• Korean Journal of Ichthyology
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• v.34 no.3
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• pp.208-217
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• 2022

We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.269-276
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• 2010

Total mercury and methylmercury concentrations were determined in 15 commonly consumed aquatic food species using total mercury analyzer and gas chromatography with electron capture detector. The mean total mercury and methylmercury concentrations (mg/kg) were 0.088 and 0.034 in mackerel, 0.061 and 0.016 in hair tail, 0.030 and 0.005 in yellow croaker, 0.032 and 0.008 in Alaska pollock, 0.059 and 0.023 in eastern catfish, 0.110 and 0.045 in snakehead, 0.030 and 0.011 in Japanese common squid, 0.026 and 0.009 in common octopus, 0.035 and 0.008 in swimming crab, 0.009 and not detected (ND) in oyster, 0.011 and ND in shortneck clam, 0.008 and ND in mussel, 0.018 and ND in sea mustard, 0.007 and ND in nori, and 0.019 and ND in sea tangle, respectively. The total weekly dietary intakes of total mercury and methylmercury were estimated, respectively, using food consumption data from diet surveys and the concentrations of total mercury and methylmercury from this study. They were $0.178\;{\mu}g/kg$ body weight (b.w.)/week (3.57% of provisional tolerable weekly intake (PTWI)) and $0.052\;{\mu}g/kg$ b.w./week (3.34% of PTWI) respectively, and all were within their respective PTWI set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA). Therefore, considering that the main contributor to mercury intake in the diet is aquatic foods and that the 15 aquatic food species examined in this study are highly consumed, it is concluded that the mercury levels in the foods measured in this study do not present a concern for consumer health.

• Journal of fish pathology
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• v.9 no.1
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• pp.21-31
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• 1996

Concerned to the Ichthyophthiriasis of aquacultural fishes, the developmental features of Ichthyophthirius multifiliis were studied in cultured Korean catfish, Silurus asotus, and rainbow trout, Oncorhynchus mykiss. In the morphological observation, the parasite developed on two kinds of parasitic and non-parasitic phases with 6 developmental stages termed phoront, trophont, protomont, tomont, tomite, and therone. The $60{\times}40-100{\times}70{\mu}m$ fusiform or spherical phoront for the invading stage has 34-38 meridional kineties and begins to develope buccal apparatus. The 80-$800{\mu}m$ spherical or amoeboid trophont for the vegetative stage has a horseshoe shape macronucleus, a inconspicuous cytostome and developmental contractile vacuoles. The 200-$800{\mu}m$ spherical protomont for the encysting stage has a inconspicuous macronucleus, abundant contractile vacuoles; and a fine gelatinous exocyst is exuded, the baccal apparatus begins to resorb. The tomont for the encysted dividing stage has a thick cyst wall, and the buccal apparatus is resorbed completely. A small 35-$50{\mu}m$ spherical tomite for each daughter cell has a cytostome end the conspicuous oral apparatus. The $25{\times}20-60{\times}40{\mu}m$ fusiform theront for the infective stage possesses a perforatorium in the anterior end, a cytostome in the mid-point respectively and has 34-38 meridional kineties. In the experiments of the reproductive, the excysted time is related to water temperature. Tomitogenesis takes 10-14 hours at $28^{\circ}C$, 12-15 hours at $26^{\circ}C$, 16-18 hours at $22^{\circ}C$, 24-28 hour at $18^{\circ}C$, 26-51 hours at $14^{\circ}C$, and 129 hours at $9^{\circ}C$ respectively.

• Korean Journal of Ecology and Environment
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• v.47 no.1
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• pp.53-61
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• 2014

