Procedures for in vitro embryo production in cattle have not been optimized. In the current experiment, we utilized a 3 × 3 factorial design to test whether the proportion of embryos becoming blastocysts in culture and the pregnancy rate after embryo transfer are affected by type of serum in the medium [no serum; 3% (v/v) KnockOut Serum Replacement (SR); 3% (v/v) fetal bovine serum (FBS)] and addition of specific embryokines [vehicle; 10 ng/mL colony stimulating factor 2 (CSF2); 100 ng/mL dickkopf related protein 1 (DKK1)] at day 5 of culture. Embryos were produced using abattoir-derived ovaries and Y-sorted semen from two Angus sires. The percent of putative zygotes and cleaved embryos becoming blastocysts was improved by SR and FBS. Pregnancy rate at day 30 was determined for 1426 Nelore recipients and calving rate for 266 recipients. In the absence of CSF2 or DKK1, pregnancy rates were lower for embryos cultured with SR or FBS. CSF2 and DKK1 reduced pregnancy rate for embryos cultured without serum but had no detrimental effect in the SR or FBS groups. Indeed, CSF2 blocked the negative effect of FBS on pregnancy rate. Data on birth weights were available for 67 bull calves. There were no effects of treatment. The sire used to produce embryos had significant and large effects on development to the blastocyst stage, pregnancy rate at day 30, calving rate and pregnancy loss between day 30 and calving. Results indicate that (1) SR and FBS can improve embryonic development in vitro while also compromising competence of embryos to survive after transfer, (2) actions of CSF2 and DKK1 depend upon other characteristics of the embryo production system, and (3) sire can have a large effect on embryonic development before and after transfer.

The aim of this study was to quantify the pregnancy rate after implantation of two embryos after FTET protocols, as well as to monitor pregnancy losses until parturition, evaluating, mainly, if this strategy results in more number of animals born. Therefore, 423 multiparous recipients were selected, standardized in terms of body score, who had high-quality corpora lutea. Animals were randomly divided into two groups according to one or two embryos transferred (1 embryo = Control, n = 237; 2 embryos = Group 1, n = 186). All recipients received the same hormonal treatment, which consisted of administering, on Day 0, 2 mL of estradiol benzoate (Gonadiol, ZOETIS) + 1.9 g multidose 1st use progesterone implant (CIDR, ZOETIS); on Day 8 the implants were removed + injected 0.4 mL of estradiol cypionate (E.C.P, ZOETIS) + 1.5 mL of eCG (Novormon, ZOETIS) + 1 mL of dinoprost tromethamine (Lutalyse, ZOETIS). The animals were evaluated by ultrasonography at 30 and 60 days after embryo transfer, to diagnose the success rate and embryo losses during this period. Furthermore, information was collected on births, length of gestation, number of twin births, number of childbirth assistance and the weight of the calves. The results showed that Group 1 had better success than the Control, with higher conception rates at 30 days (68.3% vs. 53.2%, P<0.001) and at 60 days (62.9% vs. 52.3%; P<0 .05). The number of animals born was also higher for Group 1 (53.3% vs. 43.3%, P<0.01). The percentage of twins born in Group 1 was 17.9%, and the animals had lower weight compared to the Control (34.29 + 7.36 vs 37.63 + 5.73, P<0.05). The length of pregnancy and the number of assistances were similar between groups. In conclusion, the strategy adopted in this experiment suggests a considerable increase in the calf birth rate, but losses during pregnancy and their mechanisms need to be elucidated.