• Journal of Aquaculture
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• v.7 no.1
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• pp.41-54
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• 1994

In order to evaluate the availability of lacZ as a reporter gene for producing transgenic mud loach, foreign DNA, bacterial \beta-galactosidase$ gene (lacZ) was microinjected into mud loach eggs and its insertion and expression were examined. X-gal based histochemical assay, fluorimetric analysis of \beta-galactosidase$ with 4-methylumbelliferyl-$\beta$-D-galactoside (MUG) and molecular biological examination using polymerase chain reaction (PCR), dot blot, southern blot and sequence analysis of PCR products were carried out to analyze both microinjected group and non-injected controls. The results are disccussed in this paper.

• Journal of Aquaculture
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• v.8 no.4
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• pp.327-341
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• 1995

A Study was conducted by using a histological method to examine gonadal development in diploid and triploid mud loach, Misgurnus mizolepis. Shape of ovaries from both groups looked like a single mass however, size of triploid ovary was significantly smaller than that of diploid ovary. Testis of triploid was also similar that of diploid in shape, but it was smaller than that of diploid. Ovarian development and oogenesis in diploid were significantly more rapid than those process in triploid, and first matured eggs were observed 100-days after hatching (SL, 5.68cm). Triploid ovary from hatching to 9-month-old had external appearance of undeveloped gonad and a few of oocytes of perinucleolus stage. However, normal matured eggs like those of female diploid were observed in one 5year-old triploid fish ovary examined. Although testicular development and sperrnatogenesis of diploid were prosperous sexually, testicular development of triploid were rather retarded compare to their male diploid counterpart in spermatogenesis.

• Korean Journal of Ichthyology
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• v.28 no.2
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• pp.72-78
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• 2016

The age and spawning characteristics of Iksookimia pacifica were investigated in this study to provide baseline data for its ecological characteristics in the Bukcheon (stream), Goseong-gun, Gangwon-do, Korea from January to December, 2013. Age groups of I. pacifica (female) estimated by the frequency distribution of total length in the spawning season indicated that the 30~49 mm group is 1 year old, the 50~74 mm group is 2 years old, the 75~99 mm group is 3 years old, the 100~124 mm group is 4 years old, and the 130~160 mm group is $5{\leq}$years old. The sex ratio (♂/♀) was 0.63, and females were 30~60 mm larger than males. The lamina circularis at the base of pectoral fins in males as a secondary sexual character was created at the 13th month after hatching. The spawning season according to the gonadosomatic index (GSI) was July to August (main spawning season in July) with water temperatures of $20{\sim}25^{\circ}C$. The average number of eggs was $2,503{\pm}1,337$ with a diameter of $1.11{\pm}0.04mm$ in mature eggs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.1
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• pp.16-23
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• 1987

The cyprinid loach, Misgurnus mizolepis $G\ddot{u}nther$, a freshwater teleost, is a member of the family Cobitidae and is known to distribute in Korea and China. This fish is considered to be an excellent food fish in Korea. This report is a description of the gross morphology of the embryos and larvae up to the development of juvenile period of life. The eggs were obtained by injecting gonadotropin and fertilized by wet method in the laboratory on July 10 in 1986. The inseminated eggs were rinsed with tap water and incubated at $25^{\circ}C$. Live, freshly laid eggs were separative, demersal, adhesive, light yellow and average 1.1mm in diameter. The hatching took place in about yellow and average 1.1mm in diameter. The hatching took place in about 24 hours after fertilization. The newly hatched larvae were average 2.7 mm in total length possessing yolk sac and 35-36 myotomes. Ten hours after hatching, the prelarvae attained 3.5 mm in total length. The first heart pulse and melanophores at the side of the body were observed at this stage. Yolk sac was completely reserved at four days after hatching. The larvae that was cultured 24 days after hatching, 15.4-16.4 mm in total length, entered the juvenile period of life. All the un rays were already peresent; D. III 6, A. III, 5, P.10, V. 4-5, C.14.

