We analyzed the mechanism for feline herpesvirus type 1 (FHV-1) immediate early (IE) gene expression. We demonstrated that (i) the transcription initiation site of the FHV-1 IE transcript lies in the putative FHV-1 ICP4 binding site, (ii) the FHV-1 IE transcript is spliced to remove 906 bp intron from the leader region, (iii) cis-acting elements of the FHV-1 IE promoter map both down- and up-stream of the transcription initiation site, and (iv) a deletion of a 58 bp sequence which includes the putative FHV-1 ICP4 binding site in the IE promoter resulted in significant induction of promoter activity by a FHV-1 IE gene product although the FHV-1 IE gene product slightly stimulates the FHV-1 IE promoter, indicating that the FHV-1 IE gene product down-regulates its own promoter via the region.

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Medical records of fifty-three cats diagnosed as malignant mammary tumor from 1982 to 1993 were reviewed. The mean age of the cats at diagnosis was 11.1 years, and Japanese domestic and Siamese breeds were predominant. Survival rates after 1 and 2 years of diagnosis were 31.8% and 17.7%, respectively, suggesting poor prognosis of this malignancy. The survival time was significantly associated with tumor size or WHO clinical stage, but not with breed, age, or gender. Twenty-nine cats with pulmonary metastasis died within 5 months following metastatic detection. Postoperative prognosis was significantly related to the tumor size at surgery, but not with type of surgery and adjuvant chemotherapy using cyclophosphamide and/or vincristine.

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Circulating immune complexes (CIC) in the sera from 102 clinically healthy dogs and from 16 Dirofilaria immitis (D.immitis) infected dogs were measured by the immune adherence hemagglutination (IAHA) method. The level of CIC in the sera from healthy dogs was 28.2±29.1 μg/ml. There was no significant difference in the levels of CIC regarding sexes or ages. On the other hand, the level of CIC in the sera from D.immitis infected dogs was 230.3±117.1 μg/ml, which was significantly (p<0.01) higher than that of healthy dogs. The results of this study confirm that IAHA method is one of the reliable means of detecting and/or diagnosing immune complex mediated discases in dogs.

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We constructed plasmids expressing mRNA7 coding the nucleocapsid (N) protein of JHM strain of MHV (JHMV) under the control of Rous sarcoma virus (RSV) LTR or human elongation factor (EF) la promoter, referred to as pRSV-mRNA7 and pEF-mRNA7, respectively. Although only a slight level of cytolysis was observed by the spleen cells from C57BL/6 mice injected intramuscularly with pEF-mRNA7, the spleen cells from the mice administered with pRSV-mRNA7 showed a significant level of cytolytic activities against the cells expressing the viral N protein. The difference in the level of specific cytolysis might have been mainly due to a difference in the expression levels of the N protein in the muscles between the mice injected with pEF-mRNA7 and pRSV-mRNA7, since the specific activity of chloramphenicol acetyltransferase (CAT) in muscles from the mice injected with plasmid DNA expressing CAT gene directed by RSV LTR was significantly higher than that in those administered by the plasmid DNA directed by EF-la promoter.

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We attempted to measure the qualitative saliva progesterone concentration in the sow using a commercial bovine milk progesterone qualitative test EIA kit (qualitative kit), which can measure the progesterone concentration in approximately 10 min, and investigated the possibility of applying this method of progesterone concentration measurement to early pregnancy diagnosis in the sow. The accuracy of pregnancy diagnosis at 17-24 days after last mating for 138 sows was 91.3% (105/115) for positive cases and 100% (6/6) for negative cases. The overall pregnancy diagnosis accuracy, including 17 indeterminable cases, was 80.4% (111/138). A comparison of the diagnoses based on progesterone concentrations measured by the qualitative kit and the saliva progesterone concentrations of identical samples measured by the quantitative kit showed close agreement: 6 cases diagnosed as negative pregnancy by the qualitative kit all had a progesterone concentration of less than 5 ng/ml, while 111 out of 115 cases diagnosed as positive pregnancy by the qualitative kit all showed a progesterone concentration over 5 ng/ml. Thus, the results of this study show that qualitative measurement of the saliva progesterone concentration in the sow using a bovine milk progesterone qualitative test EIA kit is a practical method for early pregnancy diagnosis.

