The pattern of cerebrovascular substance P (SP) and calcitonin gene-related peptide (CGRP) immunoreactive (-IR) innervation was investigated in the newt. SP-IR nerves supplying the cerebral arterial tree and choroid plexus were positive for CGRP, but negative for vasoactive intestinal polypeptide or neuropeptide Y. It is suggested that cerebrovascular SP- and CGRP-IR axons are sensory in nature. The supply of SP- and CGRP-IR nerves to the major cerebral arteries is relatively poor. Nevertheless, numerous SP- and CGRP-IR axons, which are contained in the fiber bundles on the cerebral carotid artery and the basilar artery, spread widely over the microvascular-epithelial regions of the choroid plexuses. It must be considered in relation to the significant role of SP-and CGRP-IR neuronal mechanisms responsible to the microcirculation, cerebrospinal fluid (CSF) production and transport action within the choroid plexus in the nutrition of the newt brain via the CSF.
Mitochondrial capsule selenoprotein (MCS) has been known as a structural protein of the mitochondrial sheath in spermatozoa. In this study, to determine the expression pattern of MCS mRNA in the mouse testis after puberty, in situ hybridization using digoxigenin-labeled RNA probes for MCS was performed in the testes of 8- and 20-week-old ICR mice. In the testes of both ages, MCS mRNA first appeared in step 3 round spermatids, gradually increased during early spermiogenesis, and persisted a high level until step 14 spermatids. After the step 14 spermatids, the signal began to decline and was weakly detected in steps 15-16 spermatids. On the other hand, compared with that in the testes of 8-week-old mice, MCS mRNA level in the testes of 20-week-old mice increased over 2-fold at stages VI-III, while it slightly increased at stages IV-V. These findings suggest that MCS gene transcription may be up-regulated after puberty in the mouse testis.
A filter immunobinding (FIB) method was developed for the detection and identification of mycoplasmas. Type strains of a total of 18 avian and bovine mycoplasma species propagated in broth media were diluted and immobilized on a nitrocellulose membrane as antigens for investigating the specificity with rabbit hyperimmune sera. Non-specific FIB reactions were easily eliminated by the procedure of absorbing rabbit hyperimmune sera in the broth. Absorbed rabbit hyperimmune sera exhibited clear species-specificity with mycoplasma antigens by the FIB. These specific reactions always agreed with the results of identification by tests of biochemical properties and growth inhibition for the isolates of M. bovirhinis, M. bovis, M. columbinum, M. columborale, M. gallisepticum and M. synoviae. Some bovine mycoplasma species, which were impossible to identify by growth inhibition test, because of their strong production of film and spots on the agar, specifically reacted with absorbed rabbit hyperimmune sera against M. bovis in the FIB. The detection limit of mycoplasmas by this method was about 104-105 colony-forming units/ml, which is lower than that of colony determination on agar. The FIB seems to be a useful technique for rapid detection and simultaneous identification of mycoplasmas.
Twenty 2nd specific pathogen-free pigs were divided into 4 groups: Group A were infected with porcine reproductive and respiratory syndrome (PRRS) virus at 6 weeks of age and treated with available swine erysipelas and swine fever combined vaccine (vaccinated) at 7 weeks of age; Group B were vaccinated at 7 weeks of age and infected with PRRS virus at 8 weeks of age; Group C were vaccinated at 7 weeks of age: Group D were neither vaccinated nor infected with PRRS virus. All pigs were challenged to Erysipelothrix rhusiopathiae C42 strain at 10 weeks of age. No clinical signs appeared after vaccination of group A and B pigs, thus confirming that the safety of the vaccine was not influenced by infection with PRRS virus. None of the pigs in Groups A and C developed erysipelas after challenge exposure to E. rhusiopathiae. In contrast, fever and/or urticaria appeared transiently in all pigs of Group B after challenge exposure. At the time of challenge exposure to E. rhusiopathiae, the PRRS virus titer was high in sera of Group B, but was low in those from Group A. However, vaccination of pigs with attenuated E. rhusiopathiae was effective in dual infection with PRRS virus and E. rhusiopathiae, because the clinical signs were milder and the E. rhusiopathiae strain was less recovered from these pigs compared to pigs of group D.
