A. L. R. Rodrigues

Semen cryopreservation protocols of Mangalarga Marchador stallions

The effect of the utilization of three semen protocols (Inra 82®, Merck Gema and Botu-crio®) and two filling techniques (0.25 and 0.50 mL straws) in Mangalarga Marchador stallions were studied in this experiment. Sperm parameters were assessed during processing and post-freezing. No interactions between the protocols and type of filling were observed, so they were assessed separately. Sperm parameters were not altered when the extender was added to the centrifugation; however, there was reduction of motility and strength when freezing extenders were added. The Botu-crio® protocol preserved the parameters of total and progressive sperm motility, smoothed path velocity (µm/s), straight line velocity (µm/s), track velocity (µm/s) and the average and fast spermatozoa percentage better than the others. No difference between the extenders for the percentage of sperm integrity was observed. There was no difference in the responses studied on the filling techniques. The stallions presented better freezing with the use of the Botu-crio® protocol. The best post-freezing viability results were found for semen frozen using the Botu-crio® protocol and there were no differences concerning the sperm quality comparing 0.25 and 0.50 mL straws.

Evaluation of the simulated physiological oocyte maturation (SPOM) system on F1 Gyr × Holstein oocytes and embryos

The present study aimed to evaluate the effect of the simulated physiological oocyte maturation (SPOM) system on F1 Gyr × Holstein oocytes and embryos by evaluating the meiotic arrest, embryo production rates, total number of cells and lipid score. Three experiments were conducted and the following three experimental groups were formed according to in vitro maturation (IVM) treatments: CONTROL 1 (TCM 199 medium without FBS), CONTROL 2 (commercial medium) and SPOM (TCM 199 medium with forskolin and 3-isobutyl-1-methylxanthine (IBMX) in pre-IVM and extended IVM with cilostamide). In the first experiment (ovum pick-up), a significant (P 0.05) difference in total number of cells among the groups. No difference (P > 0.05) was found on lipid score among the groups at Day 7 of development, in both Experiments 2 and 3. At Day 9 (Experiment 2), only the CONTROL 2 showed a significant increase (P > 0.05) compared with the other treatments. It was concluded that under our conditions, the SPOM system was efficient in prolonging meiotic arrest on Gyr × Holstein oocytes, offering the oocytes in vitro conditions more similar to those found in vivo; however, it adversely affected embryo production rates and promoted no beneficial effect on the total number of cells and the lipid score.