• Korean Journal of Ichthyology
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• v.31 no.1
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• pp.23-29
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• 2019

The embryonic development and hatchability of the artificially fertilized eggs of convict grouper (Hyporthodus septemfasciatus) ♀${\times}$giant grouper (Epinephelus lanceolatus) ♂ (CGGG) were compared to those of the maternal pure species (convict grouper ♀${\times}$♂, CG) to establish a novel grouper hybrid for aquaculture industry. The fertilized eggs were divided into nine 5-L beakers (3,000~5,000 eggs/beaker) filled with UV sterilized seawater and incubated at a temperature range of $23.5{\sim}24.8^{\circ}C$ (32.1~32.8 psu). The percentages of fertilization and hatching of CGGG were $69.4{\pm}1.5%$ and $59.0{\pm}5.1%$, respectively and were significantly lower than those of CG (p<0.05). The CGGG proceeded normal embryonic development similar to that of CG, but showed an irregular cleavage, immature embryonic body and spinal deformity in hatched larvae. The incubation time from fertilization to hatching of CGGG was 31 hrs, which was approximately 2 hrs slower than that of CG. Our study provided the possibility of mass production of grouper hybrid CGGG larvae.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.123-128
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• 1998

Implantation rates remain low following human in vitro fertilization (IVF). Suboptimal culture conditions may limit the ability of embryos to hatch as blastocysts, and artificial opening of the zona pellucida has been proposed as a means to promote subsequent hatching (assisted hatching). In this study, assisted hatching (AH) by zona drilling using acidic Tyrode's solution was performed in 320 patients, due to their age of more than 38 years (group A), the thick zona pellucida (group Z; $ZP\geq0.18{\mu}m$), and failures in implantation more than 3 times in previous IVF-ET trial (group P). This study was designed firstly, to study the effects of AH on the outcomes of IVF-ET according to the indications and secondly, to verify the appropriate application of AH. The results were as follows; 1. There was no difference in pregnancy rate between AH group (26.6%) and non-AH group (26.5%). 2. Assisted hatching (AH) showed significantly higher pregnancy rate of the patients with thick zona pellucid a than those of the patients with age factor and with the history of repeated implantation failure. But in the patients with age factor only, AH resulted in higher pregnancy rate. 3. Interestingly, the patients with complex factors including zona factor (Z: 33.9%; ZA: 30.4%; ZP: 31.6%; ZAP: 21.4%) showed higher pregnancy rates than other complex factors excluding zona factor (A: 24.4%, P: 0%; AP: 10.8%). From these results, AH is more helpful to the patients with thick zona pellucida rather than patients with older age and/or previous repeated implantation failure.

• Journal of Aquaculture
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• v.8 no.3
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• pp.241-249
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• 1995

Sperm from mud loach (Misgurnus mizolepis) were electroporated in the presence of plasmid DNA, pRSV/luc or pMT/hGH over a range of field strength of 0-1,625 V/cm with capacitance from 0 to 1,000 ${\mu}F$, and the effects of electroporation on fertilization, hatching, early survival, and efficiency of gene transfer were investigated. Average fertilization rate, hatching rate and early survival rate up to yolk sac absorption of all experimental groups were not significuntly different (P>0.05). The proportion of fish carrying pRSV/luc based on the polymerase chain reaction (PCR) analysis was ranged from 0 to $20\%$, however, the values of gene transfer efficiency from the different eledctroporation conditions were not significantly different. PCR analysis of pMT/hGH transferred groups revealed that screening of pMT/hGH transferred fish by PCR was difficult because of significant nonspecific amplifications resulted from the homologous sequences in the genome of mud loach.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.247-252
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• 1994

