• 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..

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.1
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• pp.1-7
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• 2009

The toxicity of two species of puffer fish, Takifugu poecilonotus (Heuinjeombok) and T. vermicularis (Gukmaeribok) collected from the coastal regions of Korea was determined using a mouse bioassay. In the T. poecilonotus collected in Jeju and Tongyeong, the proportion of toxic specimens containing ${\ge}10$ mouse units (MU) per gram exceeded 95% for the skin, liver, ovary, and fin, and approximately 30% for the testis and muscles. In each of the organs, the highest toxin levels were 79 MU/g in the muscle, hundreds (158-365) of MU per gram in the fin, intestine, testis, and gallbladder, but thousands (1,147-2,406) of MU per gram in the skin, liver, and ovary. In T. vermicularis collected from Incheon and Gunsan, the proportions of toxic specimens were 100% for the gallbladder, and 56-68% for the skin, fin, liver, and intestine however, no toxic muscle specimens were noted. The highest toxin scores were below 10 mouse units (MU) per gram in the muscle, 20-94 MU/g in the skin and fin, 319 MU/g in the intestine, and thousands (1,548-4,624) of MU per gram in the liver, gonad, and gallbladder. The toxicity in the muscle of T. vermicularis was deemed acceptable for human consumption, whereas the toxicities in the muscle of T. poecilonotus and the skin of both species of puffer fish were significantly high, such that special attention may be required when the fish is intended for human consumption.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.3
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• pp.353-358
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• 1998

Experiments were performed to find out the physico-chemical properties of milt, and morphological changes of cryopreserved spermatozoa in tiger puffer, Takifugu rubripes. The average number of sperm and spermatocrit in milt stripped were $9.81{\pm}0.34{\times}10^{10}/m{\ell}$ and $97.8{\pm}0.8$, respectively. While total lipid concentration from seminal fluid was higher than that from sperm, total protein concentration from sperm was higher than that from seminal fluid, Na and K concentrations in sperm and those in seminal fluid were similar each other, However, glucose from sperm and seminal fluid were not detectable. Spermatozoon of tiger puffer was consisted of head, middle Piece and tail. Size of head showing horseshoe shape was $0.65{\pm}0.10{\mu}m$ in diameter and $1.35{\pm}0.30{\mu}m$ in length. The head fully containing chromatin did not have acrosome. Mitochondrion in middle piece was $0.2{\mu}m$ in average diameter and flagellum showed 9+2 structure. A few of cryopreserved spermatozoa showed morphologically loose or swollen plasma membranes.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.168-172
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• 2007

In the present study, attempts were made to find out the physico-chemical properties of milt and the sperm motilities in various osmotic conditions using Panther puffer, Takifugu pardalis. The average concentration of sperm in the milt was $12.1{\pm}3.2{\times}10^9/mL$. pH and osmolality of seminal plasma were $8.2{\pm}0.3$, $385.5{\pm}12.5mOsm/kg$, respectively. Spermatozoa were immotile when the milt was mixed with solutions (electrolyte or non-electrolyte) of lower osmolality than the average seminal plasma osmolality ($385.5{\pm}12.5mOsm/kg$), but became motile after mixing milt with hyperosmotic solutions.

• Journal of Aquaculture
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• v.10 no.3
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• pp.273-279
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• 1997

Condition for fresh storage of tiger puffer in the liquid state were investigated in several experiments. When marine fish ringer solution and 1% NaCl were used as the diluent for the short-term preservation method, spermatozoa activity index (SAI) and survival rate showed the best result among the various diluents tested. The dilution rate for the shortterm preservation of spermatozoa was suitable between 3 and 5 times with the 1% NaCi diluent. The appropriate range of temperature for the short-term preservation showed between 0 and $5^{\circ}C$. In order to keep high SAI and survival rate of spermatozoa, antibiotic addition (800 ppm neomycin) could be suggested. These results indicated that the short-term preservation method could be employed in tiger puffer spermatozoa.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.4
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• pp.404-410
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• 2018

Takifugu rubripes, T. chinensis, T. pseudommus and an unidentified species were morphologically and genetically studied. Morphology was compared using external morphology and the genetic analysis was conducted using mitochondrial DNA cytochrome oxidase I (COI) and cytochrome b (Cyt b). The morphological characteristics of T. rubripes, T. chinensis and T. pseudommus were confirmed, as reported previously. The unidentified species was confirmed to have the characteristics of T. rubripes and T. chinensis. The COI sequences of the four species were 99-100% similar; genetic distance was d=0.0000 and Cyt b genetic distance was d=0.0000-0.00834. Species in the phylogenetic tree belonging to the same group could not be classified genetically. In conclusion, the unidentified species was considered to be a hybrid between T. rubripes and T. chinensis.

