The objective of this study was to investigate the prevalence of tick-borne pathogens in the Korean water deer (Hydropotes inermis argyropus). Pathogens were identified using PCR which included Anaplasma, Ehrlichia, Rickettsia, and Theileria. Rickettsia was not detected, whereas Anaplasma, Ehrlichia, and Theileria infections were detected in 4, 2, and 8 animals, respectively. The most prevalent pathogen was Theileria. Of the 8 Theileria-positive animals, 2 were mixed-infected with 3 pathogens (Anaplasma, Ehrlichia, and Theileria) and another 2 animals showed mixed-infection with 2 pathogens (Anaplasma and Theileria). Sequencing analysis was used to verify the PCR results. The pathogens found in this study were identified as Anaplasma phagocytophilum, Ehrlichia canis, and Theileria sp. To the best of our knowledge, this is the first report identifying these 3 pathogens in the Korean water deer. Our results suggest that the Korean water deer may serve as a major reservoir for these tick-borne pathogens, leading to spread of tick-borne diseases to domestic animals, livestock, and humans. Further studies are needed to investigate their roles in this respect.
Anaplasma/isolation & purification ; Animals ; Bacterial Infections/epidemiology/microbiology/*veterinary ; Cluster Analysis ; Coinfection/epidemiology/microbiology/veterinary ; DNA, Bacterial/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Ehrlichia/*isolation & purification ; Korea/epidemiology ; Molecular Sequence Data ; Phylogeny ; Prevalence ; RNA, Ribosomal, 16S/genetics ; Rickettsia/*isolation & purification ; Ruminants/*microbiology ; Sequence Analysis, DNA ; Theileria/*isolation & purification

Species identification using DNA sequences is the basis for DNA taxonomy. In this study, we sequenced the ribosomal large-subunit RNA gene sequences (3,037-3,061 bp) in length of 13 Chinese Theileria stocks that were infective to cattle and sheep. The complete 28S rRNA gene is relatively difficult to amplify and its conserved region is not important for phylogenetic study. Therefore, we selected the D2-D3 region from the complete 28S rRNA sequences for phylogenetic analysis. Our analyses of 28S rRNA gene sequences showed that the 28S rRNA was useful as a phylogenetic marker for analyzing the relationships among Theileria spp. in ruminants. In addition, the D2-D3 region was a short segment that could be used instead of the whole 28S rRNA sequence during the phylogenetic analysis of Theileria, and it may be an ideal DNA barcode.
Animals ; Base Sequence ; China ; DNA, Ribosomal/chemistry/genetics ; Molecular Sequence Data ; Phylogeny ; RNA, Ribosomal, 28S/genetics ; Ruminants ; Sequence Alignment ; Sequence Analysis, DNA/veterinary ; Theileria/*classification/genetics/isolation & purification ; Theileriasis/*parasitology

In this study, peste des petits ruminants virus (PPRV) was detected in frozen pooled tissue samples from a dead Asiatic lion (Panthera leo persica). The samples were negative for canine distemper virus and positive for PPRV nucleic acids when tested with one-step RT-PCR using the appropriate virus-specific primers. Subsequent amplification, cloning, and sequencing of the partial nucleocapsid, matrix, and fusion genes confirmed the presence of PPRV nucleic acid. Comparative sequence and phylogenetic analyses of the structural genes of the isolated virus confirmed that the virus belonged to Asian lineage IV and was closely related to PPRV circulating in India.
Animals ; Cloning, Molecular ; *Lions ; Peste-des-petits-ruminants virus/*genetics/*isolation & purification ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction/veterinary

Peste des petits ruminants (PPR) diagnosis from suspected samples from sheep and goats was carried out. Buffy coat, tissues, and oculo-nasal swabs were analyzed using nucleoprotein (NP3/NP4) and fusion protein (F1/F2) gene primers, respectively. Analysis of the sample types and primer set revealed that buffy coat are the best type of samples for PPR diagnosis and the use of two set of primers will increase the number of positives.
Animals ; DNA Primers/analysis ; Eye/virology ; Goat Diseases/blood/*diagnosis/epidemiology/virology ; Goats ; Hair/virology ; Nose/virology ; Nucleoproteins/analysis ; Peste-des-Petits-Ruminants/blood/*diagnosis/epidemiology/virology ; Peste-des-petits-ruminants virus/genetics/*isolation & purification ; Pigmentation ; RNA, Viral/genetics/*isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction/*methods/standards/veterinary ; Sheep ; Sheep Diseases/blood/*diagnosis/epidemiology/virology ; Uganda/epidemiology