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Although many reports are available to indicate the increase in number and size of peroxisomes by oral administration of hypolipidemic reagents such as clofibrate and bezafibrate, the decrease in number and size of proliferated peroxisomes even during administration of these chemicals has not been well documented. The present study was aimed to detect when the decrease in number and size of peroxisomes would occur. In the present study, ultrastructural changes in hepatocytes of male Wistar rats during 3 to 90 days of oral administration of bezafibrate were analyzed by morphometric and cytochemical methods. By 3-day administration, the weight of the liver and the number and size of peroxisomes were significantly increased. These parameters began to decline from the time 14 days of administration when the weight of the liver and the size of peroxisomes attained the peak. Heterogeneous catalase staining was most remarkable by 7-day administration. Mitochondria were another target cell organelle by bezafibrate. The total area of mitochondria was decreased by 3-day administration. However, by subsequent 7 to 90-day administration, it recovered almost the level of controls. By 90-day administration, many mitochondria had expanded cristae which contained filamentous substances.

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Various conditions were evaluated and modified to enhance the sensitivity of the neutralization (NT) test for detecting antibody in swine infected with porcine reproductive and respiratory syndrome (PRRS) virus. Higher NT antibody titers were consistently obtained by the addition of 10% (v/v) complement, fresh guinea pig serum, to the virus diluent and by the incubation of serum-virus mixture at 4°C for 24 hr. The appearance and persistence of antibodies detected by the modified NT test showed a similar pattern to those of antibodies detected by the indirect fluorescent antibody (IFA) assay, although the antibody titers obtained by the former method were consistently lower than those obtained by the latter method. Slow-reacting complement-requiring NT antibody was detected in sera from pig 2 weeks after infection with PRRS virus. The slow-reacting complement-requiring NT antibody in the early serum samples was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting complement-requiring NT antibody in the late serum samples was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which serum-virus mixtures were incubated at 4°C for 24 hr with complement; this gave an improved sensitivity over the previous incubation at 37°C for 60 min.

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The present study was designed to examine the interaction between the effects of lasalocid and Ca²⁺ on the growth and structure of Selenomonas ruminantium HD-4. Lasalocid, at a dose of 10 μM, inhibited cell growth almost completely after 12 hr incubation in the presence of relatively high extracellular concentrations of Ca²⁺ (from 5 to 50 mM), but only slightly reduced cell growth in the presence of 0.2 mM Ca²⁺. With Ca²⁺ alone, cell growth was also inhibited at 12 hr as a function of the concentration of Ca²⁺ over the range 5 to 50 mM. In cultures at mid-exponential phase, growth was markedly inhibited by the simultaneous addition of 10 μM lasalocid and 15 mM Ca²⁺, but only moderately inhibited by lasalocid in the absence of Ca²⁺. However, there was no significant effect on bacterial growth at the mid-exponential phase when Ca²⁺ alone was added to the incubation medium. In thin sections of cells treated with lasalocid in the presence of 15 mM Ca²⁺, abnormal cells were found with cytoplasmic voids and with an outer membrane detached from the inner membrane layer; this change in the outer membrane was also found in cells treated with lasalocid alone. There was no visible abnormality in the outer membrane in thin sections of Ca²⁺-treated cells, while most of these cells showed only a slight contraction of cytoplasmic material or a loss of cytoplasmic contents. These results indicate the presence of a synergistic effect between the actions of lasalocid and Ca²⁺ on cell growth, presumably due to induced cytoplasmic alterations.

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Cardiorespiratory effects of capsaicin (CAPS) solution instilled into the larynx and the reflex mechanisms were investigated in rats with spontaneous breathing or under artificial ventilation. The first challenge with CAPS (100 μg/ml, 20 μl) markedly inhibited spontaneous breathing due to a considerable prolongation of expiration time (T_E) (1785% of control) in all rats. Circulatory changes such as hypertension (mean systolic blood pressure=210 mmHg) and bradycardia (10.5% decrease in heart rate) were also elicited by the 1st challenge with CAPS. These changes were largely reduced by the second challenge of CAPS; but not abolished in 3 of 5 rats by bilateral section of the superior laryngeal nerves (SLNs) and recurrent laryngeal nerves (RLNs). The bradycardia and hypertension after the CAPS-instillation were able to be elicited to the same extent in rats in the absence of apnea under artificial ventilation. The bradycardia was entirely abolished by pretreatment with atropine injection, while the hypertension was largely inhibited by phentolamine and propranolol. These results demonstrated that the laryngeal and/or pharyngeal noxious stimulus could induce marked cardiorespiratory reflexes, where the circulatory changes could be elicited in the absence of apnea, suggesting the presence of eliciting mechanisms of circulatory changes independent on the occurrence of apnea. In addition, it was also suggested that those cardiorespiratory responses were mediated by the afferents such as unmyelinated C-fiber endings and thin myelinated fibers and by the efferents, both the parasympathetic and sympathetic nervous systems.