The origin of particulate organic matter (POM) and food web structure were investigated in Shingu reservoir based on stable isotope analysis from pre-monsoon (July) to post-monsoon (September) 2007. According to the depth in Shingu reservoir, the $^{13}C$ and $^{15}N$ values of POM for pre-monsoon period were nonsignificant distinction, while it was significant variation after rainfall period. The ${\delta}^{13}C$ values of POM in premonsoon period ranged from -25.1‰ to -26.1‰ in whole water column, but the ${\delta}^{13}C$ values of POM in post-monsoon period showed relatively wide range between -23.2‰ and -27.5‰. The apparently lighter values (average -27.5‰) in near bottom water (7 m water depth) demonstrate that POM in high turbid water in post-monsoon period may be derived from the outside terrestrial plants (allochthonous) through heavy rainfall during the summer monsoon period. After rainfall period, $^{13}C$ and $^{15}N$ values of D. brachyurum showed -23.3‰ and 12.2‰, respectively, while B. longirostris showed -27.1% and 8.7%, respectively. It suggested that D. brachyurum mainly feed on POM in autochthonous organic matter pool, but B. longirostris mainly consumed POM in allochthonous organic matter pool after rainfall period. Carbon and nitrogen stable isotope ratios were markedly different among secondary consumers. The carp (C. carpio) and catfish (S. asotus) were in the higher trophic level and crucian carp (C. auratus) and mudfish (M. mizolepis) were in the lower trophic level. $^{13}C$ and $^{15}N$ values of Z. platypus didn't significantly changed between before and after rainfall period. But P. parva and C. auratus apparently changed the $^{13}C$ and $^{15}N$ values after rainfall period. It is suggested that P. parva and C. auratus seem to feed allochthonous food source while Z. platypus depend on autochthonous food source.

• 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 Ichthyology
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• v.35 no.2
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• pp.91-100
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• 2023

The early life history of Silurus microdorsalis living in Jahocheon Stream was studied by observing egg and morphological development. Live fish were captured in June 2018, then reared in a circulating filtration system under a 14L : 10D photoperiod with a water temperature of 18℃. To artificially induce spawning, females were injected with 0.5 mL of Ovaprim (Syndel, Nanaimo, BC, Canada) per kg of body weight, and males were injected with 10,000 IU/kg body weight of human chorionic gonadotropin. Approximately 15 h later, eggs were artificially inseminated by the dry method. Mature eggs were light pale yellow, which separated them from immature eggs. Fertilized eggs were 2.16±0.06 mm (n=8) in diameter and fully hatched at 181 h after fertilization. The fertilization rate was 63.1±2.2%, and 10.0±3.7% of the embryos were malformed at 18℃. The rates of development were 181 h at 18℃, 109 h at 21℃, and 76 h at 24℃. The larval size immediately after hatching was 4.64±0.22 mm (n=8), and the larvae displayed negative phototaxis at 1 day after hatching. The total larval length on 7 days after hatching was 12.47±0.53 mm, with 25~30 basal anal fin rays and 14~16 basal caudal fin rays observed. The total larval length was 14.13±0.51 mm on 9 days after hatching, and approximately 90% of the black endoplasmic reticulum was deposited on the head and body. The dorsal fin had formed, and a single basal body was observed. On 15 days after hatching, the total larval length was 16.69±0.31 mm; the number of basal caudal fin rays (18 poles) was an integer because 2 dorsal fin basal rays and 60~63 anal fin basal rays were observed. The total larval length was 28.96±1.10 mm on 50 days after hatching; the numbers of caudal fins (n=18), dorsal fins (n=3), pectoral fins (n=11), and anal fin basal rays (n=67~73) were integers.

• Korean Journal of Ichthyology
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• v.35 no.4
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• pp.224-235
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• 2023

South Korea is the only divided nation globally, marked by a military demarcation line establishing demilitarized and civilian control zones, ensuring national security. Consequently, these areas exhibit relatively minimal ecological disruption compared to other regions. However, the threat to safety persists due to the presence of unexploded ordnances and landmines, imposing significant constraints on ecological research. To address this, we conducted a comparative study utilizing eDNA analysis as a supplementary and alternative approach within three points of the "Road of Peace" - Inje, Yanggu, and Hwacheon courses, located within the civilian control zone. Direct surveys and indirect eDNA sampling were carried out in May, July, and September of 2022. Genetic material obtained from the samples underwent amplification, library preparation, MiSeq sequencing, and subsequent ASV generation for indirect analysis. These results were then compared with the findings of direct surveys. Our findings revealed the detection of eDNA for both observed species at the Yanggu-1 point, and for two out of four species at Yanggu-2. Hwacheon-1 displayed the detection of eDNA for one out of one observed species, whereas Hwacheon-2 yielded seven out of twelve, Hwacheon-3 showed four out of six, and all one observed species at Hwacheon-4 exhibited eDNA detection. Consequently, approximately 69% of the fish species identified through direct surveys were confirmed by indirect eDNA analysis. It is necessary to verify if certain fish species, such as the continental trout and catfish, have genetic information registered in the NCBI database. Additionally, it is believed that further marker development research utilizing different genetic sequences is essential. Given the limitations imposed by the hazardous nature of the surveyed civilian control zone, eDNA analysis proves to be a suitable supplement for fish research in the area.