The ascarids, Toxocara canis and Toxascaris leonina, are probably the most common gastrointestinal helminths encountered in dogs. In order to understand biological differences of 2 ascarids, we analyzed gene expression profiles of female adults of T. canis and T. leonina using CLC Genomics Workbench, and the results were compared with those of free-living nematode Caenorhabditis elegans. A total of 2,880 and 7,949 ESTs were collected from T. leonina and T. canis, respectively. The length of ESTs ranged from 106 to 4,637 bp with an average insert size of 820 bp. Overall, our results showed that most functional gene annotations of 2 ascarids were quite similar to each other in 3 major categories, i.e., cellular component, biological process, and molecular function. Although some different transcript expression categories were found, the distance was short and it was not enough to explain their different lifestyles. However, we found distinguished transcript differences between ascarid parasites and free-living nematodes. Understanding evolutionary genetic changes might be helpful for studies of the lifestyle and evolution of parasites.
Animals ; Dog Diseases/*parasitology ; Dogs ; Female ; *Genomics ; Molecular Sequence Annotation ; Toxascariasis/parasitology/*veterinary ; Toxascaris/*genetics/metabolism ; Toxocara canis/*genetics/metabolism ; Toxocariasis/*parasitology

Increasing evidence suggests that cancer stem cells (CSCs) are responsible for tumor initiation and maintenance. Additionally, it is becoming apparent that cyclooxygenase (COX) signaling is associated with canine mammary tumor development. The goals of the present study were to investigate COX-2 expression patterns and their effect on CSC-mediated tumor initiation in primary canine mammary tissues and tumorsphere models using immunohistochemistry. Patterns of COX-2, CD44, octamer-binding transcription factor (Oct)-3/4, and epidermal growth factor receptor (EGFR) expression were examined in malignant mammary tumor (MMT) samples and analyzed in terms of clinicopathological characteristics. COX-2 and Oct-3/4 expression was higher in MMTs compared to other histological samples with heterogeneous patterns. In MMTs, COX-2 expression correlated with tumor malignancy features. Significant associations between COX-2, CD44, and EGFR were observed in low-differentiated MMTs. Comparative analysis showed that the levels of COX-2, CD44, and Oct-3/4 expression varied significantly among TSs of three histological grades. Enhanced COX-2 staining was consistently observed in TSs. Similar levels of staining intensity were found for CD44 and Oct-3/4, but EGFR expression was weak. Our findings indicate the potential role of COX-2 in CSC-mediated tumor initiation, and suggest that COX-2 inhibition may help treat canine mammary tumors by targeting CSCs.
Animals ; Antigens, CD44/genetics/metabolism ; Biomarkers, Tumor/genetics/metabolism ; Cell Transformation, Neoplastic/*genetics/metabolism ; Cyclooxygenase 2/*genetics/metabolism ; Dog Diseases/*genetics/metabolism ; Dogs ; Female ; Immunohistochemistry/veterinary ; Mammary Neoplasms, Animal/*genetics/metabolism ; Mammary Neoplasms, Experimental/*genetics/metabolism ; Neoplastic Stem Cells/*metabolism ; Octamer Transcription Factor-3/genetics/metabolism ; Receptor, Epidermal Growth Factor/genetics/metabolism ; Retrospective Studies

The sequences of the ccrAB genes from bovine-, canine- and chicken-originating methicillin-resistant Staphylococcus (S.) epidermidis (MRSE) and bovine methicillin-resistant Staphylococcus (S.) aureus (MRSA) were compared to investigate the frequency of intra-species horizontal transfer of the staphylococcal cassette chromosome mec (SCCmec) complex. Nineteen MRSE strains were isolated from bovine milk, chickens, and dogs, and their genetic characteristics were investigated by multilocus sequence typing and SCCmec typing. Among the animal MRSE strains, the most frequent SCCmec type was type IV, which consisted of the type B mec complex and ccrAB type 2. The ccrA2 and ccrB2 genes were sequenced from the bovine, chicken and canine MRSE strains and compared with those of the bovine MRSA strains. The sequences generally clustered as MRSA and MRSE groups, regardless of the animal source. Additionally, no bovine MRSE sequence was associated with the bovine MRSA groups. Although most of the bovine MRSE and MRSA isolates possessed SCCmec type IV sequences, our results suggest that the intra-species gene transfer of the SCCmec complex between bovine S. aureus and bovine S. epidermidis strains is not a frequent event.
Animals ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/*genetics/metabolism ; Bacterial Typing Techniques/veterinary ; Cattle ; Cattle Diseases/epidemiology/metabolism ; Chickens ; Dog Diseases/epidemiology/metabolism ; Dogs ; *Drug Resistance, Bacterial ; *Gene Transfer, Horizontal ; Methicillin/*pharmacology ; Methicillin-Resistant Staphylococcus aureus/genetics/isolation & purification ; Milk/microbiology ; Multilocus Sequence Typing/veterinary ; Poultry Diseases/epidemiology/metabolism ; Prevalence ; Republic of Korea/epidemiology ; Staphylococcal Infections/epidemiology/microbiology/*veterinary ; Staphylococcus epidermidis/genetics/isolation & purification

