Bursae of Fabricius were collected from 20 chickens diagnosed with infectious bursal disease virus (IBDV) infection from 15 prefectures in 1993 to 2004. Here we report the nucleotide sequence analysis of VP2 hypervariable domain of IBDV genome detected by reverse transcription-polymerase chain reaction from these samples. Ten sequences derived from 10 prefectures in 1996 to 2003 were of the classical type and other 10 sequences derived from 6 prefectures in 1993 to 2004 were of the highly virulent type. Of the classical type sequences, 9 sequences were closely related to the sequence of classical attenuated vaccines used in Japan. Furthermore, two were identical to the sequence of B-Chi5 which represents Vaccine B passaged 5 times in chickens and was reported to be reverted the virulence during the passages. The 10 highly virulent type sequences were classified into four sequences, none of which had been previously detected in Japan. However, the deduced amino acid sequences were identical to each other and to the sequences of highly virulent IBDVs previously detected in Japan. The most common nucleotide sequences, which accounted for 6 of the sequences, were identical to 34 highly virulent type sequences detected in various countries in BLAST search. This is the first report of detection of the sequence in Japan which is identical to highly virulent strains detected in other countries. These findings show the prevalence of classical IBDVs closely related to the attenuated vaccines and highly virulent IBDVs derived from other countries throughout Japan since 1993.
Bordetella bronchiseptica pertactin (prn) is an outer membrane protein which has been implicated as both an adhesin and a protective antigen that induces immunity against atrophic rhinitis in pigs. Previous studies demonstrated extensive heterogeneity of the prn sequence within two distinct regions of amino acid repeats for B. bronchiseptica isolated from the United States and Europe. By deducing the amino acid sequences of the repeat regions of the prn gene from recent isolates from Korea, two region 1 variants and five region 2 variants were identified. Five pertactin types were distinguished based on combinations of variants of both regions. Interestingly, none of the field isolates have the same pertactin type as the B. bronchiseptica P4 strain widely used to vaccinate pigs.
Rectal swabs from 100 dogs that were admitted to the veterinary clinic and from randomly selected 100 dogs from one kennel were examined for the presence of Salmonella spp. S. Enteritidis was isolated from a household dog and the strain was susceptible to all antibiotics that were tested. S. Typhimurium was isolated from a dog from the kennel and it was sensitive to all the antibiotics except streptomycin and neomycin. Even though, the occurence of Salmonella spp. in the feces of the dogs in this study was low, attention should always be given when handling canine feces or contaminated materials.
An 11-year-old, castrated male Miniature Poodle dog was referred due to daily plant eating followed by vomiting for 7 years. Each time the dog went out for a walk, he had been munched grass, and then vomited. There were no abnormal findings on medical examinations. Through behavior examination and consultation, we diagnosed this dog as having a plant eating problem. Because we assumed that the dog had eaten plants to supply of dietary deficiency or relieve digestive problems, advised the owner to discontinue supplying previous diet and to feed high-fiber diet. On the follow up, the owner said that the dog had not eaten plants and vomited from 3 days after initial dietary change. The dog has not shown any clinical signs for 13 months. This clinical finding shows a possibility that deficient diet, especially fiber, relates to the plant eating behavior of a dog.
Epsilon-polylysine micro particles (SGEPL) and polyethyleneimine micro particles (SGPEI) were developed by the addition of a hydrophobic group and the immunological characterization of these micro particles and aluminum hydroxide (ALUM) was investigated. BALB/c mice were injected intraperitoneally with ovalbumin (OVA) as an antigen and SGEPL, SGPEI or ALUM as an adjuvant. The results showed that the mice injected with SGEPL produced a significant portion of anti-OVA antibody subclass IgG2a in the sera and suppressed interleukin (IL)-4 and IL-5, but enhanced IL-12 and interferon-γ (IFN-γ) from the spleen cells. Similar results relating to cytokines were also obtained, even without OVA. Direct stimulation with SGEPL to naïve BALB/c mouse spleen cells induced IL-12 and IFN-γ. Both spleen and purified B cells produced IgG1 and IgE after stimulation with IL-4 and the anti-CD40 monoclonal antibody. With the addition of SGEPL, the IgE production from the cells was suppressed as a result of enhanced IFN-γ production. Furthermore, IgE production was also suppressed in the purified B cells without the influence of IFN-γ or IL-12. Thus, we suggest SGEPL drives cytokine production to Th1 profile. It will be a novel promising adjuvant based on this viewpoint.
