Update of the cooling protocol for antibiotic-free storage of boar semen at 5°C improves sperm quality and maintains low bacterial counts.

Preserving boar semen at 5°C instead of the conventional storage temperature of 17°C would enable a reduction of antibiotic use in pig insemination. To protect the chilling-sensitive boar spermatozoa, holding the extended semen at a higher temperature before cooling could be beneficial and facilitate the implementation of the innovative preservation concept in practice, provided that bacterial growth is kept at a low level. The aim of this study was to introduce a holding time (HT) at 17°C before cooling and to examine the effect on sperm quality and bacterial growth compared to the original cooling protocol for antibiotic-free 5°C semen storage. A series of experiments with semen doses from eight boars extended in Androstar® Premium without conventional antibiotics revealed that sperm kinematics and the integrity of sperm plasma membranes and acrosomes were improved with HT between 16 and 24 h followed by delayed cooling with 0.04°C/min when compared to the original protocol for semen preservation at 5°C (p < 0.05). Both a shorter HT of 6 h and a faster cooling rate of 0.07°C/min reduced sperm quality (p < 0.05). The HT for 24 h did not compromise the inhibitory effect on bacterial growth during long-term semen storage at 5°C, not even in semen doses spiked with Serratia marcescens. In conclusion, semen storage at 5°C with the modified cooling protocol improved sperm quality and is antimicrobially efficient. It thus presents a ready-to-use tool for a reduction or replacement of antibiotics in pig insemination.