For determination whether strangles has invaded the Hidaka district of Hokkaido, the main racehorse-breeding area of Japan, a epizootiological survey with bacterial isolation was carried out during the breeding season in 1995. Streptococcus equi subsp. equi, which is the causative agent of strangles, was isolated from two Thoroughbred horses with submandibular lymphadenitis. Isolates were identified by serological grouping, biochemical tests and analysis of cell surface proteins by Western immunoblotting. Through this survey, it revealed that S. equi subsp. equi has invaded the Hidaka district and that strangles has become prevalent in racehorse-breeding farms in this area.
Pregnant rats were subcutaneously administered with mouse epidermal growth factor (EGF) at the concentration of 0, 100, or 200 μg/kg body weight/day from day 18 to 21 of gestation. The amino acid analysis by high-performance liquid chromatography demonstrated that the umbilical venous/maternal and fetal/maternal ratio of serum proline concentration increased in EGF dose-dependent manner accompanied by the increase in the ratios of total fetal weight and placental weight to maternal body weight gain. These results suggested that EGF regulates fetal growth by, as one of its possible mechanism, promoting placental proline supply from mother to fetus.
Hepatic oxygen supply, energy charge (EC), and histology were examined comparatively in dogs with portal vein anastomosis (PVA group), and PA in addition to PVA (PA group). The PVA group showed a lower level of hepatic oxygen supply than those of the PA group throughout the experimental period, and also showed decreases of adenosine triphosphate (ATP) and EC level after blood perfusion. In contrast, the oxygen supply and consumption were stable in the PA group. A temporary fall of ATP level was followed by recovery to the preperfusion level in the PA group. Histological examination indicated the collapse of hepatic cords with granular and vacuolar degeneration in only the PVA group. These findings suggested that PA, when supplemented to PVA, is an available technique for preventing hepatic failure caused by ischemic conditions.
A fluorometric assay for the determination of hepatic carnitine palmitoyltransferase (CPT) activity was slightly modified for use with cattle samples. With this assay, the Km value was 0.56 ± 0.10 mM with respect to L-carnitine (mean ± SD, n=4) and was 9.6 ± 2.2 μM (n=3) with respect to palmitoyl-CoA. The average hepatic CPT activity was 33.6 ± 2.0 μmol CoASH/min/g protein in 38 healthy cattle and was similar in both sexes and among breeds. Hepatic CPT activity showed no correlation with serum phospholipid, free fatty acid, triglyceride or total cholesterol concentrations.
The oscillometric procedure was used to measure the heart rate as well as the systolic, mean, and diastolic blood pressures of 152 dogs (102 in the control group, 13 in the group with renal disease, 37 in the group with heart disease) who were brought to Azabu University of Veterinary Teaching Hospital. It was demonstrated that the blood pressure and heart rate of the control group lowered and tended to become stable as the number of measurements increased. No appreciable difference was identified in the measurements of either blood pressure or heart rate in the forelimbs and tail head. With regard to gender, males showed a significantly higher value than females (p<0.05). No interaction was identified between age and blood pressure. No difference was identified in the heart rate in all groups. In an investigation of blood pressure in all groups, the renal disease group showed significantly higher values (p<0.05) than either the control or the heart disease group in all values of systolic, mean and diastolic pressures. These results indicated that dogs with renal disease can manifest hypertension.
The effects of a maternal vaccine against newborn calf diarrhea associated with group A bovine rotavirus (BRV), bovine coronavirus (BCV), bovine parvovirus and K99 Escherichia coli (E.coli) were examined on a beef cow-calf herd. After vaccination, serum or colostrum antibody titers to BRV, BCV and E. coli K99 in the vaccinated cows were significantly higher than those in unvaccinated control cows. Serum antibody titers to BRV, BCV and E. coli K99 in calves from the vaccinated cows were also significantly higher than those in calves from the control cows for 3-4 weeks after birth. These results suggested that the immunization of cows with the maternal vaccine enhanced the passive immunity levels in calves against BRV, BCV and K99 E. coli.
