Functional dyspepsia (FD) is a gastrointestinal disorder with diverse symptoms but no structural or organic manifestations. Benachio-F® (herein named ‘BF-1’) is an over-the-counter liquid digestive formulated with multiple herbal extracts, which has been reported to improve symptoms of FD. A total two experiments were conducted. First, we examined whether BF-1 can modulate the progression of FD through two experimental rat models. A total of three doses (0.3x, 1x, 3x of the human equivalent dose) were used. In the gastric emptying model, both 1x (standard) or 3x (3-fold-concentrated) BF-1 enhanced gastric emptying was compared with that of vehicle-treated animals. In a feeding inhibition model induced by acute restraint stress, treatment with 1x or 3x BF-1 led to a similar degree of restoration in food intake that was comparable to that of acotiamide-treated animals. Among the constituents of BF, fennel is known for its choleretic effect. Thus, we next investigated whether a novel BF-based formula (named ‘BF-2’) that contains an increased amount of fennel extract (3.5-fold over BF-1), has greater potency in increasing bile flow. BF-2 showed a superior choleretic effect compared to BF-1. Furthermore, the postprandial concentration of serum secretin was higher in animals pretreated with BF-2 than in those pretreated with BF-1, suggesting that the increased choleretic effect of BF-2 is related to secretin production. Our results demonstrate that BF-1 can modulate the pathophysiological mechanisms of FD by exerting prokinetic and stress-relieving effects, and that BF-2 has a better choleretic effect than BF-1.

본 연구에서는 기능성 식품원료로써 흰점박이꽃무지 유충의 이용 가능성과 흰점박이꽃무지 유충 사료자원으로써 큰느타리버섯수확후배지의 이용 가능성을 조사하기 위해서 참나무톱밥과 큰느타리수확후배지를 각각 식이한 흰점박이 꽃무지 유충 추출물의 항산화 활성을 비교하였다. 참나무톱밥과 큰느타리수확후배지를 각각 식이한 흰점박이꽃무지 유충 추출물의 총 폴리페놀 함량은 각각 57.02±1.73 mg GAE/g과 75.33±0.43 mg GAE/g이었고 플라보노이드 함량은 각각 24.6±0.28 mg/g과 25.4±0.75 mg/g이었다. DPPH에 의한 라디칼소거 활성은 추출물의 농도가 증가할수록 증가하는 경향을 나타내었으나 참나무톱밥을 식이한 흰점박이꽃무지 유충에 비해 큰느타리버섯수확후배지를 식이한 흰점박이꽃무지 유충 추출물의 DPPH 라디칼 소거능이 우수하였으며 32 mg/ml 이상의 농도에서는 두 시료 간의 유의적 차이가 나타나지 않았다. 참나무톱밥과 큰느타리버섯수확후배지를 각각 식이한 흰점박이꽃무지 유충 추출물의 농도별 환원력도 추출물의 농도가 높을수록 환원력은 증가하였으나 시료 간의 유의적인 차이는 보이지 않았다. 참나무톱밥과 큰느타리버섯수확후배지를 식이한 흰점박이꽃무지 유충 추출물의 ORAC 지수는 각각 66.55±0.99 uM TE/g과 76.32±0.48 uM TE/g로 참나무톱밥을 식이한 흰점박이꽃무지 유충 추출물에 비해 큰느타 리버섯수확후배지를 식이한 흰점박이꽃무지 유충 추출물의 ORAC 지수가 높게 나타났다. 참나무톱밥과 큰느타리 버섯수확후배지를 각각 식이한 흰점박이꽃무지 유충 추출물을 각각 0, 2, 4, 8, 16, 32, 64 mg/ml의 농도로 처리하 였을 때 RAW 264.7 세포는 90% 이상의 생존율을 나타 내었으므로 참나무톱밥과 큰느타리버섯수확후배지를 각각 식이한 흰점박이꽃무지 유충 추출물은 RAW 264.7 세포에 독성을 나타내지 않는 것으로 판단된다.