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The sequence of the hemagglutinin (HA) gene of the LA strain of rinderpest virus (RPV) has been determined by the direct sequencing method. The amino acid sequence of the protein which it encodes displays conservation of its structural determinants with the HA proteins of the other RPV strains. The LA-HA protein was shown to have three conserved potential N-linked glycosylation sites, compared with four such sites in the HA protein of L strain. The glycosylation site at position 200 on the L-HA molecule is absent from its LA-HA counterpart, due to a lysine for an asparagine substitution. The HA proteins of L and LA strains were the same molecular weight as judged by mobility on SDS-PAGE, suggesting that the site at position 200 is not used for glycosylation.

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Primary brain tumors in two dogs were surgically removed followed by postoperative radiation therapy. The two tumors were confirmed histologically to be astrocytoma and meningioma, respectively. After the surgery, the neurological status of each dog improved dramatically and a total dose of 40 Gy was delivered to the surgical site to treat residual tumor tissue. Although the dog with astrocytoma died 6 months after surgery due to unknown causes, the dog with meningioma has lived for over 24 months with a degree of neurological disorder less severe than that before treatment. These results suggested the effectiveness of this type of therapy on brain tumor in dogs and therapeutic modality should be positively planned to treat canine brain tumors.

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Chick embryo fibroblasts (CEFs) infected with three strains of Marek's disease virus serotype 1 (MDV1), GA, Md5 and JM, were subjected to indirect immunofluorescence assay with monoclonal antibodies (MAbs) against MDV1 homolog of glycoprotein D (MDV1 gD) of herpes simplex virus. By the MAbs, a number of MDV1 gD-positive cells were detected in CEFs infected with GA, whereas only a few and no positive cells were detected in CEFs infected with Md5 and JM, respectively. The MDV1 gD in GA-infected CEFs was recognized as the band of 64 kDa in immunoblot analysis using one of the MAbs. This is the first report that the MDV1 gD was detected in MDV1-infected cell cultures.

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We investigated the effect of thiamphenicol plus lincomycin (TP + LCM) and thiamphenicol plus tylosin (TP + TS) combinations using checker board method on the growth of Pasteurella (P.) multocida, P.haemolytica and Mycoplasma (M.) bovis by culculating the fractional inhibitory concentration index (FIC index). The results showed that the FIC indexes of the TP + LCM combination for P.multocida, P.haemolytica and M.bovis were 0.36±0.10, 0.72±0.09 and 0.81±0.18, respectively. The FIC indexes of the TP + TS combination for P.multocidda, P.haemolytica, and M.bovis were 0.79±0.20, 0.66±0.11 and 0.32±0.14, respectively. Thus, these combinations are assumed to have a more synergistic or additive effect on bacteria growth than a single antimicrobial agent.

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Clinical effect of the administration of thiamphenicol (TP) and tylosin (TS) on bovine respiratory disease was investigated. Group I (n=64) were administered TP (10 mg/kg) and TS (4 mg/kg), group II (n=26) were given TP (5 mg/kg) and TS (2 mg/kg). For the control, TP group. (n=25) were given 20 mg/kg of TP and ampicillin group (n=23) were given 10 mg/kg of ampicillin. As a result, improvement of clinical findings was more rapid and the cure rate was significantly higher in group I compared to those in the other 3 groups. These results showed that a combination therapy with minimal basic doses of TP and TS is very effective for some respiratory diseases in cattle.

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Immunolocalization for four steroidogenic enzymes was performed on an ovary taken from a Hokkaido brown bear during the mating season. This specimen is considered to be in the follicular phase because of the presence of large follicles. In large follicles, cholesterol side-chain cleavage (P450scc) and 3β-hydroxysteroid dehydrogenase (3βHSD) were immunolocalized in theca interna cells and granulosa cells. 17α-hydroxylase/C17-C20 lyase cytochrome P450 (P450c17) was immunolocalized in theca interna cells but not in granulosa cells, Aromatase cytochrome P450 (P450arom) was immunolocalized only in granulosa cells. In medium follicles, however, P450scc and 3βHSD were immunolocalized only in theca interna cells, and the immunoreactivity of P450arom was detected in neither theca interna cells nor granulosa cells. Immunoreactivities of P450scc, 3βHSD and P450c17 but not P450arom were detected in interstitial cells. This study suggests that estrogen biosynthesis takes place through interrelation between theca cells and granulosa cells and is explained by the so-called two-cell mechanism. Furthermore, the granulosa cells in large follicles have the capability for pregnenolone and progesterone biosynthesis, and the interstitial cell in the bear ovary is also a steroidogenic site.

