1.Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing.
Journal of Veterinary Science 2006;7(1):63-67
The present study was to construct a parentage testing system for Thoroughbred (TB) horse. A total number of 1,285 TB horse samples including 962 foals for parentage testing, 9 sires and 314 dams for individual identification were genotyped. Genomic DNA was extracted from 5 hair roots and genotyped by using 14 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS3, HMS6, HMS7, HTG4, HTG10, LEX3 and VHL20). This method consisted of multiplexing PCR procedure and showed reasonable amplification of all PCR products. Genotypes were determined by genetic analyzer. The number of alleles per locus varied from 3 to 9 with a mean value of 6.36 in TB horse. The expected heterozygosity was ranged from 0.548 to 0.831 (mean 0.699), and the total exclusion probability of 14 microstellite loci was 0.9998. Of the 14 markers, ASB2, ASB17, ASB23, HMS7 and HTG10 loci have relatively high PIC value (> 0.7). Of the 962 foals, 960 foals were qualified by compatibility according to the Mendelism. These results suggest that the DNA typing method has high potential for parentage verification and individual identification of TB horses.
Alleles
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Animals
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DNA/chemistry/genetics
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DNA Fingerprinting/methods/*veterinary
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Female
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Genotype
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Horses/*genetics
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Korea
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Male
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Microsatellite Repeats/genetics
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Pedigree
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Polymerase Chain Reaction/veterinary
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Polymorphism, Genetic
2.Molecular fingerprinting of clinical isolates of Mycobacterium bovis and Mycobacterium tuberculosis from India by restriction fragment length polymorphism (RFLP).
Sandeep Kumar SINGH ; Rishendra VERMA ; Devendra H SHAH
Journal of Veterinary Science 2004;5(4):331-335
Forty mycobacterial strains comprising clinical Indian isolates of Mycobacterium tuberculosis (28 field isolates +1H37 Rv) and Mycobacterium bovis (10 field isolates +1 AN5) were subjected to restriction fragment length polymorphism analysis (RFLP) using IS6110 and IS1081 probes. Most of these strains originated from dairy cattle herd and human patients from Indian Veterinary research Institute (IVRI) campus isolated from the period of 1986 to 2000. Our study showed presence of 8 copies of IS6110 in most of the M.tuberculosis (96.6%) strains irrespective of their origin with the exception of one M.tuberculosis strain with presence of an extra copy (3.4%). All M.bovis strains showed a single copy of IS6110 on the characteristic 1.9kb restriction fragment. RFLP analysis with IS1081 invariably showed the presence of 5 copies in all isolates of M.bovis and M.tuberculosis at the same chromosomal location. Similarity of IS6110 RFLP fingerprints of M.tuberculosis strains from animals and human suggested the possibility of dissemination of single M.tuberculosis strain among animals as well as human. It was not possible to discriminate within the isolates of either M.tuberculosis or M.bovis, when IS1081 was used as target sequence. The IS6110 RFLP is a valuable tool for disclosing transmission chain of M. tuberculosis and M. bovis among humans as well as animals
Animals
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Bacterial Typing Techniques
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Cattle
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DNA Fingerprinting/*veterinary
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DNA, Bacterial/*genetics
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Deer
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Humans
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India/epidemiology
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Mycobacterium bovis/classification/*genetics/isolation&purification
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Mycobacterium tuberculosis/classification/*genetics/isolation & purification
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Polymerase Chain Reaction/veterinary
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Polymorphism, Restriction Fragment Length
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Zoonoses/epidemiology
3.Analysis of Salmonella enterica serotype Enteritidis isolated from human and chickens by repetitive sequence-PCR fingerprinting, antibiotic resistance and plasmid profiles.
Journal of Veterinary Science 2006;7(1):37-41
A total of 22 Salmonella enterica serotype Enteritidis (S. Enteritidis) strains isolated from human and chicken were subjected to DNA fingerprinting by repetitive sequence PCR using ERIC and BOX primers, antibiotic resistance and plasmid patterns. Both ERIC and BOX PCR amplification data revealed a highly genetic homogeneity between isolates from human and chicken except one isolate, which originated from chicken and showed a different DNA band pattern from others. Eleven of 22 S. Enteritidis isolates (50%) were resistant to more than one antibiotics and characterized by 5 resistance patterns. The most common pattern was penicillin resistant (63.6%). Only one isolate from chicken showed a multiple drug resistance patterns to 4 antibiotics. All 22 S. Enteritidis isolates harbored more than two plasmids with eight different plasmid profiles including two to six plasmids with approximate molecular size ranging from 1.9 to 21 kb. A band of 15 kb size was detected in all isolates tested, however, the band sizes smaller than 15 kb were found only in isolates from chicken.
Animals
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*Chickens
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China/epidemiology
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DNA Fingerprinting/veterinary
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DNA, Bacterial/chemistry/genetics
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Disease Outbreaks/*veterinary
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Humans
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Microbial Sensitivity Tests/veterinary
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Microsatellite Repeats/genetics
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Plasmids/chemistry/genetics
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Polymerase Chain Reaction/veterinary
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Poultry Diseases/epidemiology/*microbiology
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Salmonella Food Poisoning/epidemiology/*microbiology
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Salmonella enteritidis/drug effects/*genetics/isolation&purification
4.Molecular typing of Brucella suis collected from 1960s to 2010s in China by MLVA and PFGE.
Zhen Jun LI ; Bu Yun CUI ; Hai CHEN ; Jing Diao CHEN ; Hong Yan ZHAO ; Dong Ri PIAO ; Hai JIANG ; Li ZHANG ; Xu TANG ; Chang Wen KE ; Zhen YAO ; Guo Zhong TIAN
Biomedical and Environmental Sciences 2013;26(6):504-508
5.Occurrence and Molecular Identification of Anisakis Dujardin, 1845 from Marine Fish in Southern Makassar Strait, Indonesia.
Hilal ANSHARY ; SRIWULAN ; Mark A FREEMAN ; Kazuo OGAWA
The Korean Journal of Parasitology 2014;52(1):9-19
Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica AB517571/DQ116427. The difference of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait.
Animals
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Anisakiasis/epidemiology/parasitology/*veterinary
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Anisakis/*isolation & purification
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Cluster Analysis
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DNA Fingerprinting
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DNA, Intergenic/chemistry/genetics
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Fish Diseases/*epidemiology/*parasitology
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Genotype
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Indonesia/epidemiology
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Molecular Sequence Data
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Phylogeny
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Polymerase Chain Reaction
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Polymorphism, Restriction Fragment Length
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Prevalence
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RNA, Ribosomal, 5.8S/genetics
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Sequence Analysis, DNA
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Sequence Homology, Nucleic Acid