• 한국수정란이식학회지
• /
• 제15권3호
• /
• pp.219-230
• /
• 2000

This study was carried out to determine the factors on achieving good viability of embryos biopsied fur sexing, to investigate pregnancy rate following embryo transfer(ET) with sexed embryos, and to confirm the accuracy for the calves bort following ET with sexed embryos by polymerase chain reaction(PCR). To investigate viability of Hanwoo embryos after biopsy for sexing, fresh and frozen/thawed embryos were biopsied according to different developmental day of blastocysts, different stage of blastocysts, and different biopsy grade and the embryos themselves were incubated for 2 hours in TCM199 after microsection to be evaluated morphologically for recovery as blastocyst. The results obtained were as follows : 1. The rate of oocytes cleaved in vitro and the rate of blastocyst of the cleaved oocytes were 52.5% and 21.6%, respectively. The rate of blastocyst on day 8 was 11.2%, denoting the highest rate during whole culture period posterior to in vitro fertilization(IVF) 2. After biopsy for sexing, the viability rate of blastocyst on day 7, 8 and 9 was 75.0%, 88.4%, and 100.0%, respectively and the viability of early, mid, and expanded blastocyst after biopsy was 75.0%, 88.9%, and 91.1%, respectively The viability rate of fresh and frozen/thawed embryos was 89.9%, 71.4%, respectively. And the viability of expanded, hatching, and hatched blastocyst of frozen/thawed embryos was : 75.0%, 75.0%, and 50.0%, respectively. The viability of embryos according to biopsy grade of 10∼20%, 21∼30%, and 31∼40% was 85.7%, 91.5%, and 71.4%, respectively. 3. Pregnancy rate after transfer with biopsied embryo between flesh and frozen/thawed embryos was 22.6% and 20.0%, respectively. 4. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex deterimination was 92.3% (12/13).

• 한국식품과학회지
• /
• 제6권1호
• /
• pp.12-16
• /
• 1974

진흥(振興)과 IR667의 비중선(比重選)된 현미(玄米)를 배아(胚芽)와 배유(胚乳)로 분리(分離) 비중등급별(比重等級別)로 전(全) amino산조성(酸組成)을 조사(調査)한 결과는 다음과 같다. 1. 현미단백질함량(玄米蛋白質含量)이 높은 IR667은 배아(胚芽)에서 진흥(振興)보다 단백질함량(蛋白質含量)이 적었으며 질소당(當) lysine과 필수(必須) amino산함량(酸含量)도 낮았다. 2. 두품종 모두 amino산으로의 질소회수율(窒素回收率)이 중등비중(中等比重)에서 배아(胚芽)나 배유(胚乳) 어느 경우도 높아서 비중(比重)이 높은 경우는 전분(澱粉)에 의(依)한 가수분해중(加水分解中)의 파괴(破壞)에 기인(起因)하는 것으로 보였다. 3. 배아(胚芽)나 배유(胚乳) 공(共)히 IR667에서는 Glu.>Asp.의 순(順)으로 많고 진흥(振興)에서는 Glu.>Arg.의 순(順)으로 많아 품종간(品種間) 질소대사(窒素代謝)의 차(差)를 보이는 것 같다. 4. 두 품종 공(共)히 배아(胚芽)에서는 His.

• 한국수정란이식학회지
• /
• 제13권2호
• /
• pp.107-115
• /
• 1998

The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.0$\pm$11.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.7$\pm$1.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

• 한국수정란이식학회지
• /
• 제12권2호
• /
• pp.195-202
• /
• 1997

The present study was carried out to evaluate the effect of superovulation treatments on ovarian responses, oocyte recovery rates and grades of collected oocytes using an ultrasound-guided transvaginal approach in Korean native cows. Superovulation in cows was induced with two different regimenes: 1) FSH-decreasing dose(n=8): the cows were received twice per day for three days of the total dose of 400 mg of FSH-p, 2) FSH-single dose(n=9): the cows were administrated a single dose of 400 mg of FSH-p in 25% PVP. The Observation of visible follicles and collection of oocytes were performed 12 hours following the last FSH in FSH-decreasing dose group and 48 hours after the FSH-single dose injection. All visible follicles larger than 6 mm were punctured and aspirated with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use. The mean number of visible follicles(> 6 mm) was significantly(P<0.05) higher in the FSH-decreasing dose treatment (22.811.9) and FSH-single dose treatment (20.612.0) groups than the non-treatment group(7.0$\pm$8). The mean recovery rate of oocytes was not significantly(P<0.05) different between the treatment and control groups, but the mean number of collected oocytes was significantly(P<0.05) higher in the FSH-decreasing dose treatment( 12.611.5) and FSH-single dose treatment (11.813.6) groups than the non-treatment group(3.7$\pm$0.5). In conclusion, the FSH-single dose treatment at superovulation in cows for ultrasound-guided aspiration might increase the number of aspiratable follicles and the recovery rate of follicular oocytes as the FSH-decreasing dose treatment.

