• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.2
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• pp.160-163
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• 2001

This study was performed to investigate the effect of vitamin $B_{12}$ addition on the production and hatching rate of resting egg of freshwater rotifer, Brachionus calyciflorus. Different concentrations of vitamin $B_{12}$ was added into concentrated freshwater Chlorella before fed to rotifer and rotifer was cultured in 250 mL flask. The maximum density of rotifer was 138 inds./mL at 6 ppm and it increased with an increase of vitamin $B_{12}$ addition. Mixis rate and fertilization rate of rotifer were $63.0\%$ and $89.6\%$, respectively, at 6.0 ppm. The production of resting eggs per mL was 231 eggs/mL at 1.5 ppm, but there was no significant difference among vitamin $B_{12}$ addition levels. The highest production of resting eggs per 10,000 rotifers and Chlorella dry weight (g) were 11,289 eggs and 121,252 eggs, respectively, at the 6 ppm. The hatching rate of resting egg was not significantly different among vitamin $B_{12}$ addition levels, but significantly higher than the control (without addition of vitamin $B_{12}$ ). These results showed that vitamin $B_{12}$ addition could affect production and hatching rate of resting egg of freshwater rotifer, B. calyciflorus.

• Environmental Analysis Health and Toxicology
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• v.24 no.2
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• pp.119-125
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• 2009

This study was carried out to investigate the toxic effect of WSF (water soluble fraction) in crude oil on the hatch, survival rate, abnormality and physiological activity of fertilized eggs and early larvae in Paralicthys olivaceus. The time required in hatching the fertilized eggs exposure to crude oil was 50.8${\sim}$53.2 hours both in control group and experimental group, showing no significant difference(p>0.05). The hatching rate in the control group was more than 80% in 1.9 ${\mu}g/mL$ WSF concentration, but hatching rate was less than 55.7% in below of 3.9 ${\mu}g/mL$ WSF concentration, showing significant difference (p<0.05). The survival rate on the 3rd day of early larval stage was 61.96% with 1.9 ${\mu}g/mL$ WSF, 11.1% with 3.9 ${\mu}g/mL$ WSF, and they all died with other concentration levels. The oxygen consumption rate in experimental groups was lower than control group. The heart rate (no./min) was 47.4${\sim}$52.8 before hatching and there was no significant difference between control group and experimental group, but heart rate reduced with the decrease of WSF concentration after hatching. The abnormality was 1.1% in the control group whereas 36.7% with 3.9${\mu}g/mL$ WSF, and abnormality was mostly comprised of incomplete spinal formation and spinal curvature. The results of this study suggest that even low concentration to WSF affects the early development of the flounder, Paralicthys olivaceus.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.4
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• pp.193-200
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• 2017

Objective: This study was conducted to investigate the efficacy of laser-assisted hatching (LAH) and various vitrification times for embryonic development and blastocyst cell numbers. Methods: First, 2-cell and 8-cell embryos were collected by flushing out the oviducts. In the control groups, they were vitrified for 8 or 10 minutes without LAH. The LAH groups underwent quarter laser zona thinning-assisted hatching before vitrification (4, 6, and 8 minutes or 4, 7, and 10 minutes, respectively). After incubation, double-immunofluorescence staining was performed. Results: The hatched blastocyst rate 72 hours after the 2-cell embryos were thawed was significantly higher in the 2LAH-ES8 group (33.3%) than in the other groups (p< 0.05). In the control-8 group ($22.1{\pm}4.6$), the cell number of the inner cell mass was higher than in the LAH groups (p< 0.05). The number of trophectoderm cells was higher in the 2LAH-ES6 group ($92.8{\pm}8.9$) than in the others (p< 0.05). The hatched blastocyst rate 48 hours after the 8-cell embryos were thawed was higher in the 8LAH-ES4 group (45.5%) than in the other groups, but not significantly. The inner cell mass cell number was highest in the 8LAH-ES7 group ($19.5{\pm}5.1$, p< 0.05). The number of trophectoderm cells was higher in the 8LAH-ES10 group ($73.2{\pm}12.1$) than in the other groups, but without statistical significance. Conclusion: When LAH was performed, 2-cell embryos with large blastomeres had a lower hatched blastocyst rate when the exposure to vitrification solution was shorter. Conversely, 8-cell embryos with small blastomere had a higher hatched blastocyst rate when the exposure to vitrification solution was shorter.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.235-245
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• 2001

