in chicken feces. The biotin-labeled DNA prepared from the whole genome DNA of C. jejuni subsp. jejuni ATCC 33560 reacted well with homologous DNA and slightly with C. coli DNA. The method with the probe was found to be sensitive enough to detect a small number (10² CFU/g) of C. jejuni in chicken feces which contained a large number of background flora. It was suggested that this simple and sensitive method was useful for a wide survey of C. jejuni.

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The distribution of lymphocytes in lymphoid organs of germfree (GF) and conventional (CV) chickens was compared immunohistochemically with several monoclonal antibodies. The lymphoid follicles in the cecal tonsils of GF chickens were more poorly developed than in those of CV chickens, and IgG and IgA positive cells were also not found in GF chickens. The number of both B and T cells in the villous region of the cecal tonsils in GF chickens was less than that in CV chickens. In the spleen of GF chickens, the distribution of B cells was similar, but T cells were distributed more widely in CV chickens than in GF chickens. There was no clear difference in the distribution of lymphoid cells in the bursa of Fabricius and the thymus. These results suggest that the development of gut-associated lymphoid tissues is affected by intestinal flora.

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A 3-year-old Japanese domestic cat with a diagnosis of lymphocytic leukemia showed severe generalized seizures in the course of chemotherapy after leukemic condition was improved clinically. Computed tomography (CT) and magnetic resonance (MR) imaging of the brain were carried out. Both contrast procedures disclosed enhancements at the falx cerebri and the margin of cerebral cortex. Among these procedures contrast MR imaging demonstrated the lesion most clearly. Cytological examination of cerebrospinal fluid obtained by spinal puncture showed the infiltration of malignant cells and the diagnosis of meningeal syndrome associated with lymphocytic leukemia was defined. Intrathecal administration of cytosine arabinoside partially improved the neurologic dysfunction. Astopsy and histopathological examination confirmed the infiltration of leukemic cells in the areas of meningeal lesion demonstrated with contrast CT and MR imaging. Thus these imaging techniques, especially contrast MR imaging, are useful tools for rapid and precise diagnosis of meningeal syndrome.

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Foal diarrhea due to serotype 3 rotavirus broke out on a foal-raising farm in the years 1987 and 1989. In 1989, all of the foals, regardless of whether or not they suffered from diarrhea, received bovine colostral immunoglobulin (Ig) powder orally for 3 to 5 days during the epidemic. The morbidity of the diarrhea was lower than that observed in 1987, when the Ig powder was not administered to foals. These data suggested that the administration of Ig powder might partially prevent foal diarrhea with rotavirus infection.

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Chordoma occurred in the sacrococcygeal region of the vertebral column of a 97-week old male control F344 rat. Macroscopically, it was a mass (20 mm in diameter) protruding into the retroperitoneum and the cut surface was white to dark-reddish and showed a gelatinous appearance. Histologically, the tumor was composed of physaliphorous and stellate cells growing in nests or cords and divided into lobules by a thin fibrous stroma. The tumor cells locally invaded adjacent tissues, but no metastasis to other organs was noted. Centrally, the tumor often had islands of bone that entrapped physaliphorous cells and the bone tissue consisted of well-differentiated osteocytes. Based on these findings, this tumor was diagnosed as chordoma with osseous element.

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We have observed the ultrastructure of a canine leukocyte infected with Hepatozoon canis by transmission electron microscope. The infected leukocytes contained many small vesicles and a few typical granules in the cytoplasm. They also contained a few rough surfaced endoplasmic reticula, mitochondria and a segmented nucleus with condensed chromatin. Cytochemical analysis indicated that the infected leukocytes with H. canis contained little alkaline phosphatase, peroxidase, naphthol AS-D chloroacetate esterase or alpha naphthol acetate esterase. These results suggested that the infected leukocytes were neutrophils or monocytes which lost their typical characteristics.

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Recombinant baculovirus-infected insect cells which expressed recombinant protein, analogous to the nucleocapsid protein (NP) of Seoul type hantavirus, strain SR-11 (rNP-SR-Sf9) were applied to the indirect immunofluorescent antibody (IFA) test. The rNP-SR-Sf9 reacted with anti NP MAb clones which recognized strain specific or hantavirus common epitopes in the IFA test. The recombinant antigen was detected by antibodies to 3 serotypes of hantaviruses (Hantaan 76-118, SR-11, Puumala). Antibody titers of a group of experimentally infected mouse and urban rat sera using rNP-SR-Sf9 and strain SR-11-infected Vero cell antigens were mutual correlative. These results indicated that the rNP-SR-Sf9 were an effective and safety substitute for Vero E6 cells in the IFA test for serosurveys of hantavirus infection.

