Intravascular accumulation of blood cells after brain ischemia-reperfusion can cause obstruction of cerebral blood flow and tissue hypoxia/ischemia as a consequence. In the present study, we examined temporal and topographic changes of tissue hypoxia/ischemia after occlusion of the middle cerebral artery (MCA) for 60 min in rats with immunohistochemical staining for hypoxia (2-nitroimidazole hypoxia marker: hypoxyprobe-1 adducts). Our results showed that tissue hypoxia expressed as positive staining for hypoxyprobe-1 adducts preceded neuronal degeneration. Platelets and granulocytes were detected close to the hypoxyprobe-1 adducts positive area. These results suggested that the hypoxic environment could persist even after reperfusion of MCA, because of vascular obstruction with accumulation of platelets and granulocytes.
Decreases in egg production and increased incidence of abnormal eggs due to malformation of egg shells were observed in specific pathogen free (SPF) 173-day-old laying hens inoculated intravenously with an avian metapneumovirus (aMPV) strain PLE8T1. This strain was derived from an isolate from broiler birds exhibiting swollen head syndrome (SHS). Some SPF birds inoculated with the virus showed, slight diarrhea without any respiratory symptoms. Thus, the PLE8T1 strain was used as a challenge virus to evaluate efficacy of aMPV vaccines. SPF chickens which received a live attenuated aMPV vaccine (NEMOVAC; Merial) at 7 or 77 days old and an inactivated aMPV vaccine (OVO-4; Merial) at 105 days old were protected against poor egg production caused by the challenge with the PLE8T1 strain. Thus, aMPV, the PLE8T1 strain passaged 22 times after isolation, from birds exhibiting SHS, could induce a drop in egg production in laying hens accompanied by malformation of egg shells. It was suggested that this challenge system could be applied to evaluate the efficacy of aMPV vaccine.
Because of their unsurpassed potency in presenting antigens to naive T cells, dendritic cells are considered to be an important candidate in the development of immunotherapeutic strategies. Despite the high potential of dendritic cell-based immunotherapy, as a so-called dendritic cell vaccination, few clinical approaches using dendritic cell vaccination have been performed in the dog because of very limited information regarding the generation of canine dendritic cells and their functional properties. We therefore established a protocol for the efficient generation of dendritic cells from canine bone marrow cells using recombinant feline granulocyte-macrophage colony-stimulating factor and canine interleukin-4. Dendritic cells were generated efficiently: a yield of 1-9 × 106 cells per approximately 0.5 ml of canine bone marrow aspiration was achieved. These dendritic cells showed features shared with mouse and human dendritic cells: dendrite morphology, expression of surface markers MHC class II and CD11c, and up-regulation of molecules related to antigen presentation (MHC class II, B7-1, and B7-2) by activation with lipopolysaccharide. Moreover, the dendritic cells demonstrated phagocytic activity, processing activity of pinocytosed proteins, and activation of allogeneic T cells far more potent than that by macrophages. Our findings suggest that the bone marrow-derived dendritic cells are functional for the capturing and processing of antigens and the initiation of T cell responses.
To clarify the behavioral profiles of 56 pure breeds of dogs in Japan, 96 small-animal veterinarians participated in a questionnaire survey using the same criteria as in preceding studies conducted in the United States and the United Kingdom. We found significant differences among breeds in all behavioral traits examined. In addition, gender differences were revealed in terms of aggression to dogs, territorial defense, excitability, general activity, dominance over owner, destructiveness, watchdog barking, and snapping at children, which were all rated higher in males than females, whereas obedience training and housebreaking ease were rated higher in females. No gender differences were evident in playfulness, excessive barking, or affection demand. Using factor analyses, "aggressiveness", "reactivity", and "trainability" were determined to be consistent with results found in the US and UK surveys. On the basis of these factor scores, seven groups of breeds were determined by cluster analysis to compare to the US survey; 22 of the 38 breeds common with the US survey were categorized into the same groups as those in that survey. The results demonstrated differences in canine behavioral predisposition among breeds and between genders. The similarity in the results between our study and previous surveys, which involved distinct geographical locales, suggests that the genetic basis of breed-specific temperamental traits is manifested irrespective of the cultural or regional identities of the owners.