This study was carried out to evaluate the prevalence and clinical characterizations of gastric Helicobacter spp. infection of dogs and cats in Korea. The prevalence of Helicobacter spp. infection of dogs and cats determined by urease test was 78.4% and 64%, respectively, although Helicobacter genus-specific PCR assay showed that it was 82.3% and 84%. Urease mapping results based on urease test showed that total positive rate of tested tissues from clinically abnormal dogs was significantly higher than that from clinically normal dogs (p=0.0018; Odds ratio = 6.118; 95% Confidence Interval = 1.96~19.103). These findings were consistent with the results of Helicobacter genus-specific PCR assay which showed that positive rate of the fundus (100%) and the antrum (100%) of clinically abnormal dogs was significantly higher than that of same gastric regions of clinically normal dogs (77.5 and 67.5% respectively). In comparison of gastric regions between clinically normal dogs and abnormal dogs, positive rate of urease test for the fundus (100%) and body (90.9%) in clinically abnormal dogs was significantly higher than that of abnormal dogs (72.5% and 57.5% respectively; p<0.05). The results of urease mapping in dogs and cats also indicated that Helicobacter colonization in the fundus was more dense compared with the density in the body and antrum. In Helicobacter species-specific PCR assay for dogs, 32 of 42 fundic tissues (76.2%) were positive for H. heilmannii and two (4.8%) were positive for H. felis. In cats, 18 of 21 fundic tissues (85.7%) were positive for H. heilmannii and 2 (9.5%) were positive for H. felis. Gastritis scores of fundic tissues from clinically abnormal infected dogs were similar to that from noninfected dogs and evidence of upregulation of IL-1beta, IL-8, and TNF-alpha mRNA was not detected in gastric fundic tissues from clinically abnormal infected dogs. This study suggested that Helicobacter spp. infection in domestic dogs including private owned pet dogs and cats is highly prevalent usually with no clinical sign but high density of colonization can be related to gastrointestinal signs
Animals ; Cat Diseases/enzymology/*epidemiology/microbiology ; Cats ; Cytokines/genetics ; DNA, Bacterial/analysis/genetics ; Dog Diseases/enzymology/*epidemiology/microbiology/pathology ; Dogs ; Gene Expression Regulation ; Helicobacter/classification/genetics/isolation&purification ; Helicobacter Infections/*epidemiology/microbiology/pathology/*veterinary ; Korea/epidemiology ; Polymerase Chain Reaction ; Prevalence ; RNA, Messenger/genetics/metabolism ; Species Specificity ; Stomach/microbiology ; Stomach Diseases/enzymology/*epidemiology/microbiology/*veterinary ; Urease/metabolism

We concentrated ourselves to evaluate the prognostic significance of the p53 gene mutations, its protein expression and MIB-1 index as a proliferative marker in canine mammary tumors. In the present study, a total of 20 cases were examined, among which there were 5 malignant mixed tumors, 4 mammary gland adenocarcinomas, 1 papillary adenocarcinoma, 8 benign mixed tumors and 2 mammary gland adenomas. Positive immunostaining for p53 with PAb240 antibody was found in 2 benign (20%) and 3 malignant (30%) tumors. However, PAb421 antibody did not give positive result at all. In Western blot analysis, the p53 expression in benign and malignant tumors was detected in 4 and 3 cases, respectively. p53 mutations were found in 6 cases out of the cases with detected p53 protein expression. The MIB-1 index in benign and malignant tumors were 17.6+/-20.8% and 29.0+/-27.2%, respectively and there was no significant difference between tumor types. There was a significant correlation between p53 mutations and p53 overexpression (correlation coefficient = 0.5, p < 0.05). In Kaplan-Meier survival analysis, the p53 index was associated with significantly shortened survival time (p < 0.01). In multivariate analysis, p53 overexpression was only an independent factor for indicator of worse prognosis in canine mammary tumors (p = 0.01). These results demonstrated that p53 gene mutations and protein overexpression using the PAb240 anti-p53 antibody were useful predictors of increased malignant potential and poor prognosis in canine mammary tumors.
Animals ; Antibodies, Antinuclear/metabolism ; Antibodies, Monoclonal/metabolism ; Blotting, Western/veterinary ; Dog Diseases/genetics/*metabolism ; Dogs ; Female ; Genes, p53/*genetics ; Immunohistochemistry/veterinary ; Ki-67 Antigen/metabolism ; Mammary Neoplasms, Animal/genetics/*metabolism ; *Mutation ; Predictive Value of Tests ; Proportional Hazards Models ; Tumor Suppressor Protein p53/*biosynthesis/genetics

