• Journal of Food Hygiene and Safety
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• v.33 no.4
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• pp.280-288
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• 2018

In this study, an approach for the analysis of the five cephalopod species (octopus, long-arm octopus, squid, wet-foot octopus, beka squid) consumed in the Republic of Korea is developed. The samples were collected from the Southeast Asian countries Thailand, Indonesia, Vietnam, and China. The SYBR-green-based real-time qPCR method, based on the mitochondrial DNA genome of the five cephalopods was developed and validated. The intergroup variations in the mitochondrial DNA are evident in the bioinformatic analysis of the mitochondrial genomic DNA sequences of the five groups. Some of the highly-conserved and slightly-variated regions are identified in the mitochondrial cytochrome-c-oxidase subunit I (COI) gene, 16s ribosomal RNA (16s rRNA) gene, and 12s ribosomal RNA (12s rRNA) gene of these groups. To specify each five cephalopod groups, specific primer sets were designed from the COI, 16s rRNA and 12s rRNA regions. The specific primer sets amplified the DNA using the SYBR-green-based real-time PCR system and 11 commercially secured animal tissues: Octopus vulgaris, Octopus minor, Todarodes pacificus, Dosidicus gigas, Sepia esculenta, Amphioctopus fangsiao, Amphioctopus aegina, Amphioctopus marginatus, Loliolus beka, Loligo edulis, and Loligo chinensis. The results confirmed by a conveient way to calculate relative amplification levels between different samples in that it directly uses the threshold cycles (Ct)-value range generated by the qPCR system from these samples. This genomic DNA-based molecular technique provides a quick, accurate, and reliable method for the taxonomic classification of the animal tissues using the real-time qPCR.

• Journal of Life Science
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• v.24 no.6
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• pp.626-632
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• 2014

The genetic structure and phylogenetic relationship were investigated in Korean red spotted grouper populations using the nucleotide sequence polymorphisms of the mitochondrial DNA (mtDNA) cytochrome c oxidase subunit I (COI) gene. The COI gene was sequenced showed 99.1-99.8% identity with the EF607565 sequence previously reported. A total of twenty haplotypes were found, and the Korean population showed nineteen haplotypes. Among those, Hap_03 and Hap_08 showed Jeju-do and China-specific COI sequences, respectively. However, Hap_07 had twelve COI sequences from South Korea and records from Hong Kong and Taiwan. Neighbor-joining (NJ) trees constructed from the phylogenetic analyses based on the polymorphisms of the COI haplotypes showed a monophyletic branching pattern within the genus Epinephelus. This indicated that the red spotted grouper populations had evolved from common maternal ancestors. In addition, the Hap_08, which had the COI sequence recorded only from China Sea, was found in the middle of the NJ tree nearby Hap_07 and showed a close relationship with Hap_07. This indicates that Chinese red spotted grouper is also maternally related to other populations in East Asia. Consequently, East Asian red spotted grouper populations are maternally related, as well as sharing the same evolutionary history, and are still affected by the East Asian ocean current (Kuroshio). These findings help to explain the genetic structure and phylogenetic relationship of red spotted grouper and also contribute to research on artificial breeding and industrialization.

• Journal of Food Hygiene and Safety
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• v.34 no.5
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• pp.502-507
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• 2019

Cephalopods are one of the most important fishery resources in the world because of their desirable taste and nutritional value. In south Korea, one of the countries in which a large amount of seafood is consumed, cephalopods (e.g., octopus, squid, and cuttlefish) have an annual consumption rate of over 400,000 metric tons. In this study, octopus and squid products (n=28) sold on the market were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). For species identification, the NCBI BLAST database was screened with the sequences and analyzed as a query. In this BLAST search, twelve squid products showed 99-100% sequence identity to Dosidicus gigas (n=3) and Todarodes pacificus (n=9). In the case of the other 16 products that were declared using octopus as raw materials on the labels, six products were identified as Cistopus taiwanicus (n=1), Amphioctopus marginatus (n=1), Scaeurgus unicirrhus (n=1), and Dosidicus gigas (n=3). Monitoring results indicated that a significant percentage (37.5%) of mislabeling was present in octopus products sold on the South Korean market.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.64-70
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• 2019

Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.240-246
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• 2021

The Chironomidae is a benthic macroinvertebrate commonly found in freshwater ecosystems, along with Ephemeroptera and Trichoptera, which can be used for environmental health assessments. There are approximately 15,000 species of Chironomidae worldwide, but there are limited studies on species identification of domestic Chironomidae larvae. In the present study, we carried out species classification of the Chironomidae larvae that found in Jeju's tap water purification plants using morphological characteristics and genetic identification based on cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA. Body shape, mentum, antenna, mandible in the head capsule, and claws were observed in the larvae for morphological classification. Analysis of 17 larvae collected from faucets and fire hydrants of domestic tap water purification plants revealed the presence of two species, including 14 Orthocladius tamarutilus and 3 Paratrichocladius tammaater. These results will aid the use of the criteria information about species classification of the Chironomidae for water quality management in water purification plants and diversity monitoring of freshwater environments.

