Abstract
Our aim was to evaluate the effects of three thermal environments over time on kinetics, functionality and in vitro fertility of cryopreserved bovine spermatozoa. Four ejaculates from five bulls (n=20) were cryopreserved. After thawing, semen was evaluated (0hr), incubated for 4hr in T36.0 (36.0°C), T38.0 (38.0°C) and T39.5 (39.5°C), and analysed every hour (1hr, 2hr, 3hr, 4hr). In vitro production of embryos was performed at 0hr and 4hr. Sperm motility and cell kinetics (Computer-Assisted Sperm Analysis) were impaired after 2hr at T38.0 and T39.5 (p<0.05). Flow cytometry revealed an increase in the cells with injured plasma membrane to 39.5°C and a general reduction in the mitochondrial potential over time (p<0.05). In vitro fertility was impaired in all temperatures after 4hr, but there was no difference between 36.0°C and 38.0°C. Our results suggest that the ex situ resilience of semen at 36.0°C after thawing with no major damage to the quality is limited to 3hr. In normothermia or in thermal stress, sperm cells present a gradual reduction of movement and functionality, which were more significant after 1hr of incubation. The in vitro production of embryos is impaired when the semen is kept in a thermal environment ≥36.0°C for 4hr.
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