Monoclonal antibodies (MAbs) reported here were produced against the porcinophilic foot-and-mouth disease virus (FMDV) that caused the devastating swine disease on 1997 in Taiwan. A panel (25) of MAbs were found to react with VP1 of O/Taiwan/97 (O/97) by ELISA with various potencies. The biological identities of these VP1 reacting MAbs, such as neutralization activity, isotype and capability to distinguish between two serotype O FMDVs, O/97 and O/Taiwan/KM1/99 (O/99), were further analyzed. Eleven out of the total eighteen O/97 neutralizing MAbs were able to neutralize heterologous O/99. Eight O/97 neutralizing and five non-neutralizing MAbs could differentiate two serotype O FMDVs by immunofluorescence assay (IFA) implied that these thirteen MAbs recognized O/97 specific epitope(s). Furthermore, reactivities of the VP1 reacting MAbs with a 29 amino acids synthetic peptide (P29) representing the βG-βH loop of VP1 were analyzed by ELISA and fourteen were found positive. MAb clone Q10E-3 reacting strongest with VP1 and P29, neutralizing both but not differentiating two serotype O viruses suggested that the antibody binding site might involve the RGD motif and its C terminal conserved region on βG-βH loop. MAbs with diverse characters presented in this study were the first raised against porcinophilic FMDV. The complete set of MAbs may be used for further studies of vaccine, diagnostic methods, prophylaxis, etiological and immunological researches on FMDV.
The purposes of the present study were to determine the systolic blood pressure (SBP) of clinically normal and conscious cats, and to set up reference values of feline SBP for subsequent clinical application. SBPs were measured in 53 healthy cats using an ultrasonic Doppler device. The mean SBP was 133.6 ± 16.0 mmHg (range, 110.0-180.0 mmHg). The distribution of SBP values was not significantly affected by factors such as breed, body condition score, or age (P>0.05), but SBP values of female cats were significantly lower and more variable than those of males (t test, P=0.004; F test, P<0.001). Feline SBP between 114.3 mmHg and 149.5 mmHg was considered indicative of normotension. SBP values higher than 159.3 mmHg were defined as hypertension, and those less than 104.5 mmHg were determined as hypotension.
A 5-year-old female border collie presented with erythematous skin lesions at the axillae, groin, mucocutaneous junctions, and pinnae. Biopsy revealed lymphocytic interface dermatitis with hydropic degeneration of basal cells and keratinocyte apoptosis. Based on gross and histological features, diagnosis of erythema multiforme was made. The disease was resolved by treatment with azathioprine, prednisolone, and a hypoallergenic diet. Finally, the skin lesion was controlled without drug therapy but recurred easily every time commercial foods except the hypoallergenic diet were used, suggesting that food substances triggered this outbreak.
The clinical utility of various specimens was examined for the early diagnosis of canine distemper (CD). Seven healthy dogs at 17 weeks of age were experimentally infected with a field isolate of canine distemper virus. The RT-PCR was carried out to detect CDV NP gene. Dogs showed mild fever and leukopenia, however, typical clinical signs of CD were not seen through the experimental period. CDV amplicons were detected more, earlier and for longer period in the conjunctival swabs than in the other samples employed. These results suggested that conjunctival swab samples, which are easy to obtain and non-invasive, would be the most suitable and practical specimen for the early antemortem diagnosis of CDV infection.
Molecular cloning of feline lung resistance-related protein (LRP) was performed to evaluate the relationship between its expression level and drug resistance against chemotherapeutics. The nucleotide sequence of the coding region of feline LRP cDNA was found to be 2670-bp long and to show 84.2-92.6% homology to its human, mouse, and rat counterparts. The expression level of feline LRP mRNA was relatively high in lung, jejunum, and colon. An adriamycin (ADM)-resistant feline lymphoma subline, FT-1/ADM, showed a high level of MDR1 mRNA expression compared with parental FT-1 cells. However, no relationship was observed between the drug-resistant phenotype and the LRP mRNA expression level. Although no direct contribution of LRP to the development of the drug-resistant phenotype was observed, further investigation is advisable.
Hepatozoon sp. infections were detected in two species of Japanese wild cat, Iriomote wild cat (Felis iriomotensis) and Tsushima leopard cat (Felis bengalensis euptilura), between April 1993 and October 2005. The prevalence was 56.7% (17/30) and 14.3% (6/42), respectively. The most affected organ was the heart; all infected animals had organisms in their hearts. The parasitizing form was schizont and various developmental stages were observed. The size of schizont and merozoite was 22.3 ± 3.1 × 15.3 ± 2.2 μm and 6.1 ± 0.6 × 2.3 ± 0.2 μm, respectively. Few inflammatory reactions against the parasites were observed. Electron microscopically, organisms were located in parasitophorous vacuoles of unidentified host cells, and mature schizonts consisted of numerous merozoites. This is the first report of hepatozoonosis in Japanese felids.