• Journal of Food Hygiene and Safety
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• v.38 no.6
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• pp.496-507
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• 2023

This study investigated species identification and labeling compliance of 48 shrimp products sold in the Korean online markets. Species identification was conducted using the standard DNA barcoding method, using the cytochrome c oxidase subunit I gene. The obtained sequences were compared with those deposited in the NCBI GenBank and BOLD Systems databases. Additionally, phylogenetic analysis was performed to further verify the identified shrimp species. Consequently, 16 shrimp species were identified, including Penaeus vannamei, Pandalus borealis, Palaemon gravieri, Leptochela gracilis, Penaeus monodon, Pleoticus muelleri, Metapenaeopsis dalei, Euphausia pacifica, Lebbeus groenlandicus, Trachypenaeus curvirostris, Argis lar, Metanephrops thomsoni, Metapenaeopsis barbata, Alpheus japonicus, Penaeus chinensis, and Mierspenaeopsis hardwickii. The most prevalent species was Penaeus vannamei, found in 45.8% of the analyzed products. A significant mislabeling rate of 72.9% was found; however, upon excluding generic names such as shrimp, the mislabeling rate reduced to 10.4%. The mislabeling rate was higher in highly-processed products (89.3%) compared with that in minimally-processed products (50%). No correlation was found between the country of origin and mislabeling rate. The results of this study provide crucial data for future monitoring of shrimp products and improving the labeling of shrimp species in Korea.

• Korean Journal of Ecology and Environment
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• v.55 no.3
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• pp.212-220
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• 2022

An estuary is a water ecosystem with a high abundance of the species diversity, due to a variety of complex physicochemical factors of the area where freshwater and ocean mixed. The identification of Corbicula species in the estuary environments is difficult because of various morphological characteristics. In this study, we provide taxonomic information on Corbicula species with taxonomic difficulties using morphological and genetic analysis. This study was conducted on clams from the Seomjin River-Gwangyang Bay, one of the major production area of marsh clam in Korea. As a result, we characterized Cytocrome C Oxidase subunit I (COI) sequences of the Corbicula. The 636 bp nucleotide sequences of COI have 98% homology among Corbicula species collected from 2 sites of Seomjin River-Gwangyang Bay. The phylogenetic analysis with 17 species of Corbicula indicated that most of the species collected from Seomjin River-Gwangyang Bay were brackish water clam (Corbicula japonica), and only one Asian clam (Corbicula fluminea). The evolutionary distance between C. japonica and C. fluminea was less than 0.003. Therefore, it was confirmed that C. japonica is phylogenetically closely related to C. fluminea. In 9 species of Cyrenidae, phylogenetic tree was classified into three lineages. These results will be used as an important data for an identification of clam species by providing genetic information for Corbicula species with a morphological diversity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.208-213
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• 2016

The re-identification of two Korean tonguefishes, Cynoglossus interruptus and Symphurus orientalis, was carried out using eight specimens collected from Korean waters in 2007 and 2013. C. interruptus is characterized by having a single row of scales between rows connected to the supraorbital line and the middle lateral line, 107–113 dorsal fin rays, 86–89 anal fin rays, and 53–55 vertebrae. S. orientalis is characterized by having a 1-2-2-2-2 ID pattern, 97–100 dorsal fin rays, 83–89 anal fin rays, and 52–55 vertebrae. Molecular analysis using mitochondrial DNA Cytochrome Oxidase subunit I sequences showed that specimens of the two species corresponded well to Japanese C. interruptus and Taiwanese S. orientalis, respectively. Therefore, although several reports have raised questions regarding the distribution of C. interruptus and S. orientalis in Korean waters, morphological and molecular data confirm that these two species are indeed distributed in these waters.

• Fisheries and Aquatic Sciences
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• v.17 no.4
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• pp.491-496
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• 2014

A total of six specimens (57.03-100.72 mm in standard length) belonging to the family Diodontidae were collected around Jeju Island, Korea between July and August 2013. On the Basis of morphological and molecular analyses, we identified the specimens as Cyclichthys orbicularis (Bloch, 1785), a new record to the Korean fish fauna. Morphologically, the species is characterized by nine caudal fin rays, immovable spines on the head and body except the caudal peduncle, and small black spots dorsally. Some morphometric characters changed disproportionately with growth, which was a new finding. Caudal peduncle length, body width and preanus length to standard length tend to grow at a rate different from that of overall growth. When the 601 base pairs of mitochondrial DNA cytochrome c oxidase subunit I sequences of our specimens were compared with those of other diodontid fishes, our specimens clustered tightly around C. orbicularis (d = 0.000-0.002) in an NJ tree, with a high bootstrap value (99%). We herein describe the morphological and molecular traits of the specimens, and propose a new Korean name, "gud-ga-si-bok-sok" for the genus Cyclichthys, and "dung-geun-gud-ga-si-bok" for C. orbicularis.

• Animal Systematics, Evolution and Diversity
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• v.35 no.1
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• pp.33-36
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• 2019

The marine interstitial annelid Pharyngocirrus uchidai(Sasaki, 1981) has been only known from Japan. In this study, we report the occurrence of P. uchidai for the first time in four localities along the eastern coast of Korea: Bukmyeon, Gamchu, Gase, and Oeongchi. Species identification was confirmed by comparison of DNA barcoding sequences with morphological examination from the type locality, Oshoro, Japan. We generated a total of 25 sequences of a partial segment (580 bp) of the cytochrome c oxidase subunit I gene (COI), representing five specimens from each locality. Maximum intra-specific variation was 1.2% in terms of Kimura two-parameter (K2P) distance, observed between two individuals each from Gamchu (i.e., between two specimens from the single locality), Gamchu and Oeongchi, Gamchu and Oshoro, and Oeongchi and Oshoro. On the other hand, an identical haplotype was found in all the five localities, substantiating our species identification for the Korean populations. Inter-specific K2P distance between P. uchidai and an unidentified Saccocirrus sp. from Canada (based on public database entries) was 22.4-23.4%.