Quality and in vitro fertility of Uruguayan Criollo bulls’ semen cryopreserved in two commercial diluents
Abstract
We evaluated the quality and in vitro fertility (IVF) of semen from the Uruguayan Criollo cattle bank of the Veterinary Faculty. We thawed semen from 8 bulls (0850, 0853, 1026, 1035, 3733, 5834, 9701, 9714), diluted in Triladyl® (T) and Andromed® (A) (with/without egg yolk) and evaluated by CASA: concentration, motility, kinetics, morphology and performed a hyposmotic test, after 2h we repeated motility and kinetics. T-test (5% significance) for time and diluents was applied and percentiles to categorize the 16 samples. The best samples were: T-9714, T-1026, A-9701, A-9714, T-0853, T-9701; except bull 0850 (A and T), the others were acceptable. For IVF we aspirated 404 A-quality oocyte-cumulus complexes (OCC) from bovine ovaries from a slaughterhouse, washed and matured them in TCM with 5% BCS and hormones, in 100μl drops, covered with mineral oil, in incubator (38.5ºC, 5% CO2 and 95% humidity), 22h. 8 groups of OCC were inseminated with selected semen: 1026, 3733, 5834 and 9701 (A and T), which was capacitated by centrifugation in Percoll® gradients (90/45%), in Talp-Sperm and adjusted with Talp-Fert to 2x106sperm/ml, drops were formed, covered with mineral oil and placed the OCC inside, 18h. The zygotes were denuded and cultured in CR1aa with BCS (5%). We evaluated cleavage (48h) and embryo development (ED) on day 7 (IETS standards). Results were analyzed with Chi square (5% significance). There were no significant differences in cleavage between diluents (A:143/207; T:128/197), nor ED (69 and 68 respectively) (p>0.05). There were no significant differences in cleavage between the 1026 (73/109) bull's semen and 9701 (68/101) (p>0.05), nor between them and 5834 (77/102) and 3733 (53/92); although, 5834 had higher cleavage rate than 3733 (p=0.008). There were no significant differences in ED between bulls (p>0.05). This work was important to assess the quality and fertility of the semen bank.
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