• Journal of fish pathology
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• v.36 no.2
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• pp.403-408
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• 2023

Mullet is an important marine aquaculture fish species in Korea, with a total of 7,237 tons produced as of 2022, making it the 5th most produced marine aquaculture fish species. In this study, ectoparasites presumed to be isopods were discovered in the fins of farmed flathead grey mullet (average weight 550 g), and the characteristics of the parasites were confirmed. The length of the parasite was 5 to 18 mm, and 3 to 7 parasites were infected per fish. To analyze the characteristics of the parasites, molecular biological identification and phylogenetic analysis were performed using the cytochrome c oxidase subunit I (COI) gene, and it was confirmed to be most closely related to Nerocila japonica in the Cymothoidae family. To confirm the parasite control effect, a direct exposure drug sensitivity test was conducted on five types of aquatic drugs and fresh water, trichlorfon was confirmed to be effective.

• 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.

• 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%.

• The Plant Pathology Journal
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• v.40 no.2
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• pp.171-191
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• 2024

Identification of Helicotylenchus species is very challenging due to phenotypic plasticity and existence of cryptic species complexes. Recently, the use of rDNA barcodes has proven to be useful for identification of Helicotylenchus. Molecular markers are a quick diagnostic tool and are crucial for discriminating related species and resolving cryptic species complexes within this speciose genus. However, DNA barcoding is not an error-free approach. The public databases appear to be marred by incorrect sequences, arising from sequencing errors, mislabeling, and misidentifications. Herein, we provide a comprehensive analysis of the newly obtained, and published DNA sequences of Helicotylenchus, revealing the potential faults in the available DNA barcodes. A total of 97 sequences (25 nearly full-length 18S-rRNA, 12 partial 28S-rRNA, 16 partial internal transcribed spacer [ITS]-rRNA, and 44 partial cytochrome c oxidase subunit I [COI] gene sequences) were newly obtained in the present study. Phylogenetic relationships between species are given as inferred from the analyses of 103 sequences of 18S-rRNA, 469 sequences of 28S-rRNA, 183 sequences of ITS-rRNA, and 63 sequences of COI. Remarks on suggested corrections of published accessions in GenBank database are given. Additionally, COI gene sequences of H. dihystera, H. asiaticus and the contentious H. microlobus are provided herein for the first time. Similar to rDNA gene analyses, the COI sequences support the genetic distinctness and validity of H. microlobus. DNA barcodes from type material are needed for resolving the taxonomic status of the unresolved taxonomic groups within the genus.

• Korean Journal of Ichthyology
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• v.29 no.4
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• pp.244-251
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• 2017

A total of 15 larvae [3.53~19.49 mm standard length (SL)] belonging to the family Bothidae collected from the southern sea of Korea in 2016 were identified as Psettina tosana based on 434 base-pair sequences of mitochondrial DNA cytochrome c oxidase subunit I. Larvae of Psettina tosana have anterior-most two elongated dorsal fin rays. Uniserial melanophores present on the dorsal and anal fin base, whereas melanophores on the body absent. An inflection point in the relative growth of head length and head depth against SL was shown between 9.93 mm and 10.73 mm SL. The examined larvae of Psettina tosana are clearly distinguished from the most similar species, Psettina iijimae in having no melanophore patches in the proximity of dorsal and anal fin base.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.6
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• pp.829-836
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• 2017

This study investigated the species composition and abundance of floating fish eggs to determine the timing and location of spawning of fish inhabiting the coastal waters of Sagye, Jejudo Island. Eggs were collected with a Bongo net bimonthly from May 2009 to February 2010. Identifications were based on nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Eggs were determined to belong to 43 distinct taxa, 35 of which were identified to the species level. The assemblage spanned eight orders, 23 families, and 32 genera. The number of taxa collected varied from month to month, with 14 taxa (12 species) found in June 2009, 11 taxa (10 species) in October 2009, 10 taxa (nine species) each in August 2009 and February 2010, eight taxa (six species) in April 2009, and five taxa (four species) in December 2009. Five abundant species (Branchiostegus japonicus, Engraulis japonicus, Pseudolabrus sieboldi, Goniistius zonatus, and Halichoeres tenuispinis) together represented 52.8% of the total number of eggs collected during the study.

• Development and Reproduction
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• v.23 no.4
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• pp.367-375
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• 2019

Pufferfish (Takifugu spp.) are economically important edible marine fish. Mistakes in pufferfish classification can lead to poisoning; therefore, accurate species identification is critical. In this study, we used the mtDNA cytochrome c oxidase subunit I gene (COI) to design specific primers for six Takifugu species among the 21 domestic or imported pufferfish species legally sold for consumption in Korea. We rapidly and simultaneously identified these pufferfish species using a highly efficient, multiplex polymerase chain reaction (PCR) system with the six species-specific primers. The results showed that species-specific multiplex PCR (multiplex species-specific polymerase chain reaction; MSS-PCR) either specifically amplified PCR products of a unique size or failed. MSS-PCR yielded amplification fragment lengths of 897 bp for Takifugu pardalis, 822 bp for T. porphyreus, 667 bp for T. niphobles, 454 bp for T. poecilonotus, 366 bp for T. rubripes, and 230 bp for T. xanthpterus using the species-specific primers and a control primer (ca. 1,200 bp). We visualized the results using agarose gel electrophoresis to obtain accurate contrasts of the six Takifugu species. MSS-PCR analysis is easily performed and provides identification results within 6 h. This technique is a powerful tool for the discrimination of Takifugu species and will help prevent falsified labeling, protect consumer rights, and reduce the risk of pufferfish poisoning..

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.317-324
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• 2012

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

• Journal of Life Science
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• v.27 no.11
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• pp.1331-1339
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• 2017

DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

• 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.