• Title/Summary/Keyword: Embryo collection

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Effects of Collection Time, Culture Time and Activation Treatment of Canine Oocytes on the IVM Rates

  • Lee, B.K.;Kim, S.K.
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.219-222
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    • 2007
  • These study was carried out to investigate the effects of the collection time, culture time and activation of canine oocytes on in vitro maturation rates. The activated oocytes were cultured in 10% FCS+TCM-199 media containing hormonal supplements (10 IU/ml HCG, 10 IU/ml PMSG, 10 ug/ml gonadotropin) at 5% $CO_2$, 95% air, $38^{\circ}C$. 1. IVM rate of in vitro cultured cumulus-attached oocytes recovered from ovaries that collected at follicular and luteal stages of the reproductive cycles were 11.4% and 5.7%, respectively. IVM rate of oocytes recovered from ovaries that collected at follicular stages of the reproductive cycles was significantly higher than that of luteal stage (p<0.05). 2. When IVM was carried out at different periods of 40, 48, and 70 hrs, the IVM rates of oocytes matured in vitro were 2.9%, 8.6%, 5.7%, respectively. These results indicate that the IVM time between $48{\sim}70$ hrs gives the highest maturation rate for the oocytes matured at the different stages. 3. IVM rate of oocytes matured in vitro for 10 hrs after single and combined activation treatment by ET, IP and CH and Ca+DMAP, CH+DMAP, ET+CH were $11.5{\pm}1.2%,\;10.8{\pm}1.0%,\;9.6{\pm}1.2%\;and\;12.4{\pm}1.5%,\;11.8{\pm}1.5%,\;11.2{\pm}1.4%$ respectively. This was higher than that in both single and combined stimulated groups compared to control group ($6.2{\sim}7.2%$).

Long-term Colchicine Prophylaxis on Operative Adhesion Formation in Embryo Transfer Donor Ewes and the Cytogenetic Evalution of Therapy (Embryo Transfer Donor Ewe에 생기는 수술상의 Adhesion 형성에 대한 장기간의 Colchicine 치료와 그에 따른 세포유전학적 분석)

  • 박석천
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.63-70
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    • 1994
  • Thirty ewes received typical trauma to their oviducts and uterine horns from surgical embryo collection procedures. Ten percent Dexamethasone was used as an irrigant on the exposed abdominal tissue prior to closing the incision. The treatment group received 17mg colchicine Om! lewe) and the control group was administered a 1.0ml placebo(PSS). Fifteen ewes that were initially treated with 17mg /im colchicine showed acute colchicine toxicity within 2-5 days after initial treatment and were removed from the study. Due to acute colchicine toxicity at 17mg, the colchicine level was lowered to 8, 4 and 2mg(4 ewes/group). Treatments consisted of daily injections of colchicine. One ewe in the 8mg group developed toxicity on day 5. Therefore, ewes were then administered colchicine every other day from day 6 to day 14 postsurgeryat 4 and 2 mg. the second laparotomy was performed 9 weeks after first treatment. Following second laparotomy, the treatment group(n=5) received 4 mg colchicine every day for 14 days and there was no clinical symptoms of colchicine toxicity. The third laparotomy was performed by the same operators 5 weeks after final treatment and the adhesions scored. Adhesion grading was based on a scale of 0-4, with 4 being the most severe. The results of adhesion grading(> 3) at second laparotomy were not significantly different(P>0.05)between the two groups. Adhesion formation observed at third laparotomy showed a reduced, but not significant reduction (P>0.05) in the colchicine-treated ewes when compared with the controls. Ten ewes(5 control and 5 treatment)were examined cytogenetically by bone marrow analysis five days post-treatment. There was no difference(P>0.05)in the incidence of numerical or structural aberrations between the two groups.

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Toxicity Assessment of Ocean Dumping Wastes Using Fertilization and Embryo Development Rates in the Sea Urchin (Hemicentrotus pulcherrimus) (말똥성게 (Hemicentrotus pulcherrimus)의 수정 및 배 발생률을 이용한 해양배출 폐기물의 독성평가)