Feline immunodeficiency virus (FIV)-infected cells have been shown to undergo apoptosis by treatment with tumor necrosis factor alpha (TNF-α). This study detected a soluble factor which enhanced the apoptosis induced by TNF-α treatment. The sensitivity to TNF-α in the induction of apoptosis in a feline fibroblastoid cell line (CRFK) cells was significantly enhanced when the culture supernatant of FIV-infected CRFK cells or plasma samples from FIV-infected cats was added to the culture. These findings suggested that FIV infection induces production of a soluble factor which enhances CRFK cells sensitivity to TNF-α induction of apoptosis both in vitro and in vivo. This factor may contribute to the loss of lymphocytes in cats infected with FIV.
The inhibitory effect of chlorpromazine (CPZ), pentoxifylline (PTX) and dexamethasone (DEX) was investigated in a model of endotoxin shock in Holstein calves following an intravenous administration of Esherichia coli endotoxin (LPS). Initial correlations with its effects on the levels of tumor necrosis factor (TNF), a pivotal mediator of endotoxin shock, and clinical signs were obtained. The pretreatment of CPZ or DEX significantly decreased the serum levels of TNF, and reduced endotoxic shock. But the pretreatment of PTX hardly reduced the increase of serum TNF levels and endotoxin shock. The levels of serum endotoxin were not significantly different a minute of postinjection of LPS in calves. The results of this study indicate that pretreatment of CPZ or DEX inhibit various biological effects on endotoxin in calves.
Insertion of a 12-nucleotide repeat in c-myb gene exon 9 was observed in about 15% of sporadic bovine T-lymphomas. The 12-nucleotide repeat in the T-lymphoma cells showed deletion and insertion of the repeat units during cultivation of the cells. To know whether deficiency in DNA loop repair is involved in the instability of the repeat, abilities to bind and correct the loop structure in nuclear extracts were examined. The nuclear extracts of all examined cells had ability to bind and correct the loop structure. These data suggest that instability of the 12-nucleotide repeat in bovine T-lymphoma cells might be independent of deficiency of DNA loop repair function.
Susceptibility of five species of Cyprinidae and Salmonidae freshwater fish to the early third-stage larvae (EaL3) and advanced third-stage larvae (AdL3) of Gnathostoma nipponicum infection were examined. Two fish species inoculated orally with EaL3 were infected, and AdL3 were recovered from them with rate of 21.0% in Tribolodon hakonensis and 0.5% in Cyprinus carpio at 30 days postinoculation (PI). Attempts to infect five fish species with AdL3 were all successful. The recovery rate of AdL3 was 69.0% in T. hakonensis, 47.5% in Carassius auratus subsp., 35.0% in C. carpio, 53.0% in Oncorhynchus masou, and 32.0% in O. mykiss at 10 days PI. These results confirmed that the Cyprinidae and Salmonidae fish species reported here were susceptible to larval G. nipponicum infection and AdL3 had higher infectivity to them than the EaL3.
Decrease of spleen weight from peak to control levels together with a corresponding decline of serum IgG level at the onset of gastric hyperplasia in Taenia taeniaeformis-infected euthymic rats, may indicate that a form of down regulation of host immune response during the course of larval T. taeniaeformis infection could facilitate the occurrence of gastropathy in rats. Gastric hyperplasia developed in T. taeniaeformis-infected athymic nude rats, indicating that occurrence of gastropathy associated with larval T. taeniaeformis infection in the rat is T cell independent. Apparently, gastric hyperplasia appeared early in nude rats which suggests that absence of T cell-mediated response could have facilitated its early occurrence.
The interdigital glands of a Japanese serow infected with parapoxvirus had severe papular and nodular lesions that completely occupied the sac and duct of the gland. The lesions were characterized by acanthosis with hyperkeratosis. Intracytoplasmic inclusion bodies were detected in the vacuolated prickle cells. By electron microscopy, mature and immature viral particles were present in the cytoplasm. These glands act as scent glands, and lesions in this organ probably affected the ecological adaptation of this individual.