본 연구에서는 기능성 식의약품 및 화장품 소재로써 흰색 느티만가닥버섯(Hypsizygus marmoreus)의 이용 가능성을 조사하기 위해서 흰색 느티만가닥버섯 열수 추출물과 메탄올 추출물의 항산화 활성 및 tyrosinase 저해 효과를 비교하였다. 열수 추출물과 메탄올 추출물의 총 폴리페놀 함량은 각각 8.4±3.27 mg GAE/g과 7.3±2.85 mg GAE/g이었고 플라보노이드 함량은 각각 4.8±3.81 ug/mg과 2.5±1.95 ug/mg이었으며 총 폴리페놀과 플라보노이드 함량 모두 메탄올 추출물보다 열수 추출물에서 높게 나타났다. Tyrosinase 저해 활성은 추출물의 농도에 따라 증가하는 경향을 나타내었으나 양성 대조구로 사용한 2% 알부틴 (arbutin)비해 40 mg/ml의 높은 농도에서도 열수 추출물은 69.72%, 메탄올 추출물은 52.67%의 낮은 저해 활성을 나타내었다. 항산화 활성은 DPPH에 의한 라디칼소거 활성을 측정하여 확인하였으며 열수 추출물과 메탄올 추출물의 DPPH 라디칼 소거능은 40 mg/ml의 농도에서도 각 각 80%와 74%로 낮게 나타났다. 추출물의 세포독성은 WST-1 assay (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H- 5-tetrazolio]-1,3-benzene disulphonate)를 이용하여 추출물의 처리농도에 따른 B16BL6 melanoma cell의 세포생존율로 확인하였으며 열수 추출물과 메탄올 추출물을 각각 0-40 mg/ml의 농도로 처리하였을 때 B16BL6 melanoma cell은 90% 이상의 생존율을 나타내었으므로 열수 추출물과 메탄올 추출을 모두 B16BL6 melanoma cell에 독성을 나타내지 않는 것으로 판단된다.

Twelve strains of bacteria with cellulase and xylanase activities were isolated from spent mushroom substrates collected from button mushroom cultivation farm, Buye, Chungcheongnam-do in Korea. Among them, one strain, designated NO12, with higher cellulase and xylanase activities was selected by agar diffusion method. The strain NO12 was identified to be a Bacillus sp. by biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA gene sequence analysis showed that strain NO12 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99.2%. Based on its physiological properties, biochemical characteristics, and phylogenetic distinctiveness, strain NO12 was classified within the genus Bacillus, for which the name Bacillus subtilis NO12 was proposed. The cellulase and xylanase activities of B. subtilis NO12 were slightly increased according to bacterial population from exponential phase to stationary phase in the growth curve for B. subtilis NO12. The xylanase activity continuously increased from the beginning of the exponential phase and exhibited maximum activity in the middle of the exponential phase.

In this study, we investigate the potential use of persimmon peels (PP) in mushroom culture medium for the production of functional mushrooms. Pleurotus eryngii was cultivated in medium supplemented with PP (SMPP) at the following concentrations: 0% SMPP (control), 5% SMPP, 10% SMPP, 15% SMPP, 20% SMPP, or 30% SMPP. The total polyphenol content, DPPH radical scavenging ability, ABTS cation scavenging ability, and reducing power of P. eryngii cultivated in SMPP were investigated. P. eryngii cultured in 20% SMPP produced the highest values for all four measurements. Total polyphenol content, DPPH radical scavenging activity, ABTS cation scavenging ability, and reducing power all increased upon the addition of PP. Based on our results, we can conclude that persimmon peels are a highly valuable supplement for functional mushroom culture medium.