• 한국수정란이식학회지
• /
• 제31권3호
• /
• pp.185-190
• /
• 2016

During the ovary preservation in low temperature, the cumulus oocyte complexes(COCs) lose their developmental competences after in vitro fertilization. We used phosphate-buffered saline (PBS) as a basic solutions of at various temperatures (25, 15 or $5^{\circ}C$) and supplemented them with 1mM glucose and 0.5mM glutamine as a source of carbohydrate metabolites. After recovery of COCs and in vitro fertilization, a significantly higher number of oocytes developed into blastocysts. The developmental competence of embryos that were originated from ovaries preserved at $15^{\circ}C$ was increased compared to those of 25 or $5^{\circ}C$. The maturation rate of oocytes was not differed between 24 and 36 h at $15^{\circ}C$ but showed lower than control group (71% versus 78%). In vitro-fertilized oocytes from ovaries stored at $25^{\circ}C$ for 24 h or at $5^{\circ}C$ for 24 h had a significantly decreased developmental potentials, but at $15^{\circ}C$ did not (27% versus 29% of blastocysts to develop into day 8). With these results, bovine ovaries can be preserved at $15^{\circ}C$ for 36 h without decreasing developmental capacity of in vitro-fertilized oocyte at least to the blastocyst stage. This information provides valuable information of preserving ovaries for embryo transfer or in vitro embryo production.

• 한국수정란이식학회지
• /
• 제6권2호
• /
• pp.37-46
• /
• 1991

A field trial was performed to evaluate the effects of hormone treatment on estrus induction, ovulation, embryo transfer and reproductive performance in post-weaning sows. This trial involved 61 mixed breed sows of varying parity on a commercial pig farm. Sows were allocated to one of five trials: control group involved 25 sows that were treated with a single intramuscular injection of 5 ml physiological saline, 6 sows received 1,500 IU PMSG on the day of weanning and 500 IU HCG at the onset of estrus in trial I, 7 sows received 750 IU PMSG on the day of weanning and 500 IU HCG at the onset of estrus in trial II, 5 sows were treated with the same as trial II on day 28 after weanning in trial III. and 18 sows were treated with 10 mg PGF$_2$$\alpha$ plus 2 mg estradiol benzoate on day 31 after weanning in trial IV. Ovarian responses were checked by laparotomy and ova were recovered by oviducal flushing between 40 and l00hrs after mating. Fertilized ova were transferred into the oviduts of recipient sows synchroni- zed. The results obtained were summarized as follows: 1. Percentages of sows detected in standing estrus following treatment were 86~100% among trial groups. The interval from treatment to standing estrus(6l.7$\pm$0.5lhrs) in lOmg PGF$_2$$\alpha$ and 2mg estradial henzoate treated group was significantly earlier than in other trial groups(P<0.05). 2. Average number of ovulations was 11.5~37.8 among trial groups. The ovulation rate in 1,500 IU PMSG and 500 IU RCG treated group (37.8$\pm$ 19.87) was significantly different from other trial groups(P<0.05). 3. Ova were recovered by oviducal flushing between 40~ l00hrs after mating and recovery rates of ova wore 91.4% between 40~59hrs. 4. Fertilized ova were transferred into the oviducts of 8 recipient sows synchronized with 7 to 17 ova per animal. Three of the recipients were pregnant and delivered 25 piglets. 5. Four of the donor sows in those embryo collection was not successful were pregnant following oviducal flushing and delivered 23 piglets. 6. Recurrence of estrus and farrowing performance of experimental sows were observed following the experiment was no difference among trial groups, respectively.

