The aim of this study is to evaluate the effects of vitamin or mineral supplements on the conception rates of dairy heifers when replacing the last injection of GnRH with hCG in ovsynch protocol (experiment 1) and also to investigate whether the estrus synchronization treatment in the heifer stage affects the conception rates after 1st parturition (experiment 2). In experiment 1, 50 heifers were randomly assigned into 3 groups: 20 heifers each in groups 1 and 2, and 10 in group 3. All three groups were treated with an intramuscular injection of GnRH on day 0 (day 0 = the day of program start), PGF2α on day 7 and hCG on day 9, and were inseminated on day 10, 12~16h after hCG injection. In group 1 (vitamin group), the heifers were treated with an intramuscular injection of 5 ml of vitamin-ADE 500Ⓡ, and group 2 (mineral group) was treated twice with an intramuscular injection of 30 ml of mineral supplement-LAPTOVETⓇ on a one-week interval beginning on the day of hormone treatment (day 0 and day 7 respectively). Group 3 (control) was treated only with hormones. Pregnancy diagnosis was performed by ultrasonography through a rectal probe. First service conception rates (FSCR) and average services per conception (ASPC) were recorded for all subjects. Of the total 50 heifers, 6 (2 in group 1, 3 in group 2, and 1 in group 3) heifers were eliminated due to accidents during experiment 1. FSCRs were 58.8% (10/17), 66.7% (12/18) and 44.4% (4/9) in groups 1, 2 and 3, respectively. ASPCs were 1.53±0.72, 1.27±0.59 and 1.63±0.74 in groups 1, 2 and 3, respectively. Although there were no significant difference between the groups, relatively good results (higher FSCR and lower ASPC) were obtained in both group 1 and 2. In experiment 2, 11 primiparous cows from group 2 of experiment 1 in heifer stage which had been treated both with the hormones for estrus synchronizing and mineral supplements (ES group), and 12 primiparous cows treated only with minerals (non-ES group) were compared to examine the effects of estrus synchronization program on conception rates after 1st parturition. Following the examination, postpartum ASPCs were 1.55±0.82 and 2.17±1.47 in ES group and non-ES group, respectively. The postpartum average days open (ADO) were 116±56 and 197±93 in ES group and non-ES group, respectively. Although there were no significant difference between the two groups, desirable results (lower ASPC and shorter ADO) were found in ES group after 1st parturiton. In conclusion, experiment 1 indicates that vitamin or mineral supplement with ovsynch protocol may have some positive effect on FSCR and ASPC of dairy heifers, and in experiment 2, ES program in heifer stage had a positive effect on ASPC and ADO following 1st parturition.

Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot (20 mg × 6 / 300 IU) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ± SEM) observed which developed to larger than 2 mm in diameter were 1.6 ± 2.5, 4.3 ± 3.1, 5.5 ± 4.2, 6.6 ± 2.1 and 8.8 ± 7.8, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + 10 μg/ml FSH + 1 μg/ml 17β-estradiol for 27 h at 39℃ in 5% CO2 in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6- DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.

본 연구는 복강경을 이용하여 한국흑염소의 난소를 연속적으로 관찰하여 그 형태적 변화를 확인함으로써 한국흑염소의 성주기를 판정하고 그 결과를 한국흑염소의 수정란 이식시에 활용하고자 하는 목적으로 수행하였다. 제거 2일 후부터 복강경 수술을 통해 난소의 형태적 변화를 관찰하였다. 난소의 크기와 황체의 변화를 관찰하였으며, 황체는 그 성숙과 퇴축 과정에 따라 corpus hemorrhagicum(CH), corpus luteum(CL), corpus albi

Sodium carboxymethylcellulose(SCMC)와 Hyaluronic acid(HA)가 수정란 이식 및 채란을 위한 복강 수술 후 생식기의 유착 방지 효과 여부를 확인하기 위하여 본 실험을 수행하였다. 실험 동물로 성숙한 암컷 한국 흑염소 20두를 대조군, saline 투여군, SCMC 1%, SCMC 2% 및 HA 0.4% 투여군에 각각 4마리씩 배치하였다. 수정란 채취를 위한 통상적인 방법의 수술을 시행한 후 약물 주입용 카테터를 절

