Influence of post-thawing thermal environment on bovine sperm characteristics and in vitro fertility

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 (0 hr), incubated for 4 hr in T36.0 (36.0°C), T38.0 (38.0°C) and T39.5 (39.5°C), and analysed every hour (1 hr, 2 hr, 3 hr, 4 hr). In vitro production of embryos was performed at 0 hr and 4 hr. Sperm motility and cell kinetics (Computer-Assisted Sperm Analysis) were impaired after 2 hr 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 4 hr, 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 3 hr. In normothermia or in thermal stress, sperm cells present a gradual reduction of movement and functionality, which were more significant after 1 hr of incubation. The in vitro production of embryos is impaired when the semen is kept in a thermal environment ≥36.0°C for 4 hr.