• 한국수정란이식학회지
• /
• 제27권1호
• /
• pp.63-69
• /
• 2012

The objective of this study was to investigate the effectiveness of cryopreservation methods for the effect of various vitrification containers, such as EM-grid, OPS, or cryo-loop on the survival and developmental rate of vitrified mouse pronuclear embryos, and mouse cleavage embryo, at 21, 24, 27 and 30 hr after hCG injection. Post-thaw cleavage was similar among treatments, while the developmental rates of mouse blastocyst and hatched blastocyst were higher ($p$ <0.05) in 27 hr and 30 hr than 21 hr. The developmental rate of hatched blastocyst at vitrified cleavage mouse embryos in cryo-loop was significantly higher than vitrified pronuclear embryos of control group as well as EM-grid and OPS ($p$ <0.05). The developmental rate using cryo-loop was higher than EM-grid, but in case of OPS at vitrified cleavage and mouse pronuclear embryos, no significant difference was noticed. These results of our study show that the developmental rates of mouse embryos were unaffected by various vitrification containers, but in case of mouse embryos and hatched blastocysts at late vitrified pronuclear embryos the developmental rates were higher than early vitrified pronuclear embryos. Moreover, the developmental rate of hatched blastocyst at vitrified cleavage mouse embryos was significantly higher than vitrified pronuclear embryos. For better execution of this study, it will be mandatory to include improvement of vitrification containers, cryopreservation methods and conditions, higher survival rate, safe preservation, contamination and embryo loss.

• 한국동물생명공학회지
• /
• 제38권2호
• /
• pp.84-88
• /
• 2023

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.

• 한국수정란이식학회지
• /
• 제31권3호
• /
• pp.215-219
• /
• 2016

Value of excellent breeding animals is important in livestock industry, but their economic life time is limited. And, many countries have been trying procuration of genetic resource in good animals. Therefore, this study was conducted to determine embryo production and to test efficiency of embryo transfer via non-surgical artificial insemination (AI) in different breed of superior sows. A total of 17 sows were used in this experiment (Duroc, n=10; Landrace, n=4; Yorkshire, n=3). The sows were artificially inseminated by semen of same breed boars. After 4 or 5 days following the AI, the embryos were obtained from the sows and then transferred to Landrace and Yorkshire recipients (n=3, respectively) by non-surgical method. The corpora lutea tended to be increased in Yorkshire and Landrace than Duroc(28 and 26 vs. 17, respectively). The recovery of embryo was 78.8% in Landrace, 65.4% in Duroc and 51.4% in Yorkshire. Duroc showed lower morulaes and early blastocyst embryos than 2, 4, 8 and 16 cell. The morula in Yorkshire was higher (P<0.05) than that of Duroc (4.7 vs. 3.4). Similarly, the morulaes and early blastocyst embryos presented greater (P<0.05) in Landrace compared with other breed sows. The recipient sows were pregnant in a Landrace only. This reason may be due to little embryos inserted in the recipients. In addition, pregnancy results were limited because of the little sows. In conclusion, ovulated ovum in sows can be affected by different breed. Also, further study needed pregnant test by using the many embryo in each breed.

• 한국임상수의학회지
• /
• 제27권1호
• /
• pp.46-49
• /
• 2010

In this study, we investigated the number of follicles, oocyte recovery rate and oocyte competence after in vitro maturation according to the size of follicle. And equine oocyte competence after in vitro maturation was investigated in terms of the diameter of follicle with criteria of maturation: nuclear stage after Hoechst staining. The average number of follicles per ovary with middle size (11-20 mm, 2.68) was higher than those of small (5-10 mm, 0.74) and large size follicle (> 21 mm, 1.63), therefore medium follicle (53.1%) had higher proportion than other size of follicles. The average numbers of follicle per ovary was 5.05. The rate of oocyte recovery in small (54.5%) and middle follicle (50%) was higher than that in large follicle (40.9%). After culture for 48 h in Medium 199, 50%, 45.5%, and 44.4% of oocytes from the follicles with diameters of 5-10, 11-20, > 21 mm, respectively reached the metaphase II stage. This is the first report showing number of follicle, oocyte recovery rate according to follicular size, and in vitro oocyte maturation in Jeju mare in Korea. To fulfill in vitro equine embryo production, further studies such as the seasonal effect, in vitro fertilization etc is need.