Fecal survey of Eimeria oocysts and parasite eggs was conducted for 219 fecal samples of free-ranging Grus japonensis in Kushiro district in Hokkaido in April 2003. Positive rate and mean oocysts (or eggs) per gram in positive samples were 26% (57/219) and 8.8 (0.2-136) in oocysts of Eimeria reichenowi, 18.3% (40/219) and 320 (100-1,000) in trematode eggs, 0.1% (2/219) and 0.2 (0.2-0.3) in eggs of Nematoda A, and 4.1% (9/219) and 0.8 (0.2-3.6) in eggs of Nematoda B, respectively.
Trichoblastomas, trichoepitheliomas, and squamous cell carcinomas in the skin of dogs were analysed by immunohistochemistry for the nuclear expression of p27, p21 and proliferating cell nuclear antigen (PCNA). High levels of p27 were present in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. Detectable p21 was found in trichoepitheliomas and squamous cell carcinomas, but trichoblastomas had low level of p21 nuclear reactivity. Low levels of PCNA were detected in trichoepitheliomas and trichoblastomas compared with squamous cell carcinomas. The results suggested that nuclear p27 acts as a cyclin-dependent kinase (CDK) inhibitor in trichoepitheliomas and trichoblastomas. Nuclear p21 expression is involved in the induction of epithelial differentiation and seems to be unrelated to CDK inhibition.
This paper describes a nephroblastoma with transcoelomic metastasis in a three-year-old Japanese black bull. At necropsy, a huge, oval neoplastic mass containing the residual right kidney was found. Moreover, severe transcoelomic metastasis occurred throughout the abdominal and thoracic cavities. Histologically, the mass was mainly composed of sheets, nests, islands and cords of polygonal blastemal cells with trabeculae of fibrous stroma. In some areas, epithelial elements composed of tubules and winding duct-like structures were also observed. Glomeruloid structures were scattered in these epithelial elements. Metastatic nodules were composed of blastemal and stromal elements, which were similar to those in the mass.
Two hundred and twenty strains of Campylobacter jejuni (70 human, 51 canine and 99 chicken strains) were isolated from September 2003 to September 2004 in northern Taiwan. These strains were subtyped by PCR-RFLP analysis of the flagellin (FlaA) gene. On the basis of restrictive digest, six types were identified with AfaI, seven types with MboI and five types with HaeIII. With the combination of these three enzymes, 47 distinct PCR-RFLP patterns were observed-25 each from human and chicken isolates, and 9 from canine isolates. In human strains, the most frequently occurring types were Cj-28 (14.3%), Cj-17 (10%), Cj-16 (8.6%), Cj-37 (7.1%) and Cj-46 (7.1%). In canine strains, the most prevalent types were Cj-1 (33.3%), Cj-26 (19.6%), Cj-3 (15.7%), Cj-2 (9.8%) and Cj-10 (9.8%). In chicken strains, the most frequently occurring types were Cj-46 (40.4%), Cj-29 (9.1%), Cj-45 (7.1%) and Cj-41 (5.1%). The results suggest that poultry is a source, but not the sole source, of C. jejuni infection in humans. Two RFLP types, Cj-17 and Cj-37, frequently occurring in human isolates in this study have also been found to be prevalent in human isolates in Japan, China and the Czech Republic, indicating a possible international clonal spread.
Antimicrobial susceptibility of 183 Salmonella isolates from apparently healthy food-producing animals obtained during the period from 2000 to 2003 throughout Japan was examined. Of 29 serovars identified, Salmonella Infantis (37.7%) was the most prevalent, followed by S. Typhimurium (19.7%). Salmonella bacteria resistant to dihydrostreptomycin (77.6%) were about 10% more prevalent than those resistant to oxytetracycline (67.8%), though the nation-level veterinary use of tetracycline antibiotics is much greater than that of streptomycin in Japan. In seventeen isolates (9.3%) resistant to nalidixic acid, single point mutations were detected at 84 or 87 in the quinolone resistance-determining region of the gyrA gene.
