Immunolocalization of the secretory form of carbonic anhydrase isoenzyme, CA-VI were studied using a specific canine CA-VI antiserum, and CA-VI mRNA signals were also investigated using the reverse-transcriptase polymerase chain reaction (RT-PCR) in canine nasal mucosal epithelia and glands. Immunoreactivity to CA-VI was positive throughout the mucosal epithelial cells and in the cytoplasm of serous acinar and ductal epithelial cells of the nasal mucosa and glands, including the vestibule of the nose, but the mucous acinar cells of the glands were immunonegative. We detected CA-VI gene transcripts in the same regions as the CA-VI immunoreactivity. The physiological roles of CA-VI in the nasal mucosal epithelium and glands might maintain bicarbonate levels in nasal secretions and protect the mucosa against acid.
This study examined the branching pattern of the aortic arch and its major branches in Korean water deer (Hydropotes inermis argyropus, Heude, 1884). Silicone casts were taken from the vessels of 23 carcasses (male 14, female 9) with body weights ranging from 1.3-16.0 kg through a retrograde injection into the abdominal aorta. The findings were compared with those from other domestic ruminants. Only the brachiocephalic trunk (Bct) branched from the aortic arch in all carcasses. In 19 of the 23 cases, the Bct branched into the left subclavian artery (LSb), the left common carotid artery (LCc), and then trifurcated into the right common carotid artery (RCc), right costocervical trunk (RCct) and right subclavian artery (RSb). The subclavian artery (Sb) branched into the costocervical trunk (in left), internal thoracic artery (It), and superficial cervical artery (Sc) in that order, and continued as the axillary artery. Instead of separated carotid arteries, the bicarotid trunk from the Bct was observed in only three males and one female. Two of these males had different branching orders of the It and Sc from the Sb in one or both sides. The other male had a RCct from the RSb. The left costocervical trunk (LCct) arose from the LSb in all cases, and branched into the highest intercostal artery, the dorsal scapular artery, and the deep cervical arteries in that order, and continued as the vertebral artery. In 22 cases, the RCct branched directly from the Bct at the same point in which the RCc (or bicarotid trunk) and RSb separated. The artery branching pattern from the RCct was similar to that of the LCct. These results suggest that the Korean water deer has a Bct with different branching patterns from those of domestic ruminants.
To clarify the involvement of excitatory and inhibitory amino acids in canine necrotizing meningoencephalitis (NME), glutamate, aspartate, taurine and gamma-aminobutylic acid (GABA) were determined in the cerebrospinal fluids (CSF) from eight NME cases and ten healthy controls. NME dogs exhibited significantly higher concentrations of glutamate and aspartate than those in controls (p<0.001 and p<0.001, respectively), while there was no difference in taurine or GABA between the two groups. When fetal canine astrocytes were cultured for 24 hr in the presence of NME-CSF, supernatant concentrations of glutamate, aspartate and taurine were significantly elevated. Simultaneously, expression of excitatory amino acid transporter 2 (EAAT2) mRNA was significantly reduced in the astrocytes without change in EAAT1 mRNA. Hence, reduced expression of EAAT2 and impaired glutamate homeostasis may contribute to the pathogenesis of NME.
Two cats showing chronic vomiting, diarrhea and weight loss were found to have leukocytosis with marked eosinophilia. Both cats were diagnosed with hypereosinophilic syndrome by the findings of increased eosinophils and their precursors in the bone marrow, eosinophilic infiltration into multiple organs, and exclusion of other causes for eosinophilia. Although cytoreductive chemotherapy with hydroxycarbamide and prednisolone was performed, these two cats died 48 days and 91 days after the initial presentation.
Detection of hemotropic Mycoplasma spp. infection was attempted in cats by PCR using whole blood without DNA extraction. A total 46 of 54 (85%) cats with suspected Mycoplasma spp. infection showed a positive reaction, corresponding completely with the results of standard PCR testing. The direct PCR assay was sensitive enough to detect more than 0.0061% parasitemia for `C. M. haemominutum' and 0.0075% parasitemia for M. haemofelis. These data indicate that the direct PCR assay might be sufficient for use as a tool in clinical examinations.
