Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.
Animals ; Antibodies, Protozoan/blood ; China/epidemiology ; Dog Diseases/epidemiology/*parasitology ; Dogs ; Female ; Genotype ; Liver/parasitology ; Male ; Toxoplasma/classification/genetics/immunology/*isolation & purification ; Toxoplasmosis, Animal/blood/epidemiology/*parasitology

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

Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.
Animal Structures/chemistry ; Animals ; Antibodies, Helminth/blood ; Antigens, Helminth/chemistry/*genetics/immunology/*isolation & purification ; Cloning, Molecular ; Dirofilaria immitis/chemistry/*genetics/immunology ; Disease Models, Animal ; Dogs ; Escherichia coli/genetics ; Gene Expression ; Molecular Sequence Data ; Molecular Weight ; Recombinant Fusion Proteins/chemistry/genetics/immunology/isolation & purification ; Sequence Analysis, DNA ; Tumor Markers, Biological/chemistry/*genetics/immunology/*isolation & purification

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees C, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
Animals ; Blood/*parasitology ; Brugia/classification/genetics/*isolation & purification ; Cats ; Culicidae/*parasitology ; Dirofilaria immitis/classification/genetics/*isolation & purification ; Dogs ; Humans ; Male ; Parasitology/*methods ; RNA, Helminth/genetics ; RNA, Ribosomal, 5S/genetics ; Real-Time Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Transition Temperature ; Wuchereria bancrofti/classification/genetics/*isolation & purification

BACKGROUNDInflammation and coagulation are two intimately cross-linked defense mechanisms of most, if not all organisms to injuries. During cardiopulmonary bypass (CPB), these two processes are activated and interact with each other through several common pathways, which may result in subsequent organ dysfunction. In the present study, we hypothesized that the addition of nitric oxide, prostaglandin E1 (PGE1), and aprotinin to the systemic circulation, hereby referred to as blood hibernation, would attenuate the inflammation and coagulation induced by CPB.

METHODSThirty adult mongrel dogs were equally divided into five groups, anesthetized and placed on hypothermic CPB (32 degrees C). Each group received respectively the following treatments: (1) inhalation of 40 ppm nitric oxide; (2) intravenous infusion of 20 ng x kg(-1) x min(-1) of PGE1; (3) 80,000 kallikrein inhibitor units (KIU)/kg of aprotinin; (4) the combination of all three agents (blood hibernation group); and (5) no treatment (control group) during CPB. Activation of leukocyte, platelet, endothelial cell, and formation of thrombin were assessed after CPB.

RESULTSAs compared with the other four groups, leukocyte counts were higher, while plasma elastase, interleukin-8, CD11b mRNA expression, myeloperoxidase activities and lung tissue leukocyte counts were lower in the blood hibernation group (P < 0.05 versus other four groups after CPB). Plasma prothrombin fragment (PTF)1+2, and platelet activation factors were lower, while platelet counts were higher in the blood hibernation group (P < 0.05 versus other four groups at 6 and 12 hours after CPB). Electron microscopy showed endothelial pseudopods protrusion, with cell adherence in all four groups except the blood hibernation group where endothelial cells remained intact.

CONCLUSIONBlood hibernation, effected by the addition of nitric oxide, PGE1 and aprotinin to the circulating blood during extra-corporeal circulation, was observed to attenuate the inflammation and coagulation induced by cardiopulmonary bypass, most likely by inhibiting the important common intermediates between the two cross-linked processes.

Alprostadil ; pharmacology ; therapeutic use ; Animals ; Aprotinin ; pharmacology ; therapeutic use ; Blood Coagulation ; drug effects ; CD11b Antigen ; genetics ; Cardiopulmonary Bypass ; adverse effects ; Dogs ; Inflammation ; drug therapy ; etiology ; Male ; Nitric Oxide ; pharmacology ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the molecular genetic basis of the B3 variant of ABO blood group system with mixed-field hemagglutination in Chinese.

