Improving the quality of rooster semen frozen in straws by screening the glycerol concentration and freezing rate

ABSTRACT

• 1. This study examined different glycerol concentrations (GC) and freezing rates to improve the quality of rooster spermatozoa frozen in straws, and to determine the effect of varying GC on post-thawed spermatozoa quality, as evaluated by fertility and hatchability.

• 2.The experiment included two tests. In test 1, rooster semen straws containing 2, 4, 6, 8 and 11% glycerol were put in a rack (nine tiers with a 1 cm interval between every two tiers, 1 to 9 cm above liquid nitrogen (LN) source), and gradually frozen. The semen straws located in different tiers experienced different temperatures and freezing rates. The straws were then thawed and live sperm numbers determined. In test 2, rooster semen straws containing 2, 4, 6, 8 and 11% glycerol were put on optimal tiers (identified in test 1) for freezing, and stored at −196°C. Hens were inseminated with the frozen semen (post-thawed and glycerol removed, about 4.0 × 10⁸ sperm per hen), and eggs incubated.

• 3. The numbers of live sperm in the 11% glycerol group was higher than that in 2, 4 or 6% glycerol group (P < 0.05) for the semen straws on tiers 1 to 9, while that on tiers 1 to 5 was lower than that on tier 6 to 8 (P < 0.05). GC, freezing rate and the interaction between GC and freezing rate had a significant effect on live sperm numbers (P < 0.01). The highest fertility was in the 6% glycerol group and occurred on day 5 after insemination. The lowest fertility occurred in the 2% glycerol group on day 10 after insemination.

• 4. The optimal combination was 11% glycerol in straws located 6 cm above the LN surface (on tier 6). The 6% glycerol group achieved the highest fertility (77.6%), which surpassed that reported in recent years.

KEYWORDS:

• Gradual freezing
• liquid nitrogen
• rack
• rooster semen
• straws

Acknowledgments

The authors express their gratitude to all the staff of the funding supported institutions (see Funding) and would like to appreciate the valuable suggestions from reviewers and the section editor (Dr Glenn Baggott), which improved the manuscript greatly.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The work was funded by Star Innovation Field [2016NK3109]; The Guiding Scheme of Agricultural Technology Innovation in Hunan Province and the Science and Technology Planning Project in Changsha City [KQ1706150].