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• W624079464 abstract "The main objective of this research was to determine the effects of oocyte grading andinsemination duration on cleavage rate of embryos obtained from in vitro fertilisation(IVF) in bovine and caprine species. In addition, the effects of oocyte grading in bovineand caprine on production of parthenogenetic embryos as well as an attempt tocryopreserve embryos using vitrification technique were also evaluated. Bovine oocyteswere obtained from abattoir while caprine oocytes were retrieved through laparoscopicoocyte pick-up (LOPU) or abattoir source. For laparoscopic oocyte pick-up goats,gonadotrophin injections were involved prior to surgery, which were Estrumate (125μg), Pregnant Mare’s Serum Gonadotrophin (PMSG) (1500 IU) and Ovidrel (250 IU).Following the washing in Phosphate Buffer Saline (PBS) (for laparoscopic oocyte pickupoocytes) or TL-Hepes medium (for abattoir/ovariectomy oocytes), cumulus oocytecomplexes (COCs) were washed with in vitro maturation (IVM) medium. Subsequently,the cumulus oocyte complexes were cultured according to the grades in the droplets ofin vitro maturation medium which was pre-incubated overnight in carbon dioxide (5%)incubator at 38.5°C for 18 to 21, 24 to 27 and 22 to 24 hours, for laparoscopic oocytepick-up caprine oocytes as well as abattoir/ovariectomy caprine oocytes and bovineoocytes, respectively. The grades of oocytes were based on the cumulus layers and thematuration of oocytes was based on the presence of the first polar body. For in vitrofertilisation (IVF), oocytes were partially denuded and co-incubated with post-thawedsperm (1x106 sperm/ml). In vitro culture (IVC) of presumptive zygotes was performedafter 8 to 14 or 18 to 24 hours after fertilisation. The fertilisation rate was assessed bythe presence of the second polar body. The cleavage rates of the embryos were thenobserved and recorded. For parthenogenetic activation (PA), matured oocytes werecompletely denuded and washed with 3 droplets of calcium ionophore, subsequently dimethylaminopyridine (6-DMAP) and incubated in it for 5 hours. After being washedwith 3 droplets of preincubated in vitro culture droplets, the oocytes were cultured andthe cleavage rates were recorded daily. In an attempt of vitrifying embryos, embryoswere placed into holding medium (1 minute), followed by VS1 (3 minutes) andsubsequently VS2 (45 seconds) before being plunged into liquid nitrogen. The vitrifiedembryos were devitrified by being immersed into TS (5 minutes), DS (5 minutes), andfinally two holding medium (5 minutes each), stepwise. After being washed thrice inpre-incubated in vitro culture droplets, the oocytes were cultured and the survival rateswere recorded daily. The data were analysed by using Analysis of Variance (ANOVA)and Duncan Multiple Range Test (DMRT). In bovine in vitro fertilisation, thematuration rates of Grade A (56.78±6.50%) and Mixed grade (74.69±6.68%) oocyteswere significantly (P<0.05) higher than other grades of oocytes (Grades B:46.58±5.92% and C: 31.29±7.11). The fertilisation rates of Grade A (70.49±6.47%) andMixed grade (68.18±7.43%) oocytes and the insemination duration of 8 to 14 hours(77.37±6.52%) were significantly (P<0.05) higher. The cleavage rates of Grade A andMixed grade oocytes were significantly (P<0.05) higher than that of other groups.However, no significant difference (P>0.05) was observed in both inseminationdurations of 8 to 14 and 18 to 24 hours. In caprine in vitro fertilisation, the maturationrate of Grade A (74.19±5.79%) oocytes was significantly higher than Grades B(54.66±7.42%) and C (42.50±7.20%) oocytes. The fertilisation rate of Grade A oocytes(40.54±8.23%) was significantly higher than that of Grade C (16.26±5.99%) oocytes.The cleavage rates of Grades A (38.39±8.89%) and B (35.90±9.23%) oocytes weresignificantly higher than Grade C (10.83±5.10%) oocytes; however, no significantdifferences (P>0.05) were found in the fertilisation and cleavage rates among all gradesof oocytes as well as with both insemination durations. In parthenogenetic activation, the cleavage rate of bovine was significantly (P<0.05) higher than that of caprine withthe percentages of 63.90±7.30 and 26.77±9.75%, respectively. For the developmentalcompetence of caprine embryos exposed to vitrification solution (toxicity screening) atblastocyst, 75.00% of survival rate for blastocyst up to hatched blastocyst was obtained.The survival rate of 33.33% was achieved in the vitrification of blastocyst. Inconclusion, in vitro fertilisation protocols for both bovine and caprine species have beensuccessfully developed, producing satisfactory development of embryos in vitro.However, intrinsic and extrinsic factors (especially those pertaining to specificlaboratory situation of a country) that influence the developmental competence ofembryos after in vitro fertilisation should be studied in detail, to ensure optimumoutcomes of subsequent cleavage, pregnancy and birth." @default.
• W624079464 created "2016-06-24" @default.
• W624079464 creator A5005738452 @default.
• W624079464 date "2011-01-01" @default.
• W624079464 modified "2023-09-23" @default.
• W624079464 title "Production of caprine and bovine in vitro-fertilised as well as parthenogenetic embryos and an attempt to vitrify in vivo- and in vitro-derived embryos / Tan Wei Lun" @default.
• W624079464 hasPublicationYear "2011" @default.
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