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Qualitative and quantitative analyses showed that there was iron (Fe) in a blackish lesion removed from a cat with severe corneal sequestration. Chromatic compound did not extract. The Fe content of the lesion was about 100 times higher than that of a normal cornea, as measured by X-ray fluorescence. Results of other chemical analyses indicated that the Fe in the blackish cornea was not derived from blood in the neovascularized limbus.

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Bovine embryos with the intact zona pellucida were exposed in vitro to K99⁺ Escherichia coli (K99 E. coli). The recommended procedures for washing and treating embryos were then evaluated for their effectiveness in removing or killing the adherent bacteria. After 10-step washing, bacteria were recovered not only from the embryos exposed to E. coli suspensions but also from those treated with trypsin during washing. On the other hand, no bacteria were recovered from any embryos treated with antibiotics (gentamicin; 50μg/ml) in culture medium before washing, indicating that the recommended washing procedures with appropriate antibiotics assure that zona pellucida-intact bovine embryos are free from E. coli.

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A small DNA virus was newly isolated from the small intestines of a pig with diarrhea in 1987, in Japan. Concerning the physicochemical properties, hemagglutination and susceptibility of cell culture to the virus, the virus was identical to a formerly isolated small DNA virus, the H-45 strain and also physicochemically similar to the parvovirus group. In a serological test however, the virus was distinctly, antigenically different from the H-45 strain as well as each of porcine, bovine and canine parvoviruses.

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A 7-year-old male Yorkshire terrier was examined for multiple plaques and nodules on the skin. The clinical, cytological and histopathological features indicated a malignant histiocytosis. Cytoreductive chemotherapy produced moderate clinical improvement, but died at day 90 after the first admission. Pathological examination revealed the neoplastic histiocytes in the skin as well as in the myocardium and skeletal muscles.

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Clinical and pathological examinations were performed on a 3-year-old Holstein-Friesian cow with scirrhous adeno-carcinoma of the rectum. Hemorrhagic diarrhea causing rectal stenosis was a principal clinical manifestation. Microscopically, cancerous transformation of the intestinal epithelium was observed in the rectal crypts. Irregular glandular arrangements containing PAS positive and eosinophilic mucus were observed in the lumen, which were surrounded by scirrhous collagenous fibers. The neoplastic epithelial cell had round or ovoid nucleus and slightly eosinophilic cytoplasm. Metastatic lesions were localized in the marginal and medullary sinuses of the anorectal lymph nodes.

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Canine distemper virus (CDV) was readily isolated at high rate with marked cytopathic effect (CPE) in B95a cells, a marmoset lymphoid cell line, from the peripheral blood leukocytes, cerebrospinal fluid cells and brain of dogs. Difference in type of CPE, i.e. syncytium type and round-cell one, among the virus isolates indicate the presence of heterogeneity of virus populations in prevalent CDV. Thus, this cell system is expected to be useful for ecological studies on CDV in the field.

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A neurological disorder was noted in 5 cows on a farm. Histopathological examination in 2 of these 5 animals revealed an infiltration of eosinophils with some macrophages in the meninges and around arterioles of the cerebral cortex. The endothelial cells of the reactive blood vessels were swollen. From these observations, the brain lesions were diagnosed as "eosinophilic meningitis". In one case, pulmonary arterioles were also infiltrated with eosinophils and macrophages in a granulomatous tissue. No bacteria, fungi, protozoa, or parasitic worms were detected in the present cases.

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Bengalees (a variety of Lonchura striata) in an aviary in Nagano Prefecture died of severe diarrhea in 1991. Pathological examination revealed slight enlargement of the liver and spleen. Histopathologically, necrotic granulomatous lesions were seen in the liver. Salmonella Typhimurium was isolated from the main organs and fecal samples. No evidence of Newcastle disease or chlamydiosis was detected. The disease was finally diagnosed as S. Typhimurium infection in bengalees. In spite of some countermeasures, 198 out of 268 birds died within 6 months, a mortality rate of 74%. Epizootiological survey revealed that the infection was transmitted by birds introduced from another prefecture. This is the first report of S. Typhimurium infection in bengalees.

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In male rats gonadectomized at birth, the blast formation reaction of lymphocytes was significantly lower at 114 weeks of age, than that of non-gonadectomized controls, whereas the lymphocyte subpopulation was similar. The thymus of neonatally castrated rats of age 114 weeks was histologically normal, decreasing in weight to 68% of that of non-castrated rats.

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