• Korean Journal of Veterinary Research
• /
• v.38 no.1
• /
• pp.209-216
• /
• 1998

The purpose of present study is to improve the efficiency of fusion and the developmental rates of nuclear transplanted embryos to produce genetically identical twins from Korean native cattle. The diameter of aspirated follicles had no significant effect on the recovery rates of oocytes collected by ovum pick-up technique. The fusion rates of nuclear transplanted embryos were significantly higher in 50 and $100{\mu}s$ DC duration groups(73.3 and 72.0% ; respectively) than that in $30{\mu}s$ group(55.6% ; p<0.05). The cleavage rates of nuclear transplanted embryos appeared to be significantly higher in donor nuclei derived from in vivo (65.0%) than in those from in vitro (50.5% ; p<0.01), but the developmental rates to morulae and blastocysts were not significantly different between them(13.7 vs 10.9%, respectively).

• Korean Journal of Veterinary Research
• /
• v.38 no.3
• /
• pp.652-658
• /
• 1998

The efficiency of nuclear transfer using donor embryos originated from ovum pickup(OPU) and activated recipient cytoplasts were examined for induction of twinning in Korean native cattle(KNC). After aspiration of follicle by OPU, regardless of the vacuum applied, we obtained same result in proportions of recovered cumulus-oocyte complex (COCs) with compact cumulus. Under electric stimulation(1.0kV/cm DC for $40{\mu}s$), most of activated oocytes proceed to anaphase II/telophase II within 3h(84.7%). In the treatment of oocyte activation, the preactivation which was performed before fusion had significant effect on the developmental rates to morula/blastocyst stage(9.4 vs 4.0%). In embryo transfer of nuclear transferred embryos, we obtained 2 twins from KNC recipients and 1 twin from a Holstein recipient. Our results showed that it is possible to obtain twins using nuclear transfer technique in KNC.

• Korean Journal of Veterinary Research
• /
• v.37 no.4
• /
• pp.917-924
• /
• 1997

Estrus cycle of cow, season and follicldar stimulating treatment, having an effect on the number of follicle, are investigated for the oocyte recovery rate in ovum pick-up(OPU). The number of follicle aspirated and oocyte collected on the different days of estrus cycle(D 4~5, D 9~10 and D 14~15) were not significantly different among the groups. The higher number of viable oocytes were produced on Jan-May(79.0%) than Jun-Aug(33.3%) by OPU in cow. The number of follicle and aspirated oocyte in cows treated FSH or PMSG combined with bovine somatotropin(bST) were 1.2~1.5 times higher than in cows treated alone follicular stimulating hormone(FSH) or pregnant mare serum gonadotropin(PMSG). In conclusion, OPU can be repeatedly practiced 2 or 3 times in an estrus cycle. In addition, the high environmental temperature is not good for ovarian function of cow and the bST co-treatment with FSH or PMSG is increasing the number of aspiratable follicle.

• Journal of Embryo Transfer
• /
• v.12 no.2
• /
• pp.211-218
• /
• 1997

This experiment was carried out to develop the anesthetic methods for ultrasonography and a new simplified disposable needle guidance device for ovum pick-up(OPU) in cows. Three different anesthetic methods were applied as. 1) epidural analgesia only with 2% lidocaine(20~30 ml), 2) epidural analgesia with 2% under general sedation with xylazine, 3) epidural analgesia with 2% lidocaine under general sedation with detomidine. We evaluated the anesthetic effects with items such as relaxation of anal sphincter, tail movement and rectal wall, retractability of both ovaries, additional anesthesia and possibility of OPU. Through this experiment, the above three anesthetic methods were applicable to OPU, but the epidural anlagesia under general sedation with detomldine was most effective for OPU. We developed a new disposable needle guidance device with stainless steel tube. With this, disposable needles can be easily attatchable to any other intravaginal probes. And also, it was found to he practical, economic and effective for OPU with the recovery rate of 51.2%.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.21 no.10
• /
• pp.433-439
• /
• 2020

This study, which was designed to improve and increase the utilization of female elk, examined the possibility of collecting their embryos during the non-reproductive period using ultrasound-guided Ovum Pick-Up (OPU) technology. Once follicles in the ovaries of an elk were observed, the embryos were retrieved through the absorption of the follicles using an OPU ultrasound probe. Fifty-seven embryos were retrieved from 85 follicles, giving a retrieval rate of 67.1%. Morphological evaluation of the recovered embryos revealed 14.0% embryos to be in grade A, 19.2% in grade B, 15.7% in grade C, and 50.8% in grade D. The developmental efficiency of the retrieved embryos was also investigated using in vitro maturation, in vitro fertilization, and in vitro culture. After cultivating 28 embryos through in vitro fertilization, 19 embryos were found to be fertilized; the fertilization rate was 67.9%. Four embryos developed to the blastocyst stage, indicating a 14.3% development rate. This study confirmed that the production of fertilized embryos from a seasonal breeder, such as elk, is possible via the ultrasound-guided OPU method. If the efficiency of in vitro fertilization and in vitro culture can be improved through further research, it will help improve the efficiency of elk embryo production through the transplantation of their fertilized embryos.

