• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.365-373
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• 2000

The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P＜0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P＜0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

• Journal of Aquaculture
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• v.18 no.2
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• pp.107-114
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• 2005

Clownfish are important and very popular fish in the ornamental aquarium industry. Demand for the fish is increasing dramatically. The present study was conducted to verify methods of broodstock management, patterns of spawning, rates of egg hatching and estimates of larval growth fur the saddleback clownfish, Amphiprion polymnus. Spawning occurred 8 times between August 2002 to June 2004 with 2 females and 1 male participating. Fertilized eggs were separated by an adhesive matrix and were oval in shape. The eggs were $2.46{\pm}0.13mm$ in size as measured along the longest axis. The percentage of fertilized eggs was 96.7%. Hatching was observed seven days post-spawning and hatching rate was 85.5%. The sizes of the newly-hatched larvae were $4.58{\pm}0.21mm$ TL (total length). Larvae had an open mouth and anus, and an oval yolk sac. At the 1 st day after hatching, the sizes of the larvae were $4.90{\pm}0.35mm$ TL. The larvae began to eat rotifers after complete yolk absorption. On the 5th day post-hatch, larvae were $5.88{\pm}0.31mm$ TL with complete fins and the survival rate was 48.6%. At 8 days after hatching, a band began to appear on head and back of the larvae indicating the beginning of metamorphosis. Metamorphosis was completed at an average TL of $15.00{\pm}2.12mm$ on the 23rd day after hatching. By the 45th day after hatching, juveniles averaged $22.76{\pm}3.22mm$ TL and survival rate was 28.4%.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.2
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• pp.125-131
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• 2007

Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.

• Journal of The Korean Society of Grassland and Forage Science
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• v.22 no.4
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• pp.265-272
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• 2002

Forage breeding laboratory of National Livestock Research Institute, R.D.A. has made interspecific hybrids of Lolium multiflorum $\times$ L. pratensis and intergeneric hybrids of Lolium $\times$ Festuca since 1984, and has collected ecotypes of Italian ryegrass since 1991. Growth characteristics of these hybrids and ecotypes were researched, and then these clone lines were named. Among these clone lines, the several clones that have polen fertility, high cold-tolerance, and similar heading time were used for making synthetics, Naehan 6, 7, 8, 9, with polycrossing method in 1997. Field experiments were carried out to compare the mophological and agronomical characteristics and forage productivity and quality of the synthetics with those of Italian ryegrass varieties, Barmultra and Hwasan 101. in Suwon and Yonchun from 1999 to 2000. Heading time of the synthetics were 22th to 24th May that belong to late-mature types to be similar to that of Barmultra and Hwasan 101 in Suwon. The synthetics were 101 to 106 c3n in plant length, medium or thick in thickness of stem, dark peen in leaf color, broad and long in flag leaf, strong in lodging resistance, and excellent in regrowth. Winter survivals of the synthetics were no different from that of Barmultra or Hwasan 101 in Suwon, but better than that of Barmultra or Hwasan 101 in Yonchun where was -10 to -12$^{\circ}C$ of minimum average air temperature in January or February. Dry matter(DM) yields of the synthetics were similar to DM 8,238kg per ha of Barmultra in Suwon, but in Yonchun, were more 7 to 13% than DM 7,291kg per ha of Barmultra. Forage qualities, IVDMD, ADF, NDF and TDN of the synthetics were lower than those of Hwasan 101, but higher than those of Barmultra.

• The Korean Journal of Malacology
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• v.31 no.2
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• pp.103-112
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• 2015

Bay scallop (Argopecten irradians) has been farmed only in the South Sea of Korea. East Sea Fisheries Research Institute (ESFRI) has developed bay scallop aquaculture technologies to extend its aquaculture area to the Southeast Sea of Korea. For the artificial spawning, the water temperature was maintained at $23^{\circ}C$. Over 100,000,000 eggs were spawned through artificial spawning inductions, such as air exposure and thermal shock by rising the water temperature. The fertilization rate was over 91% with nearly 94,000,000 fertilized eggs. The shape of fertilized eggs was spherical with an average diameter of $61.7{\pm}0.05{\mu}m(54.1-67.4{\mu}m)$. Five days after fertilization, the eggs developed into prodissoconch shell, and continuously grew into umbo stage and then umbones stage. After 8 days of fertilization, the size of larva became $179.7{\pm}8.4{\mu}m$ on average ($150.4-204.8{\mu}m$), and the larva formed a foot and an eye spot. The larvae grew to $235.4{\pm}9.7{\mu}m$ in 10 days and attached to adherence material, becoming juvenile bay scallop. The shells grew from 22.71 mm to 72.40 mm in 6 month (June-December). The total weight increased from 2.0 g to 32.7 g at the same period. The daily growth rates of young scallop were $0.35mm\;d^{-1}$ (Apr. to Jun.) and $0.41mm\;d^{-1}$ (Jun. to Aug.), which were comparable to those found in the South Sea. These findings suggest that the bay scallop aquaculture may be suitable in the Southeast Sea of Korea and may provide an additional crop to aquaculturists.