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Effect of Collective Methods on the Collection Efficiency, Blastocyst and Pregnancy Rate after IVP Embryo Transfer in Hanwoo (한우 난소의 채란방법이 회수율, 배발달율 및 수태율에 미치는 영향)

  • Lee J. W.;Jung S. Y.;Son B. H.;Han K. H.;Oh I. S.;Seo H. J.;Kong I. K.
    • Journal of Embryo Transfer
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    • v.20 no.1
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    • pp.55-62
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    • 2005
  • This study was undertaken to access the effect of collection methods on the collection efficiency, blastocyst rate and pregnancy rate after IVP embryo transfer. The ovaries of Hanwoo were obtained from an abattoir and kept on 25 to $28^{\circ}C$ and transported to laboratory within 4 hrs. The oocytes were collected by aspiration of follicles $(2\~6\;mm)$ with or without slicing of ovaries after aspiration. The oocytes were matured in vitro (IVM) for 20 to 24 hrs in TCM-199 supplemented with $10\%$ fetal bovine serum at $39^{\circ}C$ under $5\%\;CO_{2}$ in air. Following routine IVM/IVF procedure, the oocytes and presumed zygotes were cultured for three day in CRlaa medium with BSA. The cumulus cells at 2 to 8-cell stage of embryos removed then the embryos and were cultured in CRlaa medium containing $10\%$ fetal bovine serum in $5\%\;CO_{2}$ at $39^{\circ}C$. The fresh blastocysts cultured for 7 to 9 days were transferred into recipients. The numbers of oocytes recovered form two different methods, the aspiration and slicing after aspiration, were compared to know what. The number of oocytes per ovary was 8.2 and 6.5 in aspiration combining slicing, and aspiration groups, respectively (p<0.05). The cleavage rate in aspiration method are significantly (p<0.05) high than those in slicing post aspiration $(27.9\%)$, and aspiration $(25.5\%)$. The pregnancy .ate in aspiration method $(62.5\%)$ was high than that in slicing method after aspiration $(54.4\%)$. The pregnancy rates of aspiration method and slicing method after aspiration in nullipara $(58.1\%\;vs\;68.2\%)$ was high than that in pluripara $(49.5\%\;vs\;53.2\%)$. The results obtained that the increased number of oocytes per ovary in slicing method after aspiration could be better than that in aspiration method. Pregnancy rate in aspiration method was slightly higher in than that in slicing method after aspiration.

Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

Serial Ultrasonographic Appearance of Postpartum Uterine Involution in Cocker Spaniel Bitches (Cocker Spaniel 개에서 분만 후 자궁수복의 연속적 초음파상)

  • Hwang S. S.;Oh K. S.;Kim B. S.;Jo Y. T.;Park C. H.;Kim J. T.;Park I. C.;Park J. T.;Park S. G.;Son C. H.
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.271-277
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    • 2005
  • This study was undertaken to determine the normal appearance of the postpartum uterine involution. Postpartum changes in uterine shape, architecture, echogenicity and diameter were monitored with ultrasonography in 8 Cocker spaniel bitches. The excretory period of vaginal discharges in 8 normal bitches of uterine involution was finished completely at 23.20$\pm$2.77 days (Mean$\pm$SD) postpartum. The short axis shape of the uterus was varied from polygonal to circular. This lasted until 1600$\pm$2.12 days Postpartum, during which time the short axis uterine shape gradually changed to circular. Also, the long axis shape of the uterus was created a beaded appearance of the horns until 25.60$\pm$2.51 days postpartum. The uterine diameter was decreased not only in the placental sites from 24.20$\pm$2.06mm at 1 day to 13.18$\pm$0.84mm at 7 days postpartum, but also in the interplacental sites 14.26$\pm$2.22mm at 1 day, 9.81$\pm$0.7mm at 7 days postpartum. There was a general trend of decreasing uterine diameter, which occurred more rapidly at the placental sites. In conclusion, normal postpartum uterine involution in Cocker spainel bitches appeared to be completed around 68 days postpartum by gross findings such as vaginal discharges, and by ultrasongraphic findings, uterine shape and echogenicity.

