• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.4
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• pp.479-485
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• 2020

We evaluated the hatchability of fertilized eggs from six hybrid combinations of highly valued grouper species inhabiting temperate and warm waters, with the goal of establishing a novel hybrid with enhanced growth and viability during the culturing period in the temperate waters of Korea. Hybrid combinations with red-spotted grouper females exhibited high hatchability with high a fertilization and hatching rate of fertilized eggs and a low deformity rate of hatched larvae. Conversely, hybrid combinations with kelp grouper females had very low hatching rates and very high deformity rates; commercial production of seed from such crosses would be difficult without improving hatchability. The hatchabilities of convict grouper ♀×giant grouper ♂ and kelp grouper ♀×red-spotted grouper ♂ were lower than those of maternal purebreds, but these two hybrid combinations were expected to produce potentially large quantities of hatched larvae. In the above evaluation, promising hybrid combinations were identified for commercial production of seed. For these hybrids to contribute to the development of Korea's mariculture industry, mass production of fertilized eggs and seeds is necessary, along with the development of advanced rearing techniques, such as the identification of a suitable rearing temperature.

• Korean Journal of Animal Reproduction
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• v.18 no.1
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• pp.47-54
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• 1994

The objective of this study was to produce Korean native calves following transfer of in vitro matured, fertilized and cultured embryos into Holstein cows. The ovaries of Korean native cows or heifers were obtained from an abattoir and kept on 25 to 28$^{\circ}C$ and transported to laboratory within 2 hrs. The oocytes were matured in vitro (IVM) for 24 hrs in TCM-199 supplemented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, 1$\mu\textrm{g}$/ml estradiol-17$\beta$ and granulosa cells at 39$^{\circ}C$ under 5% CO2 in air. They were fertilized in vitro (IVF) by epididymal spermatozoa treated with heparin for 24 hrs., and then the zygotes were co-cultured in vitro (IVC) with bovine oviductal epithelial cells for 7 to 9 days. Late morulae and blastocysts produced in vitro were nonsurgically transferred to recipient cows by unilaterial. Recipients were monitored for estrus and for pegnancy by rectal plapation in 60 days after embryo transfer. One of them was pregnant to term and produced a female weighing 42.5kg at birth.

• Korean Journal of Animal Reproduction
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• v.18 no.1
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• pp.7-13
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• 1994

To verify in vivo viability of IVF-derived bovine embryos, morula and blastocysts that developed from in vitro matured and fertilized ova were transferred to the uteri of recipient cows and normal calves were produced. To produce IVF-derived bovine morula or blastocysts, ova matured and fertilized in vitro were cultured in culture medium for 7~8 days at 39$^{\circ}C$ under the humicified atmosphere of 5% CO2. Two different culture systems, a co-culture system with TCM-199 and bovine epithelial cells (BOEC) and CR1aa without somatic cell support, were compared. Cleavage rates to 2~8 cell stage and developmental rates of IVF-derived bovine embryos to blastocyst stage were not different between co-culture system (51.3 and 14.0%) and CR1aa medium (60.4 and 22.1%), respectively. Embryos were classified into three grades by embryo quality and then one or two embryos in higher quality(A and B grades) were transferred to the uterus of recipients. In this study Korean Native calf was first born after transfer of IVF-derived embryos. Total four live calves were normally developed to term from IVF-derived bovine blastocysts and one female fetus was still-born approximatedly 8 months of gestation, but there was no pregnancy after transfer of morula. Therefore, normal calves could be produced after transfer of IVF-derived bovine embryos cultured in CR1aa medium without somatic cell support. In addition, our results suggest that in transfer of IVF-derived bovine embryos blastocyst stage is better than morula.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.9-18
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• 1997

These studies were carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained in studies on the factors influencing in vivo embryo production were as follows : 1. There were no significant differences in the number of total recovered, fertilized, transferable and freezable embryos among the hormone doses(FSH-P, 28∼34mg; SUPER-OV, 75IU) used for superovulation. However, over 30mg doses of FSH-P showed a slightly higher effect than others. 2. There were slight decrease in the number of fertilized, transferable and freezable embryos in 3 times repeated superovulation. But there were no significant differences among 1, 2 and 3 times repeated superovulations. 3. Age of donors did not affect the number of transferable and freezable embryos, but the number of fertilzed embryos were highest in 2∼3 years old donors and were lowest in 8∼9 years old donors(P<0.05). 4. Season had a significant effect on the production of embryos(P<0.05). the embryo production, and the number of fertilized, transferable and freezable embryos were most effective in summer and follow by spring, autumn and winter. 5. The number of transferable and freezable embryos was highest in donors flushed at 7∼8 days after estrus(P<0.05).

