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Retinol Supplemented to a Maturation Medium Stimulated In Vitro Development of Porcine Oocytes after In Vitro Fertilization (돼지 체외 수정란 생산 효율 제고를 위한 레티놀 첨가 효과)

  • Kim Y.S.;Song S.H.;Cho S.K.;Kwack D.O.;Kim C.W.;Park C.S.;Chung K.H.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.101-107
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    • 2006
  • The objective of this study was to investigative the effects of retinol supplement to IVM and/or IVC medium on maturation, fertilization and development of pig oocytes. North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (pFF) was used as base medium. Each 1 uM, 5 uM and 10 uM concentration of retinol was supplemented to IVM and /or IVC medium. When the retinol was supplemented to maturation medium, the maturation rates were not different (p>0.05) among treatment groups ($66.7{\pm}6.0{\sim}69.2{\pm}5.3%$), but the developmental rate to blastocyst stage was higher (p<0.05) in $5{\mu}M$ group ($20.4{\pm}2.6%$) than in 0 uM ($13.6{\pm}2.1%$) and 10 uM groups ($9.7{\pm}1.7%$). Moreover, total cell number was significantly greater (p<0.05) in the 5 uM group ($37.0{\pm}1.6$) than in the other groups ($29.8{\pm}1.0{\sim}33.2{\pm}1.0$). Retinol supplement to maturation medium did not significantly affect the rates of fertilization and polyspermy (p>0.05). When the retinol was supplemented to culture medium or both maturation and culture medium, the rates of cleavage, and develop to morula and blastocyst stage were not affected, while those of 10 uM group were significantly decreased (p<0.05). These results indicate that 5 uM retinol supplement in maturation medium significantly stimulates embryo development, also improves the total cell number of blastocyst stage in pig.

Production of In-Vitro Fertilized Embryos and Result of Transfer with Fresh or Frozen Embryos for Hanwoo and Holstein Cattle (한우 및 젖소에서 체외 수정란 생산과 신선 및 동결 수정란 이식 결과)

  • Kim Y. J.;Kim H. C.;Seo S. H.;Jeong K. N.;Kim Y. S.;Lee H. R.;Shin D. S.;Jo S. W.;Kim S. H.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.79-87
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    • 2005
  • The ovaries from Hanwoo and Holstein were collected from labattoir and transferred to laboratory. Oocytes were aspirated and incubated in $CO_2$ incubator for 24 hours for maturation. Oocytes were coincubated with the sperms for 5 hours. Cleaved oocytes were selected 48 hours after coincubation and half of the medium was changed newly every 48 hour until blastocyst formation. Cleavage rate and blastocyst rate were investigated according to different breeds and different status of cumulus cells surrounding the oocytes. Blastocysts were either transferred to the recipients or frozen until use. The result of embryo transfer with fresh or frozen embryos was investigated. The rate of male offspring following embryo transfer was also investigated. The rate of cleavage was $66.4\%$ for Hanwoo and $62.4\%$ for Holstein oocytes. The rate of cleavage according to status of oocyte was shown highest in the oocytes completely surrounded with cumulus cells and lowest in denuded oocytes for both Hanwoo and Holstein oocytes. The rate of blastocyst from cleaved oocytes was $40.6\%$ for Hanwoo and $36.9\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with fresh IVF embryos was $57.2\%$ for Hanwoo and $53.3\%$ for Holstein. The rate of pregnancy/delivery following embryo transfer with frozen IVF embryos was $40.9\%$ for Hanwoo and $36.4\%$ for Holstein. The rate of male calf produced by embryo transfer was $63.6\%$ for Hanwoo and $50.0\%$ for Holstein.

Changes of Frozen-Thawed Semen Characteristics in Miniature Pig and Duroc (Miniature Pig와 Duroc 종간의 동결-융해 후 정액 성상 비교)