Lymphocytic-plasmacytic enteritis (LPE) is a type of canine inflammatory bowel disease (IBD). One of its most probable causes is a defect in the mucosal permeability barrier. In the present study, intestinal permeability in LPE dogs was examinated to evaluate its clinical value. Twenty-nine dogs with LPE diagnosed by clinical and histological examinations were included in this study. Intestinal permeability was evaluated by measuring the ratio of the concentrations of two sugars (lactulose (L) and rhamnose (R)) with different molecular weights in urine samples after oral administration of a solution containing them. Biopsy specimens of duodenum were evaluated according to histological criteria. The urinary L:R ratio in the 29 LPE dogs (1.68 ± 1.17, mean ± SD) was significantly higher than that in the 10 healthy control dogs (0.75 ± 0.38, P<0.01). In the LPE dogs, a weak correlation was observed between the histopathological grading score of the duodenum and the urinary L:R ratio (r=0.408, P<0.05). The urinary L:R ratio in the 20 dogs showing hypoalbuminemia (< 2.5 g/dl) was significantly higher than that in the 9 dogs with normal serum albumin levels > 2.5 g/dl (P<0.01). In conclusion, permeability of the intestinal mucosa as determined by the urinary L:R ratio could be a useful laboratory parameter for evaluating intestinal damage in LPE dogs.
A 4-year-old, male Labrador retriever, weighing 27 kg, presented with abrupt clinical signs including mental retardation, circling and head pressing. The dog never ingested water by choice. An adipsia of the dog was persisted and developed to hypernatremia with artifactual hyperchloremia. Serial endocrine results and image findings were suggestive of a hypothyroidism. The dog revealed the presence of antithyroid antibodies in the cerebrospinal fluid and serum. With the administration of levothyroxine sodium, his neurologic signs were alleviated within the first week of treatment and adipsia was also resolved.
The hemodynamic effects of vasopressin of high/low doses on dogs were investigated using experimentally induced hemorrhagic shock model. Experimental groups were categorized according to administered doses of vasopressin (0.1, 0.4 and 1.6 IU/kg) and hemodynamic parameters were measured before and after the graded-dose administration of vasopressin. Administration of high- and middle-dose vasopressin (0.4 and 1.6 IU/kg) showed superior increase in blood pressure and systemic vascular resistance, compared with those of low-dose one (0.1 IU/kg). Results of systolic arterial pressure and mean arterial pressure in 1.6 IU/kg-administered group revealed lower efficacy than that in 0.4 IU/kg group in spite of administration of higher dose. This study demonstrates that 0.4 IU/kg of vasopressin can be used as the most effective dose for improving hemodynamic condition in the decompensatory phase of hemorrhagic shock in dogs.
Female reproductive organs are mainly regulated by estrogen and progesterone. Specifically, the uterus, vagina and mammary gland show organ-specific mitosis and morphological changes during proliferative events, such as estrous cycle, gestation and lactation. The mechanism underlying these organ-specific estrogen-dependent events is still unknown. We examined, therefore, global gene expression in the mature uterus, vagina and mammary gland of ovariectomized adult mice 6 hr after an injection of 5 μg/kg 17β-estradiol (E2) using a microarray method in order to identify primary E2-responsive genes. Half of the E2 up-regulated genes in the uterus were similar to those in the vagina. E2 up-regulated the expression of Insulin-like growth factor 1 (Igf-1) genes in the uterus and vagina. In the vagina, E2 up-regulated the expression of IGF binding proteins (Igfbp2 and Igfbp5). In the mammary gland, unlike the uterus and vagina, no gene showed altered expression 6 hr after the E2 exposure. These results suggest that expression of Igf-1 and morphogenesis genes is regulated by E2 in an organ-specific manner, and it is supported by the results of BrdU labeling showing E2-induced mitosis in the uterus and vagina except the mammary gland. The differences in organ specificity in response to E2 may be attributed by differences in gene expression regulated by E2 in female reproductive organs. The candidate estrogen-responsive genes in the uterus and vagina identified by profiling provide an important foundation understanding functional mechanisms of estrogen regulating morphogenesis and maintenance of each reproductive organ.