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.311-317
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• 2000

This study was carried out to compare the temperature and time of heat shock, and the effect of heat shock on development of embryos after in vitro maturation and fertilization in bovine oocytes. The results obtained were as follows. 1. The optimum temperature and time of heat shock were 41$^{\circ}C$ and 30sec on in vitro development of embryos from 4~8 cell to blastocyst. 2. The rates of cleavage on zygotes produced on in vitro were significantly increased by heat shock after IVM than before IVM(P<0.05). 3. When the oocytes were treated heat shock after IVM and 5 days cultured, developmental rates to blastocyst were increased than other experimental treatments.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.1
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• pp.31-37
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• 2021

The mass production of fertilized eggs of the convict grouper Hyporthodus septemfasciatus was studied from 2013 to 2020 for industrial aquaculture. The experiment was divided into two groups. Group 1 broodstock was raised from wild-caught fry and used from 2013 to 2020. Group 2 broodstock was raised from artificially propagated fry and used from 2019 to 2020. Males used to collect sperm for artificial insemination weighed more than 7 kg. The effects of various hormones on artificial ovulation were investigated from 2013 onward. Among these, luteinizing hormone-releasing hormone analogue (LHRHa) at 100 ㎍/kg body weight showed the most effective results and was used for artificial egg collection from 2014 onward. In Group 1, the average total egg production per year, average egg production per individual, fertilization rate, and hatching rate were 26,143 mL, 609.7 mL, 93.3%, and 91.8%, respectively, and in Group 2, were 2,750 mL, 316.5 mL, 92.1%, and 90.4%, respectively. Based on these results, we showed that a large number of fertilized eggs for artificial seeding could be produced consistently. Moreover, the mass production of fertilized eggs in Group 2 establishes a foundation for the complete aquaculture cycle of H. septemfasciatus.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.58 no.3
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• pp.205-213
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• 2022

The experimental fish transplanted from China in 2015 was used after seedling production and cultivated. Breeding management for experiment was carried out from October 2020 to February 2021. Also, it succeeded in inducing artificial maturation three to four months earlier than wild broodstock and secured good quality fertilized eggs. The average size of fertilized eggs was 1.22 mm, at 20℃ Blastodisc (15 minutes post-fertilization), 2 cell (50 minutes), 4 cell (1 hours), 8 cell (2 hours), 16 cell (2 hours and 30 minutes), 32 cell (2 hours and 50 minutes), morula (3 hours), blastula (8 hours), gastrula (15 hours), skull formation (20 hours), organ formation (30 hours) and hatching yolk larvae stage (35 hours). The total length of the just hatched larvae were 2.50 ± 2 mm, and then gain growth of 42.5 mm by 60 days, reaching 45 ± 5 mm.

• Korean Journal of Poultry Science
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• v.31 no.1
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• pp.9-15
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• 2004

This study was conducted to evaluate the effects of dietary herbal plant mixture on the performance in laying hens under heat stress. One hundred ninety two 54-weeks-old ISA Brown commercial layers, were used in 56 d experimental assay. Dietary treatments included CON (control; basal diet), HPM0.05 (basal diet ＋ 0.05% herbal plant mixture), HPM0.1 (basal diet ＋ 0.1% herbal plant mixture), and HPM0.2 (basal diet ＋ 0.2% herbal plant mixture). For overall period, the hens fed with HPM0.1 and HPM0.2 diets showed lower in the hen day egg production than the hens fed with CON diet(P＜0.05). At the end of the experimental period, egg weight was heavier in HPM 0.1 treatment than in CON (P＜0.05). There were no significant differences among the treatments in egg shell breaking strength, egg shell thickness, Haugh unit, and yolk color unit. Total cholesterol concentration of yolk tended to decrease as the level of herbal plant mixture in the diet increased. Total protein of blood was higher in the hens fed with herbal plant mixture than in the hens fed with CON diet (P＜0.05). Albumin concentration of blood was increased in HPM0.05 and HPM0.1 treatments compared with CON(P＜0.05). Red blood cell (RBC) and white blood cell (WBC) concentrations in serum were increased in HPM0.1 and HPM0.2 treatments compared with CON treatment (P＜0.05). In conclusion, dietary herbal plant mixture in laying hens under heat stress adversely affected egg production but increased total protein, albumin, RBC and WBC in blood.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.89-95
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• 2007