In order to isolate thermophilic bacteria with high activity of CMCase and xylanase, spent mushroom substrates was collected from an oyster mushroom cultivation farm in Jinju, Gyeongnam, Korea. Among the isolates, one strain designated as YJ09 was selected by agar diffusion method. The isolate YJ09 was identified as a member of Bacillus licheniformis based on biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA sequence analysis showed that isolate YJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus licheniformis with sequence similarity of 98.9%. Based on its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the isolate YJ09 was classified as Bacillus licheniformis. The CMCase and xylanase activity of B. licheniformis YJ09 was slightly increased corresponding to the bacterial population from exponential phase to stationary phase in the growth curve of B. licheniformis YJ09.

This study was conducted to investigate the effects of Kojongsi persimmon peels as a mushroom substrate on antioxidant activity of Pleurotus eryngii. The Pleurotus eryngii was growth on the medium with 0, 5, 10, 15, 20, 30, 40, 50% Kojongsi persimmon peels. The Pleurotus eryngii was extracted into ethanol and their antioxidant activities were analyzed. The total polyphenol content was highest in the 20% Kojongsi persimmon peels treatment (40±0.38 mg of gallic acid equivalents/100 g) than 0% Kojongsi persimmon peels treatment (21 ±0.18 mg of gallic acid equivalents/100 g). The 20% Kojongsi persimmon peels treatment showed better DPPH scavenging activity (36.81±0.23% at 1 mg/ml), ABTS scavenging activity (39.03±0.10% at 1 mg/ml) and reducing power (0.3±0.10% at 1 mg/ml) than 0% Kojongsi persimmon peels treatment and another extract ones. Overall, total polyphenol content and antioxidant activities of extracts from Pleurotus eryngii was increased dose dependently and 20% Kojongsi persimmon peels treatment was showed better total polyphenol content and antioxidant activities than 0% Kojongsi persimmon peels treatment and another extract ones.

Bottom mushroom (Agaricus bisporus) is an edible basidiomycete mushroom in Europe and North America. The wheat straw, wheat brain and chicken manure were used for bottom mushroom cultivation. Bottom mushroom substrates for cultivation was collected from bottom mushroom farm in Buyeo. About 16 bacteria were isolated from fresh mushroom substrates on TSA medium. Among of the isolates, isolate 3HS12 possessed high cellulase and mannase activity was selected as a hydrolase-producing bacteria. The isolate 3HS12 was identified as members of the Bacillus subtilis by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that isolate 3HS12 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 98.8%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the 3HS12 isolate was assigned to the Bacillus subtilis. The cellulase and mannase activity of Bacillus subtilis 3HS12 was increased according to the bacterial density at 40°C.

The Kojongsi persimmon peels is byproduct after production of dried persimmon. This study was conducted to investigate optimal mixing ratio of Kojonsi persimmon peels as a substrate for cultivation of Pleurotus eryngii. The primary nutritive material of substrate for Pleurotus eryngii was corncob, soybean meal, beet pulp, ground corn and bran. Five to forty percent of persimmon peels were mixed with primary nutritive materials as new substrate. The chemical properties of substrate mixed with Kojongsi persimmon peels was similar to the mixture without Kojongsi persimmon peels in T-C, T-N, C/N ratio, and other nutrients. Mycelial growth time on medium with 5 to 30% persimmon peels was similar to the control without persimmon peels as 27 days but that on medium with 40% and 50% persimmon peels was delayed for 10 days. The time of pinhead formations in medium with 5 to 30% persimmon peels was 12 days, the growth time to harvest was 6 days. These results were similar to those of the control without persimmon peels. The sizes of pileus of treatments with 5 to 30% persimmon peels were tend to be smaller compared with the control. The length of stipe of 10% persimmon peels treatment was 80 mm which was shorter than that of the control with 120 mm.

