The present study describes the genotypic distribution of rotaviruses (RVs) in an Indian bovine population with unexpectedly higher proportions of G3 alone or in combination of G8/G10. PCR-genotyping confirmed that 39.4% (13/33) of the prevalent RVs were the G3 type while 60.6% (20/33) were dual G3G10 or G3G8 types. P typing revealed that 93.9% (31/33) of the samples were P[11] while 6.1% (2/33) possessed a dual P[1]P[11] type. Sequence analysis of the VP7 gene from G3 strains viz. B-46, 0970, and BR-133 showed that these strains had sequence identities of 90.5% to 100% with other bovine G3 strains. The highest identity (98.9% to 100%) was observed with RUBV3 bovine G3 strains from eastern India. The G3 strains (B-46, 0970, and BR-133) showed 97.5% to 98.8% sequence homologies with the Indian equine RV strain Erv-80. Phylogenetic analysis demonstrated that G3 strains clustered with bovine RUBV3 and J-63, and equine Erv-80 G3. Overall, these results confirmed that the incidence of infection by RVs with the G3 genotype and mixed genotypes in the bovine population was higher than previously predicted. This finding reinforces the importance of constantly monitoring circulating viral strains with the G3 genotype in future surveillance studies.
Animals ; Cattle ; Cattle Diseases/epidemiology/*virology ; Desert Climate ; Feces/virology ; Genotype ; India/epidemiology ; Molecular Sequence Data ; Phylogeny ; RNA, Viral/genetics ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Rotavirus/classification/*genetics/isolation & purification ; Rotavirus Infections/epidemiology/*veterinary/virology ; Sequence Analysis, Protein/veterinary ; Sequence Analysis, RNA/veterinary ; Sequence Homology ; Tropical Climate

The research of p53 is being conducted to find the mechanisms of tumorigenesis and to treat various cancers. Homeodomain-interacting protein kinase2 (HIPK2) is an important factor to regulate p53 and to increase the stability of p53. Activation of HIPK2 leads to the selective phosphorylation of p53, resulting in growth arrest and the enhancement of apoptosis. In this study, the canine HIPK2 cDNA fragments were obtained, and their overlapping regions were aligned to give a total sequence of 3489 bp. The canine HIPK2 cDNA (GenBank accession number; AY800385) shares 93% and 90% sequence identity with those of human and mouse HIPK2, respectively. The canine HIPK2 cDNA contains an open reading frame encoding 1163 amino acid residues and the predicted amino acid sequence has 98% and 96% identity with those of human and mouse, respectively. The deduced amino acid sequence of canine HIPK2 has also all domains' sites compared with human and mouse HIPK2. Therefore, these structural similarities suggested that the canine HIPK2 shares the basic biological functions that HIPK2 exhibit in other species.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary/chemistry/genetics ; Dogs/metabolism/*physiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; Protein-Serine-Threonine Kinases/*genetics ; Sequence Alignment ; Sequence Analysis, DNA

The glycoprotein 3 (GP3) of type II porcine reproductive and respiratory syndrome virus has the characteristic domains of a membrane protein. However, this protein has been reported to be retained in the endoplasmic reticulum (ER) rather than transported to the plasma membrane of the cell. In this study, we performed confocal laser scanning microscopy analysis of variants of GP3 and foundthat the signal sequence of the GP3 led to confinement of GP3 in the ER, while the functional ortransmembrane domain did not affect its localization. Based on these results, we concludedthat the signal sequence of GP3 contains the ER retention signal, which might play an important role in assembly of viral proteins.
Animals ; Cell Line ; Cell Membrane/*metabolism/virology ; Cricetinae ; Endoplasmic Reticulum/*metabolism/virology ; Microscopy, Confocal/veterinary ; Plasmids/genetics/metabolism ; Porcine respiratory and reproductive syndrome virus/*genetics/metabolism ; *Protein Sorting Signals ; Sequence Analysis, Protein/veterinary ; Viral Envelope Proteins/chemistry/*genetics/metabolism