A 15-year-old, neutered-male pony presented with a history of weight loss during 4 months. Clinical evaluation revealed severe bradycardia and complete atrioventricular block. At necropsy, a lobulated mass in the anterior mediastinum and moderate enlargement of the superficial cervical lymph nodes were observed. The vagus nerve and the brachiocephalic trunk were embedded in this anterior mediastinal tumor. Histologically, the mass was composed of sheets of neoplastic lymphoid cells expressing CD3, with a low mitotic rate. To the best of our knowledge, this is the first reported case of mediastinal lymphoma associated with complete atrioventricular block in horses.
Glutamate decarboxylase (GAD) is the primary enzyme in the brain that catalyzes the synthesis of γ-aminobutyric acid (GABA), the main inhibitory neurotransmitter. There are two isoforms named according to their molecular weights, GAD67 and GAD65, which are encoded by GAD1 and GAD2, respectively. To investigate the association between GAD genes and temperament in domestic dogs, Canis familiaris, we sequenced the full lengths of the coding regions of these genes and identified three single nucleotide polymorphisms (SNPs) in GAD1 and one in GAD2. When comparing genotype and allele frequencies of SNPs among five breeds with different behavioral traits, statistically significant interbreed differences were observed for three SNPs in GAD1. These results suggest that GAD1 SNPs may be useful for behavioral genetic studies in dogs.
Tryptophan hydroxylase (TPH) serves as the rate-limiting enzyme in the biosynthesis of serotonin, and two forms of TPH genes, TPH1 and TPH2, have been reported with specific nucleotide sequences and expression patterns. We determined the sequences of these two genes and identified polymorphisms within their coding regions. We identified five novel single nucleotide polymorphisms in the TPH2 gene, one of which resulted in a non-synonymous mutation and the remaining four in synonymous mutations. Comparison of the genotype frequencies of the polymorphisms among five popular breeds with different behavioral traits revealed statistically significant interbreed differences for three polymorphisms. These results suggest that these polymorphisms would be useful genetic markers when performing behavior studies in dogs.
In this study, several cytokine responses were investigated during Mycoplasma hyopneumoniae (Mhp) infection using a gnotobiotic infection model. We found that several inflammatory cytokines (IL-1β, IL-8, IL-18, and TNF-α) and an anti-inflammatory cytokine IL-10 were induced from peripheral blood mononuclear cells (PBMC) of germ-free (GF) piglets stimulated with heat killed Mhp whole antigens, but no IFN-γ and IL-4 were induced by Mhp. After the intranasal infection of Mhp, IL-1β, IL-8, IL-18, and IFN-γ were also detected in the broncho-alveolar lavage fluids (BALF). The antigen-specific IFN-γ and IL-10 responses after infection of Mhp were gradually suppressed during Mhp infection as well as non-specific immune response to concanavalin A (ConA) and lipopolysacchalide (LPS) at early stage of infection. These results suggested that Mhp infection modulates the immune response of pigs by inducing several cytokines, and causes immuno-suppression of pigs in a gnotobiotic condition.
Cyclosporine A (CsA) has been widely used for suppression of transplant rejection and controlling pruritus in allergic dermatitis in humans, dogs and cats. CsA is known to suppress the expression of inflammatory cytokines, including IL-2, IL-4, IFN-γ and TNF-α in humans, dogs and experimental mice. However, little is known about the immunomodulating effect of CsA in cats. The aim of this study was to evaluate the effects of CsA on the expression of inflammatory cytokines in feline peripheral blood mononuclear cells (PBMC). Real-time PCR analyses with Concanavalin A (ConA)-stimulated PBMC obtained from 5 cats revealed that the expression of mRNAs for IL-2, IL-4, IFN- γ and TNF-α was inhibited by CsA in a dose-dependent manner. Moreover, an enzyme-linked immunospot (ELISPOT) assay, which is capable of detecting IL-2 secreting cells as single spots, revealed that the frequency of IL-2 secreting cells in ConA-stimulated feline PBMC was significantly reduced in the presence of CsA. These results might provide an explanation for the mechanisms of action of CsA in the suppression of transplant rejection and the control of pruritus in cats.