• Korean Journal of Ichthyology
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• v.25 no.2
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• pp.65-73
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• 2013

Egg development and early life history of the Korean spine loach, Iksookimia pumila was investigated to provide basic information regarding biological characteristics and restoration. Adult fish were sampled using spoon net in the Baek Stream, Sangseo-myeon, Buan-gun, Jeollabuk-do, Korea, 7 July 2010. Eggs and sperms were obtained from the females and male with Ovaprim injecting (0.5 mL/kg) and then fertilized using the dry method in the laboratory. Number of spawned eggs were 1,107 (352~1,440). Spawned eggs were slightly adhesive, light yellowish coloring and measured $1.3{\pm}0.04$ mm (mean${\pm}$SD) in diameter. Spawned eggs hatched out 52 (47~55) hours after fertilization at water temperature of $23^{\circ}C$, and newly hatched larvae an average were $4.7{\pm}0.14$ mm in total length. At 5 days after hatching, larvae averaged $7.1{\pm}0.20$ mm in total length and their yolk sacs had been completely absorbed. Beginning at 17 days after hatching, fish entered the juvenile stage and reached $11.0{\pm}0.50$ mm in total length. At 100 days after hatching, the band patterns and external form of juvenile fish were similar to those of adults, and they averaged $31.3{\pm}3.98$ mm in total length.

• Korean Journal of Ichthyology
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• v.26 no.2
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• pp.89-98
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• 2014

Eggs development and early life history of the endangered Korean dwarf loach, Kichulchoia brevifasciata (Teleostei: Cobitidae) was investigated to provide basic information regarding biological characteristics and restoration. Adult fish specimens were sampled using a spoon net at Geurnsan-myeon, Goheung-gun, Jeollanam-do, Korea from June to July 2011. Since, spawning characteristics were analyzed, and females were induced to spawn by injecting Ovaprim (0.5 mL/kg) and their eggs were artificially fertilized with sperms by the dry method in the laboratory. Total length of mature female were 46~76 mm with GSI $9.6{\pm}3.77%$, and total length of mature male was 42~52 mm with GSI $3.5{\pm}1.04%$. Sex ratio (♂/♀) was 0.10, and there were no secondary sexual characteristics. The number of mature eggs was averaged $60{\pm}28.7$ per female. The lemon yellow eggs were slightly adhesive $1.46{\pm}0.07mm$ in diameter. The embryo hatched approximately 66 h after fertilization at $25^{\circ}C$, and the hatched larvae were averaged $5.5{\pm}0.07mm$ in total length (TL). At 6 days after hatching, the larvae averaged $9.0{\pm}0.29mm$(TL) and their yolk sac was completely absorbed. At 17 days after hatching, they entered the juvenile stage and reached $12.6{\pm}0.24mm$ (TL). At 80 days after hatching, the band patterns and external form of the juveniles were similar to those of adults, and they averaged $33.0{\pm}2.19mm$(TL).

• Korean Journal of Environment and Ecology
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• v.35 no.3
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• pp.285-293
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• 2021

This study was conducted to elucidate spawning period characteristics and early life development of eight barbel loach, Lefua costata (Balitoridae) at the Jusucheon stream, Okgye-myeon, Gangneung-si, Gangwon-do, Korea between January and December in 2018. The spawning period was estimated to be from May to August, considering the change in the gonad-somatic index, the appearance of young fry, and frequency distribution in egg diameter. It was a multi-spawning type. The gender ratio was 1:0.79 with 1,117 females and 879 males collected. The egg size was 0.24-0.93 mm, with mature and immature eggs found during the spawning period. The size of mature eggs was 0.71±0.02 mm, and the average number of fecundity was 1,786±818 (n=31). Observation of the egg development showed that the fertilized eggs were the sticky, gray, segregated, and demersal type with 0.76±0.03 mm in diameter. The hatching of the embryos began at about 34 hours (hatching rate 50%) after fertilization underwater temperature of 25℃. The average length of the newly hatched pre-larvae was 2.7±0.11 mm. The average length of pre-larvae at 4 days after hatching was 4.5±0.16 mm, and the yolk sac was completely absorbed and entered the post-larvae stage. At 20 days after hatching, the average length of post-larvae was 11.5±0.67 mm, and their fin rays were formed before they transitioned to the juvenile stage. At 100 days after hatching, the average length reached 49.8±2.60 mm, and the appearance and the lateral sideband patterns were similar to those of the adult fish.

