• Journal of Embryo Transfer
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• v.25 no.3
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• pp.141-144
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• 2010

In vivo embryo produced from Hanwoo donor cows were collected and transferred to Hanwoo recipients. Cows, at random stages of the estrous cycle, received Progesterone Releasing Intravaginal Device (CIDR-plus, InterAg, New Zealand) together with injection of 1 mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment began 4 day later. For superovulation, a total of 28 mg FSH was intramuscularly injected twice a day in the way of decreasing doses 4 day (5, 5, 4, 4, 3, 3, 2 and 2 mg). Twenty one Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered 7 days after the second insemination by flushing the uterus with Embryo Collection Medium. The results obtained were as follows: The rates of transferable embryos were 50.3%, and 78 fresh embryos at morulae and blastocysts stage were transferred into Hanwoo recipients on day 7 of estrus cycle. The pregnancy rates were first embryo transfer 55.6%, 2nd 62.9% and 3rd 57.9%, respectively. In conclusion, These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos. Also, since it seems the condition of recipient cows greatly affect pregnancy rate, it is very important to evaluate recipient for effective cattle production.

• Journal of Embryo Transfer
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• v.22 no.2
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• pp.101-105
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• 2007

본 연구는 한우와 홀스타인 젖소에서 연속적인 다배란 처리 시 효율성 및 부작용에 대한 문제점을 연구하고자 다른 종간 다배란 처리효과를 비교, 조사하였다. 연속적인 다배란 처리를 위하여 estradiol benzoate(EB)와 CIDR plus, FSH 호르몬을 이용하여 한우에서는 17두, 젖소에서는 9두를 2회 이상 반복 이용하였다. 다난포 발생을 위해서는 FSH 호르몬을 이용하고 호르몬 처리 동기화를 위해서는 CER plus 기구를 질 내 삽입하였으며, 주요 조사 항목으로는 번식기관의 문제점, 배란 효율, 회수된 수정란 수, 우수 수정란 수 등을 분류하고, 종합적으로 종간 연속적인 다배란 효과를 비교, 조사하였다. 한우에서는 17두 중 4두(23.5%)에서 자궁경관의 협착이 발생하였고, 자궁 내에 손상이 발생하여 채란이 되지 않는 경우도 4두(23.5%)의 발생이 조사되었으나, 상대적으로 젖소에서는 9두 중 4두에서 일시적으로 배란이 전혀 일어나지 않는 경우가 있었다. 연속 처리 후 채란된 수정란의 수를 종합해 보면, 젖소에서는 평균 $4.3{\pm}3.4$개, 한우에서는 $2.7{\pm}2.5$개였으며, 종간 호르몬 효율과 연관된 배란율 조사에서는 한우에서는 89.7%, 젖소에서는 80.4%의 성적을 보였다. 또한, 3회까지 연속 다배란 처리를 한 결과, 한우나 젖소에서 채란된 수정란의 수는 비슷한 수준으로 채란되었다. 이와 같은 성적을 종합해 볼 때 EB제나 CIDR plus, FSH를 이용하여 1개월 주기로 연속적인 수정란 채란을 위한 다배란 처리가 소의 종에 무관하게 가능하다고 사료되지만 일시적으로 배란이 일어나지 않는 소에 대한 문제나 번식기관의 손상, 채란되는 수정란의 수가 적다는 것에 더 많은 연구가 필요하다고 사료된다.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.311-315
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• 2005

This study was conducted to investigate the effects of disregarding of heat detection on embryo production in superovulation of Hanwoo cows. Donors which showed 1 or 2 times of normal heat and had no abnormality of reproductive tract were selected The superovulation was performed injection of 2.5 mL FSH (Antorin R-10, Japan) 2 times on 5 days before next heat and continuously with the reduction of dose to 0.5 mL of first injection 2 times in a day for next 3 days. Otherwise, the donors of CIDR group were inserted CIDR plus (with the capsule of estradiol benzoate 10 mg) on Day 10 from standing heat for 9 days. On 6 days from insertion of CIDR, FSH was injected above same manners. The response according to the natural heat and CIDR were $82.2\%,\;89.7\%$, respectively. There were no difference between both treatments. The recovery rates of embryos were 7.7, 10.5, respectively and transferable embryos were 3.4, 3.8. There showed significant difference between both treatments (p<0.05). These results suggested that disregarding of heat detection in superovulation could be produced transferable embryos for embryo transfer and preserve the donors from the excess hormonal administration and maintain the economical lift span of genetically available Hanwoo donors.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.15-20
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• 2012