When a solution of xylitol was rapidly administered intravenously (bolus infusion) to healthy cattle or those with ketosis, different results were obtained. In healthy cattle, a temporary surge in insulin secretion was observed, whereas in ketotic cattle no such surge was found, but instead a moderate level of secretion continued for a lengthy period. No significant difference in the areas under the insulin curve (AUC) was found between healthy cattle and ketotic cattle up to 120 min after xylitol infusion. These results clearly demonstrated that a bolus infusion of xylitol solution in ketotic cattle does not cause a temporary surge in insulin secretion unlike in healthy animals, but rather results in a continuous, gradual rise in secretion.
This study was performed to examine normal brainstem auditory evoked potential (BAEP) data for adult Japanese Black cattle and to evaluate whether differences exist in the peak latencies, interpeak latencies (IPL) and waveforms of BAEP between Japanese Black and Holstein cattle. The peaks were detected as major waves I, II, III and V in each group. The threshold of the BAEP waves in the Holstein cattle was 65-75 dB nHL, but the threshold in the Japanese Black cattle was 75-85 dB nHL. The I-III and I-V IPLs were significantly shorter in the Japanese Black compared with the Holstein cattle at an intensity of 105 dB nHL. The present findings suggest that the IPL and wave threshold of BAEP are influenced by bovine breed.
The growth-retarded (grt) mouse has an autosomal recessive hypothyroidism and the female shows lifelong infertility. We previously reported that these mutant phenotypes are caused by a deficiency in the enzymatic activity of tyrosylprotein sulfotransferase-2 (TPST2), and severe thyroid hypogenesis and consequent dwarfism are mainly due to the impairment of the tyrosine sulfation of thyroid-stimulating hormone receptor (TSHR) by TPST2. Although TPST2 is ubiquitously expressed and many proteins are predicted to be tyrosine sulfated and involved in many biological processes, the functional roles of tyrosine sulfation in the reproductive organs remain unclear. These findings tempted us to hypothesize two possible mechanisms underlying the infertility; a deficiency in TPST2 activity in the reproductive organs might cause the infertility in grt mice, or a significant decrease in serum thyroid hormones might impair the normal development of reproductive organs. When mutant female mice were fed a diet supplemented with sufficient thyroid powder to correct their growth retardation, the rate of copulation, pregnancy, and parturition was completely restored. Therefore, we concluded that the infertility in grt female is due to a thyroid hormone deficiency.
Group A consisted of chickens infected with a single dose of Ascaris suum and group B of chickens infected with two successive doses. At days 1, 3, 7, 14 and 21 after the first or second infection dose, six chickens from each group were sacrificed. In both groups, larvae were recovered from the livers on days 1, 3, and 7 and lungs on days 3 and 7. No larvae were detected in chickens on day 14. Clear white lesions were noticed only on the livers from chickens of group B at day 7 but had disappeared at day 14. A comparison with group B showed mild histological changes that developed relative to the livers from group A.
A 1-year-and-3-month-old, male standard poodle dog with intrahepatic portosystemic shunt (PSS) was autopsied. Nineteen regions of the liver were prepared for detailed examination, and the distribution of hepatic lesions caused by PSS was studied in the liver of this dog. Histopathologically, the liver revealed a variety of hepatic lesions including lipogranulomas in the hepatic parenchyma, and a ductular reaction and microvascular proliferation in portal areas. The distribution of the lesions was not significantly different among liver regions. It is concluded that, in the present case, hepatic lesions caused by PSS are independent of shunt location, and are distributed equally in the liver.