Intracytoplasmic sperm injection(ICSI) was known as effective method in treatments of couples who unable to be helped by conventional in vitro fertilization. In 78 treatment cycles of 78 infertile couples using ICSI performed at our infertility clinic between May and August 1994 were analyzed. These patients were classified two groups, andrological factor(AF) and non-andrological factor(non-AF) group. The AF group, which had abnormal sperm physiology, included oligozoospermia, asthenozoospermia, oligoasthenoteratozoospermia(OATS) and microsurgical epididymal sperm aspiration(MESA) patients. The non-AF group, which had abnormal oocyte physiology, included abnormal zona pellucida, poor quality of oocyte and immune factor infertile patients. A single spermatozoon was injected into the ooplasm of 776 metaphase II oocytes. The fertilization rate was 44.6%(346/776) and 319 embryos were transferred. After 73 embryo transfers(93.6% of treatment cycles) 23 pregnancies were estabilshed, i. e. pregnancy rate of 29.4% per started cycle and 31.5% per embryo transfer. Fertilization rate of AF and non-AF group was 46.2% and 35.8%, pregnancy rate was 34.5%(20/58) and 20.0%(3/15), respectively. In order to increase the pregnancy rate, assisted hatching(AHA) has done after lCSl in 47 treatment cycles. Pregnancy rate of ICSI with AHA and without AHA group was 34. 0% (16/47) and 26.9%(7/26), respectively. ICSI was more effective in andrological factor infertility and the pregnancy rate was increased by ICSI with AHA procedure.

• Development and Reproduction
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• v.22 no.3
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• pp.289-295
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• 2018

Large quantity of eggs fail to be fertilized and many of fertilized eggs are unable to hatch in the eel, Anguilla japonica. Larvae of eel absorb egg yolk up to 8 days after hatching but the majority of hatched larvae die before they reach the stage of first feeding in this species. Genes of key enzymes for yolk processing (cathepsin B, D, L and lipoprotein lipase - abbreviated as ctsb, ctsd, ctsl and lpl, respectively) could be associated with egg quality. In this study, we investigated differences in the expression of these genes between floating eggs and sinking eggs, and also the relationship between the gene expressions of the enzymes and fertilization rates in the fertilized eggs obtained from artificially matured female eels. Expressions of yolk processing enzyme genes did not show significant difference between floating and sinking egg groups. Expression of ctsb decreased when fertilization rate was high. Expression of ctsd, ctsl and lpl, however, did not show any significant differences. These results suggest that ctsb expression could be an indicator of egg quality, and that some proteins prone to be digested by ctsb could be very important in the process of fertilization and normal cleavage in this species. Further study should identify these critical proteins to improve our understanding on the quality of fish eggs.

• Journal of Aquaculture
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• v.8 no.3
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• pp.159-170
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• 1995

Triploid abalone, Haliotis discus hannai, was induced by the cold $(0^{\circ}C\;and\;3^{\circ}C)$ or the heat $(35^{\circ}C\;and\;40^{\circ}C)$ shock procedure with fertilized eggs, 12 min. or 32 min. post fertilization with the various time intervals of shock duration. Fertilization rate of each experimental group was not significantly different from that of corresponding diploid control (P>0.05). However hatching rates and normality rates of triploid larvae were significantly different from those of corresponding diploid control (P <0.05). In heat shock groups at $40^{\circ}C$, fertilization rate of eggs was extremely low $(0\~2.7\%)$ and hatched larvae were not detected in these treatment groups. Incidence of triploidy was confirmed by chromosome count and the highest rates of triploid $(84.0\%)$ revealed in cold $(3^{\circ}C)$ shock with 15 min. treatment duration 12 min. after fertilization. The number of diploid chromosome was 2n: 36, and that of tiploid was 3n=54.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.1
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• pp.2-8
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• 2021

Dimethachlor is a synthetic herbicide, belonging to the chloroacetanilide group, that inhibits the undesirable growth of weeds via the suppression of very long-chain fatty acid synthesis. Although dimethachlor has been shown to run off from agricultural fields into aquatic ecosystems, the toxicity of dimethachlor on aquatic invertebrates and vertebrates is unknown. In our study, we assessed the toxicity of dimethachlor on developing zebrafish embryos by analyzing viability, hatching ability, and phenotypic changes. Embryonic viability decreased from 48 h post-fertilization (hpf) at the highest concentration of dimethachlor. Decreased hatching ratio, shortened body length, and pathological changes in the eye, heart, and yolk sac were observed at sub-lethal concentrations. Additionally, dimethachlor increased the number of apoptotic cells and level of reactive oxygen species 120 hpf. Our results indicate that dimethachlor may act as an anti-developmental toxicant when accumulated in an aquatic environment.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.4
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• pp.497-502
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• 1996

