Because multiple ovulation embryo transfer (MOET) in cattle includes several benefits such as wide spreading of genetically superior offspring for long distance, this biotechnological method has been widely applied to Hanwoo. When the recipients are not stayed close after embryo recovery from donor, the embryos are moved to other farms via several vehicles (car, train, and airplane). However, air travel induces lesser oxygen level, increased vibration, lower air pressure, higher noise, and increased exposure of cosmic radiation to living things than ground level. It was still unknown that fresh embryos obtained from multiple ovulation of Hanwoo could maintain their fertility after being transported via air plane, the present case report introduced a clinical case of MOET in Hanwoo after shipping fresh embryos via air transportation. The donor was multi-ovulated via follicle-stimulating hormone series of injection, which was followed by a gonadotrophin-releasing hormone injection and artificial insemination twice. The embryos were recovered by the uterine flushing, packed in ministraws, transported to recipients for 6 h including 1 h air flight, and then transferred to the synchronized recipients. During pregnancy diagnosis of early gestation period, 5 of 7 recipients (71.4%) presented no heat signs and showed fetal sacs with fluid under transrectal ultrasonography. After normal gestation period, all recipients naturally delivered healthy calves (male n = 2 and female n = 3) without abortion, stillbirth, and premature birth. The present case report indicated that transportation of fresh embryos for MOET via domestic flight in Korea did not affect to their fertility.
Until recently, there have been many researches about the freezing methods and several methods of cryopreservation. Hypothermic preservation has been used to complement the embryo freezing technology. There is a study to show the successful results for long-term hypothermic preservation. For that reason, FBS and BSA are commonly added to the culture medium to support embryo development. We investigated the effectiveness of hypothermic preservation method at 4℃ according to embryonic developmental stages for Hanwoo embryos and evaluated the effect of FBS and BSA on Hanwoo embryos as a supplemental reagent in hypothermic preservation medium after recovering preserved embryos from hypothermic preservation. The present study reported that survival and hatching rates of embryos at morula stage following storage at 4℃ is Day 7 group was significantly higher (p < 0.05) compared than those of other groups (p < 0.05). As a result, the survival and hatching rates of embryos at the blastocyst stage following storage at 4℃ result is showed that significantly higher (p < 0.05) survival rates than those of other groups an Day 6. The result showed that hatching rate at Day 6 and 7 were significantly lower (p < 0.05) compared with other groups. The result regarding the survival and hatching rates of bovine embryos following storage at 4℃ for 72 h in various concentrations of BSA are shown The results showed that survival rate of 1% BSA group was not significantly different (p < 0.05) compare with control (FBS) group. Also, the results showed that hatching rate of control (FBS) and 1% BSA were significantly different (p < 0.05) compared with other groups. In conclusion, our result demonstrated that the hypothermic preservation did not effect on the survival and hatching rates of embryos after recovering. In addition, the supplementation of BSA in preservation medium showed no difference in the embryo developmental competence after hypothermic preservation compared to FBS treatment. With that, BSA can be an alternative reagent for the hypothermic preservation medium as an energy source and pH buffer.
본 연구는 한우의 전장유전체 내에 존재하는 단일염기 다형성(SNP)에 대해 DNA chip 분석을 통해 각 개체의 유 전자형을 파악하고, SNP 표지인자간 연관불평형의 크기를 알아보기 위해 실시하였다. 강원도에서 사육되고 있던 한 우 거세우 139두의 조직을 채취하여 DNA를 추출하였으 며, Bovine SNP50K BeadChip 분석을 이용하여 분석에 사 용 가능한 최종 35,769개의 SNP 표지인자를 얻어 연관성 분석을 실시하였다. 분석에 이용된 SNP 표지인자의 총 길 이는 2499.26Mb이었고, 전장 유전체에서 평균 대립유전자 빈도(MAF)는 0.273이었으며, SNP 표지인자간의 평균 거 리는 0.060과 0.082 사이에 분포하였다. 강원지역 내 한우 거세우를 이용하여 확인한 본 실험 결과는 기존의 전국의 한우를 대상으로 한 연구결과와 유사한 패턴을 보였다. 50Kb 이하로 인접한 거리를 갖는 SNP 표지인자들의 연관 불평형 값(r2)이 0.2 초과인 인자가 34%로 나타났다. 또한 SNP 표지인자간 거리가 멀수록 측정값이 떨어지는 지수형 식의 그래프를 보였다. 염색체별로 구분한 40kb 이하로 인 접한 SNP표지인자의 연관불평형 값(r2)은 0.2 이하로 나타 난 것이 총 7개(13, 15, 19, 23, 25, 28, 29번 염색체)였다. 종 합하여 볼 때, 본 연구 결과는 강원한우의 유전적 개량을 이 용한 기초자료로서 활용될 수 있을 것이며, 유전적 개량을 이용한 육종시스템에 도움을 줄 수 있을 것으로 기대된다.