The mannanase-producing bacteria, designated YJ17, was isolated from spent mushroom (Flammulina velutipes) substrates. The isolate YJ17 was a facultative anaerobic and was grown at temperatures ranging from 20oC to 50oC with an optimal temperature of 40oC. The DNA G+C content of the YJ17 was 44 mol%. The major fatty acids were anteiso-15:0 (38.9%), 17:0 (7.6%), and iso-15:0 (36.5%). The 16S rRNA gene sequence similarity between the isolate YJ17 and other Bacillus strains was from 98% to 99%. In the phylogenetic analysis based on these sequences, the isolate YJ17 and Bacillus amyloliquefaciens clustered within a group together and separated from other species of Bacillus. Based on the physiological and molecular properties, the isolate YJ17 was classified within the genus Bacillus as B. amyloliquefaciens YJ17. The optimal pH and temperature for mannanase activity of B. amyloliquefaciens YJ17 were pH 7.0 and 50oC, respectively.

본 연구는 지역 특산물인 감과피를 이용한 버섯배지 개 발을 위한 기초자료로서 감과피의 이용 가능성을 검토하 고자 수행되었으며 감과피는 경상남도 산청지역에서 곶감 을 생산하고 남은 것을 수거하여 건조한 후 사용하였고 공시균주는 ASI 2312를 사용하였다. 대조구는 혼합배지:콘코브:톱밥(220:65:15, v/v)을 혼합한 기본배지를 사용하 였고 기본배지 중 혼합배지를 5%, 10%, 15%, 20%, 30%, 40%, 50%의 건조된 감과피 처리구로 대체한 처리 구를 시험구로 사용하였다. 대조구인 기본배지에 비해 시 험구인 5%, 10%, 15%, 20%, 30%, 40%, 50% 감과피 처 리구의 T-N 함량은 감과피 첨가량이 증가할수록 감소하 는 경향을 나타내었으며 T-C 함량은 큰 차이가 없었고 C/ N율은 감과피 첨가량이 증가할수록 증가하는 경향을 나 타내었다. 감과피 첨가량이 균사생장에 미치는 영향을 조 사한 결과 대조구보다 감과피가 첨가된 모든 시험구에서 균사생장이 느리게 나타났다. 감과피 첨가량에 따른 배양 일수는 대조구인 혼합배지에 비해 감과피를 30%까지 첨 가하여도 배양일수와 초발이 소요일수에는 뚜렷한 차이는 없었지만 감과피 첨가량이 증가할수록 배양일이 길어지는 경향을 보였으며 혼합배지에 비해 감과피 첨가비율이 증 가할수록 갓의 직경은 감소하고 대의 길이는 길어지고 직 경은 감소하는 경향을 나타내었다. 수확 후 동결 건조한 큰느타리버섯의 수분, 단백질, 조회분 함량은 대조구와 시 험구에서 유의성 있는 차이를 나타내지 않았지만 조지방 함량은 감과피 첨가량이 증가함에 따라 증가하는 경향을 나타내었고 β-glucan 함량은 대조구에 비해 10%와 15% 감과피 첨가구에서 높게 나타났다. 따라서 대조구와 비교 했을 때 15% 감과피 첨가구는 자실체 수량도 높게 나타 나고 β-glucan 함량도 높게 나타났기때문에 감과피 첨가 비율은 15%가 가장 적합한 것으로 사료된다.

Cellulase와 xylanase 분비능이 우수한 부숙촉진 세균을 분리하기 위하여 진주시 집현면 소재의 느타리버섯 재배 농장으로부터 느타리버섯 수확후배지를 수집하였다. 느타 리버섯 수확후배지로부터 19종의 균주를 분리하였으며 이 중 cellulase와 xylanase을 동시에 분비하는 균주를 최 종 선발하여 CA105로 명명하였다. Bacillus ID kit와 VITEK 2 system를 이용하여 분리균 CA105의 생화학적 특성을 조사한 결과에서도 분리균 CA105은 B. subtilis와 유사한 특징을 나타내었으며 16S rRNA 염기서열 분석 결과에서는 B. subtilis와 98.9%의 상동성을 나타내었다. 이와 같은 결과를 종합하여 분리균 CA105은 Bacillus subtilis CA105로 동정되었다. 분리균이 분비하는 cellulase와 xylanase 활성은 분리균이 증식함에 따라 대수 증식기 중반부터 급격히 증가하였고 정지기에 진입하면 효소활성이 더 이상 증가하지 않는 것으로 나타났다.