In a four-year-old male cat, a subcutaneous phyma about 3.5 cm in diameter was surgically removed from the left inframandibular region. Histopathologically, the phyma was found to be the swollen medial retropharyngeal lymph node containing an actinomycotic abscess. The filamentous organisms in the abscess stained positively by the Gram's, Grocott's and periodic acid-Schiff methods, and were negative by the Ziehl-Neelsen method. By the immunoperoxidase method, the organisms were specifically identified as Actinomyces viscosus serotype 2 by its antiserum absorbed with A. viscosus serotype 1 antigen.
The incidence of Campylobacter jejuni and Campylobacter coli in broiler farms was 33.9% (19/56). C. jejuni-positive flocks accounted for 20.0% (17/85) and C. coli-positive ones was 4.7% (4/85). There were 14 patterns (fla type) of restriction fragment length polymorphism (RFLP) of flagellin A gene among these 22 strains of C. jejuni and C. coli including the standard strain C. jejuni ATCC 33560. Different fla types of Campylobacter were isolated from broilers in different growing cycles on the same farms. Four strains of C. jejuni were isolated from four breeder farms and four fla types of C. jejuni were detected from their progenies reared on growing farms. Three fla types of C. jejuni detected from the progenies were different from those of each breeder. Also, the other three fla types of C. jejuni were detected from different progenies of each growing farm during the next growing cycle. These findings indicate that the RFLP analysis may contribute to epidemiological studies of C. jejuni and C. coli contamination of broilers and suggest the risk of contamination with different types of Campylobacter in every growing cycle of broilers on the farm even on the same farm. They also supported that there was little likeliness of the vertical transmission of C. jejuni and C. coli from breeders to broilers.
65Zn is one of the induced radioactive nuclides which are generated in power reactors. In the present experiment, several parameters of 65Zn metabolism were studied in mice maintained on diets with various zinc contents from 45 to 4,500 mg/kg to evaluate the efficacy of the dilution method for radiation protection against internal contamination with 65Zn. Gastrointestinal absorption of 65Zn was suppressed and its excretion accelerated as the dietary zinc content increased over a wide range. Clearance of 65Zn from tissues was generally accelerated by feeding mice a high-zinc diet, but that from the femurs was not affected by dietary zinc content. Zinc concentrations in tissues were regulated homeostatically up to a dietary zinc content of 1,350 mg/kg. Although a significant accumulation of zinc occurred in the liver, pancreas, kidneys, and femurs when mice were given 4,500 mg/kg diet, the concentrations except in the femurs recovered within a few days after switching to a normal-zinc diet. These results suggest that oral administration of zinc is effective for preventing the absorption and for enhancing the excretion of 65Zn to protect the body from internal radiation exposure with this isotope.
The mutagenicity of 15 household dog urine specimens were measured by the combination of blue rayon extraction and ultramicro forward-mutation method with Salmonella Typhimurium TM677 strain. A good dose-response relation was observed between the urine volume and mutation frequency. The minimum amount of urine required was 20 ml or less. The specific mutation frequency of urine greatly varied from one dog to another. The average specific mutation frequencies in the presence and absence of S9 mix were 28.7 ± 51.5 (× 10-4) and 12.0 ± 13.3 (× 10-4), respectively, and there was no significant difference between them. The mutation frequency markedly increased after the ingestion of broiled fish. Ten human urines specimens showed a similar level of specific mutation frequency to that of the dog urine specimens in both the presence and absence of S9 mix.
The inhibitory effect of glucocorticoids (GCs) on proliferation of canine mast cell tumor (MCT) was studied using two types of MCT cells; JuMC cells and LuMC cells derived from spontaneous canine cutaneous and intestinal MCT, respectively. In in vitro study, growth of JuMC cells was significantly inhibited with more than 1 nM GCs and apoptotic-like cell death was seen, while that of LuMC cells was never inhibited even with 10 μM GCs. Growth rate of masses in nude mice developed by inoculation of JuMC cells was reduced in a dose-dependent manner by administration of GC, while growth inhibition of masses developed by inoculation of LuMC cells was minimal with increasing GC doses. Competitive binding studies and Scatchard analysis demonstrated the presence of high-affinity, low capacity GC receptors in both JuMC and LuMC cells. Kd was estimated to be 1.30 nM in JuMC cells and 0.45 nM in LuMC cells, respectively. It is concluded that canine cutaneous MCT cells responded to GCs in vitro and in vivo, whereas intestinal MCT cells did not, though both types of cells had specific GC receptors.