Canine ovarian tumors (epithelial tumor, sex-cord stromal tumor, germ cell tumor) classifying into 9 histological types were examined immunohistochemically using placental alkaline phosphatase (PLAP), cytokeratin7 (CK7), desmin, S100, AE1/AE3, inhibin α, vimentin, and alfa feto-protein (AFP). The papillary and tubular types observed in epithelial tumors were immunoreactive for desmin and AE1/AE3. The papillary type was also immunoreactive for PLAP and CK7. The solid type, nest type, cord type, palisade type, cystic type and spindle type, which were observed in sex-cord stromal tumors, showed a positive immunoreaction for S100 but little or no positive immunoreaction for inhibin α with an exception of positive result in the palisade type. Most of the sex-cord stromal tumors were AE1/AE3-positive except for the palisade type. In the cobblestone type observed in germ cell tumors, only vimentin and AFP were positive. The present study elucidated the detailed histological and immunohistochemical characteristics of canine ovarian tumors.
A 20-year-old thoroughbred mare had a mass in the right kidney. The mass was encapsulated with fibrous capsule and composed of variably-sized papillary projections lined by a single layer of flattened to cuboidal neoplastic epithelial cells with no cytological and nuclear atypia. Immunohistochemically, the neoplastic cells were broadly positive for cytokeratin AE1/AE3 and granular staining for alpha-1-antitrypsin was focally detected; this immunohistochemical property was similar to that of the normal distal nephron. From these results, this case was diagnosed as papillary renal adenoma of distal nephron differentiation.
We previously reported that no Kupffer cells reacted with the antibody against lysozyme, and Ito cells contained a large cytoplasmic vacuole in the feline liver. In this report, we further examined the characteristics of porcine and bovine hepatic non-parenchymal cells. In the liver of both animals, Kupffer cells were positive for lysozyme, and cytoplasmic vacuoles in Ito cells were small. The histopathological characteristics of porcine and bovine hepatic non-parenchymal cells were different from those of the feline liver.
This paper describes a neuroendocrine (NE) tumor of the lung that was observed during the necropsy of a 14-year-old female black spider monkey (Ateles paniscus) with sudden death. Grossly, multifocal firm and coalescing nodular masses were observed in the lung. The histological examination showed the tumor to be an typical NE tumor with polygonal cells grouped in small solid aggregates, with regularly sized, spherical, centrally placed nuclei with modest, lightly granular cytoplasm suspended in a fibrovascular stroma. The immunohistochemical examination revealed the tumor to be positive for cytokeratin, chromogranin A and synaptophysin, and negative for CD56. To the best of our knowledge, this is the first report of NE tumor in the lung of the black spider monkey.
Concern has been growing about the cardiac toxicity of antimalarial drugs. Artemisinin, a unique type of antimalarial drug originating from a Chinese medicinal plant, has minimal adverse effects, but it has been reported to inhibit delayed rectifier potassium current, a voltage-gated potassium current. However, no studies have been published concerning the effect of artemisinin on ligand-gated potassium currents. Therefore, in the present study, we examined the influence of artemisinin on the acetylcholine receptor-operated potassium current (IK.ACh), a ligand-gated potassium current, in guinea pig atrial myocytes using a patch clamp technique. Artemisinin (1 to 300 μM) inhibited IK.ACh induced by extracellular application of both carbachol (1 μM) and adenosine (10 μM) and that induced by intracellular loading of GTPγS (100 μM) in a concentration-dependent manner. Artemisinin inhibited carbachol-induced, adenosine-induced, and GTPγS-activated IK.ACh within almost the same concentration range. In left atria, artemisinin (1 to 100 μM) partially reversed the shortening of action potential duration induced by carbachol in a concentration-dependent manner. Carbachol-induced negative inotropic action in left atria was also inhibited by artemisinin (10 to 300 μM). In conclusion, we suggest that the anticholinergic action of artemisinin is mediated through inhibition of IK.ACh via inhibition of the muscarinic potassium channel and/or associated GTP-binding proteins.