  • Lee, Y.S.;Choi, W.C.;Lee, S.H.;Cheong, H.T.;Lee, S.Y.;Yang, B.K.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.263-271
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    • 2006
  • The purpose of this study was undertaken to compare ability of frozen-thawed sperm characteristics between two strains (miniature pig and Duroc). The semen was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle. The semen was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. The frozen-semen straw were thawed at 20, 37 and $50^{\circ}C$ for 1 min, 45 sec and 10 sec within water-bath. The semen sample were evaluated at 0, 3, 6, 9, and 12 h after incubation at $37^{\circ}C$ for analysis of sperm ability. Abnormality of spermatozoa in miniature pig was significantly (p<0.05) higher than that in Duroc at 0, 9 and 12 h of post-thawing incubation after frozen-thawing. The percentage of F-patterned spermatozoa in miniature pig was significantly (p<0.05) lower, while the percentage of AR (acrosome reacted spermatozoa) pattern was higher in the miniature than in the Duroc. On the other hand, there was no significant difference in the viability of spermatozoa thawed at different temperature ($20^{\circ}C\;and\;37^{\circ}C$) between two species, but the viability in miniature pig was higher (p<0.05) than in Duroc when sperm was thawed at $50^{\circ}C$. In conclusion, this study suggest that suitable freezing method for miniature pig semen is required for increasing post-thawing viability and fertilization capacity.

In Vivo Embryo Production and Embryo Transfer in Hanwoo and Jeju Black Cattle Using CIDR (CIDR를 이용한 제주 한우 및 흑우의 체내 수정란 생산과 이식)

  • Kim, Y.H.;Koo, J.C.;Oh, C.W.;Kang, S.Y.;Yang, B.S.;Oh, S.J.;Kim, C.N.;Song, J.Y.;Kim, I.H.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.191-198
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    • 2006
  • The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

Treatment of Reproductive Dysfunctions and Reproductive Monitoring Using Ultrasonography in Dairy Cow (초음파 검사에 의한 젖소 번식 검진과 번식 장애 치료)

  • Lim, W.H.;Oh, K.S.;Seo, G.J.;Hwang, S.S.;Kim, B.S.;Bae, C.S.;Kim, S.H.;Kim, J.T.;Park, I.C.;Park, S.G.;Son, C.H.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.217-223
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    • 2006
  • This study was carried out principally to obtain the basic data for the improvement of the reproductive performance and production using plasma progesterone assay and ultrasonography in dairy cow. The results obtained from this studies were as follows. The results of reproductive examination in 85,983 cows were ovarian diseases 40,399 (47.0%), uterine diseases 11,912 (13.9%), pregnancy or pregnant failures 26,587 (30.9%), adhesion of reproductive tracts 172 (0.2%), freemartin 8 (0.01%), and others 6,905 (8.3%), respectively. The treatment status of reproductive dysfunction in 30,241 cows were silent heat or error of estrus detection 14,909 (49.3%), follicular cysts 3,750 (12.4%), luteal cysts 907 (3.0%), inactive ovaries 665 (2.2%), granulosa cell tumor of ovary 3 (0.01%) and endometritis 6,986 (23.1%), respectively. The indices of reproductive efficiency after the periodical examination of reproductive status were as follows; the mean intercalving inteual was reduced from 475 days at the first examination to 381 days at the last examination of reproductive status, the mean interval calving to conception was reduced from 186 to 98 days, the mean interval calving to first service was reduced from 106 to 66 days, the cows showing heat by 60 days postpartum were increased from 32 to 90%, the mean conception rate to first service was increased from 42 to 64%, and the mean service per conception was reduced from 2.6 to 1.8 times, respectively.

Effect of Lipopolysaccharides Treatment on Uterine Involution in Dairy Cows with Retained Placenta (후산정체 젖소에 있어서 Lipopolysaccharide의 처리가 자궁 회복에 미치는 영향)

  • Baek, K.S.;Park, S.J.;Park, S.B.;Lee, W.S.;Kim, H.S.;Jeong, G.Y.;Ki, K.S.;Suh, G.H.;Jeon, B.S.;Ahn, B.S.;Kim, T.I.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.233-239
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    • 2006
  • This study was carried out to investigate the effect of intra-uterine infusion of lipopolysaccharides (LPS) on uterine involution in dairy cows with retained placenta. The LPS isolated from Bacteroides helcogenes and Fusobaoterium varium was infused at the rate of 100 ug (n=11) and 200 ug (n=11) with 30 ml of phosphate buffer saline in each cow at 20 days post-partum. The cows in conrol group (n=11) with retained placenta were infused with 30 ml phosphate buffer saline in each cow at 20 days postpartum. Cross-sectional ultrasonography was performed at days 40 postpartum to examine uterine involution and exudate in the all dairy cows in study. The cows in control group have shown 45.5, 27.3 and 27.3% gravid horn with less than 30 mm (normal), $31{\sim}50mm$ (medium) and more than 51 mm (large) in diameter. Respective average values of gravid horn diameter in cows treated with LPS 200 ug were noticed 72.7, 18.2 and 9.1%. However, the gravid horn diameter in all the cows treated with 100 ug of LPS was less than 30 mm. The 18.2, 63.6 and 54.6% cows in the control, LPS 100 ug and LPS 200 ug, respectively, have shown no exudate in uterine cavity. However, the respective rates of small amount of exudate retention in uterine cavity were 45.5, 0.0, and 9.1%. Approximately 36.4% cows in all 3 treated groups have shown minute quantity of exudate. In conclusion, the LPS intra-uterine infusion promoted postpartum uterine involution in dairy cows with retained placenta.