The Staphylococcus (S.) intermedius group (SIG) has been a main research subject in recent years. S. pseudintermedius causes pyoderma and otitis in companion animals as well as foodborne diseases. To prevent SIG-associated infection and disease outbreaks, identification of both staphylococcal exotoxins and staphylococcal cassette chromosome mec (SCCmec) types among SIG isolates may be helpful. In this study, it was found that a single isolate (one out of 178 SIG isolates examined) harbored the canine enterotoxin SEC gene. However, the S. intermedius exfoliative toxin gene was found in 166 SIG isolates although the S. aureus-derived exfoliative toxin genes, such as eta, etb and etd, were not detected. SCCmec typing resulted in classifying one isolate as SCCmec type IV, 41 isolates as type V (including three S. intermedius isolates), and 10 isolates as non-classifiable. Genetic relatedness of all S. pseudintermedius isolates recovered from veterinary staff, companion animals, and hospital environments was determined by pulsed-field gel electrophoresis. Strains having the same band patterns were detected in S. pseudintermedius isolates collected at 13 and 18 months, suggesting possible colonization and/or expansion of a specific S. pseudintermedius strain in a veterinary hospital.
Animals ; Bacterial Toxins/genetics/metabolism ; Cat Diseases/epidemiology/*microbiology ; Cats ; Chromosomes, Bacterial/genetics/metabolism ; Dog Diseases/epidemiology/*microbiology ; Dogs ; Electrophoresis, Gel, Pulsed-Field/veterinary ; Enterotoxins/genetics/metabolism ; Exfoliatins/genetics/metabolism ; Exotoxins/*genetics/metabolism ; Hospitals, Animal ; Humans ; Medical Staff, Hospital ; Molecular Sequence Data ; Pets/microbiology ; Polymerase Chain Reaction/veterinary ; Republic of Korea/epidemiology ; Staphylococcal Infections/epidemiology/microbiology/*veterinary ; Staphylococcus/genetics/isolation & purification ; Staphylococcus intermedius/*genetics/*isolation & purification

Amplification of the 16S rRNA gene from a blood sample obtained from a dog in southeastern Brazil was used to confirm a naturally acquired Ehrlichia (E.) canis infection. Following isolation and culturing of the new bacterial strain called Uberlandia, partial sequences of the dsb and p28 genes were obtained. The dsb partial sequence of the novel strain was 100% similar to dsb gene sequences of E. canis obtained from different geographic areas around the world. Conversely, the p28 partial sequence for the E. canis Uberlandia strain differed at several nucleotides from other sequences available in GenBank. To confirm the antigenic profile of the Uberlandia strain, an indirect immunofluorescence assay against E. canis antigens was performed using dog sera collected from two different areas in Brazil (Uberlandia and Sao Paulo). The results suggest that both antigens were able to identify animals seropositive for E. canis in Brazil since these Brazilian strains appear to be highly conserved.
Animals ; Antigens, Bacterial/blood/*diagnostic use ; Bacterial Outer Membrane Proteins/genetics/metabolism ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; Brazil ; Dog Diseases/diagnosis/*microbiology ; Dogs ; Ehrlichia canis/*genetics/*immunology/isolation & purification ; Ehrlichiosis/diagnosis/microbiology/*veterinary ; Fluorescent Antibody Technique, Indirect/veterinary ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics/metabolism ; Sequence Alignment/veterinary