We examined the influence of propofol infusion on cardiovascular system at the rate of 0.14, 0.20 and 0.30 mg/kg/min in six adult Thoroughbred horses. The cardiovascular parameters were heart rate (HR), mean arterial pressure (MAP), mean right atrial pressure (MRAP), stroke volume (SV), cardiac output (CO), systemic vascular resistance (SVR), pre-ejection period (PEP) and ejection time (ET). In order to keep the ventilation conditions constantly, intermittent positive pressure ventilation was performed, and the partial arterial CO2 pressure was maintained at 45 to 55 mmHg during maintenance anesthesia. SV showed a significant dose-dependent decrease however, CO did not show significant change. SVR decreased significantly at higher dose. PEP was prolonged and PEP/ET increased significantly at the highest dose. From these results, it became clear that SV decreases dose-dependently due to decrease of cardiac contractility during anesthesia with continuous propofol infusion in horses. On the other hand, since MAP and CO did not show significant changes, total intravenous anesthesia with propofol was suggested to be suitable for long-term anesthesia in horses.
Retinoids are well recognized as promising antitumor agents in humans. However, there have only been a few reports about the effect of retinoids in canine cancers. To investigate the antitumor effect of retinoids on mast cell tumors (MCT), inhibitory effect on cell growth and induction of apoptosis were examined in vitro. Although sensitivity of these cells differed among the cells, the growth of three MCT cell lines (CoMS, CM-MC and VI-MC) were inhibited dose dependently when they were treated with retinoids. FACS analysis of PI-stained nuclei revealed an apoptotic fraction in CM-MC cells about 30% when treated with retinoids, while those of control cells were less than 5%. Caspase-3 activation was observed after retinoid treatment in CM-MC cells. This was confirmed by inhibiting the retinoid-induced apoptosis using the pan-caspase inhibitor, ZVAD-FMK. Both retinoid receptors, RARs and RXRs, were detected by immunoprecipitation followed by western blot analysis in all the three MCT cells. These data suggests that retinoids inhibit the growth of MCTs partly through apoptosis, and this growth inhibition by retinoids may be mediated by RARs and RXRs. We conclude that retinoid may be a potential adjunctive chemotherapeutic agent for the treatment of canine MCT.
The incidence of primary and secondary glaucoma in dogs was investigated. A total of 1244 dogs received ophthalmologic examinations, including tonometry and gonioscopy. Goniophotographs were taken using a goniolens to evaluate the iridocorneal angle (ICA) as well as pectinate ligament (PL). The anterior width of the ciliary cleft and the total distance from the origin of the PL to the anterior corneal surface were measured from the goniophotographs. Glaucoma was diagnosed based on the cupping of the optic nerve head, clinical signs, ocular changes, and high IOP, and it was synchronized with gonioscopic grades to differentiate between primary and secondary glaucoma. We investigated 1244 dogs of 29 breeds, including the mixed breed; among these, glaucoma was diagnosed in 127 dogs (162 eyes). Of 162 eyes, primary glaucoma was diagnosed in 129 eyes and secondary glaucoma in 33 eyes. Shiba Inu dogs (42 dogs, 33%) showed the highest incidence of glaucoma, followed by Shih-Tzu (21 dogs, 16.5%). Furthermore, all the glaucomatous Shiba Inu dogs had primary glaucoma with abnormal ICA grades and dysplastic PLs. The findings of our study reveal that the Shiba Inu breed in Japan may have a hereditary predisposition to glaucoma.
We investigated the effect of olprinone on canine myocardial pump function and myocardial damage after ischemia-reperfusion injury. Three dogs of the experimental group were given olprinone (Olprinone group) and another 3 dogs were served as control (Intact group). All animals were occluded left anterior descending artery for 60 min, followed by 6 hr of reperfusion. In the experiment, hemodynamics, infarct area, creatine kinase and troponin-I were measured. Olprinone infusion induced significantly high cardiac output value and significantly low values in left ventricular end diastolic pressure and systemic vascular resistance index after reperfusion. Also, olprinone tend to attenuate the infarct area, creatine kinase and troponin-I.
A 14-year-old intact male Maltese dog was presented with a history of pain and swelling of the left upper forelimb and lameness for 3 weeks. Hematological and radiographical examinations showed regenerative anemia and osteolysis of the humerus. Fine needle aspiration biopsy detected epithelioid- and fibroblast-like anaplastic cells with blood components. A hemorrhagic and osteolytic malignant tumor was suspected, and the affected forelimb was amputated. Histopathologically, the dog was diagnosed with primary hemangiosarcoma of the humerus. Thereafter, metastatic lesions appeared in the skin, and the dog was euthanized 1 month after the operation.
