OBJECTIVETo demonstrate a noninvasive large mammalian genetic sampling method using blood meal obtained from a tabanid fly.

METHODSBlood meal was recovered from the abdomen of an engorged tabanid fly (Haematopota sp.) which was captured immediately after biting a Sumatran rhino in captivity. The blood was applied on to a Whatman FTA(®) blood card. Subsequent laboratory work was conducted to extract, amplify and sequence the DNA from the sample. Validation was done by sampling the hair follicles and blood samples from the rhinoceros and subjecting it to the same laboratory process.

RESULTSBLAST search and constructed phylogenetic trees confirmed the blood meal samples were indeed from the rhino.

CONCLUSIONSThis method could be used in the field application to noninvasively collect genetic samples. Collection of tabanids and other haematophagous arthropods (e.g. mosquitoes and ticks) and other blood-sucking parasites (e.g. leeches and worms) could also provide information on vector-borne diseases.

Animals ; Diet ; veterinary ; Diptera ; genetics ; physiology ; Endangered Species ; Female ; Food Chain ; Indonesia ; Insect Bites and Stings ; blood ; veterinary ; Molecular Sequence Data ; Perissodactyla ; Phylogeny ; Polymerase Chain Reaction ; veterinary ; Sequence Analysis, DNA ; veterinary

Polymorphisms of the prion protein gene (PRNP) havebeen detected in several cervid species. In order toconfirm the genetic variations, this study examined theDNA sequences of the PRNP obtained from 33 captivesika deer (Cervus nippon laiouanus) in Korea. A total ofthree single-nucleotide polymorphisms (SNPs) at codons100, 136 and 226 in the PRNP of the sika deer wereidentified. The polymorphic site located at codon 100 hasnot been reported. The SNPs detected at codons 100 and226 induced amino acid substitutions. The SNP at codon136 was a silent mutation that does not induce any aminoacid change. The genotype and allele frequencies weredetermined for each of the SNPs.
Animals ; Base Sequence ; DNA/chemistry/genetics ; Deer/*genetics ; Genetic Variation ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; Polymorphism, Single Nucleotide ; Prions/*genetics ; Sequence Analysis, DNA

This study examined the occurrence of Anaplasma spp. and hemoplasma infection in leopard cats, Prionailurus bengalensis euptilurus, in Korea. Twenty-nine biological samples were tested by molecular analysis. Two (6.9%) and eight (27.6%) tested specimens were positive for Anaplasma bovis and hemoplasma infection, respectively. Based on our results, Anaplasma/Ehrlichia spp. and hemoplasma are regularly infecting leopard cat populations of Korea. Considering their endangered status, regular monitoring of infection by arthropod-borne pathogens known to cause clinical symptoms in feline hosts such as Anaplasma/Ehrlichia spp. and hemoplasma would be crucial as part of ongoing conservation efforts.
Anaplasma/*isolation & purification ; Anaplasmosis/*epidemiology/microbiology ; Animals ; DNA, Bacterial/genetics ; *Felidae ; Molecular Sequence Data ; Mycoplasma/*isolation & purification ; Mycoplasma Infections/epidemiology/microbiology/*veterinary ; Phylogeny ; Polymerase Chain Reaction/veterinary ; RNA, Ribosomal, 16S/genetics ; Republic of Korea/epidemiology ; Sequence Analysis, DNA/veterinary

Natural habitat fragmentation and reducing habitat quality have resulted in an increased appearance of Japanese macaques, Macaca fuscata (Gray, 1870), in suburban areas in Japan. To investigate the risk of zoonotic infections, a coprological survey of helminth eggs passed by wild Japanese macaques was carried out in 2009 and 2010 in Shiga Prefecture, Japan. Microscopic examination found helminth eggs in high prevalence, and nucleotide sequencing of DNA extracted from the eggs identified Oesophagostomum cf. aculeatum and Trichuris trichiura. A fecal culture also detected infective larvae of Strongyloides fuelleborni. These zoonotic nematodes pose a potential health issue to local people in areas frequented by Japanese macaques.
Animals ; DNA/chemistry/genetics ; Feces/*parasitology ; Japan ; Macaca ; Molecular Sequence Data ; Oesophagostomiasis/parasitology/*veterinary ; Oesophagostomum/classification/*isolation & purification ; Primate Diseases/*parasitology ; Sequence Analysis, DNA ; Strongyloides/classification/isolation & purification ; Strongyloidiasis/parasitology/veterinary ; Trichuriasis/parasitology/*veterinary ; Trichuris/classification/*isolation & purification