METHODSSerological techniques were performed to characterize the erythrocyte phenotype of two discrepant samples. A sequential agglutination method and 13 short tandem repeat (STR) loci were tested to exclude the possibility of exogenous or endogenous DNA chimera. Mutations in exons 6 and 7, including partial intron of the ABO gene, were screened by polymerase chain reaction and DNA sequencing. Haplotypes of the two individuals were also analyzed by sequencing.

RESULTSA mixed-field hemagglutination of RBCs with anti-B and anti-AB antibodies was detected in the two unrelated individuals. Exogenous ABO-incompatible RBC transfusion and endogenous genetic chimera were excluded by sequential agglutination method and STR. The ABO phenotypes of the two individuals were classified as A1B3 according to the ABO subgroup definition. The sequence region from intron 5 to 3'-UTR of the B allele was identical to that of ABO*B101 allele, except for a T to C substitution at nucleotide position 425 in exon 7. This substitution resulted in an amino acid change of M142T in the B glycosyltransferase.

CONCLUSIONA novel B allele with 425T>C substitution resulting in B3 subgroup was identified in two Chinese individuals.

1,4-alpha-Glucan Branching Enzyme ; genetics ; ABO Blood-Group System ; genetics ; Alleles ; Amino Acid Sequence ; Amino Acid Substitution ; Animals ; Asian Continental Ancestry Group ; genetics ; Cattle ; DNA Mutational Analysis ; Dogs ; Humans ; Methionine ; genetics ; Mice ; Molecular Sequence Data ; Mutation ; Phenotype ; Rats ; Sequence Alignment ; Sequence Analysis, DNA ; Threonine ; genetics

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dog is the definitive host for N. caninum and can infect dairy cattle. The aim of this study is to determine the prevalence of Neospora oocysts in feces of dogs from dairy farms. A total of 174 fecal samples was collected from 89 farm dogs and 85 household dogs during 2006 and 2008. Fecal samples of dogs were microscopically examined for detecting Hammondia Neospora-like oocysts (HNLO) by Mini Parasep(R)SF fecal parasite concentrator. HNLO were microscopically detected in 4 fecal samples (2.2%). The fecal samples with HNLO were examined by N. caninum-specific PCR. Two of the samples were positive for N. caninum. The 2 positive fecal samples were selected for inoculation to calves. Two inoculated calves were seronegative by ELISA for 4 months post-infection. This is the first report of finding N. caninum DNA in feces of farm dogs in Mashhad area, Iran.
Animals ; Antibodies, Fungal/blood ; Cattle ; Cattle Diseases/immunology/parasitology ; Coccidiosis/epidemiology/parasitology/*veterinary ; DNA, Fungal/genetics/isolation & purification ; Dog Diseases/*epidemiology/*parasitology ; Dogs ; Enzyme-Linked Immunosorbent Assay/methods ; Feces/*microbiology ; Iran/epidemiology ; Male ; Microscopy/methods ; Neospora/*genetics/*isolation & purification ; Oocysts/cytology ; Polymerase Chain Reaction/methods ; Prevalence

OBJECTIVETo investigate the patency efficacy of small diameter artificial vascular graft with eNOS gene transfected endothelial cells in canine.

METHODSBy using the two steps high pressure injection, eNOS gene transfected endothelial cells were planted on artificial vascular graft with a diameter of 3 mm. The artificial vascular grafts were transplanted into canine femoral artery, and the patency of the artery was observed through digital subtracted angiography (DSA) and electron telescope 1, 4, 12 and 24 weeks after the operation.

RESULTSThe adherence rate of eNOS gene transfected endothelial cells to artificial vascular grafts was up to 90%. For 10 days, the cells extended and formed a continuous intima. One week after the operation, 3/9 grafts were obstructed in the group of simple artificial vascular grafts; 4 weeks after, all of grafts (9/9) were obstructed in the group of simple artificial vascular grafts and almost half grafts (5/10, 4/9) in un-transfected groups were obstructed; 12 weeks after, all of the grafts were obstructed in the group of simple artificial vascular grafts and in un-transfected groups (9/9, 9/10); 24 weeks after, all of the grafts were obstructed in the groups of un-transfected artificial vascular grafts, while nearly all of grafts (8/10) were open in eNOS groups. Electron microscope scanning showed that endothelial cells of artificial vascular grafts in eNOS transfected groups arranged closely and formed a continuous intima; only few red blood cells, leucocytes, platelets deposited at the surface of endothelial cells in the artificial vascular grafts.

