The aim of this study was to test the effects of five different concentrations (0, 10−3, 10−4, 10−5, and 10−6 M) of resveratrol (Res) supplementation in bull sperm washing and fertilisation medium on levels of reactive oxygen species (ROS), phosphatidylserine (PS) externalisation, mitochondrial membrane potential (Δψm), ATP and malondialdehyde (MDA), acrosomal integrity, blastocyst rate, and blastocyst quality after in vitro fertilisation (IVF). The results for sex-sorted sperm from three bulls showed: (1) ROS and MDA levels in 10−3 M and 10−4 M Res groups were significantly lower than those of controls (P < 0.05); (2) the percentage of viable sperm, percentage of sperm with high Δψm, and the ATP content in 10−3 M and 10−4 M Res groups were significantly higher than those of controls (P < 0.05); (3) the percentage of viable sperm with acrosomal integrity, and the blastocyst percentage and quality of the 10−4 M Res group were significantly higher than those of controls (P < 0.05). In conclusion, 10−4 M Res supplementation in washing and fertilisation medium of sex-sorted bull sperm significantly decreased ROS, PS externalisation, and MDA, and protected mitochondrial function and acrosomal integrity, thereby increasing blastocyst percentage and quality following IVF.

The fertilization capacity of sex‐sorted sperms is seriously decreased, which inhibits its wide application. However, little information is still available about the effect of vitamin C (VC) and lycopene (Lyc) on the fertilization capacity of sex‐sorted bull sperm. In this study, the washing medium and fertilization medium of sex‐sorted sperm from three bull individuals were supplemented with different concentrations of VC (0, 1 × 10–3, 1 × 10–4, 1 × 10–5, 1 × 10–6 M) or Lyc (0, 1 × 10–4, 1 × 10–5, 1 × 10–6, 1 × 10–7). After washing twice and incubation for 1.5 hr, the malondialdehyde (MDA) level, phosphatidylserine (PS) translocation, membrane potential (Δψm) and IVF (in vitro fertilization) ability of sex‐sorted sperm were investigated. For the sex‐sorted sperm of bulls A, B and C, 1 × 10–3 M VC or 1 × 10–4 M Lyc treatment significantly decreased their MDA levels and PS translocation and increased their Δψm levels and cleavage rates after IVF. When blastocysts were concerned, 1 × 10–4 M Lyc significantly improved the blastocyst rates and their IFN‐tau expression of bulls A and C. In conclusion, supplementation of 1 × 10–3 M VC or 1 × 10–4 M Lyc in washing and fertilization medium contributed greatly to improving the fertilization capacity of sex‐sorted bull sperm during IVF procedure.