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Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Nutritional Components and Their Antioxidative Protection of Neuronal Cells of Litchi (Litchi chinensis Sonn.) Fruit Pericarp (리치 과피의 영양화학 성분 및 항산화성 신경세포 보호효과)

  • Jeong, Hee-Rok;Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Kim, Yeon-Su;Jeong, Chang-Ho;Kim, Dae-Ok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.481-487
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    • 2010
  • The nutritional components, antioxidant, and neuroprotective effects of water and a 50% methanol extract from litchi fruit pericarp were investigated. The most abundant mineral, amino acid, and fatty acid were K, proline, and palmitic acid, respectively. In addition, the total water phenolics and 50% methanol extracts were 8.02 and 12.28 mg/g, respectively. The DPPH, ABTS radical scavenging activities and ferric reducing antioxidant power of the water and 50% methanol extracts showed dose-dependent antioxidant activity. In a cell viability assay using MTT, almost all extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase leakage was also inhibited by the pericarp extracts. In particular, the 50% methanol extract showed a higher cell membrane protective effect than the water extract at the highest concentration. Consequently, these data suggest that litchi fruit pericarp can be utilized as an effective and safe functional food substances for natural antioxidants and may reduce the risk of neurodegenerative disorders.

Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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Development of Leaf Protein Concentrates II. Extraction of Leaf Protein Concentrates of Some Plants Growing in Korea (잎 단백질(蛋白質)(Leaf Protein Concentrates)의 개발(開發)에 관한 연구(硏究) II. 한국산(韓國産) 각종 식물(植物)로 부터의 잎 단백질(蛋白質)의 추출(抽出))

  • Choe, Sang;Kim, Geon-Chee;Chun, Myung-Hi;Kim, Kil-Hwan
    • Korean Journal of Food Science and Technology
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    • v.2 no.2
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    • pp.17-25
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    • 1970
  • Juice were extracted from fresh leaves of 70 species of plants growing in Korea by mincing and pressing the resulting pulp through a cotton cloth. Leaf protein concentrates could be prepared from many species of land and water plants that are at present economically unimportant. The choice of plants is of considerable important. Total-N, protein-N and pH determinations were made on the extracts, and total-N remaining in the fibre were calculated. Leaf protein concentrates were precipitated from the extracts at $75{\sim}80^{\circ}C$, and analysed total-N as protein-N of products. The present paper deals with the calculated yields of leaf protein concentrates from various plants, relations between yield of leaf protein concentrates and total-N of leaves, or pH of extracts, and the amino acid compositions of leaf protein concentrates. Results are summarized as follows. 1. Spinach and radish were the best sources of easily extractable, but good results were also obtained with indian mustard, kail, chenopod, red bean, cucumber, squash, houndberry, white flowered gourd, potato, Humulus japonicus, arrowroot and soybean as a good resources for the production of leaf protein concentrates. 2. In general, the greater the protein content of leaves the greater the yield of leaf protein concentrates. However, there are some plants difficult to make a adequate protein extraction by a simple mechanical process. 3. It was to be expected that leaf protein concentrates would be more extractable with the higher pH of extracts. There were a poor yield of the leaf protein concentrate in the pH values lower than 5.50 of the first extracts. 4. Protein content of the leaf protein concentrate shows marked differences, depending on species and season. It ranged between 29 to 80% of protein contents. However, the majority of plants yielded products containing more than 50% of protein. Products containing more than 75% of protein were obtained from two species of radish and indian mustard. Cabbage and Digitaria sanguinalis cilialis (summer) made products containing 29 to 32% of protein. 5. The amino acid composition of leaf protein concentrates was not greatly altered by species of plants. On an amino acid compositional basis, the leaf protein concentrate has a favorable balance of essential and non-essential amino acids, the only exception being methionine, which was usually low in all cases.

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Changes of Clinical Findings of Acute Poststreptococcal Glomerulonephritis (소아에서 연쇄상 구균 감염 후 급성 사구체 신염의 임상양상의 변화)