The technique of SCNT is now well established but still remains inefficient. The in vitro development of SCNT embryos is dependent upon numerous factors including the recipient cytoplast and karyoplast. Above all, the metaphase of the second meiotic division (MII) oocytes have typically become the recipient of choice. Generally high level of MPF present in MII oocytes induces the transferred nucleus to enter mitotic division precociously and causes NEBD and PCC, which may be the critical role for nuclear reprogramming. In the present study we investigated the in vitro development and pregnancy of White-Hanwoo SCNT embryos treated with caffeine (a protein kinase phosphatase inhibitor). As results, the treatment of 10 mM caffeine for 6 h significantly increased MPF activity in bovine oocytes but does not affect the developmental competence to the blastocyst stage in bovine SCNT embryos. However, a significant increase in the mean cell number of blastocysts and the frequency of pregnant on 150 days of White-Hanwoo SCNT embryos produced using caffeine treated cytoplasts was observed. These results indicated that the recipient cytoplast treated with caffeine for a short period prior to reconstruction of SCNT embryos is able to increase the frequency of pregnancy in cow.
After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q -ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.
포유동물 수정란의 동결보존기술은 최근 기후 변화에 따른 생물종 다양성을 보존하기 위해서 중요하게 여겨지는 연구 분야이다. 따라서 멸실 위험에 처한 동물의 개량과 증식, 보존과 복원 및 생명공학의 분야에 이르기까지 응용 기술은 다양하게 이용되어진다. 본 연구에서는 한우 수정란의 동결 후 생존성 향상을 위해서 동결 방법에 따른 체내 외수정한의 내동성을 조사하였다. 완만동결에 따른 체내 외수정란의 동결 융해 후 수정란의 재확장률은 89.6%와 81.5% 그리
This study was performed to examine the effects of recombinant bovine somatotropin (rBST) and fermented alcoholic feedstuff(FAF) on blood metabolites, carcass characteristics and profitability. Sixteen Hanwoos(bulls; 8, steers; 8) with average 143㎏ of body weight were randomly assigned to bull control group(BC), steer control group(SC), bull group(BrA) treated with rBST and FAF, and steer group(SrA) treated with rBST and FAF. Sustained release rBST(0.03㎎/㎏/body weight) was administered biweekly and dose of rBST was adjusted monthly according to body weight increase. The blood glucose content was higher in the groups treated with rBST and FAF than control groups. The blood cholesterol contents tended to be lower in the treatment groups (BrA, 120.5 mg/dl; SrA, 104.0 mg/dl) than in the control groups (BC, 141.0 mg/dl; SC, 125.5 mg/dl). No significant difference between the groups of bulls and steers were found in the blood triglyceride concentrations as well as the albumin contents. The rib eye area of BC and BrA were 70 and 72 cm2, respectively, which are slightly larger than the area of SC and SrA. The treatment did not affect the backfat thickness both in bulls and steers. The marbling scores of BC and BrA appeared as No. 1 and No. 2, respectively, and No. 3 and No. 4 were obtained in SC and SrA. Lower non saturate fat concentrations were observed in the treatment groups (BrA, 64.30%; SrA, 59.59) than in control groups (BC, 58.98%; SC, 58.09%). Saturated fatty acids and amino acids contents were slightly more improved in the treatment than in the control. Average auction prices of BC and BrA were 7,900 and 8,089 wons per 1kg of carcass, respectively, whereas the prices of SC and SrA were 9407 and 9866 wons per 1kg of carcass, respectively. It is concluded that the treatment with rBST and FAF can improve the growth performance such as marbling scores and rib eye area and increase the auction prices.