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As a rapid and sensitive method to detect canine distemper virus (CDV), an immunocapture enzyme-linked immunosorbent assay (ELISA) was performed. The sensitivity and specificity of the immunocapture ELISA were considered to be high enough. Virus neutralizing (VN) test was also established using the immunocapture ELISA. By using this test, the different cross VN titers between sera of dogs experimentally infected with the Onderstepoort strain and those with a field isolate of CDV were observed.

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The LEC strain of rats (LEC rats), originally developed as a model for hereditary fulminant hepatitis, is highly sensitive to whole-body X-irradiation when compared to WKAH strain of rats (WKAH rats). The present results showed that frequencies of certain types of chromosome aberrations induced by in vitro X-irradiation in the fibroblasts of LEC rats were higher than those of WKAH rats. In particular, frequencies of chromatid gaps and chromosome exchanges in LEC cells were higher approximately 4- to 5-fold and 6- to 8-fold, respectively, than those of WKAH cells.

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Rumen ciliates species and composition of the sheep, Friesian-cattle and water buffaloes in Egypt were surveyed. As a result, 7 genera with 18 species and 6 formae in sheep, 10 genera with 28 species and 11 formae in cattle and 12 genera with 29 species and 7 formae in water buffaloes were detected. Twenty-two species were common in both cattle and buffaloes, while 12 species in sheep were common with the other ruminants. Entodinium spp., such as E.simplex, E.nanellum and E.exigum, appeared most frequently in every host. In general, Egyptian domestic ruminants had the rumen ciliate composition similar to that of domestic ruminants in temperate zone, though the water buffaloes had several tropical species such as Entodinium longinucleatum forma spinonucleatum. The ciliate density was estimated as 10⁵/ml in every host species.

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Plasma iothalamate (IOT) disappearance rates were measured after a single-injection of IOT (113.8 mg/kg, IV) in cats with experimentally induced renal failure. The disappearance rates especially fitted into the one compartment model. The mean value of plasma disappearance rates of IOT in these cats with induced renal failure (2.16±0.240×10^-3 μg/ml/min) was markedly lower than that of clinically healthy cats (4.10±1.00×10^-3 μg/ml/min). These results demonstrate that IOT clearance is available for evaluation of renal function in cats.

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The serological property and replication in swine alveolar macrophages (SAM) and MARC-145 cell cultures of 35 porcine reproductive and respiratory syndrome (PRRS) viruses isolated in SAM were investigated. All the isolates were reacted almost equally with antisera against three Japanese isolates including EDRD-1 strain (American type) in immunoperoxidase monolayer assay (IPMA), but weakly or did not react with antiserum against Lelystad virus (European type) indicating that the Japanese isolates are more closely related to an American type virus. Twenty two of 35 isolates replicated with CPE both in SAM and MARC-145 cells, whereas remaining 13 isolates did not show CPE in MARC-145 cells. The antigenicity of the isolates did not relate to the virus origin and the replication in the cell cultures.

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Development of enteric lesions in closed jejunal and ileal loops inoculated with Aujeszky's disease virus (ADV) was examined in four 6-week-old SPF pigs. A large number of ADV antigens were detected first in necrotic foci in the subepithelial areas, and subsequently in the epithelial cells, lymphoid follicles in Peyer's patches and neuronal cells of Meissner's and Auerbach's plexuses.

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The growth and fertility of a female calf obtained from a frozen-thawed bovine oocyte was assessed. The birth weight of the calf was lower than the mean birth weight of calves from in vitro fertilized embryos (IVF-controls) and calves obtained by artificial insemination (AI-controls). The growth rate of the calf up to 6 months was slower than that of the IVF-controls, but similar to that of the AI-controls. When the calf developed into a heifer (200 kg), she was inseminated with frozen semen and 280 days later delivered a male calf. The chromosoms of this cow were normal. These findings suggest that the growth and fertility of the calf derived from the frozen oocyte are normal.

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A seven-year-old female mongrel dog showed incurable skin lesions for about a year. The lesions were at nasal bridge and periocular area, and were composed of crust formation and scaling. Biopsy specimens from the skin lesions possessed with multiple intraepidermal pustules containing many acantholytic keratinocytes. Direct immunoperoxidase stain using paraffin-embedded section showcd IgG deposition at the intercellular area of upper epidermis and follicular infundibula. IgG was not detected at basement membrane zone. Clinical and pathological findings of the present case were identical to those of pemphigus foliaceus, an uncommon pustular autoimmune skin disease.

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