• Journal of Fisheries and Marine Sciences Education
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• v.29 no.2
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• pp.447-452
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• 2017

A closely associated set of characteristics was analyzed using both classical and truss dimensions to discriminate sexual dimorphism between spawning season and non spawning season in grass puffer, Takifugu niphobles. In non-spawning season, $1{\times}10/Ls$ of classical dimension was significantly different between male and female (P<0.05). In spawning season, $1{\times}9/Ls$ and $1{\times}10/Ls$ of classical dimension, $3{\times}8/Ls$, $3{\times}9/Ls$, $3{\times}10/Ls$, $4{\times}8/Ls$, $4{\times}9/Ls$ and $8{\times}9/Ls$ of truss dimension and $7{\times}9/HL$ of head part dimension were significantly different between male and female (P<0.05). The results of this study may be useful as basic information of other fish species to compare the change of sexual dimorphism between spawning season and non spawning season.

• Journal of fish pathology
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• v.20 no.2
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• pp.129-137
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• 2007

Skeletal deformity in hatchery-reared river puffer (Takifugu obscurus) fingerlings occurred. The vertebral abnormality was recognized with naked eyes 80 days after hatching. Affected fish had a good appetite but no clinical signs were found except the vertebral abnormality. As more than 90% of the hatchery-reared fish exhibited vertebral abnormality, the fingerlings could not be used for commercial seeds any more. Morphological changes in the vertebrate and the swim bladder were observed with a soft X-ray. Histopathological changes on the caudal muscle around the deformed vertebrae, gill, and spleen were also examined to clarify the cause of the deformity. Spinal curvature occurred between vertebrae 10 and 12, but any malfunction of the swim bladder was not found. The diameters of the muscle fibers around the deformed vertebrae were much smaller and more irregular than those in normal fish, and the gabs between the fiber bundles were greatly enlarged. No evidence of inflammation response was found in the muscle layer. In the hatchery, feed was putting at the position of water inflow, which might attract the fingerings to move toward the feed in spite of incomplete development of their caudal muscle fibers. From these results, it is suggested that the high speed of water current in rearing aquaria might be associated with the development of vertebral abnormality.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.3
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• pp.243-246
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• 1999

To obtain the basic data for the preservation of river puffer (Takifugu obscurus) sperm, experiments were carried out on the physico-chemical properties and cold storage of milt. The average number of sperm and spermatocrit in milt stripped were $1.13\pm0.34\times10^{10}/ml$ and 64.8$\pm$1.4, respectively. Osmolality of seminal fluid was 266$\pm$2 mOsm/kg, Total protein and total lipid from sperm were higer than that from seminal fluid. $Ca^{2+}$ and $Na^{+}$ concentrations were higher in the seminal fluid than in the sperm, while $Mg^{2+}$ and $K^{+}$ concentrations were lower in the seminal fluid. When sperm of river puffer were preserved in $0\pm0.5^{\circ}C$ with various diluents for 16 days, fertilization rate was $0\~0.7\%$. It suggested that cold storage of river puffer sperm was detrimental to sperm fertility.

• Journal of fish pathology
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• v.11 no.2
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• pp.119-127
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• 1998

Two species of the parasitc copepods, Clavellopsis hugu (Lemaeopodidae; Siphonostomatoida) and Taeniacanthus yamagutii (Taeniacanthidae; Poecilostomatoida) from cultured Korean fish, Takifugu obscurus, is described and reported for the first time in Korea. The parasite was recovered from the buccal cavity, nose and skin. Clavellopsis hugu is easily identified by the body shape, 2nd antenna, mandible and 2nd maxillae. In female, cephalothorax flexed dorsally, second antenna biramose, mandible with serrate blade, second maxillae completely fused at base of cephalothorax. In male, cephalothorax and trunk fused into an ovoid, two pairs of maxillipeds chelate. The most distinctive features of Taeniacanthus yamagutii are terminal process of second maxilla stout, maxilliped claw with conspicuous digitiform process at base and a setiform element at the tip of each exopod spine of legs 2-4. Both parasitic copepod species are new to the Korean fauna. The korean fish, Takifugu obscurus should be considered as an additional host of this species because these parasite were recovered from the western coast of Korea (Boryung Hatchery).