Japanese horse chestnut (Aesculus turbinata) seed extract inhibits the activity of cyclooxygenase (COX), but its active constituents have not been identified. In the present study, COX inhibitors were isolated from the hexane extract of this seed by means of 4 steps of liquid chromatography and were identified by gas chromatography/mass spectrometry and nuclear magnetic resonance. The COX inhibitors in the extract of Japanese horse chestnut seeds were identified as linoleic acid, linolenic acid, and oleic acid. Their efficacies were in the following order: linolenic acid = linoleic acid > oleic acid. These active constituents are C18 unsaturated fatty acids; stearic acid, a C18 saturated fatty acid, had no activity. Linolenic acid and linoleic acid had high selectivity toward COX-2 (selectivity index = 10), whereas oleic acid had no selectivity. Considering the efficacy and yield of each fatty acid, linoleic acid may be the principal COX inhibitor in this seed.
In excretory urography, the osmolarity of contrast media has rarely been treated as important in veterinary medicine. In this study, the contrast effect of two contrast media (monomeric iohexol and dimeric iodixanol) in the renal cortex and aorta were compared using computed tomography (CT). Five beagle dogs were used and the study employed a cross-over method for each contrast media. The results showed that there was no difference between the media in the aorta, but iodixanol showed higher CT value and a longer contrast effect than iohexol in the renal cortex, in spite of having the same iodine dosage. It is believed that iodixanol, with its low osmolarity, is diluted less by osmotic diuresis than monomeric iohexol. It is important to consider the osmolarity of the contrast media when evaluating the contrast effect, and it is essential to use the same contrast media for each examination, or the renal excretory speed will be under/overestimated.
A one-year-old female cat was unable to stand. Magnetic resonance imaging was performed, and an enlargement of the lateral, third, and fourth ventricles and syringomyelia were detected. The cat was diagnosed with an isolated fourth ventricle (IFV) with syringomyelia. The serum isoantibody test for the feline infectious peritonitis (FIP) virus was 1:3,200. After the cat died, a pathological examination revealed nonsuppurative encephalomyelitis. We suspected that the IFV, detected in the cat, was associated with FIP encephalomyelitis. To our knowledge, there has been no report on IFV in veterinary medicine.
The incidence of canine rabies has been widely reported in Brazil, and new rabies virus (RV) variants, genetically similar to canine RV, have recently been isolated from foxes. In order to derive the epidemiological characteristics of Brazilian Carnivora RV, Brazilian RVs isolated from dogs, cats, and foxes were genetically analyzed. Brazilian Carnivora RV isolates were divided into 2 main lineages. The predominant lineage was found in dogs and cats, which included the Argentinean and Bolivian Carnivora RV isolates, and was extensively distributed throughout Brazil and surrounding countries. The other lineage consisted of three sublineages containing Brazilian dog and fox RV isolates, with the dog sublineages located on an internal branch of 2 fox sublineages, suggesting that RV transmission events might have occurred between foxes and dogs in the past. These results suggest that contact between dogs and wildlife has the potential to generate new rabies variants and that it is important to control RV infection cycles in both dogs and wildlife to prevent spread of rabies infection.
To compare the molecular and growth properties of two newly isolated canine distemper virus strains in the Asia 1 and 2 groups with clinico-pathological findings in dogs, nucleotide and predicted amino acid sequence comparisons of genes H and P were performed together with comparative growth profiling. The predicted amino acid sequences of the H gene contained 12 cysteine residues that were conserved among the examined Asia 1 and Asia 2 viruses. The hydrophobic region in the H gene of the Asia 2 isolates was one amino acid longer than that of the Asia 1 group. The H gene of the Asia 1 group had nine putative asparagine (N)-linked glycosylation sites, while there were eight sites in the Asia 2 group. The titers of the cell-associated viruses for the Asia 1 strains were higher than those of the released viruses and were opposite to those of the Asia 2 strains in a previous study. The molecular and growth properties of the Asia 1 and Asia 2 groups seem to vary, although no significant differences were observed in the clinical signs and pathological findings between the two groups.