  • Choi, Byung-Ho;Chu, Mi-Ae;Hong, Eun-Hui;Hwang, Hyun-Hee;Cho, Min-Hyun;Ko, Cheol-Woo
    • Childhood Kidney Diseases
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    • v.12 no.2
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    • pp.157-163
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    • 2008
  • Purpose : Several studies have reported the recent increase in the incidence of acute poststreptococcal glomerulonephritis(APSGN). The objective of this study is to see changes of clinical findings/manifwstation in children with APSGN. Methods : Medical records of 63 children who were diagnosed with APSGN in the deparment of Pediatrics, Kyungpook National University Hospital, between January 1992 and December 2006 were reviewed retrospectively. We analyzed various clinical characteristics such as age, sex, degrees of proteinuria, degrees of hematuria, and presence or absence of histories of systemic antibiotic use in children with APSGN, and compared the children with APSGN who were diagnosed between 1992 and 2000 to those who were diagnosed between 2001 and 2006. Results : Age of the patients ranged from 2-14 years(median 7.11 years) at the time of disease onset. Study patients consisted of 41 boys and 22 girls. APSGN followed infection of the throat in 87% of cases. Patient developed an acute nephritic syndrome 12 days after an antecedent streptococcal pharyngitis. Forty patients presented with gross hematuria. Fortyone patients had hypertension at the time of diagnosis. Hypertension disappeared within 7.8$\pm$8.2 days, gross hematuria within 11.3$\pm$17.2 days and microscopic hematuria within 3.5$\pm$3.9 months from the disease onset. Patients in 2001-2006 had significantly higher increase of antistreptolysin O(ASO) titer. However, no significant differences in clinical characteristics were observed. Age, sex, severity of proteinuria, gross or microscopic hematuria, antibiotic therapy did not affect the clinical manifestations of glomerulonephritis. In other words, hypertension, duration of hematuria, recovery of serum C3 level are not different between the two time periods. Conclusion : Our data indicates that patients in 2001-2006 had significantly higher level of ASO titer. However, they did not show significant clinical differences. To evaluate the causes of the resurgence of APSGN, a national epidemic is needed.

Pedological Characteristics of Asian Dust in Korea (한국에 강하한 황사의 토양학적 특성)

  • Zhang, Yong-Seon;Kim, Yoo-Hak;Sonn, Yeon-Kyu;Lee, Gye-Jun;Kim, Myung-Sook;Kim, Sun-Kwan;Weon, Hang-Yeon;Joa, Jae-Ho;Eom, Ki-Cheol;Kim, Sang-Hyo;Kwak, Han-Kang;Kim, Han-Myeong
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.6
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    • pp.301-306
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    • 2005
  • Asian dust was collected in Korea and soils in the arid area of northern China were analysed for its physical and chemical properties, and mineral compositions for in order to interpret the origin of Aeolian soils and estimate the effect of dust wind on the soil environment in Korea. Asian dust was collected at Suwon in Korea from 2002 to 2004. Soil samples were collected from the desert and Loess plateaus around Gobi desert in China. As a result of analysis of desert soil distributed on northern region and Loess soils in China, it was observed that soil pH was about 9, organic matter 11 to $23g\;kg^{-1}$, and CEC 7.1 to $18.4cmolc\;kg^{-1}$, showing a high spatial variation among different sampling locations. About 62 to 80% of particles were composed of quartz and feldspars, 2 to 14% calcite ($CaCO_3$) and dolomite [$Ca{\cdot}Mg(CO_3)_2$], and trace other clay minerals. All the dust particles in Korea were below 50 m in diameter, and the mineral compositions were quartz, mica, feldspar and some clay minerals. Major components of clay mineral of Asian dust was mainly illite as compared to the kaolin of soils in Korea. The base saturation of exchangeable Ca, Mg, K and Na in the Asian dust was above 250% due to the high content calcite. Most of upland soil in Suwon was thin and sharp type, but Asian dust in Korea was the spherical shape. Asian dusts in Suwon, Korea, did not show a definite mineralogical variation of the dust during the collection period. Difference between the Asian dust collected in Korea and the soils in arid area of China was observed in the physical and chemical properties, especially for particle size distribution, cations such as Ca, Mg, K and Na. However, some similarities were found on the mineral compositions and chemical properties between Asian dust collected in Korea and the loess of China.

Ecological Characteristics of Red Rice (Local Name "SARE," Oryza sativa L.) and Factors Affecting Its Competition with Rice (적미(赤米) (자생도(自生稻), 속명(俗名) "사레")의 생태적(生態的) 특성(特性) 및 벼와의 경합요인(競合要因))