Inherited copper toxicosis in Bedlington Terriers (CTBT) is a copper associated hepatopathy caused by an autosomal recessive genetic defect of gene involving copper metabolism. To compare clinical and histopathological findings with previous reports and to expand our knowledge for future genetic studies, 18 terriers were clinically and histopathologically examined in this study. Pedigree information and dietary history were obtained from the owners before a thorough clinical examination was undertaken. Following the examination, a blood sample was collected for haematology, biochemistry and genetic analysis and a urine sample for urinalysis. Seven dogs were also liver biopsied for histopathology, histochemistry and electron microscopy. In this study, plasma alanine transaminase (ALT) activity was highly concordant with DNA marker test results and was the most reliable and sensitive biochemical test measured. Also clinical and biochemical copper toxicosisaffected states were noticed in a genotyped carrier dog. Histopathological and electron microscopy findings showed that the severity of the lesion was more closely correlated to the presence of clinical signs than to hepatic copper concentration. In addition, the involvement of apoptosis and p53 gene was observed in electron microscopy. The general findings related to CT-BT in this study was similar to those previously reported except few differences in histopathology and electron microscopy.
Alanine Transaminase/blood ; Animals ; Biopsy/veterinary ; Blood Chemical Analysis/veterinary ; Copper/*metabolism ; Dog Diseases/*genetics/*metabolism ; Dogs ; Female ; Histocytochemistry/veterinary ; Leukocyte Count/veterinary ; Liver/metabolism/pathology/ultrastructure ; Male ; Metal Metabolism, Inborn Errors/*genetics/metabolism/pathology/*veterinary ; Microscopy, Electron/veterinary ; Urinalysis/veterinary

We investigated the distribution of commensal staphylococcal species and determined the prevalence of multi-drug resistance in healthy cats and dogs. Risk factors associated with the carriage of multi-drug resistant strains were explored. Isolates from 256 dogs and 277 cats were identified at the species level using matrix-assisted laser desorption ionisation-time of flight mass spectrometry. The diversity of coagulase-negative Staphylococci (CNS) was high, with 22 species in dogs and 24 in cats. Multi-drug resistance was frequent (17%) and not always associated with the presence of the mecA gene. A stay in a veterinary clinic in the last year was associated with an increased risk of colonisation by multi-drug resistant Staphylococci (OR = 2.4, 95% CI: 1.1~5.2, p value LRT = 0.04). When identifying efficient control strategies against antibiotic resistance, the presence of mechanisms other than methicillin resistance and the possible role of CNS in the spread of resistance determinants should be considered.
Animals ; Anti-Bacterial Agents/*pharmacology ; Bacterial Proteins/genetics/metabolism ; Cat Diseases/epidemiology/*microbiology ; Cats ; Coagulase/genetics/metabolism ; Dog Diseases/epidemiology/*microbiology ; Dogs ; *Drug Resistance, Multiple, Bacterial ; Female ; Male ; Prevalence ; Risk Factors ; Seasons ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/veterinary ; Staphylococcal Infections/epidemiology/microbiology/*veterinary ; Staphylococcus/classification/genetics/*isolation & purification ; Switzerland/epidemiology

Due to the therapeutic potential of gene therapy for neuronal injury, many studies of neurotrophic factors, vectors, and animal models have been performed. The presumed dog beta-nerve growth factor (pdbeta-NGF) was generated and cloned and its expression was confirmed in CHO cells. The recombinant pdbeta-NGF protein reacted with a human beta-NGF antibody and showed bioactivity in PC12 cells. The pdbeta-NGF was shown to have similar bioactivity to the dog beta-NGF. The recombinant pdbeta-NGF plasmid was administrated into the intrathecal space in the gene therapy group. Twenty-four hours after the vector inoculation, the gene therapy group and the positive control group were intoxicated with excess pyridoxine for seven days. Each morning throughout the test period, the dogs' body weight was taken and postural reaction assessments were made. Electrophysiological recordings were performed twice, once before the experiment and once after the test period. After the experimental period, histological analysis was performed. Dogs in the gene therapy group had no weight change and were normal in postural reaction assessments. Electrophysiological recordings were also normal for the gene therapy group. Histological analysis showed that neither the axons nor the myelin of the dorsal funiculus of L(4) were severely damaged in the gene therapy group. In addition, the dorsal root ganglia of L(4) and the peripheral nerves (sciatic nerve) did not experience severe degenerative changes in the gene therapy group. This study is the first to show the protective effect of NGF gene therapy in a dog model.
Amino Acid Sequence ; Animals ; Base Sequence ; CHO Cells ; Central Nervous System Diseases/chemically induced/therapy/*veterinary ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Cytomegalovirus ; Dog Diseases/*chemically induced/therapy ; Dogs ; Female ; Gene Therapy/*veterinary ; Genetic Vectors ; Male ; Molecular Sequence Data ; Nerve Growth Factor/genetics/*metabolism/*therapeutic use ; Pyridoxine/*toxicity