Selection of Promising Forage Crops and Variety for Forage Production in Paddy Field 1. Middle region(Suwon) (권역별 답리작 사료작물 최대 생산을 위한 적작목(품종) 선발 1. 중부지방(수원)을 중심으로)

  • Seo, Sung;Kim, W.H.;Kim, J.G.;Choi, G.J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.24 no.3
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    • pp.207-216
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    • 2004
  • This study was carried out to select the promising forage crops for forage production in paddy field of NLRI, RDA, Suwon from 1999 to 2001. The species of forage crop used in this experiment were barley(3 varieties), wheat(2), rye(3) and Italian ryegrass(IRG, 3). Stages of heading, milk and yellow ripe of barley were 1 May, middle May and late May, respectively. In milk to yellow ripe stage, dry matter(DM) and total digestible nutrient(TDN) yield of barley were $7.30{\sim}9.58$ MT and $4.75{\sim}6.24$ MT per ha, and Albori was the most promising variety among the barley. The growth of wheat was a little late compared to that of barley. but it seemed to be crop having higher forage yield and nutritive value. In milk to yellow stage, DM and TDN yield of wheat were $8.17{\sim}10.82$ MT and $5.33{\sim}7.31$ MT per ha. Heading stage of rye was 27 to 29 April, and it seemed to be suitable for harvest at heading to flowering stage because of heavy lodging after flowering. In flowering stage, DM and TDN yield of rye were 10.18 and 6.03 MT per ha. Heading stage of early maturing IRG was 7 May and it seemed to be good for harvest at flowering stage of middle May in cropping system. In flowering stage, DM and TDN yield of early type IRG were 4.48 and 2.96 MT per ha. The results demonstrated that the promising forage crops for forage production in paddy. field were rye and barley(Albori) in Middle region, and rye harvesting at early May was recommended when considered middle May of rice transplanting period, and rye and barley considered late May of rice planting period. Also early maturing and high yielding IRG variety was strongly needed.

Selection of Promising Forage Crops and Variety for Forage Production in Paddy Field 2. Honam region(Iksan) (권역별 답리작 사료작물 최대 생산을 위한 적작목(품종) 선발 2. 호남지방(익산)을 중심으로)

  • Seo, Sung;Kim, W.H.;Lee, J.H.;Park, T.I.;Chun, C.K.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.24 no.3
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    • pp.217-224
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    • 2004
  • This study was carried out to select the promising forage crops for forage production in paddy field of HARI, RDA, Iksan from 1999 to 2001. The species of forage crop used in this experiment were barley(5 varieties), wheat(2), rye(3) and Italian ryegrass(IRG, 3). Stage of heading, milk and yellow ripe of barley were 26 $April{\sim}l$ May, middle May and late May, respectively. In milk to yellow ripe stage, dry matter(DM) yield of barley was $6.64{\sim}10.47$ MT per ha, and averaged ADF and NDF contents were $31.6\%\;and\;62.2\%$ respectively. Albori was the most promising variety among the barley). The growth of wheat was a little late compared to that of barley, but it seemed to be crop having higher forage yield md nutritive value. In milk to yellow stage, DM yield of wheat was $7.51{\sim}11.96$ MT per ha, and nutritive value of wheat was high(ADF $30.6\%$ and NDF $60.1\%$). Heading stage of rye was $25{\sim}27$ April, and it seemed to be suitable to harvest at heading to flowering stage because of heavy lodging after flowering. In flowering stage, DM yield of rye was 17.26 MT per ha, and nutritive value of rye was the lowest among 4 species(ADF $38.6\%$ and NDF $65.5\%$). Heading stage of early maturing IRG was 30 April and it seemed to be suitable for harvest at flowering stage of middle May in cropping system. In flowering stage, DM yield of early type IRG was 10.77 MT per ha, and averaged ADF and NDF contents were $31.6\%\;and\;59.2\%$, respectively. The results demonstrated that the promising forage crops for forage production in paddy field were rye, barley, early maturing IRG and wheat in Honam region. In Southern region, rye harvesting at early May was recommended when considered middle May of rice transplanting period, and barley(Albori) and early maturing IRG harvesting at middle May were recommended when considered late May of rice planting period. And also, wheat with high nutritive value and possibility for harvesting at late May were recommended in case of transplanting period at early June.

