• Journal of Life Science
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• v.26 no.9
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• pp.1007-1014
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• 2016

A highly efficient, rapid, and reliable multiplex polymerase chain reaction based method for distinguishing ten species of genus Cynoglossus (C. senegalensis, C. abbreviates, C. macrolepidotus, C. arel, C. semilaevis, C. interruptus, C. joyneri, C. lingua, C. robustus, and C. monodi) is described. The species-specific primer sets were designed base on the cytochrome oxidase subunit I gene (1,500 bp). The optimal PCR conditions and primers were selected for ten of Cynoglossus species to determine target base sequences using single PCR. Multiplex PCR using the ten pairs of primers either specifically amplified a DNA fragment of a unique size or failed, depending on each species DNA. The length of amplification fragment of 208 bp for C. senegalensis, 322 bp for C. abbreviates, 493 bp for C. macrolepidotus, 754 bp for C. arel, 874 bp for C. semilaevis, 952 bp for C. interruptus, 1,084 bp for C. joyneri, 1,198 bp for C. lingua, 1,307 bp for C. robustus, and 1,483 bp for C. monodi with the species-specific primers, visualized by agarose gel electrophoresis, allowed perfectly distinction of the Cynoglossus species. The multiplex PCR assay can be easily performed on multiple samples and attain final results in less than 6 hours. This technique should be a useful addition to the molecular typing tools for the tentative identification of Cynoglossus species.

• Korean Journal of Ichthyology
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• v.29 no.1
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• pp.1-12
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• 2017

A natural hybrid of interspecific between the Coreoleuciscus splendidus and C. aeruginos (Cypriniformes: Cyprinidae) was captured in the Hwang-ji Stream, a tributary of the Nakdong River basin in Korea. An interspecific hybrid between C. splendidus and C. aeruginos was genetically identified based on morphological characteristics and the sequence analysis of nuclear recombination activating gene 1 (RAG1) gene (1,334 bp) and mitochondrial cytochrome c oxidase subunit 1 (CO1) gene (1,551 bp). As a result of morphological variations, the natural hybrid appeared to have an intermediate character between two parental species (C. splendidus and C. aeruginos) in three variations of black array (s) on dorsal, caudal and anal fin rays. Phylogenetic analysis inferred from RAG1 and CO1 sequence data revealed that Coreoleuciscus populations from Hwang-ji stream consist of two pure Coreoleuciscus species and a hybrid individual group. The individuals were clearly identified the cross and reciprocal hybrid by CO1 gene analysis. In RAG1 gene, 13 nucleotide variation loci were detected and the hybrid individuals displayed the double peaks of sequence chromatograms at the 9 diagnostic positions. In this study, molecular data and morphological variations were clearly demonstrated that hybridization did occur between C. splendidus and C. aeruginos. However, F2 hybrid generation and reproductive capacity of F1 hybrid individuals were not demonstrated.

• Korean Journal of Ichthyology
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• v.32 no.4
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• pp.199-209
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• 2020

In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.

• Parasites, Hosts and Diseases
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• v.58 no.1
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• pp.93-97
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• 2020

The cestode Taenia hydatigena uses canids, primarily dogs, as definitive hosts, while the metacestode larval stage cysticercus infects a range of intermediate hosts, including domestic animals such as goats, sheep, and pigs. Cysticercosis due to T. hydatigena has large veterinary and economic drawbacks. Like other taeniids, e.g., Echinococcus, intraspecific variation is found among the members of the genus Taenia. In Africa, few studies are available on the epidemiology and distribution of T. hydatigena, and even fewer studies are available on its genetic variation. In this study, we molecularly identified 11 cysticerci from sheep in Sudan and demonstrated the genetic variation based on the NADH dehydrogenase subunit 1 (nad1) and cytochrome c oxidase subunit 1 (cox1) mitochondrial genes. The isolates were correctly identified as T. hydatigena with more than 99% similarity to those in the GenBank database. Low diversity indices and insignificant neutrality indices were observed, with 3 and 2 haplotypes for the nad1 and cox1 genes, respectively. The results suggest the presence of unique T. hydatigena haplotypes in Sudan, as haplotypes with 100% similarity were not found in the GenBank database. With few available studies on the genetic variation of T. hydatigena in Africa, this report represents the first insights into the genetic variation of T. hydatigena in Sudan and constitutes useful data.