Dogs receiving anterior cruciate ligament transection (ACLT) were treated with either intravenous (IV) or intraarticular (IA) administration of hyaluronan (HA), and differences in appearance of chondrocyte apoptosis of the stifle joint were investigated. Chondrocyte apoptosis was detected using flow cytometry as well as by staining with TdT-mediated dUTP nick end labeling (TUNEL). The percentage of apoptotic chondrocytes in dogs with ACLT was significantly higher than that in intact (non-ACLT) dogs. Dogs treated with IA or IV injection of HA after ACLT had fewer apoptotic chondrocytes than non-treated dogs after ACLT. It was suggested that ACLT-induced apoptosis of chondrocytes was suppressed by HA administration of either IA or IV.
Preweaning mortality risks, recorded death reasons and related factors for preweaning mortality were studied in 105 breeding herds. Preweaning mortality risk at the herd level was calculated as the difference between the number of pigs born alive in farrowed litters and the number of weaned pigs divided by the number of pigs born alive in litters that farrowed and weaned. The mean of annual preweaning mortality risk was 10.7%. In regression analysis, higher mortality risks were associated with higher parity at farrowing, greater numbers of pigs born alive, and longer lactation length. The period from July to September had a higher mortality risk than that from April to June. The means of cause-specific proportional mortality ratios (PMR) in trauma with low viability and scours were 80.4 and 6.2%, respectively. Sows with pig age 0-1 day during lactation had the highest daily PMR. Sows with pig age 0 to 7 days had higher PMR due to trauma and low variability than those with pig age 8 days older. Sows with pig age over 7 days had higher PMR due to scours than those with pig age 0-7 days. Careful management at farrowing and in early lactation on high parity sows with large litters should be considered to prevent piglets from death due to trauma and low viability, and appropriate herd health programs should be implemented for reducing preweaning mortality due to scours during late lactation.
The treatment time needed for high-intensity focused ultrasound (HIFU) ablation might be decreased substantially by using the split-focus approach, so we made a prototype 4.2-MHz split-focus therapeutic transducer combined with a small 6.5-MHz imaging ultrasonic probe for transrectally treatment of canine prostatic cancer and used it to experimentally evaluate the feasibility of using split-focus transrectal HIFU to ablate canine prostatic tissue without injuring surrounding tissues. The prostates of 5 dogs were transrectally treated with split-focus ablation at a peak intensity in the water of 1.7 kW/cm2 for 4 s (4 shots) under the guidance of ultrasonic B-mode imaging. After ultrasonic exposure, the prostates became stiff because of thermal effect of HIFU. For the first 3-5 days after treatment, dogs were catheterized daily for urinary management and treated with oral antibiotics to prevent urinary tract infection. The dogs were able to urinate normally by a week after. Within two weeks a large centrally located cystic cavity had formed in the prostate by replacing the necrotic parenchyma around the prostatic urethra. Necropsy three months after treatment found the rectum and prostate capsule to be normal grossly and histologically. The 4 shots of split-focus HIFU destroyed the prostatic parenchyma and created a prostatic cavity 0.34-0.45 cm3 in volume without injuring surrounding tissues. These results suggest that split-focus HIFU ablation could be used for noninvasive treatment of prostatic cancer in dogs.
The purpose of present study was to determine annual changes in serum progesterone (P4) concentrations and to clarify basic reproductive characteristics, such as breeding season, estrous cycle, and puberty in female bharals (Pseudois nayaur). Blood was collected from 9 female bharals once or twice weekly for approximately one year. Serum P4 concentrations were determined by radioimmunoassay. Serum P4 concentrations showed remarkable and cyclic changes between November/December (winter) and May/June (late spring). The mean estrous cycle was 24.9 ± 0.5 days. Chasing insistently to other females and discharge of mucus from the vulva were observed around the time when the serum P4 concentrations began to increase. The chasing behavior and discharge of mucus were considered to be external indicators of estrus in female bharals. Serum P4 concentrations of a pregnant female had non-cyclic changes, and the values remained high. In this study, all 37 deliveries were between April and September, and about 70% of these were concentrated in May and June. The conception month determined on the day of birth was between October and April for all animals, and the most common month was in December (54%). This month corresponded to an early stage of the period when the serum P4 concentrations changed cyclically. These results indicate that many female bharals become pregnant at the beginning of the breeding seasons and, if they do not become pregnant, the estrous cycle, about 25 days in length, is repeated.