Objectives: The purpose of this present study was to compare mouse embryo development in 3 commercial media and hatching competence of mouse embryo with or without enzymatic treatment. Methods: Collected 375 mouse embryos were divided into three groups, and then cultured in IVF-20 (G2), Medicult IVF (M3), P-1 (blastocyst M), respectively. Three day mouse morulae were cultured in G2 media treated with pronase. The results were analyzed using Chi-square test, and considered statistically significant when p<0.01. Results: The developmental rate of 2 cell mouse embryo after 72 hours was highest in IVF-20 (G2) among conventional 3 media. The hatching rate of mouse morulae was low when clultured in G2 media without pronase during 48 hours. However, it was higher when cultured in media treated with $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$ pronase, respectively. Conclusions: Using good media and digestion of zona pellucida with enzymatic treatment improve development and hatching rate of embryo. Therefore, implantation and pregnancy rate could be improved.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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• pp.106-111
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• 2016

Objective: To study the clinical outcomes of single frozen-thawed blastocyst transfer cycles according to the hatching status of frozen-thawed blastocysts. Methods: Frozen-thawed blastocysts were divided into three groups according to their hatching status as follows: less-than-expanded blastocyst (${\leq}EdB$), hatching blastocyst (HgB), and hatched blastocyst (HdB). The female age and infertility factors of each group were evaluated. The quality of the single frozen-thawed blastocyst was also graded as grade A, tightly packed inner cell mass (ICM) and many cells organized in the trophectoderm epithelium (TE); grade B, several and loose ICM and TE; and grade C, very few ICM and a few cells in the TE. The clinical pregnancy and implantation rate were compared between each group. The data were analyzed by either t-test or chi-square analysis. Results: There were no statistically significant differences in average female ages, infertility factors, or the distribution of blastocyst grades A, B, and C in each group. There was no significant difference in the clinical pregnancy and implantation rate of each group according to their blastocyst grade. However, there was a significant difference in the clinical pregnancy and implantation rate between each group. In the HdB group, the clinical pregnancy and implantation rate were similar regardless of the blastocyst quality. Conclusion: There was an effect on the clinical outcomes depending on whether the blastocyst hatched during single frozen-thawed blastocyst transfer. When performing single frozen-thawed blastocyst transfer, the hatching status of the frozen-thawed blastocyst may be a more important parameter for clinical outcomes than the quality of the frozen-thawed blastocyst.

• 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.

• Parasites, Hosts and Diseases
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• v.24 no.2
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• pp.115-120
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• 1986

This study was carried out to reveal the effect of temperature, salinity and aeration on maturation and hatching of Fibricola seoulensis eggs. The eggs were incubated and were observed daily for the appearance of eyespots and hatching. The results were summarized as follows. 1. From $4{\sim}5$ days after incubation in distilled water at $28^{\circ}C$ or at $11{\sim}26^{\circ}C$, the eyespots began to appear and the rates of eggs with eyespots were over 90% in $28^{\circ}C$ on the 7th or 8th day. However, eyespots did not appear in $5{\sim}15^{\circ}C$ or $4^{\circ}C$ by the 18th day. 2. The mature eggs began to hatch at the 8th day, and hatching rate 2 weeks after incubation was over 90% at $28^{\circ}C$, but it was below 5% at $11{\sim}26^{\circ}C$, and 0% at $5{\sim}15^{\circ}C$ and at $4^{\circ}C$. 3. Aeration did not influence the appearance of eyes pots nor hatching. 4. In salines under 0.6%, the rates of eyespots appearance were over 90% on the 7th day. The rate was 55.0% in 0.9% at 20 days, and 0% in 1.2%. 5. The hatching rates in salines below 0.3% concentration were over 90% by 14 days of incubation. However, the rate decreased to 44% in 0.6% saline and to 0% over 0.9% salinity. 6. The eggs incubated in the dark hatched in 12.5% on the 10th day, but hatching rate of mature eggs increased to 85.7% within 2 hours after exposure to light. Above results demonstrated that the best temperature for maturation and hatching of F. seoulensis eggs was $28^{\circ}C$, and the miracidia began to hatch at $8{\sim}9$ days after incubation. In the field, hatching and invasion into snails of the miracidia may occur from May to September in Korea. In salines under 0.3% concentration maturation and hatching were not influenced, but as salinity increased hatching was inhibited more than maturation was.