본 연구는 한우 난소의 난포 발달에 있어서 bFF 또는 anti-inhibin serum(AI)의 생리적 역할을 검토하기 위해 수행하였다. Saline(0.95%, control), bFF 또는 AI를 경정맥 주사 처리한 9마리의 한우 암소에서 채혈하여 혈중 estradiol-17(E2), inhibin 및 progesterone(P4) 농도를 분석하였으며, 이들 처리에 의한 난포의 발달은 초음파 진단기를 이용하여 관찰하였다. 본 연구에서 얻어진 결과는
본 연구는 과배란처리에 의한 수정란이식 시 우수한 수정란을 다량 확보하고 이식 후 수태율 향상을 위하여 수란우에 rbST처리가 수태율 및 progesterone 농도와 황체의 크기에 어떠한 영향을 미치는가를 조사하고자 실시하였다. 공란우는 Folltropin-V와 PGF₂α를 이용하여 과배란처리를 유도하여 12시간 간격으로 2 straw씩 3회 인공수정을 실시하였다. 공란우와 수란우는 대조구와 rbST 처리구로 구분하였으며, rbST (500 mg)처리는 발정발현 후 미근부에 근육 주사하였다. 과배란처리된 공란우의 수정란채란은 수정 후 7∼8일째에 비외과적인 방법으로 실시하였다. 수정란이식 후 수태율은 rbST 처리구에서 대조구보다 유의적으로 높았다 (64.0 vs. 47.1%; p<0.05). 채취한 혈액의 progesterone 농도 분석과 황체의 단면적를 조사한 결과 progesterone 농도는 대조구와 rbST 처리구에서 6일 동안 차이가 없었지만, 6일 이후부터 차이가 나타났으며 황체크기 역시 rbST 처리구에서 대조구에 비해 높게 나타났다. 본 연구결과에서 rbST처리는 이식 후 수태율의 향상시킬 수 있을 것으로 판단된다.
한우의 유전자원을 보존하기 위하여 정액동결 시험을 수행하였으며 고급육계통은 육질에 대한 육종가 상위 10% 이내의 자손에서 선발하였고 다 유계통은 어미의 이유시 체중에 대한 육종가 상위 10%이내의 자손에서 선발하였으며 2개 계통에서 종모우 총 13두를 선발하여 공시하였다. 정액채취는 인공질법으로 실시하였고 의빈대에 수소를 계류하고 채정대상우를 승가시켰으며, 3회 가승가후에 채정하였다. 채정후 10분 이내에 실험실로 옮겨와 검사항목을 조사하고 37에서
본 연구는 과배란처리에 의한 수정란이식 시 우수한 수정란을 다량 확보하고 이식 후 수태율 향상을 위하여 공란우 및 수란우에 bST처리가 수정란회수를 및 수태율에 어떠한 영향을 미치는가를 조사하고자 실시하였다. 공란우는 Folltropin-V와 PGF를 이용하여 과배란처리를 유도하여 12시간 간격으로 2 straw치 3회 인공수정을 실시하였다. 공란우와 수란우는 대조구와 bST처리구로 구분하였으며, bST(500 mg)처리는 발정발현 후 미근부에 근육주사
This study was carried out to investigate the effect of co-culture system(bovine oviduct epithelial cells; BOEC) and defined culture system (modified TALP ; mTALP) on the conception of embryos transferred, and pregnancy and twin birth rates after transfer of fresh or frozen-thawed bovine blastocysts produced in vitro were also evaluated. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol. The results obtained were as the following. The pregnancy rate after transfer was higher in co-culture group than in mTALP group, but was not signficantly different, and there is no difference between fresh embryo group and frozen-thawed embryo group in conception rate. The conception rate was not different whether 3∼4 blastocysts or 2 blatocysts transferred into a recipient, but the production rate of twin calves was significantly higher (p<0.05) when 3∼4 embryos transferred. The average birth weight of twin calves(24.38kg) was numerically, but not significantly lighter than that of single calves(26.68kg).
The objective of this study was to produce calves by transfer of embryos derived from slaughter house(SHD) and ultrasound-guided ovum pick-up (OPU). At 60 hrs after injection of 400 mg FSH dissolved in 25% polyvinylpyrrolidone(PVP) by single dose, ultrasound-guided follicular oocyte aspiration was ferformed. Day-7 and day-8 blastocysts produced by in vitro maturation (IVM), fertilization (IVF) and culture(IVC) of the oocytes derived from SHD and OPU were nonsurgically transferred into recipients. The results obtained were as follows. The cleavage rate and the development rate to blastocysts were not significantly (P<0.05) different between the oocytes obtained by SHD (72.9% vs. 34.1%) and OPU (75.9% vs. 38.4%). The oocyte recovery rate from the number of follicles by ultrasound-guided aspiration were not significantly (P<0.05) different between Holstein (61.7%) and Hanwoo (60.1%), but the rate of oocytes useful for IVF was significantly (P<0.05) higher in Hanwoo (69.3%) than Holstein (59.6%). The cleavage rate and the development rate to blastocysts was not significantly (P<0.05) different between Holstein (74.9% vs. 39.2%) and recipients on day 8 of estrus cycle resulted in 13 pregnancies (34.2%). One of them was sacrificed during gestation period due to mastitis and another was aborted spontaneous. The resulting 14 calves were morphologically normal at birth. Seventy eight fresh OPU-IVF embryos were transferred into 21 recipients on day 8 of estrus cycles, resulting in pregnancy of 12 recipients (41.4%). Two of them were sacrificed during gestation period due to mastitis and the other two were aborted. Nevertheless, the 11 OPU-calves have been born normally.