Effect of Maturity at Harvest on the Changes in Quality of Round Baled Rye Silage (수확시 숙기가 호밀 라운드베일 사일리지의 품질변화에 미치는 영향)

  • Kim, J.G.;Chung, E.S.;Seo, S.;Kang, W.S.;Ham, J.S.;Kim, D.A.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.1
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    • pp.1-6
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    • 2001
  • This experiment was conducted to evaluated the effect of maturity at harvest on the changes in quality of round baled rye silage at forage experimental field of Grassland and Forage Crops Division, National Livestock Research Institute, RDA, Suwon in 1998. The experimental design was a split-plot design with three replications. The main plots were three different harvest stages : boot, heading and flowering stages, and the subplots were days after ensiling : 1, 2, 3, 5, 10, 30, 45, and 60 days. The wilting period of boot, heading and flowering stages were 1, 0.5 and 0.5 days, respectively. The final pH of rye silage was higher in the order of flowering, boot and heading stages. And pH of flowering stage began to change at early fermentation period, but that of boot and heading stages was delayed 1~2 days. Ammonia-N content of boot stage was highest. and that was increased as fermentation progressed. But Ammonia-N of heading stage was decreased to 30 days. then that was increased after 45 days fermentation. Among fermentation periods, inside temperature of deep place was not affected by external temperature. And that of deep place was increased to 3$0^{\circ}C$ at early fermentation. then decreased as fermentation progressed. However surface temperature was affected by external temperature after 10 days. Acetic acid content was not changed with 5 days by harvest stages, but that of boot stage was increased after 10 days. Butyric acid of boot stage was increased after 5 days. but that of heading stage was increased after 10 days. However lactic acid was increased from 1~2% to 6~8%. Lactic acid bacteria (LAB) of heading and flowering stages were highest at 5 days fermentation, and that of boot stage was highest at 10 days fermentation. The results of this study indicate that fermentation of round baled rye silage occur within 5 days. Therefore, any modification should be applied with an 5 days for high quality of round baled rye silage.

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Effect of Mower Conditioner at Different Harvest Stage on the Field Drying Rate and Quality of Rye Hay (수확시기별 Mower Conditioner 처리에 의한 속성 양질 호밀 건초조제 효과)

  • Chung, E.S.;Seo, S.;Kim, J.G.;Kang, W.S.;Kim, J.D.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.19 no.3
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    • pp.251-258
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    • 1999
  • A field experiment was carried out to determine the effects of chemical /mechanical treatments at mowing on the field drying rate and hay quality of rye (Secale cereale L.). The chemical drying agent/mower conditioner ($K_2CO_3$ 2%, conditioning, $K_2CO_3$ 2% + conditioning and control) were treated at different harvest stage (late boot, heading and bloom stage) for hastening hay-making in the spring of 1996. After field dry, square bales were made by hay baler, and the dry matter(DM) loss, visual estimation and nutritive value of rye hay were evaluated after storing two months. The field drying rate of rye was higher with delayed stage of harvest, and mechanical and chemical + mechanical treatment, but the effectiveness of chemical alone was very low. With mower conditioning, the duration of field dry was shortened by 1.5 to 2 days compared with control. The DM loss of rye hay was reduced by late harvest and mechanical, and chemical + mechanical combined treatment, but the efficiency by chemical alone was very low. The visual score (Ieafiness, green color, odor and softness) of hay after storing was high in mechanical and chemical + mechanical, but the score by chemical alone was very low. The nutritive value (ADF, NDF, digestibility, and relative feed value) of hay was also high with treatment of mechanical and chemical + mechanical, but the quality by chemical alone was similar compared with control. The quality of hay was very low when harvested at bloom stage. In conclusion, mower conditioning can enhance the field drying rate of rye, but the drying effectiveness of chemical drying agent was very low. The effect of chemical/mechanical combined treatment was very similar when compared with mechanical alone. Harvest at early heading to heading stage was recommended for high quality rye hay.

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