The objective of this study was to investigate the relationship between body weight, body condition score (BCS), blood urea nitrogen (BUN), glucose, cholesterol and number of transferable embryos for the purpose of improving reproductive performance in Hanwoo donors. Seventy five cows, at random stages of the estrous cycle, received a CIDR together with injection of 1mg estradiol benzoate and 50 mg progesterone, and gonadotropin treatment begann. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU = 1 ml) administered twice daily in constant doses over 4 days. On the 3rd administration of FSH, CIDR was withdrawn and 25 mg $PGF_2{\alpha}$ was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received $100{\mu}g$ GnRH at the time of 1st insemination. Embryos were recovered 7 days after the 1st insemination. In conclusion, cows with body weight < 400, 400~450 and > 450kg had number of transferable embryos of $4.2{\pm}1.7$, $6.1{\pm}2.7$ and $4.8{\pm}2.6$, cows with BCS <2.25, 2.25~2.75 and ${\geq}2.75$ had number of transferable embryos of $4.6{\pm}1.6$, $5.7{\pm}2.4$ and $5.1{\pm}2.7$ respectively. These data indicate that a body weight and BCS for superovulation of CIDR-treated Korean native cows does not affect the embryo yield.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.265-269
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• 2009

The purpose of this study was to determine the effect of different feeding ratios of whole crop barley silage on the embryo production in Hanwoo donors. All donors were basically fed 2.5 kg concentrate daily. Donors were divided into three groups according to the different feeding of forage; hay 70% and rice straw 30% (control, n = 21), whole crop barley silage 80% and rice straw 20% (T1, n = 25), and whole crop barley silage 60% and rice straw 40% (T2, n = 23) fed based on TDN 6.70/ BW 500 kg. All Hanwoo donors received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone ($P_4$, Day 0). Four days later, they were superovulated with 28 mg FSH twice daily IM in decreasing doses over 4 days. Then donors received 2 doses of $PGF_2{\alpha}$ (25 and 15 mg) with the 5th and 6th injections of FSH on Day 6. CIDR were withdrawn at the 6th FSH injection and the donors received $100\;{\mu}g$ GnRH 36 h after the second $PGF_2{\alpha}$ injection. The donors were artificially inseminated twice, at 8 and 24 h after GnRH, and embryos were recovered 7 or 8 days after the 1st insemination. The flush rate of the donors following positive superovulation responses did not differ among groups (76.2~96.0%, p>0.05). The number of corpus luteum (CL) at embryo recovery also did not differ among groups (10.6~14.0, p>0.05). Furthermore, the mean numbers of total ova (9.4, 10.5 and 12.0) and transferable embryos (5.3, 12.0 and 6.5) did not significantly differ among the control, T1 and T2 groups, respectively (p>0.05). However, mean concentrations of serum $P_4$ of the T1 (64.2 ng/ml) and T2 groups (55.7 ng/ml) were higher than that of control group (43.3 ng/ml, p<0.01), while serum cholesterol concentrations in the control (105.8 mg/dl) and T2 groups ($96.9\;{\pm}\;mg/dl$) were significantly lower than in the T1 group (121.1 mg/dl, p<0.05). Conclusively, whole crop barley silage can be fed a good substitute for hay forage for Hanwoo donors. Furthermore the ratios of whole crop barley silage 60% and rice straw 40% might be more worthful for embryo production.

• Journal of Animal Science and Technology
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• v.65 no.4
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• pp.779-791
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• 2023

This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60-70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.

• Applied Microscopy
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• v.30 no.2
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• pp.213-232
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• 2000

The development of the knee joint was studied by electron microscopy in human fetuses ranging from 20 mm to 260 mm crown-rump length ($7\sim30$ weeks of gestational age). The appearance of the primordium of the meniscus and cruciate ligament was conspicuous as the mesenchymal cells , preceeding that of joint space at 30 mm fetus. The primitive joint cavity was first seen in the interzone from the 40 mm fetus and its intermediate layer proceeded developing as a narrow cleft which was closely incorporated with two chondrogenic layers. Poorly differentiated mesenchymal cells of the meniscus at 40 mm fetus containing predominantly free ribosomes differentiated into fibroblasts at 60 mm fetus. By 100 mm fetus, the fibroblast in inner zone of the meniscus presented as oval profiles with a short cell processes, whereas middle and peripheral zones presented as elongated cells. Differentiation of the synovial membrane coincided with clarification of the joint cavity When dilatation of the synovial cavity occurred, the two types of synovial cells were identified at 60 mm fetus. By 100 mm fetus a majority of the intimal cells were B-type. B-type cells were clearly distinguishable from A-type cells by their content of extensive rough endoplasmic reticula and well developed Golgi complexes. In contrast, A-type cells had numerous filopodia, pinocytotic vesicles, lysosomes and large vacuoles. At 260 mm fetus the B-type cells were also a majority of intimal cells. At 260 mm fetus the inner zone of the meniscus was filled with parallel oriented fascicles of collagenous fibers and oval fibroblasts. The middle zone was constituted of parallel and radially arranged fibers and fibroblasts. The outer zone was populated by elongated fibroblasts encircled by crossed collagenous fibers with the blood vessels. At 30 mm fetus the fibroblasts of the cruciate ligament contained rough endoplasmic reticula and mitochondria. Collagen fibrils were noted within narrow cytoplasmic processes which were continued with the extracellular space. Collagen fibrils of ligament were filled in the bulk of extracellular space at 100 mm fetus. By $150\sim260mm$ fetus, the cruciate ligaments were constituted of longitudinally oriented bundle of collagen fibrils with irregular rows of round cells between.