• Journal of Aquaculture
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• v.9 no.4
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• pp.329-337
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• 1996

Dietary values of phytoplanktons, concentrated Chlorella and commercial yeasts were investigated for the resting egg production of the Korean rotifer, B. plioatilis S-type. The hatching rate of the resting egg of the rotifer fed on different diets was also studied. The highest production of the resting egg was 3,760 eggs/20 ml with N. oculata. However, the hatching rate was not significant among 4 phytoplankton species. The highest production of resting eggs among 9 diet groups ($\omega$-yeast, baker's yeast, 2 refrigerated Marine Chlorella, frozen Marine Chlorella, frozen Chlorella ($70\%$) + baker's yeast ($30\%$), frozen Chlorella ($30\%$) + baker's yeast ($70\%$), 2 refrigerated freshwater Chiorella) was 283 eggs/ml in the frozen Chlorella ($30\%$) +baker's yeast ($70\%$) and the refrigerated freshwater Chlorella. The highest number of resting egg from 10,000 rotifers and 1 mg dry wieght diet was 5,566 eggs and 2,131 eggs in the frozen Chlorella ($30\%$) + baker's yeast ($70\%$). However, the highest hatching rate of the resting eggs was $67.4\%$ in $\omega$-yeast. In this study, the results suggest that the baker's yeast with a small amount of frozen Chlorella seems to be good feeding regime for the economical production of resting eggs.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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• pp.157-164
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• 1992

This stduy was carried out to obtain basic information available on ovulation, spawning, fertilization rate, hatching rate, and deformity rate after hCG injection in cyprinid loach, Misgurnus mizolepis. The results obtained in these experiments were as follows: 1. Ovulation and spawning occurred simultaneously and spawing was completed within 1 hour after ovulation. 2. More than 80% of fertilization rates appeared within 12 hours at 21$^{\circ}C$, 8 hours at $25^{\circ}C$, and 4 hours at 29$^{\circ}C$, respectively, following the onset of spawing. Afterwards, the fertilization rates of released eggs sharply decreased in three different water temperatures. 3. More than 70% of hatching rates appeared within 8 hours at 21$^{\circ}C$, 6 hours at $25^{\circ}C$, and 2 hours at 29$^{\circ}C$, respectively, following the onset of spawing. Afterwards, hatching rates of spawned eggs abruptly decreased in three different water temperatures. 4. The deformity rates of hatched larvae were high at $25^{\circ}C$, 8 hours following the onset of spawing. 5. Based on the developmental ability of oocytes, the optimum time of fertilization was 4 hours (stage 5) following the onset of spawing.

• Development and Reproduction
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• v.17 no.3
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• pp.247-255
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• 2013

The gonadsomatic index (GSI) of mottled skate was the highest in April, GSI and HSI showed a reverse phase for its reproductive cycle. The fish had one pair of egg capsules, having 1 to 7 fertilized eggs, and spawned all the year round. When surveying the reproductive characteristics of females over 63 cm in disc width, we found the spawning peak was between April to June, and the appearance ratio of egg capsules was the highest in May (32.1%). The eggs were hatched at $8^{\circ}C$, $13^{\circ}C$, $18^{\circ}C$, water temperature (12.8 to $24.2^{\circ}C$), and the best hatching temperature was $18^{\circ}C$. The number of fish hatched was 4 to 5 fish/egg capsules, and the hatching rate was 100%. The sex ratios of hatching larvae were 45.5% female and 54.5% male. Therefore this study will provide fundamental data and information for artificial reproduction of the mottled skate.