• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.9-19
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• 2014

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.

• Parasites, Hosts and Diseases
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• v.44 no.2 s.138
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• pp.117-125
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• 2006

Genetic diversity of 18 Acanthamoeba isolates from ocean sediments was evaluated by comparing mitochondrial (mt) DNA RFLP, 18S rDNA sequences and by examining their cytopathic effects on human corneal epithelial cells versus reference strains. All isolates belonged to morphologic group II. Total of 16 restriction phenotypes of mtDNA from 18 isolates demonstrated the genetic diversity of Acanthamoeba in ocean sediments. Phylogenetic analysis using 18s rDNA sequences revealed that the 18 isolates were distinct from morphological groups I and III. Fifteen isolates showed close relatedness with 17 clinical isolates and A. castellanii Castellani and formed a lineage equivalent to T4 genotype of Byers' group. Two reference strains from ocean sediment, A. hatchetti BH-2 and A. griffini S-7 clustered unequivocally with these 15 isolates. Diversity among isolates was also evident from their cytopathic effects on human corneal cells. This is the first time describing Acanthamoeba diversity in ocean sediments in Korea.

• The Korean Journal of Malacology
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• v.10 no.1
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• pp.27-37
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• 1994

한국산 담수 또아리물달팽이과(Planorbidae)에 속하는 또아리물달팽이(Gyraulus convexiusculus),수정또아리물달팽이(Hippeutis cantori) 및 배꼽또아리물달팽이(Segmentina hemisphaerula)3종에 대한 종간 유전적 변이와 이들 상호간의 분류학적 유연 관계를 생화학적 측면에서 밝히고자 하였다. 즉, 모계유전으로 자손에 유전되고 있는 미토콘드리아 DNA(mitochondrial DNA; mt DNA)의 변이를 보기위하여 제한효소(restriction enzyme)를 처리하고 잘라진 mtDNA절편들을 상호 비교하는 restriction fragmint length polymorphism(RFLP)기법을 응용하였다. 본 실험에서 10개의 제한효소 중 CIa I, Dra I, Eco RI, Hin dIII, Kpn I및 pst I의 6개 제한효소에서 좋은 결과를 얻어 종간의 공통절편(shated fragmints)을 비교하였고, 염기분화율(nucleotide divergince rate)을 각각 측정하였다. 미토콘드리아 DNA 크기(genome size)는 또아리물달팽이가 12.08 kb, 수정또아리물달팽이가 14.4 kb, 그리고 배꼽또아리물달팽이가 12.93 kb로 관찰되었다. 염기분화율(p)는 또아리물달팽이/수정또아리물달팽이 군에서 p=12.7%, 배꼽또아리물달팽이와 상기 2종군 사이의 염기분화율은 P=56.6%여서 배꼽 또아리물달팽이류는 타 2종보다 그 분화율이 매우 높음을 알 수 있었다. 이상의 결과로 보아 분류군(taxa)의 mtDNA 변이에 의한 RELP기법이 앞으로 한국산 담수 패류 연구에 널리 응용될 수 있음이 확인되었다.

• Parasites, Hosts and Diseases
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• v.46 no.3
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• pp.157-164
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• 2008

Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 188 and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in southeastern Korea.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.61-64
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• 2016

We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

• Parasites, Hosts and Diseases
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• v.52 no.4
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• pp.383-389
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• 2014

Anisakiasis, a human infection of Anisakis L3 larvae, is one of the common foodborne parasitic diseases in Korea. Studies on the identification of anisakid larvae have been performed in the country, but most of them have been focused on morphological identification of the larvae. In this study, we analyzed the molecular characteristics of 174 Anisakis type I larvae collected from 10 species of fish caught in 3 different sea areas in Korea. PCR-RFLP and sequence analyses of rDNA ITS and mtDNA cox1 revealed that the larvae showed interesting distribution patterns depending on fish species and geographical locations. Anisakis pegreffii was predominant in fish from the Yellow Sea and the South Sea. Meanwhile, both A. pegreffii and A. simplex sensu stricto (A. simplex s.str.) larvae were identified in fish from the East Sea, depending on fish species infected. These results suggested that A. pegreffii was primarily distributed in a diverse species of fish in 3 sea areas around Korea, but A. simplex s.str. was dominantly identified in Oncorhynchus spp. in the East Sea.

• Korean journal of applied entomology
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• v.53 no.1
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• pp.85-91
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• 2014

In this study, we analyzed the phylogenetic characterization of root-knot nematodes (Meloidogyne spp.) in soils from fruits and vegetables greenhouses in Korea. Soil samples were collected from 12 greenhouse fields in which tomato, cucumber, watermelon, and Oriental melon were being cultivated. Meloidogyne spp. were detected in all the soil samples at an average number of $72{\pm}6$ nematodes/300 g of soil to $2,898{\pm}468$ nematodes/300 g of soil. Phylogenetic analysis using polymerase chain reaction-restriction fragment length polymorphism was attempted for the second-stage juveniles (J2) of Meloidogyne spp. collected from the greenhouse soils. Twelve Meloidogyne spp. from the greenhouse soils were classified into two groups by using HinfI digestion of mitochondrial DNA, resulting in 900, 410, 290, and 170 bp fragments (group A) and 900, 700, and 170 bp fragments (group B). Phylogenetic analysis based on mitochondrial DNA sequences (1,483-1,521 bp) showed that nine group A isolates were identified as Meloidogyne incognita (99.73-99.93%) and three group B isolates showed 99.54-99.73% similarity to Meloidogyne arenaria.

• Parasites, Hosts and Diseases
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• v.43 no.1 s.133
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• pp.7-13
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• 2005

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of $0.2-1.1\%$ with each other, but more than $2\%$ difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group lion the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.355-360
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• 2013

Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.25-28
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• 2008

In order to distinguish Korean cattle (Hanwoo) beef from the imported beef from Australia in Korean markets, DNA markers based on PCR-RFLP from mitochondrial genes and SRY gene were applied. A total of 2,826 beef samples comprising 1,495 Hanwoo and 1,331 foreign cattle breeds were obtained in Korea. An 801 bp fragment of the SRY gene on the bovine Y chromosome, a 343 bp fragment of ND4 gene and a 528 bp fragment of ND5 gene in the bovine mtDNA were amplified by PCR and digested with three restriction enzymes, MseI, HpyCH4III and Tsp509I, respectively. The results showed that Bos taurus (T) type was the majority in Hanwoo by combining three markers (99.5%). However, 78.2% of Bos indicus (I) type was observed in the imported beef samples. These results indicated that three markers used in this study will be used as valuable markers for discriminating imported beef against Hanwoo.