A three-year-old male Siberian Husky dog was referred to the Veterinary Teaching Hospital in Osaka Prefecture University with a complaint of difficulty in expelling the stools. By rectal examination, a mass as big as a fist could be detected occupying the cavum pelvis. Radiographically the mass had a thin bony shell bulging from the pubic periosteum. In the shell, radiolucent trabeculation gave the area a "soap bubble" appearance. The cut surface of the removed mass showed a honeycomb-like pattern constituted of some small loculate bony cysts. These cysts were separated from each other by a fibrous or bony trabeculae with blood-filled vascular channels or sponge-like structures. From clinical and pathological findings, this mass was diagnosed as a pelvic aneurysmal bone cyst. After surgery, the patient completely recovered without tenesmus.
A six and half-year-old nulliparous mixed breed cat which had the complaints of vomiting, abdominal distention and depression was presented to the Veterinary Teaching Hospital of Osaka Prefecture University. She was suspected of pyometra by clinical signs and tests. By laparotomy, it was clarified that both uterine horns and vagina showed distension by the accumulation of secretions, and the vagina ended blindly leaving a tough connective tissue at the border between cranial and caudal part of the vagina. Postoperative contrast-radiograph of the remaining vagina proved it had no persistence of the hymen. From these findings, the condition was diagnosed as a feline atresia vaginalis with the transverse vaginal septum which is caused by the embryonic failure of canalization of the paramesonephric duct between the end of the Müllerian duct and the urogenital sinus.
Thermostability of sperm genome against freezing-thawing and high temperature treatments was assessed by comparing the degradation patterns of genomic DNAs from epididymal sperms and somatic tissues. Golden hamster liver, kidney, epididymal sperm, and testis were frozen and thawed repeatedly, or incubated in a hot water bath. Genomic DNAs were isolated and then separated by agarose gel electrophoresis. It was revealed that the size of sperm genomic DNA was hardly changed after freezing-thawing treatment, however, the DNA sizes of the other three tissues were gradually reduced with an increasing number of freezing-thawing cycles. In contrast, high temperature treatment appears to damage not only the genomic DNAs of somatic cells but also those of spermatozoa.
Glycoprotein D (gD) of canine herpesvirus (CHV) YP2 strain was expressed in COS-7 and insect (Spodoptera frugiperda; Sf9) cells. The gDs expressed in COS-7 and Sf9 cells reacted with a panel of monoclonal antibodies (MAbs) against CHV gD (hemagglutinin) and an MAb 25C9 against feline herpesvirus type 1 (FHV-1) gD by indirect immunofluorescence assay, and possessed a molecular weight (MW) of approximately 51-55 and 41-46 kilodalton (kDa), respectively, when examined by immunoblot analysis. After treatment with tunicamycin, the MW of the gD expressed in Sf9 cells became approximately 37 kDa. By hemadsorption (HAD) tests using canine or feline red blood cells (RBC), COS-7 cells expressing CHV gD adsorbed only canine RBC, but not feline RBC, whereas control COS-7 cells expressing FHV-1 gD adsorbed feline RBC, but not canine RBC. By hemagglutination (HA) tests, lysates of Sf9 cells expressing CHV gD agglutinated canine RBC, but not feline RBC. These HA and HAD activities were inhibited by HA-inhibition MAbs against CHV gD. Control lysates of Sf9 cells expressing FHV-1 gD agglutinated only feline RBC. Serum from mice inoculated with lysates of Sf9 cells expressing CHV gD possessed a high titer of virus-neutralizing activities against CHV infection. These results indicated that CHV gD is structurally similar to FHV-1 gD, but is functionally different from FHV-1 gD.