Background and Aim: Essential oils found frequently in plants are well known for their activities against bacteria, viruses, and fungi, and antioxidant properties. This study aimed to analyze egg yolk replacement by seed oils of Gossypium spp. (cotton), Balanites aegyptiaca (desert date), and Sesamum indicum (sesame) in semen extender, on ram sperm quality chilled at 4°C and frozen-thawed. Materials and Methods: Ejaculates were collected from adult rams and refrigerated at 4°C in a Tris-based extender containing 1.25%, 2.5%, 5%, and 10% of Gossypium spp., B. aegyptiaca, and S. indicum seed oils, to evaluate which were the two best extenders for comparison with BIOXcell, a commercial extender for deep freezing ram semen. Results: The data showed that sperm movements analyzed by the CASA system were faster in extenders supplemented with 2.5-5% of cottonseed oil and 1.25-10% of sesame oil, whereas in the extender containing B. aegyptiaca oil, all seminal parameters studied had the worst values. During the sperm-freezing process, 5% of cottonseed oil and 5% sesame seed oil were selected from the first study, with sesame oil reaching the best sperm quality. Thus, sperm motility and velocity were 44.14±13.99%, 24.44±12.6%, and 25.92±11.50%; and 20.26±9.56%, 8.76±6.38%, and 9.42±5.40%, respectively, for sesame oil, cottonseed oil, and BIOXcell. Conclusion: In summary, 2.5-10% of cottonseed oil and 1.25-10% of sesame seed oil can replace egg yolk in a Tris–egg yolk–based extender. Moreover, a Tris-based extender supplemented with 5% sesame seed oil could be an alternative for deep freezing ram semen, even though these results need to be confirmed with semen collected from rams with appropriate sexual rest.