• The Korean Journal of Malacology
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• v.32 no.1
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• pp.55-62
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• 2016

In this study, an attempt has been made to analyze the morphology of Cephalopods distributed in Korea and collected samples from South-East Asian countries including Thailand, Indonesia, Vietnam, and China. A phylogenetic analysis was performed using the mitochondrial gene, Cytochrome c oxidase subunit I (COI) to understand the genetic divergences of the species and validate their origins. For achieving the objectives, samples were collected directly from Thailand Hat Yai, Songkhla, Indonesia Medan, Vietnam Ho Chi Minh, and Vung Tau in August 2015 and from China in September 2015. A total of 23 species of Cephalopods were identified falling under three orders, four familyies and nine genus. The species were distributed under Order: Octopoda (1 family, 3 genus, and 9 species), Order: Sepiolioda (1 family, 2 genus, and 8 species), and Order Teuthoidea (2 family, 4 genus, and 6 species). 23 species which is 1 family 3 genus 9 species in Octopoda, 1 family 2 genus 8 species in Sepiolioda, 2 family 4 genus 6 species in Teuthoidea. Phylogenetic analysis using COI gene was conducted for 18 species. For the remaining 5 species sequencing results showed severe variation and hence were not considered further. The COI phylogenetic analysis for the 18 species of Cephalopods were found consistent with the morphological identification. The excluded species will be subjected for a further detailed analysis.

• Parasites, Hosts and Diseases
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• v.58 no.6
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• pp.653-660
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• 2020

Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

• Korean Journal of Ichthyology
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• v.36 no.1
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• pp.94-100
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• 2024

During ichthyoplankton survey in the southwestern sea of Korea, we collected six individuals of noodlefish larvae and juveniles between April and May 2023. They were identified as Neosalanx anderssoni by mitochondrial DNA cytochrome c oxidase subunit I or 16S ribosomal RNA sequences, and their external morphological traits were described for the first time. All six individuals have a slender and elongated body. When preflexion and flexion larval stages (10.24 mm notochord length, NL and 15.47 mm total length, TL, respectively), oval-shaped black melanophores were distributed in a row along the ventral side of the gut. However, when postflexion larval and juvenile stages (23.58~25.90 mm TL, and 29.20~31.26 mm TL, respectively), melanophores on the ventral side of the gut were disappeared, and dark spot-shaped melanophores appeared along the dorsal side of the gut in a single row. Also, from the postflexion larval stage (23.58 mm TL), two large black spots began to appear symmetrically on the caudal fin. Our results suggest that N. anderssoni may use coastal area as spawning and/or nursery ground unlike previous study (Kim and Park, 2002).

• Journal of Food Hygiene and Safety
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• v.29 no.4
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• pp.347-355
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• 2014

In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

• Korean journal of applied entomology
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• v.59 no.2
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• pp.93-107
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• 2020

Adults of seven noctuid potential pests (Spodoptera frugiperda, S. litura, S. exigua, Ctenoplusia agnata, Mythimna loreyi, Athetis dissimilis, and A. lepigone) of soybean and maize in Suwon, Korea were identified by their morphological characteristics in the wing pattern and male genitalia and partial mitochondrial DNA sequences of cytochrome c oxidase subunit 1 gene. The generation number of adults that emerge annually in six species (except A. lepigone) was estimated from the data on density fluctuations of adults caught in sex pheromone traps in 2019 and the forecasted data using temperature-associated development and reproduction models for those species. S. frugiperda adults were caught from July 27th to October 31st in 2019, and hence were initially estimated to emerge three times per year. But, it was finally expected that S. frugiperda adults could possibly emerge a total of four times per year in Suwon, considering larval emergence observed during mid- and late June in other areas. Adult emergence of S. litura, S. exigua, C. agnata, and M. loreyi in 2019 was observed from May 29th to November 6th, from May 14th to November 6th, from May 26th to October 25th, and from May 31st to November 23rd, respectively. Annual adult emergence of these four species was estimated as at least four times. Adults of A. dissimilis were caught from May 26th to September 11th in 2019, and adult emergence was estimated at only twice per annum. It was postulated that the first adult populations of five species except the two Athetis species were probably migrated from other areas.