CONCLUSIONSThe patency efficacy of eNOS gene transfected endothelial cells planting on artificial vascular graft is satisfactory. It could provide an experimental basis for further clinical application of the artificial vascular grafts.

Animals ; Blood Vessel Prosthesis ; Blood Vessel Prosthesis Implantation ; Cells, Cultured ; Dogs ; Endothelial Cells ; cytology ; metabolism ; Female ; Genetic Vectors ; Male ; Nitric Oxide Synthase ; genetics ; Random Allocation ; Transfection

OBJECTIVE: To establish an accurate, simple, quick, specific and sensitive method for species identification by amplifying 12S rRNA gene with the same reaction system. METHODS: Based on the downloaded 12S rRNA gene sequences of eleven species (human, chicken, duck, goose, pig, rabbit, rat, sheep, bull, dog and goat) from GenBank, a pair of universal primers to eleven species and three pairs of specific primers to human, chicken and duck were designed. The amplicons amplified with universal primers were used for internal controls, and the amplicons amplified with specific primers were used as identification of human, chicken and duck. DNA was extracted from various samples including blood stains, fresh or freezing muscles, heat-treated muscles and hairs. Both single DNA of human, chicken or duck and mixed DNA of any two kinds of them were amplified. RESULTS: The lengths of universal amplicons were about 400 bp. The lengths of specific amplicons were 163 bp for human, 286 bp for chicken, and 374 bp for duck, respectively. No cross amplification was observed, indicating a high specificity of the specific primers. The identification rate was 100% for human, 99% for chicken, and 100% for duck, respectively. The detection sensitivity ranged from 2.5 pg to 200 pg of DNA concentration depending on species, even in mixtures of different species DNA without interference. CONCLUSION The method established could identify different species under the same reaction system.
Animals ; Blood ; Cattle ; DNA/analysis* ; Dogs ; Forensic Genetics ; Hot Temperature ; Humans ; Polymerase Chain Reaction/veterinary* ; Poultry/genetics* ; RNA, Ribosomal/genetics* ; Rabbits ; Rats ; Sheep ; Species Specificity ; Swine

For epidemiological investigation of the rabies virus carrier rates of domestic dogs, cats and wild animals like rodent animals and bats,three kinds of regions where rabies had higher incidence (Hunan and Guizhou Provinces), lower incidence (Jiangsu Province, Wuhan City) and provisionally rabies-free (Shenyang City) were selected. Then the antigenic types, the genovariation of the isolaled viruses and the currently vaccine matching of the virus strains were analyzed. The results showed that in China the principal host of rabies is dog,the total virus carrier rate of the captured dogs was 2.56%, and the highest positive isolation rate was 20.0% in some monitoring site. However,there was no evidence about the rabies virus carrier rate in rodent animals,bats or other wild animals. The rabies vaccines which prepared from aG and CTN strains have already been produced successfully in China. The research showed that the nucleotide sequences of the newly isolated viruses were more similar with the glycoprotein gene of CTN strain. In order to evaluate the safety and the efficacy of the vaccines currently used, two groups (50 people each) were injected with vaccine of aG strain and CTN strain respectively in five surveillance points. The neutralizing antibody tested were 0.49 IU/mL-0.52 IU/mL and 6.7 IU/mL-7.53 IU/mL after the 7 and the 14 days of vaccine injection respectively. In addition, the rates of antibody positive seroconversion were 45.1%-47.9% and 100% respectively, and there was no moderate or severe adverse reactions observed. These data showed the vaccines have satisfactory effect on safety and protection.
Animals ; Antibodies, Viral ; blood ; Carrier State ; epidemiology ; veterinary ; Cats ; virology ; Cercopithecus aethiops ; Dogs ; virology ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Rabies Vaccines ; immunology ; Rabies virus ; classification ; genetics ; isolation & purification ; Vero Cells