The high genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) has been an obstacle to developing an effective vaccine for porcine reproductive and respiratory syndrome (PRRS). This study was performed to assess the degree of genetic diversity among PRRSVs from Korean pig farms where wasting and respiratory syndrome was observed from 2005 to 2009. Samples from 786 farms were tested for the presence of PRRSV using reverse transcription PCR protocol. A total of 117 farms were positive for type 1 PRRSV while 198 farms were positive for type 2. Nucleotide sequences encoding the open reading frame (ORF) 5 were analyzed and compared to those of various published PRRSV isolates obtained worldwide. Sequence identity of the ORF 5 in the isolates was 81.6~100% for type 1 viruses and 81.4~100% for type 2 viruses. Phylogenetic analysis of the ORF 5 sequences showed that types 1 and 2 PRRSVs from Korea were mainly classified into three and four clusters, respectively. The analyzed isolates were distributed throughout the clusters independent of the isolation year or geographical origin. In conclusion, our results indicated that the genetic diversity of PRRSVs from Korean pig farms is high and has been increasing over time.
Animal Husbandry ; Animals ; *Genes, Viral ; *Genetic Variation ; Lung/virology ; Lymph Nodes/virology ; *Open Reading Frames ; Phylogeny ; Porcine Reproductive and Respiratory Syndrome/virology ; Porcine respiratory and reproductive syndrome virus/chemistry/classification/*genetics/isolation & purification ; Republic of Korea ; Reverse Transcriptase Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA/veterinary ; Sequence Analysis, Protein/veterinary ; Swine

The present study was conducted to identify new target immunogenic molecules from the larval stage of the cattle tick, Boophilus annulatus (Acari: Ixodidae). Two specific larval glycoproteins (GLPs) were isolated by two-step affinity chromatography. The larval immunogens were first purified with CNBr-Sepharose coupled to rabbit anti-larval immunoglobulins, and the glycoproteins were then purified with Con-A Sepharose. These glycoproteins have molecular weights of approximately 32 and 15 kDa with isoelectric points between 6.8 and 7.2. Antibodies against the two GLPs, labeled I and II, were detected in the anti-whole tick, -whole larval, and -gut antigens through immunoblot analysis. These results suggest that these GLPs are good immunogens and can be useful in the vaccination of cattle against tick infestation.
Amino Acid Sequence ; Animals ; Cattle ; Cattle Diseases/immunology/*parasitology/prevention & control ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Glycoproteins/immunology/*isolation & purification ; Immunoblotting ; Isoelectric Focusing ; Ixodidae/chemistry/*immunology ; Male ; Molecular Weight ; Rabbits ; Sequence Analysis, Protein ; Tick Infestations/immunology/parasitology/prevention & control/*veterinary

Mutation of the p53 tumor suppressor gene has been related in the pathogenesis of numerous human and canine cancers, including breast cancers and mammary tumors. We have investigated exons 5-8 of the p53 gene for mutations in 20 spontaneous canine mammary tumors using polymerase chain reaction (PCR) with direct sequence analysis to evaluate the role of this gene in canine mammary tumorigenesis and analyzed to compare with other clinicopathological parameters including age, histology, stage, recurrence and death from tumor. Four missense (one case had two missense mutations) and one nonsense mutations were detected in 10 malignant lesions (40%), and two missense and one silent mutations were found in 10 benign mammary tumors (30%). Five of the missense mutations were located in highly conserved domains II, III, IV and V. After a follow-up period, four dogs showed a progression and three of these patients revealed death from mammary carcinoma with p53 mutation. These results demonstrated that the p53 gene mutations might be involved in the development of canine mammary tumors and contribute to the prognostic status in canine mammary carcinomas.
Animals ; Codon, Nonsense/genetics ; DNA, Neoplasm/chemistry/genetics ; Dog Diseases/*genetics ; Dogs ; Female ; Genes, p53/*genetics ; Mammary Neoplasms, Animal/*genetics ; Mutation, Missense/genetics ; Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA ; Survival Analysis ; Tumor Suppressor Protein p53/genetics

A total of 16 Taenia multiceps isolates collected from naturally infected sheep or goats in Gansu Province, China were characterized by sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The complete cox1 gene was amplified for individual T. multiceps isolates by PCR, ligated to pMD18T vector, and sequenced. Sequence analysis indicated that out of 16 T. multiceps isolates 10 unique cox1 gene sequences of 1,623 bp were obtained with sequence variation of 0.12-0.68%. The results showed that the cox1 gene sequences were highly conserved among the examined T. multiceps isolates. However, they were quite different from those of the other Taenia species. Phylogenetic analysis based on complete cox1 gene sequences revealed that T. multiceps isolates were composed of 3 genotypes and distinguished from the other Taenia species.
Animals ; China ; Cluster Analysis ; Cysticercosis/parasitology/veterinary ; DNA, Helminth/chemistry/genetics/isolation & purification ; DNA, Mitochondrial/chemistry/genetics/isolation & purification ; Electron Transport Complex IV/*genetics ; *Genetic Variation ; Goat Diseases/parasitology ; Goats ; Phylogeny ; Polymerase Chain Reaction ; Protein Subunits/genetics ; Sequence Analysis, DNA ; Sheep ; Sheep Diseases/parasitology ; Taenia/*classification/genetics/*isolation & purification