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.

Semen collection from three boars was performed twice a week, and doses (100 ml each) of semen diluted with Beltsville Thaw Solution (BTS) extender were prepared containing 1 × 109, 3 × 109 and 5 × 109 spermatozoa. Diluted semen was then stored at 17°C for a maximum of 120 h. Percentage of motile spermatozoa (PMS) and type of spermatozoa motility (TSM) were determined by microscopic examinations performed at 24-h intervals, from zero time (beginning of storage at 17°C) up to 120 h of storage, and a sperm motility index (SMI) calculated. Sows (no. = 360) were divided into nine experimental groups (40 sows per group) and a combination of number of spermatozoa per dose and storage time was used for artificial insemination in each group. All the animals were inseminated twice at 12 and 36 h after detection of standing oestrus by a teaser boar. Return and farrowing data including litter size were recorded. PMS, TSM and SMI decreased significantly (P < 0·01) from 48 h up to 120 h of storage, this being more marked in semen doses of 5 × 109 spermatozoa. When semen stored for up to 24 h was used, no significant differences (P > 0·05) regarding return rates and farrowing rates were observed among the three groups inseminated with semen containing different numbers of spermatozoa per dose. Inseminations with semen doses of 1 × 109 and 3 × 109 spermatozoa stored for 48 to 72 h have significantly lower return rates (P < 0·05) and significantly higher farrowing rates (P < 0·05) than with doses of 5 × 109 spermatozoa. Inseminations with semen stored for more than 72 h gave relatively higher return rates and lower farrowing rates. Semen storage time and number of spermatozoa per dose appeared to reduce litter size only where semen doses of 5 X109 spermatozoa were inseminated after storage for more than 48 h.