Indole-3-carbinol (I3C) is a phytochemical found in cruciferous vegetables and possesses a variety of biological and biochemical effects. Despite a wealth of data about the chemopreventive properties of I3C, its effects on gap junctional intercellular communication (GJIC), which is associated with the promotion and progression phases of the multi-stage process of carcinogenesis, has not been studied. In this study, we examined the ability of I3C to prevent H2O 2-induced inhibition of GJIC in WB-F344 rat liver epithelial cells (WB cells). The cells were preincubated with I3C for 48 hr, and then treated with 1 mM H2O2 for 1 hr. We found that I3C could prevent the H2O2-induced inhibition of GJIC through prevention of the phosphorylated state of gap junction protein connexin 43 (Cx43) phosphorylation. Prevention of GJIC by I3C was dependent upon inactivation of Akt, but not MAPK, although inhibition of GJIC by H2O2 leads to activation of both. Similar to I3C, modulation of Akt activation through the phosphoinositide-3 kinase inhibitor, LY294002, could also prevent H2O2-induced inhibition of GJIC and phosphorylation of Cx43. Our results suggest that I3C might exert its dietary chemopreventive effects by interfering with the Akt signaling pathway, which appears to be linked to modulating GJIC, a cellular mechanisms regulating cell proliferation, differentiation and apoptosis.
Genetic characterization of the Jeju horse (JH) was performed to construct a correct pedigree of the JH family. A total of 111 horses including 79 JH were genotyped using 20 microsatellite loci. The number of alleles varied from 5 to 11 (mean 7.45) in the JH. The observed heterozygosity and expected heterozygosity ranged from 0.293 to 0.891 and from 0.357 to 0.841, respectively. The polymorphic information contents (PIC) ranged from 0.335 to 0.816. AHT4, ASB2, ASB17, ASB23, CA425, HMS2, HMS3, HTG10, LEX3 and VHL20 loci had relatively high PIC values (> 0.7). The total exclusion probability (PE) of the 20 microsatellite loci was 0.9999 in the JH. These results provide basic information for developing an accurate pedigree and will be useful in making decisions regarding conservation of the JH.
The objectives of the study were to investigate the phenotypic and genotypic characterization of the persistent Salmonella Enteritidis (S. Enteritidis) isolates in two integrated broiler chicken operations, with attention focused mainly on the epidemiological approach. In the distribution of virulence genes, Salmonella enterotoxin (stn), invading host cell (invA), and Salmonella plasmid virulence (spvC) genes were widely distributed among the S. Enteritidis irrespective of their source of isolation, and Salmonella fimbrial (sefC) and plasmid encoded fimbrial (pef) genes were present in 28 and 20 S. Enteritidis strains, respectively. A total of 5 different XbaI-PFGE types were obtained from 31 S. Enteritidis isolates. Twenty-one types were divided on the basis their PFGE pattern, phage type and antimicrobial resistance pattern determined. There was a significant difference in phenotypic and genotypic characterization by two integrated broiler operations. Also, 8 isolates shown susceptible to all antimicrobials and 11 isolates with resistance to nalidixic acid were partly classified by XbaI PFGE pattern and by the phage type.
Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 μg/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 μg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR.
The Eperythrozoon suis (E. suis) antigen was purified using a Sephadex G-200 chromatograph, and thereby, a high-affinity, specific E. suis antigen was collected and confirmed with Western blotting. Using this antigen, an enzyme-linked immunosorbent assay (ELISA) system to detect the antibody against E. suis in swine was established. There was no cross-reaction with swine sera, which were affected with Mycoplasmal pneumonia, swine fever, swine colibacillosis, or toxoplasmosis. A comparison of this ELISA system with an indirect hemagglutination (IHA) test using 78 swine samples revealed that the ELISA system significantly improved the sensitivity, specificity, and stability for the serodiagnosis of swine E. suis.