Artificial fertilization of ice fish, Mypomesus transpaciticus nipponensis caught at Milyang-river and Osib-chun brook was performed in March 24, 1990, and the hatched larvae were reared for 25 days to describe the development of eggs and larvae. Fertilized eggs were spherical in shape, measuring $0.85\~1.05\;mm$ in diameter (mean: 0.97 mm) and translucent adhesive with many small-sized oil globules on the surface. Hatching in the indoor tank started from the 170 hours after fertilization under $16.5^{\circ}C$ water temperature. Newly-hatched larvae were measured $3.85\~4.25\;mm$ in total length (mean: 4.05 mm), and mouth and anus were not yet open. They had one yolk sac on the anterior part of abdomen, straight-type's notochord, and $52\~54$ myotomes. The larva of 5 days old transformed to postlarval stage and measured $5.20\~5.65\;mm$ (mean: 5.37 mm) in total length. As the yolk sac was completely absorbed, mouth and anus were open, and they fed rotifers vigorously. In 20 days after hatching, the larvae grew to 8.38 mm in TL, and the caudal notochord flex at $45^{\circ}$. In 25 days after hatching, total length reached 9.63 mm. The pan of the fin-fold of the future dorsal and anal fins became high.

• Journal of Aquaculture
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• v.10 no.4
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• pp.463-472
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• 1997

Floating and hatching rates, size and survival activity index(SAI) of spawned eggs of red spotted grouper, Epinephelus akaara were studied to understand egg qualities during the spawning periods in 1992, 1993 and 1995. The fish spawned from mid-July to mid-August when temperature ranged 23~$28^\circC$. Floating, fertilization and hatching rates were higher in the eggs from early to middle than those from late during the spawning period. SAI ranged 1.8~9.6, and it was closely related to survival rate until mouth operning time (SMOT). Egg diameter decreased gradually with the time of spawning, while diameter of oil globule was constant during the spawning periods. Induced spawnings were carried out with one injection of HCG over 5, 000 IU/kg body weight. Floating, fertilization and hatching rates of the induced spawning eggs were higher than those of untreated ones. Thus, induced spawning method with HCG was recommended for collecting fertilized eggs in bulk. Optimal time for the collection of high quality eggs was supposed to be from early to middle during the spawning periods. Considering that egg qualities were vulnerable within the period, hatchability, incidence of malformed larvae, SMOT and SAI were factors to be considered together to get good quality of eggs.

• Korean Journal of Ichthyology
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• v.22 no.1
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• pp.25-33
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• 2010

Effects of different salinity levels on spawning performance, embryonic development and early viability of a euryhaline medaka species, Oryzias dancena, were examined. O. dancena were able to spawn eggs in a wide range of salinity from 0 to 70$^{\circ}/_{\circ\circ}$, however, the spawning frequency was lowered in complete freshwater (0$^{\circ}/_{\circ\circ}$) and in highly salted water (70$^{\circ}/_{\circ\circ}$). Fertilization success was negatively affected when the environmental salinity was higher than the salt concentration found in normal seawater. Embryonic viability and hatching success were also inversely related with the salinity levels. Typical abnormality was observed in developing embryos incubated at high salinities (30, 45 and 60$^{\circ}/_{\circ\circ}$). In addition, the time to hatch was significantly delayed with increasing salinities: peak hatching occurred at 12~14 days post fertilization (dpf) in freshwater and at least at 17 to 18 dpf in 60$^{\circ}/_{\circ\circ}$. Mean survival rates of the hatched larvae up to 7 days post hatching (dph) were at least 97% in salinity levels ranging from 0 to 30$^{\circ}/_{\circ\circ}$. However, larvae reared in 45 and 60$^{\circ}/_{\circ\circ}$ experienced significant mortality, especially in the early phase, resulting in only 75% and 64% survival rates up to 7 dph, respectively.