Evaluation of Metformin Supplemented Diluent on Cryopreservation of Canine Semen

Abstract

Background: Cryopreservation of semen in canines is getting more advanced lately, however the fertility following the insemination of the frozen thawed semen is lower than other species. The higher content of PUFA and lipid peroxidation leading to production of reactive oxygen species is one of the reasons for the same. Antioxidants play a role in improving the post thaw semen quality in canines. Methods: Semen collected from 10 male dogs were further divided into two aliquots. The first aliquot was extended with TRIS-fructose egg yolk extender and the second one was extended with TRIS-fructose egg yolk extender supplemented with 50 µM of metformin. The extended semen was cryopreserved using conventional methods. Subsequently, both routine semen parameters and advanced semen parameters including viability, acrosomal integrity, DNA integrity and mitochondrial reactive oxygen species (ROS) were measured in the spermatozoa through the utilization of flow cytometry. Result: Present study indicated that, post thaw motility was significantly higher in the metformin supplemented group in comparison with control while, the mitochondrial ROS was significantly lower in metformin supplemented group. Our study concludes metformin could be effectively used to reduce the ROS produced during the process of cryopreservation of the canine semen.