261DIFFERENT TRANSVAGINAL OVUM PICK-UP STRATEGIES TO OPTIMIZE THE OOCYTE RETRIEVAL AND EMBRYO PRODUCTION OVER A FIXED PERIOD OF TIME

Abstract

The objective of this study was to compare the efficacy of different oocyte retrieval schemes over a period of 10 weeks. Fifteen multiparous Angus cows were randomly assigned (n=3/group) to the following groups: 1) OPU once/week (7-day interval), 2) OPU twice/week (3- and 4-day interval, alternately), 3) Dominant follicle removal (DFR)+OPU once/week. DFR followed by OPU 72h later, 4) DFR+FSH+OPU once/week. DFR followed 36h later by FSH (Folltropin, Bioniche, Belleville, Ontario, Canada) (120mg s.c. and 80mg i.m. administered simultaneously) followed by OPU 48h later, 5) FSH+OPU twice/week. FSH followed by OPU1 30h later and OPU2 96h after OPU1. The interval between OPU2 and next FSH was approximately 42h. The follicles were aspirated using an Aloka ultrasound scanner (Wallingford, CT, USA) and a 5-MHz probe. The COCs were selected based on morphology and matured in TCM-199, supplemented with 10% FCS, 0.01 unitsmL−1 bFSH, 0.01 unitsmL−1 bLH and antibiotics. Fertilization (Day 0) was carried out with TALP-FERT medium containing capacitation factors. Frozen semen from the same bull was used (1×106mL−1) throughout. After 18h the presumptive zygotes were cultured in SOF with 5% FCS (Holm P et al., 1999 Theriogenology 52, 683–700). The embryos were evaluated based on IETS guidelines (Grades 1 and 2 selected). The data were analyzed by chi-square test and ANOVA. In all parameters, the DFR followed by FSH and subsequent OPU once/week protocol gave the best results on a per-cow-per-week as well as total (3 cows over 10 weeks) basis. Though OPU was done only once/week, this group produced more total oocytes (303) than groups where OPU was done twice/week, either with FSH (286) or without FSH (229) and also produced more total embryos on Day 8 (71 blastocysts, 23.4% of oocytes cultured) than the latter two groups (64, 22.4% and 49, 21.4%, respectively). Among the nonstimulated groups, the OPU twice/week group had more total oocytes (236) than groups with OPU once/week, either without DFR (137) or with preceding DFR (160). However, a preceding DFR seemed to have a positive effect on oocyte quality as this group had a better embryo development rate (26.9%), producing more total embryos (42). In comparison, OPU twice/week produced total 49 embryos (21.4%) and OPU once/week produced 26 (19.4%). In conclusion, DFR coupled with single-shot FSH administration can be used effectively over a period of at least 10 consecutive weeks and can increase (P<0.05) the oocyte yield by two-fold and embryo production following IVF by two and half-fold, as compared to routine OPU-IVF done once a week. Table 1 Per cow per week performance