Survival and Development of Porcine Embryos Produced in vitro Using Open Pulled Straw Methods (돼지에서 Open Pulled Straw(OPS) 방법에 의해 동결-융해한 수정란의 생존능력)

  • Lee, S.Y.;Yu, J.S.;Sa, S.J.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.255-262
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    • 2006
  • The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

Estrus Induction Rates and Conception Rates by MGA Feeding System in Hanwoo (MGA Feeding System에 의한 발정 동기화 방법이 한우의 발정 발현율과 수태율에 미치는 영향)

  • Lee, M.S.;Jung, Y.H.;Park, S.B.;Im, S.K.;Park, J.J.;Jung, J.K.;Moon, S.J.;Kim, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.177-182
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    • 2006
  • This study was conducted to investigate the effects of melengesterol acetate (MGA) and $PGF_{2{\alpha}}$ administrations on serum progesterone level, synchrony of estrus and conception rates in Han-woo. Firstly, ten heifers and one freematin were fed 0.5 mg MGA/day for 14 days in a grain carrier, and after 19 days of MGA feeding, a single injection of 25 mg $PGF_{2{\alpha}}$ were treated. Blood samples were collected to evaluate serum progesterone concentrations from the start of feeding of MGA until the end of feeding and subsequent estrous detection and artificial insemination (AI) at 3 days intervals, and on days of $PGF_{2{\alpha}}$ injection, estrous detection, AI, and 15th and 60th days after AI. The level of progesterone in the blood began to increase from 7 days after MGA feeding, and 9 days after feeding it became 5.4 ng/ml and maintained that level thereafter. On the 33th day when the $PGF_{2{\alpha}}$ was injected, it reached the peak level of 7.6 ng/ml. However, 2-3 days after $PGF_{2{\alpha}}$ injection, it dropped to 1.4 ng/ml drastically (p<0.05). Secondly, one hundred and ninety four Hanwoo heifers or cows were divided into two groups to compare estrous induction and conception rates: the one treated with MGA and $PGF_{2{\alpha}}$, (n=104) and the other with $PGF_{2{\alpha}}$ treatment (two injections at 11 days interval, n=90). The heifers or cows treated with MGA and $PGF_{2{\alpha}}$ were identical to those used as above. The percentages of heifers or cows showed estrus were higher in the $MGA+PGF_{2{\alpha}}$ treatment (91.3%) than in the $PGF_{2{\alpha}}$ treatment (72.2%, p<0.05). Conception rates were also higher in the $MGA+PGF_{2{\alpha}}$ treatment (94.2%) than in the $PGF_{2{\alpha}}$ treatment (88.9%, p<0.05). The results of this experiment indicate that estrus synchronization using $MGA+PGF_{2{\alpha}}$ is more effective than that using $PGF_{2{\alpha}}$ (two injections) in Hanwoo.

Studies on Early Selection of Excellent Gilts for Improvement of Reproductive Efficiency I. First Estus and Litter Size of Candidate Gilts (번식효율 증진을 위한 후보 종빈돈의 조기선발에 관한 연구 I. 후보 종빈돈의 첫발정 일령과 산자수)