본 연구는 PI한 한국흑염소 수정란의 이식 결과를 통해 수란 흑염소의 수태율에 영향을 미칠 것으로 생각되는 여러 가지 요인을 분석함으로써, 높은 수태율을 얻을 수 있는 수란 흑염소의 최적 조건을 찾아낼 목적으로 수행하였다. 분석 결과, 수태율에 유의적인 영향을 주는 요인들은 발정형태, 수술 빈도, 이식 부위, 황체의 발육 단계, 수정란의 발육단계, 이식된 수정란의 수 등이었다. 자연 발정이 관찰되어 이식된 흑염소(59.1%, 13/22)들이 로 발정이

본 연구는 소 형질 전환 체세포 핵이식에서 용이한 탈핵을 위해 demecolcine을 이용할 시 탈핵율과 핵이식란의 발육능을 높이기 위한 최적의 조건을 알아보고자 실시되었다. 도축장 유래 미성숙 난자를 18시간 체외성숙 후 제1극체가 확인된 성숙 난자를 0.1, 0.2, 0.4 및 0.8 ug/ml의 demecolcine이 첨가된 배지에서 1시간 더 처리한 다음 세포막이 돌출되어 있는 난자를 체세포 핵이식에 공여하여 각 군간 배반포로의 발육능을 비교하였

본 연구는 개 동결 정액 융해 시 straw 크기 및 융해 속도가 융해 정자의 질(quality)에 미치는 영향을 조사하고 최적의 융해 조건을 조사하는데 그 목적이 있다. 정상적인 번식능을 가진 비글 수컷 5마리에서 정액을 채취하여 원심 분리하여 정장을 버리고 남은 정자에 동결보호제인 glycerol이 첨가된 tris-glucose-egg yolk extender를 첨가하여 동결하고 액체질소에 보관한 후 융해하였다. 동결 융해 조건에 따른 효과를 알아보

During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period (1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor (hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

This study was carried out to select the best cryoprotectant and to establish optimal concentration of the cryoprotectant in ultrarapid freezing of mouse 4-cell embryos and morulae, respectively. We investigated survival of ultrarapid frozen embryos according to various cryoprotectants such as glycerol, ethylene glycol, propylene glycol and dimethyl sulfoxide (DMSO). Suvival of the embryos frozen at different concentrations (3.0, 4.0 and 5.0 M) of indivisual cryoprotectant was also tested. Preimplantation developmental rate (96.3%, 83/86) of 4-cell mouse embryos treated with 4.0 M ethylene glycol after ultrarapid freezing and thawing was higher than those of other cryoprotectants (glycerol, propylene glycol and DMSO). In the ultrarapid freezing of mouse morulae, the highest developmental rate (98.8%, 89 /90) of the embryos to blastocysts was shown in the group of 5.0 M glycerol. Thus, these results demonstrate that 4.0 M ethylene glycol and 5.0 M glycerol are optimal for ultrarapid freezing of 4-cell mouse embryos and morulae, respectively.

The effects of cryoprotectants (glycerol, DMSO and ethylene glycol) and the concentrations (0, 0, 25, 0.5and 1.0 M) of sucrose in the diluent on the is vitro survival of mouse morulae froaen rapidly in liquid nitrogenvapour were examined. When the embryos were equilibrated in 1.5 M cryoprotectants +0.25 M sucrose in one-step or in 3.0 M cryoprotectants +0.25 sucrose in two-step and diluted with 0, 0.25, 0.5, or 1.0 M sucrose solution after thawing, high survival rates were obtained in ethylene glycol (48.0% to 88.2 %) or in glycerol (35.0 % to 77.8 %). These results show that 1.5 M ethylene glycol is a highly efficient cryoprotective agent for the rapid freezing of mouse morula embryos and 0.5 M sucrose was optimal concentration in the diluent after thawing.