This study was carried out to investigate optimal mixture of persimmon peels for Pleurotus eryngii cultivation. The 5-50% of persimmon peels was mixed with mushroom growing medium for Pleurotus eryngii cultivation. Mycelial growth of persimmon peels treatment were 74-165 mm (28 days) and slower than that of control (treatment without persimmon peels) as 160 mm (28 days). Mycelial growth time of 5-20% persimmon peels treatment were similar to control as 34 days but that 30-50% of persimmon peels treatment were delayed for 17 days. The time of pinhead formations in control and 5-50% persimmon peels treatment were 11 days, the growth time to harvest were 6-7 days. The sizes of pileus of 15% persimmon peels treatment was tend to be smaller compared with control. The length of stipe of 15% persimmon peels treatment was 124 mm which was longer than that of the control with 118 mm. The thickness of stipes of 15% persimmon peels treatments was 48 mm which were tend to be thicker than that of the control with 47 mm. Thirty percent of persimmon peels treatment was better than other treatment comparing with control.

This study was carried out to develope mushroom growing medium for Pleurotus eryngii cultivation using persimmon peels. Total nitrogen, carbon source and protein of commercial cultivation medium used in this study was 1.34, 53.9 and 0.09 %, respectively. The C/N ratio was 40.22. The total nitrogen was increased by increasing mixing ratio of persimmon peels. The carbon source and protein was no significant difference by increasing mixing ratio of persimmon peels. In column test, mycelial growth of 15% persimmon peels treatment was similar with control, but mycelia of 20-50% persimmon peels treatment was growed slowly compared with control. To investigate yields of fruiting body, 15% of persimmon peels was mixed with commercial cultivation medium. The fruiting body of 15% persimmon peels treatment was no significant difference yield compared with control. But, the chemical contents of 15% persimmon peels treatment were the higher than control.

본 연구는 큰느타리버섯 수확후배지 발효사료에 적합한 생균제를 개발하고 큰느타리버섯 수확후배지 발효사료 급여가 비육한우에 미치는 영향을 조사하기 위하여 수행되었다. 큰느타리버섯 수확후배지 발효사료를 제조하기 위하여 큰느타리버섯 수확후배지로부터 cellulase, xylanase 활성이 우수하면서 Asp. flavus에 대해 항균활성을 나타내는 균주를 선발하여 Bacillus subtilis CS21로 명명하고 버섯수확후배지 발효사료용 생균제로 사용하였다. 사양시험은 13개월령 비육한우 20두를 공시하여 29-30개월령까지 수행하였다. 공시동물은 성장단계(육성기, 비육전기, 비육후기)에 따라 배합비를 조절한 TMR 사료를 급여한 대조구와 30%의 큰느타리버섯 수확후배지 발효사료가 첨가된 TMR 사료(30% F-SMS TMR)를 급여한 처리구로 나누어 수용하였다. 시험기간 동안 총 증체량과 일당 사료섭취량은 대조구보다 30% F-SMS TMR 급여구에서 약간 높게 나타났다. 육질등급에서는 대조구와 30% F-SMS TMR 급여구 모두 1등급 이상 출현율이 100%를 나타내었으나 육량등급의 A등급 출현율은 대조구(57%)가 30% F-SMS TMR 급여구(41.67%)보다 높은 경향을 나타내었으며 C등급 출현율은 대조구(0%)보다 30% F-SMS TMR 첨가구(25%)에서 높은 경향을 나타내었다. 이상의 결과를 종합하면 발효공정을 거쳐 기호성과 저장성이 개선된 큰느타리버섯 수확후배지 발효사료 급여는 육질등급에서는 대조구와 비슷하였지만 증체량은 대조구보다 30% F-SMS TMR 급여구가 높은 반면 육량등급의 A등급 출현율은 낮고 C등급 출현율은 높은 경향을 나타내었으므로 육량등급을 높일 수 있도록 배합사료 조성비를 개선한다면 큰느타리버섯 수확후배지는 비육한우의 대체사료자원으로 이용될 수 있을 뿐만 아니라 사료비절감 효과를 기대할 수 있을 것으로 사료된다.