  • 손동수;이장희;최선호;연성흠;류일선;서국현;허태영;박성재;조규호
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.249-255
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    • 2003
  • These studies were performed to improve the reproductive efficiency of gilts and we investigated the effects of puberty periods, first mating time and backfat thickness and will adapt to these results for early selection of excellent gilts. The main results were as follows; 1. First heats on birth season were showed 194.14 day, 163.25 day, 160.25 day and 157.92 day at birth of spring, summer, autumn and winter, respectively and birth of spring was significantly latest among other seasons (p<0.01). 2. First service on birth season were revealed 222.05 day in spring, 193.00 day in summer, 199.20 day in autumn and 190.11 day in winter. birth of spring was significantly latest among others (p<0.01). 3. First heat period of cadidated gilt had 13∼16 mm backfat thickness was 180.32 day, 171.24 day in 17∼20 mm and 162.20 day in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness. 4. First service of cadidate gilt had 13∼16mm backfat thickness was 211.12 day, 202.43 day in 17∼20 mm and 195.43 day in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness. 5. The litter size were 9.64 in gilts under 160 day of first heat, 10.14 in 161∼180 day, 9.56 in 181∼200 day and 9.13 in over 201 day. There showed the largest litter size in 161∼180 day of first heat but was no differences. 6. The litter size in gilts under 180 day of first service was 9.13, 9.75 in 181∼200 day, 10.13 in 201∼220 day and 9.45 in over 221 day. There showed the largest litter size in 201∼220 day of first service but was no differences. 7. The litter size of gilt had 13∼16 mm backfat thickness on first service was 9.33, 9.81 in 17∼20 mm and 10.17 in 21∼23 mm and was showed delay in thin backfat gilts. There was no differences among backfat thickness.

Studies on Early Selection of Excellent Gilts for Improvement of Reproductive Efficiency II. Return of Estrus after the Parturition of Candidate Gilts (번식효율 증진을 위한 후보 종빈돈의 조기선발에 관한 연구 II. 후보 종빈돈의 분만후 발정재귀일)

  • 손동수;이장희;최선호;연성흠;류일선;서국현;허태영;박성재;조규호
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.257-262
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    • 2003
  • These studies were performed to improve the reproductive efficiency of gilts and we investigated the effects of nursing periods after parturition and backfat thickness will adapt to these results for early selection of excellent gilts. The main results were as follows; 1. The backfat thickness in sows nursing for 17∼21 days was 19.19 mm, and 16.52 mm in 22∼26 days and longer nursing period affected significantly the backfat thinner (p<0.01). 2. The recurrence of estrus in sows nursing for 17∼21 days was 5.76 day and 5.62 day in 22∼26 days and there was no difference between nursing periods. 3. The recurrence of estrus in sows had 13∼16 mm backfat thickness was 5.69 day, 5.67 day in 17∼20 mm and 5.75 day in 21∼23 mm and there was no difference among backfat thickness. 4. The concentration of plasma estradiol were 28.49 pg/$m\ell$ in parturition, 12.39 pg/$m\ell$ in weaning and 16.52 pg/$m\ell$ in return of estrous. The concentration ot plasma progesterone were 1.50 ng/$m\ell$ in parturition, 0.69 ng/$m\ell$ in weaning and 0.94 ng/$m\ell$ in return of estrous. The concentration of plasma cortisol were 57.74 ng/$m\ell$ in parturition, 43.01 ng/$m\ell$ in weaning and 47.89 ng/$m\ell$ in return of estrous. There showed the highest level of estradiol, progesterone and cortisol in parturition.

Evaluation of Future Turbidity Water and Eutrophication in Chungju Lake by Climate Change Using CE-QUAL-W2 (CE-QUAL-W2를 이용한 충주호의 기후변화에 따른 탁수 및 부영양화 영향평가)

  • Ahn, So Ra;Ha, Rim;Yoon, Sung Wan;Kim, Seong Joon
    • Journal of Korea Water Resources Association
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    • v.47 no.2
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    • pp.145-159
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    • 2014
  • This study is to evaluate the future climate change impact on turbidity water and eutrophication for Chungju Lake by using CE-QUAL-W2 reservoir water quality model coupled with SWAT watershed model. The SWAT was calibrated and validated using 11 years (2000~2010) daily streamflow data at three locations and monthly stream water quality data at two locations. The CE-QUAL-W2 was calibrated and validated for 2 years (2008 and 2010) water temperature, suspended solid, total nitrogen, total phosphorus, and Chl-a. For the future assessment, the SWAT results were used as boundary conditions for CE-QUAL-W2 model run. To evaluate the future water quality variation in reservoir, the climate data predicted by MM5 RCM(Regional Climate Model) of Special Report on Emissions Scenarios (SRES) A1B for three periods (2013~2040, 2041~2070 and 2071~2100) were downscaled by Artificial Neural Networks method to consider Typhoon effect. The RCM temperature and precipitation outputs and historical records were used to generate pollutants loading from the watershed. By the future temperature increase, the lake water temperature showed $0.5^{\circ}C$ increase in shallow depth while $-0.9^{\circ}C$ in deep depth. The future annual maximum sediment concentration into the lake from the watershed showed 17% increase in wet years. The future lake residence time above 10 mg/L suspended solids (SS) showed increases of 6 and 17 days in wet and dry years respectively comparing with normal year. The SS occupying rate of the lake also showed increases of 24% and 26% in both wet and dry year respectively. In summary, the future lake turbidity showed longer lasting with high concentration comparing with present behavior. Under the future lake environment by the watershed and within lake, the future maximum Chl-a concentration showed increases of 19 % in wet year and 3% in dry year respectively.