  • Hwang, Un-Ki;Ryu, Hyang-Mi;Kim, Sung-Gil;Park, Jong-Soo;An, Kyoung-Ho
    • Environmental Analysis Health and Toxicology
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    • v.25 no.1
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    • pp.11-18
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    • 2010
  • Toxicity assessment of ocean dumping wastes (dye waste, urban sewage, food waste) were examined in the fertilization and embryo development rates of the Sea Urchin, Hemicentrotus pulcherrimus. Spawning was induced by injecting 1 mL of 0.5 M KCl into coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. Experiments were began within 30 min the collection of both gametes. The fertilization and embryo development rates test were performed for 10 min and 64 h after fertilization, respectively. The fertilization and embryo development rates in the control condition (not including ocean dumping wastes sludge elutriate) were greater than 90%, but suddenly decreased with increasing of ocean dumping waste sludge elutriate concentrations. The fertilization and normal embryogenesis rates were significantly inhibited in all waste sludge elutriate from dye waste ($EC_{50}$=4.37; $EC_{50}$=1.76), urban sewage ($EC_{50}$=5.79; $EC_{50}$=2.00) and food waste ($EC_{50}$=7.68; $EC_{50}$=2.16), respectively. The NOEC (<3.13) and LOEC (3.13) of fertiliztion and normal embryogenesis rates very similar in all waste sludge elutriate. These results suggest that biological assay using the fertilization and embryo development rates of H. pulcherrimus are very useful test method for the ecological toxicity assessment of ocean dumping wastes.

Estrus Induction and Embryo Transfer in Post-Weaning Sows (이유후 모돈에서 발정유기 및 수정란이식)

  • 이종수
    • Journal of Embryo Transfer
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    • v.6 no.2
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    • pp.37-46
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    • 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.

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Toxicity Assessment of Ocean Dumping Wastes Using Fertilization and Embryo Development Rates in the Sea Urchin (Strongylocentrotus nudus) (둥근성게(Strongylocentrotus nudus)의 수정 및 배 발생률을 이용한 해양배출 폐기물의 독성평가)

  • Hwang, Un-Ki;Rhee, Choong-Won;Kim, Kwang-Seop;Kim, Hyoung-Choul;An, Kyoung-Ho;Park, Seung-Youn
    • Environmental Analysis Health and Toxicology
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    • v.24 no.1
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    • pp.25-32
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    • 2009
  • Toxicity of ocean dumping wastes(dye waste, urban sewage, food waste) were examined by observing fertilization and embryo development rates of the Sea Urchin, Strongylocentrotus nudus. Spawning was induced by injecting 1 mL of 0.5 M KCl into coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. Experiments were began within 30 min after the collection of both gametes. The fertilization and embryo development rates tests were performed for 10 min and 48 h after fertilization, respectively. The fertilization and embryo development rates in the control condition(not including ocean dumping wastes sludge elutriate) were greater than 90%, but markedly decreased with increasing concentrations of ocean dumping waste sludge elutriate. The fertilization and normal embryogenesis rates were significantly inhibited in all waste sludge elutriate from dye waste($EC_{50}$=5.76; $EC_{50}$=4.53), urban sewage($EC_{50}$=9.82; $EC_{50}$=9.67) and food waste($EC_{50}$=3.90; $EC_{50}$=3.27), respectively. The NOEC(>3.13%) and LOEC(3.13%) of fertiliztion and normal embryogenesis rates very similar in all waste sludge elutriate. These results suggest biological assay using the fertilization and embryo development rates of S. nudus are very useful test method for the ecological toxicity assessment of ocean dumping wastes.

In vitro embryo production from ewes at different physiological stages

  • Alfredo Lorenzo-Torres;Raymundo Rangel-Santos;Agustin Ruiz-Flores;Demetrio Alonso Ambriz-Garcia
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.10.1-10.10
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    • 2023
  • Background: The collection of ovaries from slaughterhouses is an important source of oocytes for in vitro embryo production. On the other hand, the physiological stage of slaughtered females varies and influences embryo production. Objectives: The study examined the in vitro efficiency of embryos and demi-embryos from young, non-pregnant adult, and pregnant adult ewes from a local slaughterhouse. Methods: One thousand three hundred ovaries were collected from August to October 2020. The recovered oocytes were matured, fertilized, and cultured at 5% CO2, 38.5℃, and 100% humidity. Embryo bisection was performed in 96 blastocysts (n = 32 per treatment). The demiembryo pairs were incubated for their reconstitution for 12 h. SAS was used for data analysis. Results: The number of oocytes collected from the experimental group of non-pregnant adult ewes was higher (p ≤ 0.007) than those collected from the group of pregnant adult ewes (2.67 ± 0.19 vs. 2.18 ± 0.15 oocytes/group, respectively). The blastocyst rate was higher (p ≤ 0.0001) in the non-pregnant adult group (36.39%) than in the young (17.96%). The ratio of demi-embryos that recovered the blastocoelic cavity was higher (p < 0.05) in the young group (81.25%) than in the pregnant adult group (59.38%). The diameter of the demi-embryos was higher (p < 0.05) in the non-pregnant adult group (186.54 ± 8.70 ㎛) than those in the young and pregnant adult groups. Conclusions: In conclusion, the in vitro embryo production efficiency was highest when using oocytes from non-pregnant adult ewes under the conditions of this study.