본 연구는 지역 특산물인 감과피를 이용한 버섯배지 개발을 위한 기초자료로서 감과피의 이용 가능성을 검토하 고자 수행되었으며 감과피는 경상남도 진주시에 소재한 진주단감원예조합으로부터 건조된 것을 수집하여 사용하 였고 공시균주는 큰느타리 2312를 사용하였다. 대조구는 시판혼합배지를 사용하였고 건조된 감과피 5%, 10%, 15%, 20%, 25%, 30%이 첨가된 시판혼합배지를 시험구 로 사용하였다. 대조구인 시판혼합배지에 비해 시험구인 5%, 10%, 15%, 20%, 25%, 30%의 감과피가 첨가된 시 판혼합배지의 총질소 함량은 감과피 첨가량이 증가할수록 증가하는 경향을 나타내었으며 총탄소와 유기물 함량은 큰 차이가 없었고 C/N율은 감과피 첨가량이 증가할수록 감소하는 경향을 나타내었다. 감과피 첨가량에 따른 시험 관 내에서의 균사생장을 조사한 결과 대조구인 시판혼합 배지보다 감과피가 첨가된 모든 배지에서 균사생장이 느 리게 나타났다. 시험구의 혼합배지 중 5% 감과피가 첨가 된 배지에서 균사 생장이 가장 빨라지만 감과피 첨가량이 증가할수록 균사 생장속도는 느려지는 경향을 보였다. 감 과피 첨가량에 따른 배양일수는 대조구인 시판혼합배지에 비해 감과피를 30%까지 첨가하여도 배양일수와 초발이 소요일수에는 뚜렷한 차이는 없었지만 감과피 첨가량이 증가할수록 배양일이 길어지는 경향을 보였으며 시판혼합 배지에 비해 감과피 첨가비율이 증가할수록 갓의 직경과 두께는 감소하고 대의 길이는 짧아지고 굵기는 굵어지는 경향을 나타내었다. 따라서 감과피를 배지자원으로 이용 하기 위해서는 감과피 첨가량에 따른 혼합배지의 조성비 가 필요할 것으로 생각된다.

True hermaphrodites are animals of equivocal sex in which both male and female gonads develop simultaneously. The frequency of true hermaphroditism is higher in pigs than in other domestic animals. Two Korean pigs were diagnosed with true hermaphroditism showing ovotestes, epididymes, penes, and uteri. Histomorphologically, the testicular tissues consisted of Sertoli cells that were devoid of spermatogenic germ cells and showed proliferation of interstitial cells. However, the uteri were of normal architecture and had well-developed uterine endometrial glands. The samples were 38, XX female karyotype without the sex-determining region Y (SRY) gene. The findings of this study could contribute to the understanding of true hermaphroditism in the Korean pig industry. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio- technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being.

The objective of this study was to evaluate in vitro production of bovine embryos in Hanwoo. Oocytes were collected by ovum pick up (OPU) from ovaries of genetically high-value Hanwoo or by needle puncture from ovaries of slaughtered cattle. OPU was done every 3 4 days duing experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, We compared the in vitro maturation rate in two groups (Experiment 1). 545 oocytes were recoverd from 4 females by 32 trials of OPU and then 433 oocytes were shown MⅡ stage after in vitro maturation (79.4%). In case of needle puncture group, 1905 oocytes were collected and then 1420 oocytes were matured to MⅡ stage during in vitro culture(74.5%). Second, we compared the developmental rate to blastocyst in two groups (Experiment 2). 1420 oocyte by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24 48 h after in vitro fertilization (IVF) was 88.6% and blastocyst development rate was 20.5% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (84.8%), blastocyst development rate was higher compared with needle puncture group (26.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio-technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being. This research was suppoted by Imsil-gun agricultural technology service center.