Effects of different concentration and combinations of cryoprotectants on sperm quality, functional integrity in three Indian horse breeds.

The study focused on the effects of different cryoprotectant concentrations and combinations on the quality and functional integrity of sperm, using semen samples from three Indian horse breeds. The research revealed that the cryoprotectant Di-methyl Formamide (DMF) at a 5% concentration was more effective than traditional Glycerol in preserving sperm quality.

Objective and Methodology

• The study aimed to explore the impacts of different cryoprotectant concentrations and combinations on sperm traits, specifically focusing on membrane, acrosome and DNA integrity. The main factor of comparison was between the effectiveness of traditional cryoprotectant Glycerol and another agent, Di-Methyl Formamide.
• Therefore, the research was designed to include the semen samples of three Indian horse breeds, namely Marwari, Manipuri, and Zanskari. 4-6-year-old stallions were selected for the collection process.
• Once the semen was collected, it was evaluated and extended with a semen extender containing different concentrations and combos of Glycerol and DMF. The levels used were 5% Gly, 5% DMF, 2% Gly, 2% DMF, 2.5% Gly +2.5% DMF, and 1% Gly +1% DMF.
• Post-extension, the semen was frozen and later evaluated based on various parameters such as post-thaw motility, live sperm count, hypo-osmotic swelling test results, and acrosome and DNA integrity.

Results and Conclusion

• The study revealed that semen frozen with 5% DMF saw significantly higher values in plasma membrane integrity, acrosome integrity, and DNA integrity than those with Glycerol or combined concentrations of the two cryoprotectants.
• Combinations of Glycerol and DMF at different concentrations (2.5 and 1%) were less effective than the single use of DMF at 5% concentration in improving various post-thaw seminal characteristics of indigenous stallion semen.
• The results indicate that the 5% DMF concentration gave better protection to the plasma membrane and retained the acrosome and DNA integrity of the spermatozoa.
• Based on these findings, the study concluded that DMF at a concentration of 5% could be utilized for the cryopreservation of indigenous stallions, as it effectively maintains the seminal quality.