ABSTRACT
Bovine parainfluenza virus type 3 (BPIV3) is one of the most important of the known viral respiratory pathogens of both young and adult cattle. However BPIV3 has not been detected or isolated in China prior to this study. In 2008, four BPIV3 strains were isolated with MDBK cells from cattle in China and characterized by RT-PCR, nucleotide sequence analysis, transmission electron microscope observation, hemadsorption and hemagglutination tests. Nucleotide phylogenetic analysis of partial hemagglutinin-neuraminidase (HN) gene for four isolates and the complete genome for the SD0835 isolate implicated that the four Chinese BPIV3 strains were distinct from the previously reported genotype A (BPIV3a) and genotype B (BPIV3b) and might be a potentially new genotype, which was tentatively classified as genotype C (BPIV3c). This is the first study to report the isolation and genetic characterization of BPIV3 from cattle in China.
Subject(s)
Cattle Diseases/virology , Cattle/virology , Parainfluenza Virus 3, Bovine/isolation & purification , Respirovirus Infections/veterinary , Animals , Base Sequence , Cattle Diseases/epidemiology , China/epidemiology , Genotype , HN Protein/genetics , Hemadsorption , Hemagglutination Inhibition Tests , Parainfluenza Virus 3, Bovine/genetics , Phylogeny , RNA, Viral/genetics , Respirovirus Infections/epidemiology , Respirovirus Infections/virology , Sequence Analysis, RNAABSTRACT
Eighteen bovine viral diarrhea viruses (BVDV) from cattle in China between 2005 and 2008 were genetically typed by sequencing of the 5'-untranslated region (5'-UTR) of the viral genome and for selected isolates the N(pro) region. Phylogenetic reconstructions indicated that all of the 18 BVDV positive samples examined in this work clustered within the BVDV type 1 genotype. Of the 15 previously described subgenotypes of BVDV1 (1a-1o), 12 of the samples examined in this work clustered with the Chinese BVDV ZM-95 strain of pig origin, which was the prototype of BVDV1m, while 2 samples clustered with the BVDV1b. But 4 samples formed a separate group appearing to be a potentially new subgenotype, which was tentatively typed as "BVDV1p". Based on these results there appears to be highly genetic variation within the Chinese BVDV field isolates. As well, the phylogenetic reconstructions indicate that the clustering of the Chinese BVDV1m subgenotype in the phylogenetic tree is a result of geographic isolation. The information obtained from this work will be useful when carrying out epidemiological surveys of BVDV detected in China, especially for the BVDV1m detection in Chinese cattle.
Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/virology , Diarrhea Viruses, Bovine Viral/genetics , Animals , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Cattle , China/epidemiology , Genotype , Phylogeny , Time FactorsABSTRACT
OBJECTIVE: In order to construct the recombinant bovine hepervirus-1 (BHV-1) which expressed foot and mouth disease virus (FMDV) VP1 gene, we constructed a BHV-1 gE gene transfer vector by inserting the synthetic VP1 gene of FMDV (O/China/99) under the immediate-early promoter of cytomegalovirus. METHODS: The mixtures of parental virus (BHV-1/gE(-)/LacZ+) DNA and transfer vector was transfected into bovine turbinate cells using calcium phosphate-mediated transfection. Then the propagated viruses were harvested. The recombinant BHV-1 (designated BHV-1/gE(-)/VP1) was obtained by selection for white virus plaques. RESULTS: PCR results showed that VP1 gene was successfully inserted into the genome of BHV-1/gE(-). The expression of VP1 in infected cells was proved by indirect immunofluorescence assay and Western blotting. CONCLUSION: The research provided a basis for development of BHV-1 vector vaccines for FMD and other important bovine infectious diseases.
Subject(s)
Capsid Proteins/metabolism , Foot-and-Mouth Disease/virology , Herpesvirus 1, Bovine/metabolism , Recombinant Fusion Proteins/genetics , Viral Vaccines/genetics , Animals , Antibodies, Viral/genetics , Blotting, Western , Capsid Proteins/genetics , Capsid Proteins/immunology , Cattle , Cells, Cultured , Cloning, Molecular , Cytomegalovirus/genetics , Gene Expression , Genetic Vectors , Herpesvirus 1, Bovine/genetics , Promoter Regions, Genetic , Recombinant Fusion Proteins/metabolism , SwineABSTRACT
Foot-and-mouth disease (FMD) and infectious bovine rhinotracheitis (IBR) are two important infectious diseases of cattle. Using bovine herpesvirus type 1 (BHV-1) as a gene delivery vector for development of live-viral vaccines has gained widespread interest. In this study, a recombinant BHV-1 was constructed by inserting the synthetic FMDV (O/China/99) VP1 gene in the the gE locus of BHV-1 genome under the control of immediately early gene promoter of human cytomegalovirus (phIE CMV) and bovine growth hormone polyadenylation (BGH polyA) signal. After homologous recombination and plaque purification, a recombinant virus named BHV-1/gE(-)/VP1 was acquired and identified. The immunogenicity was confirmed in a rabbit model by virus neutralization test and enzyme-linked immunosorbent assay (ELISA). The result indicated that the BHV-1/gE(-)/VP1 has the potential for being developed as a bivalent vaccine for FMD and IBR.