Developmental Competence of Oocytes Collected from Individual Ovaries of Slaughtered Korean Native Cattle with Grade of Meat Quality and Meat Yield

  • Kim, So-Seob;Kim, Ji-Sun;Park, Hum-Dae;Lee, Su-Kap;Park, Il-Kun;Lee, Dong-Won;Kim, Yun-Sik;Park, Yong-Su;Kim, Jae-Young;Park, Min-Chul;Lee, Jung-Hyung;Oh, Dae-Sik;Kim, Jae-Myeoung
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.291-298
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    • 2008
  • We separately cultured follicular oocytes collected from individual ovaries of slaughtered Korean native cows and examined both the embryonic development rate and pregnancy rate after embryo transplantation according to the meat yield and quality grades of the source beef carcass. Oocytes from meat yield grade B cows exhibited a higher fertilization rate and embryonic developmental rate to the eight-cell stage than oocytes from grade A or C animals (p<0.05), but there was no significant difference in rate of development to the blastocyst stage among meat yield grades A, Band C. The oocyte cleavage rate and development rate to the eight-cell stage from meat quality grade 3 cattle was higher than grades 1++, 1+, 1 and 2 (p<0.05). Embryos derived from grade animals displayed a development rate to the blastocyst stage of 19.4%, which was also higher than all other meat quality grades (p<0.05). Transplantation of in vitro-cultured oocytes from meat yield grade A ovaries led to a higher pregnancy rate (64.2%) than in vitro-cultured oocytes from meat yield grade B ovaries (56.5%), but there was no significant difference between the two groups in pregnancy or abortion rates. In conclusion, embryonic development rate and pregnancy rate has a close relation to meat quality grades of the source beef carcass, this results is to give information for the Korean native cows improvement of breed.

Sperm DNA fragmentation negatively influences the cumulative live birth rate in the intracytoplasmic sperm injection cycles of couples with unexplained infertility

  • Repalle, Deepthi;Saritha, Kallimakula Venkata Reddy;Bhandari, Shilpa
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.3
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    • pp.185-195
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    • 2022
  • Objective: This study aimed to determine the effect of sperm DNA fragmentation (SDF) on the cumulative live birth rate (CLBR) in intracytoplasmic sperm injection (ICSI) cycles in couples with unexplained infertility. Methods: We conducted a prospective study of 145 couples who underwent ICSI cycles for unexplained infertility. Based on the SDF rate, patients were categorized into a low SDF group (SDF ≤30%, n=97) and a high SDF group (SDF >30%, n=48). SDF was assessed using the acridine orange test on density gradient centrifugation prepared samples. The CLBR was calculated as the first live birth event per woman per egg collection over 2 years. Results: The high SDF group (SDF >30%) showed a significantly lower CLBR (p<0.05) and a significantly higher miscarriage rate (p<0.05) than the low SDF group (SDF ≤30%). No significant difference was observed in the implantation and cumulative pregnancy rates between the two SDF groups. The total number of embryo transfers was stratified further into fresh and frozen embryo transfers. In the fresh embryo transfers, there were significant differences in the implantation rates, clinical pregnancy rates, and live birth rates (p<0.05) between the low SDF and high SDF groups. However, in the frozen embryo transfers, there were no significant differences in clinical outcomes between the two groups. In the multivariable logistic regression analysis, SDF was a predictor of CLBR (p<0.05) when adjusted for possible confounding factors. Conclusion: High SDF was associated with a lower CLBR and a higher miscarriage rate in the ICSI cycles of couples with unexplained infertility.

Effect of In Vivo Embryo Production and Embryo Transfer Following Superovulation in Hanwoo (다배란 처리에 따른 한우 체내 수정란 생산 효율과 수정란이식)

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Son, Jun-Kyu;Choi, Soo-Ho;Kim, Young-Joo;Lee, Poong-Yeon;Yeon, Sung-Heum;Kim, Hyun-Jong;Son, Dong-Soo
    • 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.