The nucleotide sequences of P gene from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The P gene is 1,655 nucleotides long with two overlapping open reading frames (ORFs). The first ORF is 1530 nucleotides long and would produce P protein of 509 amino acid residues. The second ORF is 534 nucleotides long and would produce C protein of 177 amino acid residues. The first ORF produces a second mRNA transcript of 897 nucleotides long with an extra G nucleotide at position 751. Translation from this mRNA would produce V protein of 298 amino acid residues. The nucleotide and deduced amino acid sequence were compared with the homologous region of other PPRV isolates. At the amino acid level, the "China/Tib/Gej/07-30" shares homology of 86.10%-97.3%, 84.3%-94.9%, and 82.9%-96.3% for P, C, and V proteins respectively. Several sequence motifs in the P genes were identified on the basis of conservation in the PPRVs and the morbilliviruses.
Amino Acid Sequence ; Animals ; China ; Female ; Goat Diseases ; virology ; Goats ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; veterinary ; virology ; Peste-des-petits-ruminants virus ; chemistry ; genetics ; isolation & purification ; metabolism ; Phosphoproteins ; chemistry ; genetics ; metabolism ; Sequence Analysis ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics ; metabolism

Peste des petits ruminants virus is a member of Morbillivirus Paramyxoviridae. The complete genome of a Peste des petits ruminants virus (PPRV) isolate, China/Tib/07 was sequenced and molecular characteristics was analyzed. The internal sequences of the virus genome were amplified by RT-PCR with primers designed according to the published data in GenBank, while the sequences of the 3' and 5' ends of the genome were determined by RACE. Amplification products were directly sequenced,assembled and analyzed with DNAStar4.0. Results showed that China/Tib/07 genome consisted of 15 948 nucleotides in length, encoding six structural proteins and two non-structural proteins just like other known PPRV genomes. Phylogenetically, the virus genome shared homology of 91.6%-98.1% with Southwest Asian isolates among PPRV strains and the highest homology of 64.3% with rinderpest virus among morbillivirus members.
Animals ; Base Sequence ; Cercopithecus aethiops ; China ; Genome, Viral ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; veterinary ; virology ; Peste-des-petits-ruminants virus ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Alignment ; Sequence Analysis, DNA ; Sheep ; Sheep Diseases ; virology ; Vero Cells ; Viral Proteins ; genetics

To develop a reverse genetics system of Peste des petits ruminants virus(PPRV), five pairs of oligonucleotide primers were designed on the basis of the full-length genomic sequence of PPRV Nigeria 75/ 1 strain. Using RT-PCR technique, five over-lapping cDNA fragments, designated as JF1, JF2, JF3, JF4 and JF5, respectively, were amplified, followed by cloning into pcDNA3.1(+)vector. An AscI restriction enzyme site and a T7 promoter sequence were introduced immediately upstream of 5'-end, while a PacI restriction enzyme site was engineered downstream of 3'-end. Using pok12 as a plasmid vector, the full-length cDNA clone pok12-PPRV of Nigeria 75/1 was assembled by connecting the five cDNA fragments via the unique restriction endonuclease site of PPRV genome. The resultant nucleotide sequence of the PPRV Nigeria 75/1 strain in the study was compared with other members of genus morbillivirus, and phylogenetic analysis was used to examine the evolutionary relationships. The results showed that PPRV Nigeria 75/ 1 was antigenically closely related to Rinderpest virus and Measles virus. Successful construction of full-length cDNA clone of PPRV Nigeria 75/1 strain lays the basis rescuing PPRV effectively and enables further research of PPRV at molecular level.
Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genome, Viral ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; virology ; Peste-des-petits-ruminants virus ; classification ; genetics ; Phylogeny ; Sequence Analysis, DNA