• Journal of Embryo Transfer
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• v.20 no.3
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• pp.317-322
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• 2005

This study was conducted to improve the efficiency of embryo recovery and to establish the protocols of superovulation in Holstein cows. Sixteen Holstein cows were used the test the efficacy of three superovulation regimens using Folltropin. In the case of regimen 1, CIDR plus with E2 capsule was inserted in cows at the random stage of estrous cycle and the total of 400 mg Folltropin V was adminstered twice a day for 4 days(Folltropin V group). In regimen 2, CIDR was inserted and 3.0 mg estradiol benzoate was administered i.m. next day and the total of 400 mg Folltropin was adminstered twice a day for 4 days(Folltropin V+EB group). For regimen 3, CIDR insertion was same as in the regimen 2 and the total of 400 mg Folltropin diluted with $10\%$ PEG 8,000 was administered once(Folttropin V+PEG 8,000 group). In all the regimens, CIDR were removed on 12th day and 45 mg dinoprost was administered i.m. simultaneously. The heat detected donors were administered 200 ug LH-RH and inseminated twice with 2 straws of frozen semen 12 hours apart. Embryo were collected using Foley catherter in each uterine homs on 6${\~}$8 days after inseminations. The evaluation of collected embryos were according to the IETS manual. The CL responses according to the superovulation treatments were 5.8, 20.6, 24.0 in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively and there were significant different among the treatments(p<0.01). Transferable embyos collected were 3.6$\pm$2.4, 3.3$\pm$l.8 and 2.8$\pm$2.3, in the Folltropin V, Folltropin+EB and Folltropin V+PE 8,000 groups, respectively. Degenerated and unfertilized embryos in regimen 2 and 3 than regimen 1. These results indicates that superovulation treatments with both multiple injections and a single injection using PEG of Folltropin combined with CIDR insertion at the random stage of estrus cycle can be used to produce Holstein embryos.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.185-190
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• 2005

This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

• Journal of Veterinary Clinics
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• v.31 no.5
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• pp.382-388
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• 2014

The objective of this study was to determine the risk factors associated with pregnancy following 3 synchronization protocols in dairy cows. Data were collected on 1,952 cows from 22 dairy farms, including synchronization protocols ($PGF_{2{\alpha}}$ + estradiol benzoate [PG+EB], Ovsynch, and CIDR-ovsynch), cow parity, body condition score (BCS), and dates of previous calving, insemination and conception. The odds ratio (OR) for pregnancy were analyzed by logistic regression using the LOGISTIC procedure in SAS. The analysis revealed that farm (p = 0.005), cow parity (p = 0.0001), BCS (p < 0.005), and AI season (p < 0.05) significantly affected and calving to AI interval tended to affect (p < 0.1) the probability for pregnancy. Although synchronization protocols did not affect the probability for pregnancy (p > 0.05), cow parity and synchronization protocols showed a significant interaction (p < 0.005); the OR (0.60) was significantly lower (p < 0.0001) for multiparous cows compared to primiparous cows using PG+EB, whereas the OR (1.44) tended to be higher (p < 0.1) for multiparous cows compared to primiparous cows using the Ovsynch, and the probability for pregnancy did not differ between multiparous and primiparous cows using the CIDR-ovsynch (p > 0.05). Cows with BCS ${\geq}$ 3.00 were more likely pregnant (OR: 1.41) compared with cows having BCS ${\leq}$ 2.75, whereas cows inseminated during summer had a lower OR (0.73) compared with those inseminated during spring. Cows with a calving to AI interval > 150 days were more likely to be pregnant (OR: 1.20) compared with cows with a calving to AI interval ${\leq}$ 150 days. In conclusion, the OR for pregnancy following synchronization protocols in dairy cows was affected by farm, parity, BCS, calving to AI interval of the cow, and AI season, and there was a significant interaction between cow parity and synchronization protocols; the OR for pregnancy was lower for multiparous cows compared with primiparous cows using the PG+EB protocol.