A 12-year-old spayed female mixed-bred dog presented with nasal bleeding of 2 days duration and a skin nodule in the left flank. No abnormalities were found in coagulation profiles and blood pressure. Cytological evaluation of the nodule revealed numerous characteristic round organisms having a nucleus and a bar within macrophages and in the background, consistent with leishmaniasis. In vitro culture was unsuccessful but PCR of the nodular aspirate identified the organisms as Leishmania infantum, and the final diagnosis was canine leishmaniasis. No history of travel to endemic countries was noted. Because the dog had received a blood transfusion 2 years before the illness, serological screening tests were performed in all donor dogs of the commercial blood bank using the commercial Leishmania ELISA test kit, and there were no positive results. Additional 113 dogs with hyperglobulinemia from Seoul were also screened with the same kits but no positive results were obtained. To the best of the author's knowledge this is the first autochthonous case of canine leishmaniasis in Korea.
Animals ; Base Sequence ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Dog Diseases/*diagnosis/epidemiology ; Dogs ; Enzyme-Linked Immunosorbent Assay/veterinary ; Female ; Giant Cells/pathology ; Leishmania infantum/genetics/immunology/*isolation & purification ; Leishmaniasis, Visceral/diagnosis/*veterinary ; Molecular Sequence Data ; Polymerase Chain Reaction/veterinary ; Protozoan Proteins/genetics ; Republic of Korea ; Sequence Analysis, DNA/veterinary ; Serologic Tests/veterinary

In August 2008, forty dogs out of 400 developed oral warts in a breeding farm in Korea. Canine oral papilloma infection is a common disease in dogs. However, there has been no report of an outbreak of canine oral papillomavirus (COPV) in a group of dogs or in dog breeding farms in Korea, and the genetic analysis of COPV in Korea has yet to be performed. This study diagnosed canine oral papilloma from the oral samples of these dogs based on histopathological examination and immunohistochemistry. Polymerase chain reaction was applied to amplify the corresponding products using pre-existing primer sets for COPV and a universal human papillomavirus targeting L1 gene. Further genetic analysis of the major viral capsid gene L1 confirms the sequences of Korean COPV, which shows a close relationship to previously reported COPV. This study describes the histopathological and immunohistochemical characteristics of canine oral papilloma in a group of breeding dogs in Korea and discloses the complete L1 gene sequences of Korean COPV.
Animals ; Base Sequence ; Capsid Proteins/chemistry/genetics ; DNA, Viral/chemistry/genetics ; Disease Outbreaks/*veterinary ; Dog Diseases/epidemiology/*virology ; Dogs ; Immunohistochemistry/veterinary ; Korea/epidemiology ; Lambdapapillomavirus/genetics/*isolation & purification ; Molecular Sequence Data ; Mouth Diseases/epidemiology/*veterinary/virology ; Papillomavirus Infections/epidemiology/*veterinary/virology ; Polymerase Chain Reaction/veterinary ; Sequence Analysis, DNA