The present study was conducted to examine some factors affecting in vitro development and fecundity of embryos recloned with somatic cell nuclear transfer (SCNT). Fibroblast cells retrieved from the ear of a 3-week-old, cloned Korean goat (Jinsoonny) were used as karyoplast donors and serum-starvation was conducted in tissue culture medium (TCM)-199 supplemented with 0.5% FBS. Recipient oocytes were surgically collected by flushing the oviducts 35 h after hCG injection following FSH priming. The zonae pellucidae of the oocytes were partially perforated with a laser drill and a donor cell was transferred into an enucleated oocyte. The couplets were electrically fused and activated by ionomycin (5 min) and 6-DMAP (4 h). The reconstructed embryos were cultured in mSOF medium containing 0.8% BSA at $39^{\circ}C$ in an atmosphere of 5% $CO_2$, 5% $%O_2$, 90% $N_2$ for 12 to 15 h. Re-cloned embryos (2- to 4-cell stages) were surgically transferred into the oviducts of the recipients and pregnancy was subsequently diagnosed by progesterone assay and ultrasound on Days 21 and 63 of pregnancy. The fusion rate following 1st fusion pulse was higher (p<0.05) in 2nd cloning (65.9%) compared to 1st cloning (51.0%), but it was not different in the other groups. The rate of cleavage after fusion was significantly higher (p<0.05) in 1st (77.7%) than in 2nd cloning (56.0%). A total of 175 re-cloned embryos were transferred into 28 recipients. On day 21 and 60 after transfer, 11 (39.3%) and 4 recipients (17.4%) were pregnancy, respectively. In comparison of pregnancy rate by estrous synchronization, a total of 66 and 109 re-cloned embryos were transferred into 11 recipients in natural estrus and 17 recipients in induced estrus, respectively. Five (45.4%) and 2 recipients (18.2%) in natural estrus were pregnant on days 21 and 63 while 6 (35.3%) and 2 (11.8%) recipients in induced estrus were pregnant, respectively. These results show that recloning of goat can be achieved by SCNT and estrous synchronization between donor and recipient animals may be one of the major factors affecting success rate.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.1
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• pp.49-54
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• 2017

We collected and reared Theragra chalcogramma walleye pollock brood-stock for use in natural spawning tests and undertook to obtain domestic pollock via fertilized egg capture, development of fertilized eggs, and absorption of yolk sac after hatching. Whole pollock were caught with trammel and set nets and immediately placed in a deep-sea water tank. Adults were the most common pollock age group (43.0%; n = 86) among the 254 pollock captured in March 2014 with 57.9% (n = 147) being captured off Southern Gosung, Korea. The main spawning period of pollock is February (spawning phase of 91% of pollock). From the deep-sea tank, we collected 1640 mL of naturally fertilized eggs (~820,000 eggs) from 12 spawning events occurring between February 4 and 22 2015. The floating/ live eggs were maintained in deep-sea water tanks at $5.5{\pm}0.2^{\circ}C$. Egg size was $1.5{\pm}0.03mm$. Six hours after fertilization the eggs were at the 2 cell stage, and the eggs hatched approximately 340 hours after collection. At hatching, larval length and yolk sac area were $5.2{\pm}0.25mm$ and $9.5{\pm}1.00mm^2$ (100%), respectively. Four days after hatching, the yolk sac area was $2.2{\pm}0.53mm^2$ ($23.1{\pm}5.55%$). This is the first report of collection of naturally fertilized eggs from pollock and their subsequent hatching while held in an indoor deep-sea water tank. The results suggest that such collection could assist in the recovery of pollock resources and the possibility of domestic rearing of cultivated larvae.