• Korean Journal of Ichthyology
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• v.34 no.1
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• pp.25-33
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• 2022

We investigated to elucidate the early life history of siberian stone loach, Barbatula nuda (Pisces: Nemacheilidae). Adult fish were collected using net in the Sangcheoncheon Stream, Cheongpyeong-myeon, Gapyeong-gun, Gyeonggi-do, Korea, 26th April 2020. HCG 10 IU/g was injected to female and male fish and fertilized by dry method in the laboratory. The number of eggs of B. nuda was 1,308±293 (TL: 70.64~79.24 mm), and the egg color was yellowish grey. Fertilized eggs measured 1.13±0.01 mm (n=10) in diameter and showed adhesion. The fertilized eggs started hatching after 104 hours. At the time of egg development, the 15℃ group showed an average hatching rate of 80.1±1.2% and the 20℃ group showed 61.5±6.8%, so the 15℃ group was suitable (p<0.05). Newly hatched larva an average were 3.12±0.06 mm (n=10) in total length. At 4 days after hatching, the total length was 4.89±0.09 mm (n=10), and feeding began with the mouth and anus opened, and entered to preflexion larvae stage. At 21 days after hatching, the total length was 10.98±0.40 mm (n=10), and the distal part of the notochord was bent to 45°, and entered to postflexion larvae stage At 26 days after hatching, the total length was 13.47±0.37 mm (n=10), and entered the juvenile stage.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.38-38
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• 2003

Blocking the first mitotic cleavage of the zygote is a key tool for chromosome-set manipulations in fish. We developed an improved method for inducing tetraploidy by blocking the mitosis with a combination of heat shock at 40.5$^{\circ}C$ for 1, 2 or 3 min followed by cold shock at $1.5^{\circ}C$ for 30, 45 or 60 min. When applied during the first cleavage metaphase of mud loach (Misgurnus mizolepis) zygotes, the optimal combination was heat for 2 min followed by cold for 45 min. At 1 month, the frequency of 4N survivors and the yield from total eggs fertilized was 55.7% and 14.4%, respectively, compared to heat shock alone with 20.0% efficiency and 3.6% yield. The effectiveness of the procedure was confirmed by diploid mitotic gynogenesis using transgenic markers. The overall yield of homozygous diploids, 34.0%, was better than that for single heat shock, 17.3%. The tetraploids and homozygous diploids had higher early mortality than normal diploid controls. However at 1 month, the viability of the tetraploids was the same as normal diploids. For gynogenetic diploids, the survival was similar to normal diploids after 3 months. The high efficiency of this new protocol extends the opportunity to study polyploidy in basic and applied research.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.40-40
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• 2003

DNA content of the sperm of tetraploid mud loach (Misgurnus mizolepis) males and the ploidy status of progenies generated by crossing tetraploid males with diploid females are described. Reproductive performance of the induced adult tetraploid males ranged from sterility to fertility. Of 48 tetraploid males tested, 12 were sterile but the other 36 produced functional sperm. Of these 36, 26 produced haploid sperm, which on fertilizing the haploid eggs, generated diploid progenies. Seven tetraploid males were mosaics in their sperm, as indicated by the production of diploid, aneuploid and/or triploid offspring. Only 3 males produced diploid sperm rendering the production of all-triploid progenies. The DNA content of sperm of a tested tetraploid male was consistent throughout the 3 progeny tests, i.e. the haploid sperm-producing 4n males persisted to produce the haploid sperm only likewise the diploid sperm producing4n males consistently produced the diploid sperm only, when progeny testing was extended to 3 successive but alternate years. Hence, a careful and direct examination of the DNA profile of sperm from tetraploid males is a pre-requisite for reproductive containment of genetically modified fish.