• Journal of Embryo Transfer
• /
• v.15 no.3
• /
• pp.279-286
• /
• 2000

This study was conducted to develop an improved method for oocyte pick-up(OPU) with finger-sensibility using ultrasound-guidance from ovarian follicles in Holstein heifers. Oocytes were aspirated from ovarian follicles of clear-outline (>2mm), obscure-outline and invisible($\leq$ 2mm) on ultrasound images with 3 different vacuum pressure(40, 80, 120mmHg). Total number of oocytes recovered/follicles were 309/237(130.4%). 113/80(141.3%) and 107/74(144.6%) with 40, 80 and 120 mmHg of vacuum pressure, respectively. Mean number of oocytes recovered was higher in 2 OPU/week (18.3$\pm$5.3) than 1 OPU/week(14.5$\pm$4.1), but this difference was not statistical1y significant. The recovery rates were not affected by the number of OPU as 135.6%(282 oocytes/208 follicles) in 1~20 OPU, 137.7% (168/122) in 21~40 OPU and 148.4%(92/62) in 41~60 OPU, respectively. The proportions of good oocytes (Grades I) recovered were not significantly different by the number of OPU until 40 OPU(12.4% in 1~20 OPU vs 16.7% in 21~40 OPU). However, a significantly(P<0.05) lower recovery rate resulted from more than 40 OPU compared to less than 40 OPU(7.6%). These results imply that more fertilizable oocytes can be produced from invisible-immature follicles by transvaginal aspiration with finger-sensibility from Holstein heifers.

• Korean Journal of Veterinary Service
• /
• v.44 no.4
• /
• pp.227-237
• /
• 2021

Artificial insemination of Korean native cattle (KNC) is the predominant method for breed improvement. However, industrialization of embryo production and transfer is necessary to utilize the genetic potential of KNC. The aim of this study was to examine associations between KNC donor cows and ovum pick-up (OPU) conditions, in-vivo oocyte recovery, and embryo development. Oocyte recovery and blastocyst development rates were higher at 50 and 60 mmHg OPU vacuum pressure than at 40 mmHg, which was, however, not significant. Regarding follicle growth, injection of 500 ㎍ GnRH 36 hours before OPU significantly increased the number of OPU oocytes from an average of 4.6 to 7.6 (P<0.05); no significant difference in embryo development rates was observed. Significant differences were observed in the numbers of OPU oocytes, embryo development rates, and transplantable blastocysts per individual among nine KNC donors (P<0.05). Furthermore, although there was no difference in OPU oocyte recovery intervals in approximately 2~8 weeks, the number of recovered oocytes significantly decreased at the 12-week interval (P<0.05); there was no difference in embryo development rates. The number of oocytes and embryonic development rates only tended to decrease until the seventh OPU session, but decreased significantly until the eighth session (P<0.05). The average pregnancy rate after transfer of OPU-derived in-vitro embryos into recipient cows was 41.8%. To improve the efficiency of OPU egg recovery and in-vitro embryo production, considering KNC donor characteristics, vacuum pressure of 60 mmHg, GnRH pretreatment to induce follicle growth, and effective OPU egg recovery up to seven times at intervals of 2~4 weeks appears to be most suitable. This study may facilitate the industrialization of KNC embryo production and transfer using high-quality cows.

• Journal of Embryo Transfer
• /
• v.13 no.2
• /
• 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.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.3
• /
• pp.314-319
• /
• 2005

To compare two different schemes, continuous scheme (CS) and discontinuous scheme (DCS), of once-weekly ovum pick up (OPU) with ultrasound-guided follicular puncture technique, Holstein heifers were randomly divided into two groups of five. After characterization of their two normal estrous cycles, the heifers were subjected to consecutive 20 weeks of once-weekly OPU under two schemes: the CS (one week interval between continuous OPU, total 100 OPU sessions performed) and the DCS (OPU fixed to the day 3 and day 10 of each estrus). Then, the status of ovaries and artificial insemination results were observed. On oocyte yield, the total number of punctured follicles using DCS was lower than that using CS, but the mean numbers of punctured follicles and recovered oocytes per session were higher in DCS than CS group. So the total number of recovered oocytes was similar in both groups. There were also no differences in the quality of recovered oocytes, nor in the developmental ability of oocytes fertilized in vitro between groups. The heifers in the DCS group showed regular estrous cycles with stable estrous signs through the periods of before, during, and after OPU, while those in CS group showed longer estrous cycles and less estrous signs during and/or after OPU compared with before period. Furthermore, the mean number of inseminations required for obtaining pregnancy after completion of the experiments was lower in DCS than CS group. The research demonstrates that similar quantity and quality oocytes can be achieved, and the side effects on donors are lower in DCS that needs fewer OPUs than CS group, and DCS is superior to CS.

• Journal of Embryo Transfer
• /
• v.13 no.1
• /
• pp.77-86
• /
• 1998

A combined technology of transvaginal ovum pick-up(OPU) system with in vitro-oocyte manipulation technique can be used for improving reproductive efficiency in the cattle. The objective of this study was to establish a newly-conceived breeding program using OPU in the pregnant cows. The OPU trial was performed in pregnant cows every 10 days from 40 through 90 days of artificial insemination (Al), and number of follicles in ovary, number of retrieved oocytes and embryo development following in vitro-fertilization, were evaluated. Reduced number of follicles in the ovaries of pregnant cows was firstly detected from 70 days after A' and a significant (P<0.05) decrease in the follicle number (5.4 follicles /donor) was found at 90 days than at 40, 50, 60 and 80 days after Al (8.0~9.2). A similar pattern was also observed in the number of oocytes retrieved by OPU apparatus during experimental period. When retrieved oocytes were matured and inseminated in vitro with frozen bull semen, development of the oocytes to the blastocyst stage was not significantly affected by the retrieval time. Four embryos (morula or blastocyst stage) derived from oocytes retrieved from pregnant cows were nonsurgically transferred to four recipient cows on day 7 of estrus cycle. For the first time in Korea, three of four transferred embryos developed to live calves with normal physiological parameters. In conclusion, an effective breeding program employing pregnant cow can be developed by use of OPU trial and in vitro culture techniques of oocytes ; OPU system could be repeated in pregnant cows with no risk of abortion and viable offsprings were borne after transfer to the recipients.