Pioneering comparative analysis of intracellular cryoprotectants on Gaga chicken sperm quality during cryopreservation

This study aimed to determine the impact of various types of intracellular cryoprotectants on the quality of Gaga rooster sperm following chilling and freezing protocols. A completely randomized design was used, utilizing four types of cryoprotectant treatments, all at a dosage of 7%. The cryoprotectants included glycerol, dimethyl sulfoxide (DMSO), ethylene glycol (EG) and dimethylacetamide (DMA). Sperm were collected from Gaga roosters using the massage method, and samples exhibiting a minimum motility of 70% were diluted with an egg yolk lactate ringer, incorporating the aforementioned cryoprotectant. The semen was subsequently packed into frozen straws and equilibrated, followed by pre-freezing, freezing and thawing procedures. Evaluations were carried out before freezing and post-thawing, collected on several variables, including motility, recovery rate, viability, abnormality, plasma membrane integrity, DNA damage, intracellular calcium intensity and sperm ultrastructure analysis. The results indicated that the treatments significantly (P < 0.05) affected pre- and post-thawing motility, recovery rate, post-thawing viability and DNA damage in both chilled and post-thawing semen. Furthermore, it was demonstrated that glycerol and DMSO effectively maintained the quality of Gaga chicken sperm during freezing, outperforming EG and DMA.