Bacillus (B.) anthracis, the etiological agent of anthrax, is one of the most genetically monomorphic bacteria species in the world. Due to the very limited genetic diversity of this species, classification of isolates of this bacterium requires methods with high discriminatory power. Single nucleotide repeat (SNR) analysis is a type of variable-number tandem repeat assay that evaluates regions with very high mutation rates. To subtype a collection of 21 isolates that were obtained during a B. anthracis outbreak in Korea, we analyzed four SNR marker loci using nucleotide sequencing analysis. These isolates were obtained from soil samples and the Korean Center for Disease Control and Prevention. The SNR analysis was able to detect 13 subgenotypes, which allowed a detailed evaluation of the Korean isolates. Our study demonstrated that the SNR analysis was able to discriminate between strains with the same multiple-locus variable-number tandem repeat analysis genotypes. In summary, we obtained SNR results for four SNR marker loci of newly acquired strains from Korea. Our findings will be helpful for creating marker systems and help identify markers that could be used for future forensic studies.
Bacillus anthracis/*classification/*genetics/isolation & purification ; *Genetic Variation ; *Minisatellite Repeats ; Polymerase Chain Reaction/veterinary ; Republic of Korea ; Sequence Analysis, DNA/*methods/veterinary ; *Soil Microbiology

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

Five cases of orf virus infection in Korean black goats were diagnosed in our laboratory between 2010 and 2011. One orf virus (ORF/2011) was isolated from an ovine testis cell line (OA3.Ts) for use as a vaccine candidate. Sequences of the major envelope protein and orf virus interferon resistance genes were determined and compared with published reference sequences. Phylogenetic analyses revealed that orf viruses from Korean black goats were most closely related to an isolate (ORF/09/Korea) from dairy goats in Korea. This result indicates that the orf viruses might have been introduced from dairy goats into the Korean black goat population.
Animals ; Ecthyma, Contagious/epidemiology/*virology ; Goat Diseases/*epidemiology/virology ; Goats ; Molecular Sequence Data ; Orf virus/*genetics/isolation & purification/metabolism ; Phylogeny ; Polymerase Chain Reaction/veterinary ; Republic of Korea/epidemiology ; Sequence Analysis, DNA/veterinary ; Sequence Homology

Giardia intestinalis infections arise primarily from contaminated food or water. Zoonotic transmission is possible, and at least 7 major assemblages including 2 assemblages recovered from humans have been identified. The determination of the genotype of G. intestinalis is useful not only for assessing the correlation of clinical symptoms and genotypes, but also for finding the infection route and its causative agent in epidemiological studies. In this study, methods to identify the genotypes more specifically than the known 2 genotypes recovered from humans have been developed using the intergenic spacer (IGS) region of rDNA. The IGS region contains varying sequences and is thus suitable for comparing isolates once they are classified as the same strain. Genomic DNA was extracted from cysts isolated from the feces of 5 Chinese, 2 Laotians and 2 Koreans infected with G. intestinalis and the trophozoites of WB, K1, and GS strains cultured in the laboratory, respectively. The rDNA containing the IGS region was amplified by PCR and cloned. The nucleotide sequence of the 3' end of IGS region was determined and examined by multiple alignment and phylogenetic analysis. Based on the nucleotide sequence of the IGS region, 13 G. intestinalis isolates were classified to assemblages A and B, and assemblage A was subdivided into A1 and A2. Then, the primers specific to each assemblage were designed, and PCR was performed using those primers. It detected as little as 10 pg of DNA, and the PCR amplified products with the specific length to each assemblage (A1, 176 bp; A2, 261 bp; B, 319 bp) were found. The PCR specific to 3 assemblages of G. intestinalis did not react with other bacteria or protozoans, and it did not react with G. intestinalis isolates obtained from dogs and rats. It was thus confirmed that by applying this PCR method amplifying the IGS region, the detection of G. intestinalis and its genotyping can be determined simultaneously.
Sequence Analysis, DNA ; Sensitivity and Specificity ; Polymerase Chain Reaction/*methods ; Phylogeny ; Mice ; Humans ; Giardiasis/parasitology/veterinary ; Giardia lamblia/*classification/genetics/*isolation & purification ; Genotype ; Dogs ; Dog Diseases/parasitology ; DNA, Ribosomal Spacer/*analysis ; DNA, Protozoan/*analysis/isolation & purification ; Base Sequence ; Animals