Effect of genistein added to bull semen after thawing on pronuclear and sperm quality

The aim of this research was to study the effect of different genistein treatments on bull sperm after thawing on pronuclear formation after in vitro fertilization (IVF) and on different sperm quality variables. Three experiments were performed. In Experiment 1, three treatments (Control, sperm incubation for 1 h at 37 °C with or without genistein) and two sperm concentrations during IVF (1 or 3 × 106 sperm/mL) were evaluated to study the influence of genistein on pronuclear formation (PNF). Sperm incubation for 1 h before IVF reduced PNF regardless of sperm concentration. However, after sperm incubation and with 3 × 106 sperm/mL in IVF, the genistein treatment group had greater fertilization rates than the untreated group. In Experiment 2, six treatments plus the control group were performed to study the effect of genistein (presence or not) and incubation conditions (30 min at 37 °C, 1 h at 27 °C or at 37 °C) on PNF using 3 × 106 sperm/mL for IVF. When incubation time was reduced to 30 min, PNF rate from the genistein treatment group was no different from either the control group or in the group in which incubation occurred for 1 h at 27 °C. In Experiment 3, the effect of several genistein treatments (control; genistein treatment for 30 min of incubation at 37 °C; genistein treatment for 1 h of incubation at 27 °C) on sperm motility, viability and DNA fragmentation were evaluated. Genistein did not improve sperm motility and, depending on the experimental group or time, it either reduced or had no effect on sperm motility. Genistein treatment did not improve sperm viability after 5 h of incubation. However, genistein treatment for 1 h at 27 °C decreased sperm DNA fragmentation compared with the control group after 5 h of sperm incubation. In conclusion, the treatment of bull sperm with genistein for 1 h at 27 °C could decrease sperm DNA fragmentation, although PNF rate after IVF and sperm motility were reduced.