The commercial applicability of bovine artificial insemination (AI) depends on the effectiveness of diluents for maintaining sperm fertility. Challenges faced by the AI industry due to recent advances in assisted reproduction, and the limitations inherent in using fresh and frozen-thawed sperm for AI, could be overcome with the development of better semen diluents. Research into the different microenvironments of bovine sperm as they progress towards maturity, capacitation and fertilisation is revealing various mechanisms that could be exploited to improve the formulation of semen diluents. These are reviewed here. A rationale for a more detailed investigation of bovine cervical mucus for factors that may allow further progress towards this goal are also discussed.