The nucleotide sequences of M and F genes from a field strain of peste des petits ruminants virus (PPRV) ("China/Tib/Gej/07-30") was firstly determined. The M gene was 1 483 nucleotides in length with a single open reading frame (ORF), encoding a protein of 335 amino acids. The F gene was 2411 nucleotides in length, encoding a protein of 546 amino acids. The resulting nucleotide sequence and the deduced amino acid sequences were compared with the homologous regions of other PPRV isolates. The nucleotide sequences of M and F genes of the "China/Tib/Gej/07-30" was 92.4%-97.7% and 85.5%-96.1% identical to other PPRV isolates, respectively, while a homology of 97.0%-98.2% and 94.3%-98.2% could be observed at the amino acids level respectively. Several sequence motifs in the M and F genes had been identified on the basis of conservation in the PPRVs and the morbilliviruses. The 3' untranslated region of M gene was 443 nucleotides in length with 82.4%-93.5% identical to other PPRV isolates. The 5' untranslated region of F gene was 634 nucleotides in length with 76.2%-91.7% identical to other PPRV isolates.
Amino Acid Sequence ; Animals ; Base Sequence ; Molecular Sequence Data ; Peste-des-Petits-Ruminants ; veterinary ; virology ; Peste-des-petits-ruminants virus ; chemistry ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Homology, Amino Acid ; Sheep ; Sheep Diseases ; virology ; Tibet ; Viral Fusion Proteins ; chemistry ; genetics ; Viral Matrix Proteins ; chemistry ; genetics

In order to screen microsatellites for conservation genetics studies of the species, a total of 23 microsatellite loci from Korean goral (Naemorhedus caudatus), including 15 previously developed loci and 8 new loci in this study, were tested. Eleven microsatellites were screened and subjected to cross-species amplification using a test panel of four Caprinae species, Japanese serows (Capricornis crispus), Chinese gorals (Naemorhedus goral), Northern chamois (Rupicapra rupicapra) and domestic goats (Capra hircus). In addition, all eleven microsatellites (SY3A, SY12A, SY12B, SY48, SY58, SY71, SY76, SY84, SY84B, SY112, and SY129) satisfied the criteria to be a core set of microsatellites. This core set of microsatellites and cross-species amplification of Korean goral microsatellites were found to be helpful for high-resolution studies for conservation and management of Korean goral and other endangered Caprinae species.
Animals ; Conservation of Natural Resources ; Genetic Variation ; Hybridization, Genetic/*genetics ; Microsatellite Repeats/*genetics ; Republic of Korea ; Ruminants/*genetics

The purpose of the study is to construct recombinant goat pox virus (GPV) expressing Peste des petits ruminants virus (PPRV) H protein, and to evaluate the immunization effect. Recombinant GPV containing PPRV H gene (rGPV-PPRV-H) was selected and purified by gpt and eGFP utilizing plaque purification, and the final selected recombinant GPV was proved to be purified by PCR. Immunofluorescence and Western blotting showed that the recombinant virus could express H protein of PPRV while infecting lamb testis cells. Six goats were immunized with 2 x 10(6) PFU rGPV-PPRV-H through intradermal injection, and were immunized for the second time at 28 days with the same dose recombinant virus after first immunization. Serum was collected after immunization, and was analyzed for the neutralization antibodies. 21 days after first immunization, the neutralization antibodies of GPV were 40, 80, > or = 80, > or = 80, 40, > or = 80 in turn, and neutralization antibodies of PPRV were 80, 80, 80, 80, 40, 40, 10 in turn; 14 days after second immunization, the neutralization antibodies of GPV were all > or = 80, and the neutralization antibodies of PPRV were > 80, 80, > 80, 80, 80 and 40 in turn. This study established a foundation for the industrialization of the PPRV recombinant GPV vaccine.
Animals ; Capripoxvirus ; genetics ; immunology ; Goat Diseases ; immunology ; prevention & control ; virology ; Goats ; Hemagglutinins, Viral ; genetics ; immunology ; metabolism ; Peste-des-Petits-Ruminants ; immunology ; prevention & control ; Peste-des-petits-ruminants virus ; genetics ; immunology ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Vaccines, Combined ; immunology ; Vaccines, Synthetic ; immunology ; Viral Vaccines ; immunology