The purpose of this study was to compare between the injection speed and iodine delivery rate in order to establish a concept for reproducible contrast timing in contrast-enhanced computed tomography (CT) for small animals. Clinically healthy beagle dogs were administered a nonionic iodinate contrast medium at a dose of 800 mgI/kg; they were divided into 3 groups (n=5, crossover method): in one group, the injection speed was fixed at 1.0 ml/sec, and in the second and third groups, the iodine delivery rate was fixed (the injection durations were 30 and 60 sec, respectively). The variation in scatter of the time to aortic and hepatic peak enhancement in the fixed iodine delivery groups was lower than that in the fixed injection speed group. These results suggest that in contrast-enhanced CT for small animals, the contrast medium should be injected at a fixed iodine delivery rate in order to provide reproducible contrast timing.
The objective of this study was to investigate cystic ovarian disease (COD) in commercial Japanese Black cows and to evaluate the efficacy of 7-day insertion of an intravaginal progesterone insert (CIDR) combined with prostaglandin F2α (PGF2α) at CIDR removal. Experiment 1 was conducted to group cysts into 4 patterns based on alteration of plasma progesterone (P4) concentrations on d -7 and d 0 (=CIDR insertion) with 1.0 ng/ml as the cut-off level by ultrasonographic examination of 28 cows with COD that were ≥40 days postpartum and anoestrous after calving. In Experiment 2, a total of 55 cows under the same conditions as in Experiment 1 were utilized, and the same regimen as in Experiment 1 was performed without 7 days of pre-observation before treatment. As a result, 92.9% of CLs on d 21 were highly formed in Experiment 1 and 83.6% were highly formed in Experiment 2. The conception rates within 60 days after CIDR removal were also satisfactory high and were 71.4% and 54.5%, respectively. There were no differences in any overall reproductive parameters between Experiments 1 and 2 (P>0.05). The average days between CIDR removal and conception were 24.4 ± 5.3 and 24.0 ± 6.5 days, respectively (P>0.05); therefore, the conception dates of the cows in Experiment 2 were at least 7 days earlier compared with Experiment 1. In conclusion, treatment with a CIDR and PGF2α against COD could minimize the risk of incorrect treatment and provide sufficient reproductive performance in Japanese Black cows.
The morphology of spermatozoa of modern Thoroughbred stallions in Japan was investigated during the breeding season. A total of 299 semen samples were collected from the penises of 16 stallions immediately after service. The rate of abnormalities in sperm heads and tails, spermatozoa with cytoplasmic droplets and slides with medusa cells to total observed slides in each stallion were 3.9 ± 2.1%, 11.5 ± 5.9%, 2.4 ± 2.6% and 20.1%, respectively. The values for the area, length, width and aspect ratio of the stallion sperm head were 12.54 ± 1.34 μm2, 5.93 ± 0.40 μm, 2.69 ± 0.21 μm and 0.46 ± 0.05, respectively. With the exception of medusa cells, the features were significantly different among the stallions (P<0.05).
The objective of this study was to evaluate the influences of genetic and antigenic variations in field isolates of porcine reproductive and respiratory syndrome virus (PRRSV) on vaccine efficacy. Four-week-old pigs were vaccinated with a commercial modified live virus vaccine. Four weeks after vaccination, pigs in both the vaccinated group and the non-vaccinated group were challenged intranasally with 107 TCID50 of PRRSV wt-11 (Experiment 1) or PRRSV wt-7 (Experiment 2). Based on genome sequencing of ORF5 and cross neutralization test results, PRRSV wt-11 is similar to the vaccine strain, whereas wt-7 is distinct from the vaccine strain. In the vaccinated challenged groups, clinical signs were less severe, the mean rate of weight gain was greater, and gross lung lesions were less severe when compared with the non-vaccinated challenged groups in both experiments. In Experiment 1, the virus was isolated from serum at 3 days post-challenge, and the mean virus titers in broncho-alveolar lavage fluids (BALF) and tissues were lower in pigs in the vaccinated challenged groups compared with those in the non-vaccinated challenged group. In Experiment 2, virus isolation from serum, BALF and tissues showed no significant differences between the groups. These results suggest that commercial PRRSV vaccine could be effective in reducing clinical disease following a challenge with field isolates of PRRSV. However, with regards to virological protection, the efficacy of the vaccine may be affected by the nature of the PRRSV isolates.