Formulation of Bacillus amyloliquefaciens A-2 and Its Efficacy to Control Tomato Leaf Mold Caused by Fulvia fulva (길항세균 Bacillus amyloliquefaciens A-2를 이용한 토마토 잎곰팡이병 방제용 미생물 제제)

  • Kong, Hyun-Gi;Chun, Ock-Joo;Choi, Ki-Hyuck;Lee, Kwang-Youll;Baek, Joung-Woo;Kim, Hyun-Ju;Murugaiyan, Senthilkumar;Moon, Byung-Ju;Lee, Seon-Woo
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.27-34
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    • 2010
  • This study was performed to develop a formulation using an antagonistic bacterium Bacillus amyloliquefaciens A-2 to control tomato leaf mold caused by Fulvia fulva. B. amyloliquefaciens A-2 was grown in a medium with rice oil and mixed with various carrier and additives. One of the formulations, A2-MP, showed the best disease control value among the tested formulations. The disease control value of A2-MP at 100-fold and 500-fold diluted treatment was not significantly different from that of chemical fungicide triflumizole in a growth chamber. Although disease control effect was decreased by serial diluted treatment of the prepared A2-MP, 1,000-fold diluted treatment of A2-MP still showed high disease control value of 72.0%. For the green house experiments, the disease control values of A2-MP was indicated as 79.4% which is similar to that of chemical fungicide, triflumizole showing 79.6%. When the disease control activity of the formulation A2-MP was compared in tomato production conditions, disease control values of 100-fold diluted A2-MP and 3,000 fold diluted triflumizole exhibited 60%, 81.6%, respectively. The disease control efficiency by A-2MP was 73% of the disease control value of chemical fungicide. The formulation A-2MP maintained the stable bacterial viability and disease control activity when stored at $4^{\circ}C$. This result suggested that A-2MP develped from B. amyloliquefaciens A-2 could be used to control tomato leaf mold.

Survey of Asbestos-Related Disease on Chest Radiograph of Workers Exposed to Asbestos in Ship Repair (조선수리업종 석면노출 근로자들의 흉부방사선상 석면관련 폐질환 소견의 발생양상)

  • Lee Hyun Jae;Hong Young Seoub;Son Ji Eun;Lee Young Ill;Kim Sang Hoon;Im Hyoung June;Kim Jung Man;Kim Joon Youn
    • Journal of Life Science
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    • v.16 no.1
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    • pp.58-63
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    • 2006
  • There was cross-sectional study on investigating the factors related asbestos-related disease on chest radiograph of workers exposed to asbestos in ship repair. According to Occupational Safety & Health Administration asbestos standard, medical surveillance, chest radiologic study, pulmonary function test and medical questionnaire are performed in 118 subjects exposed to asbestos for 10 years or more in ship repair. The sixteen out of 118 subjects $(13.6\%)$ were observed asbestos-related disease finding on chest radiography. Significant factors related to those finding on chest radiography were abnormal pulmonary function test, cough, sputum, past history of respiratory disease, work duration. Workers with Significant factors related asbestos-related disease finding on chest radiography should be managed on a strict program. Further, factors unrelated with asbestos-related disease finding on this study should be evaluated prospectively for medical surveillance program of workers exposed to asbestos.