  • Ree, D.W.;Hong, Y.K.;Kim, J.C.;Kim, Y.H.
    • Korean Journal of Weed Science
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    • v.3 no.2
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    • pp.143-150
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    • 1983
  • Red rice (local name "Sore") as a weed has been a serious threat to rice production in direct-seeded rice culture in Ganghwagun, Gyeonggi province. In the Ganghwagun 508 ha out of 1,420 ha in the Samsanmyeon area is infested with red rice. The average lowland rice yield is about 4,300 kg/ha in the Ganghwagun, but the average upland rice yield is about 2,000 kg/ha in infested area. This study was carried out in order to clarify the ecological characteristics of red rice and factors affecting the competitive ability of five red rice varieties, collected from Samsanmyeon in 1981, with rice cultivar. Five varieties-Monggeunsare, Salsare, Ginkaragsare, Galsaegsalsare, Galsaegkaragsare-showed the same morphological characteristics of cultivated Japonica type, Chuncheongbyeo, but red rice tillers more profusely, is taller and lodges more easily than Chucheongbyeo. It shatters easily about 10-15 days after heading date, and at this time the hull is discolored in yellow or dark brown. There are many types of red rice with short or long owns on the spikelet, occasionally with or without own on the spikelet in the same hill, and the grains are short or long. In red reice leaf blast occurs more severely than in cultivated Indica/Japonica type, Teabaegbyeo, particulary serious in Monggeunsare. When red rice invaded in direct-seeded rice, number of panicles of rice became reduced more than other yield components.

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Effect of Submergence and Air Exposure of the Shoot on Growth, Nutrient Uptake and Photosynthesis in Monochoria vaginalis Presl. (물달개비 경엽(莖葉)의 침수여부(沈水與否)에 따른 생장(生長), 양분흡수(養分吸收) 및 광합성(光合成) 비교(比較))

  • Soh, C.H.;Yang, K.S.;Kwon, Y.W.
    • Korean Journal of Weed Science
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    • v.16 no.1
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    • pp.14-20
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    • 1996
  • Growth, nutrient uptake and photosynthesis as affected by submersion of shoot in pickerel weed (Monochoria vaginalis Presl.) were determined. The shoots of pickerel weeds in hydroponic culture were subjected to the submerged or emerged condition at 3- or 5-leaf stage for 8 or 10 days. Under submerged condition, growth in plant height was enhanced, but leaf number, leaf area, fresh and dry weight were reduced compared to those under the emerged condition. Similar responses in growth to submergence were obtained with the pickerel weeds rooted in the soil. Under submergence, chlorophyll content increased during the first 2 days, but thereafter remarkably decreased at 3-leaf stage and after the first 4 days at 5-leaf stage. Compared to the emerged condition, uptakes of $NH_4\;^+$-N, $NO_3\;^-$-N, $P_2O_5$ and $K^+$ were reduced, but uptakes of $Ca^{++}$ and $Mg^{++}$ increased under the submerged condition. Photosynthetic rate of shoot under water, measured by $CO_2$electrode, showed the maximum by 210 ${\mu}$moles $HCO_3\;^-$/g F.W. at the 8th day after submergence(DAS) at 3-leaf stage and 320 ${\mu}$moles $HCO_3\;^-$/g F. W. at 6 DAS at 5-leaf stage. These results indicate that pickerel weeds grow much better when the shoot is air-exposed and are less tolerable to submergence at 3 leaf-stage than at 5-leaf stage.

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Utilization of Probiotic Starter Cultures for the Manufacture of Low-fat Functional Fermented Sausages (저지방 기능성 발효소시지의 제조를 위한 복합 유산균주의 이용)

  • Kim, Young-J.;Lee, Hong-C.;Park, Sung-Y.;Park, Sun-Y.;Oh, Se-Jong;Chin, Koo-B.
    • Food Science of Animal Resources
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    • v.28 no.1
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    • pp.51-58
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    • 2008
  • This study was performed to evaluate the physico-chemical properties of fermented sausages containing probiotic starter cultures (LK-30 plus, Lactobacillus plantarum 155 and 167, and Pediococcus damnosus L12) with reduced fat levels, and to determine the optimum condition for the manufacture of these products. Although low-fat fermented sausages were reduced fat content at the amount of 90% and the ripening time by 1-2 weeks, as compared to regular-fat counterpart, they became harder and had many winkles outside due to the extreme drying. In addition, fat level in fermented sausages affected the composition and shear force values. During ripening, pH, lightness and yellowness values tended to decrease, however, microbial counts of inoculated lactic acid bacteria were increased up to $10^8-10^9cfu/g$ within 3 days and remained constant thereafter. Low-fat fermented sausages had higher microbial counts than regular-fat ones. Although the inoculated probiotic starter cultures alone had the functional properties, such as cholesterol reduction, anti-high blood pressure and antimicrobial activity, they did not have distinctive characteristics in the fermented sausages. Based on these results, the low-fat fermented sausages were successfully manufactured, but a little bit increased fat level and improved functional properties in the fermented sausages would be required to have better quality as compared to regular-fat counterparts.

Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.293-302
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    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

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