109 To cite this paper: Hasan BJ, Al-Mutar HAH, Taher JK, and Hussain SO (2025). Effects of Adding Natural Honey to Semen Extender on Ram Epididymal Sperm Quality. World Vet. J., 15 (1): 109-116. DOI: https://dx.doi.org/10.54203/scil.2025.wvj13 2025, Scienceline Publication World ’s Veterinary Journal World Vet J, 15(1): 109-116. ISSN 2322-4568 Effects of Adding Natural Honey to Semen Extender on Ram Epididymal Sperms Quality Baqer Jafar Hasan 1 * , Hayder Aabd-alkareem Hasan Al-Mutar 2 , Jawad Kadhum Taher 3 , and Souhayla Oneeis Hussain 4 1 Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq 2 Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq 3 Department of Physiology, College of Veterinary Medicine, University of AL-Shatrah, Thi-Qar, Iraq 4 Department of Surgery and Obstetrics, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq *Corresponding author's Email: Baqerhikma.iq@gmail.com ABSTRACT Numerous studies have indicated that male infertility is often associated with poor semen quality. The present study investigated the use of natural honey as an antioxidant and nutrient additive to semen extender to evaluate its effect on sperm parameters. A total of 16 fresh testes from sexually mature rams were collected for the investigation and immediately transported to the reproductive technology laboratory at the College of Veterinary Medicine, Baghdad University. For the first group, sperm was collected and diluted with an egg yolk extender mixed with 10% natural honey, while the second group consisted of epididymal sperm diluted with 0.9% normal saline. Epididymal fluid was collected and evaluated for both groups. The results showed significant differences in mean individual motility between the two groups after 48 and 72 hours, as determined by the comparison of proportions. Egg yolk plus honey diluent was significantly more effective than normal saline diluent in preserving sperm cell viability after 48 and 72 hours. The same finding applied to progressive motility; the egg yolk plus honey diluent was significantly more efficient than the normal saline diluent for the time frame after 48 and 72 hours, respectively. In conclusion, the findings demonstrated that the egg yolk extender supplemented with 10% honey was more effective in preserving ram sperm motility over time than normal saline. The addition of honey to the egg yolk extender improved the motility, the live-dead ratio, and the viability of the liquid storage of ram epididymal fluid. Furthermore, egg yolk plays a crucial role in protecting sperm from the detrimental effects of low temperatures. Keywords: Epididymal sperm, Honey, Ram, Semen extender, Sperm motility ORIGINAL ARTICLE Received: January 14, 2025 Revised: February 16, 2025 Accepted: March 12, 2025 Published: March 31, 2025 INTRODUCTION Honey may greatly raise the quality of semen in various bull breeds (Chung et al., 2019). A study by El-Sheshtawy et al. (2016) on Arab stallions in Egypt has revealed that supplementation of semen with honey not only improves its motility and morphology but also protects the sperm against free radical-induced DNA breakage and cryoprotectant damage. Another study indicated that a 15% concentration of pure honey in cryoprotectant media could maintain the quality of infertile semen (El-Sheshtawy et al., 2016). In another study, combining honey with a cryoprotectant medium increased the fertility of rabbits and enhanced the quality of their semen during conservation (Chung et al., 2019). Additionally, studies on Jersey bulls suggested that Bioxcell without additives was more effective than a 1% honey concentration for cryopreserving bull semen (Chung et al., 2019; Gulov and Laskin, 2021). Honey provides a safe, efficient, and natural alternative for enhancing semen quality and prolonging its conservation in various sheep species. Farmers and breeders choose honey due to its natural qualities (Cheepa et al., 2022). Because of its ability to support oxidative stability, protect sperm from cryoprotectant damage, and prevent DNA breakage caused by free radicals, it also presents a viable strategy for increasing the fertility of infertile semen and fostering spermatogenesis (Kotze et al., 2024). The addition of honey to semen samples, particularly in artificial insemination and cryopreservation, requires careful consideration to maintain sperm quality and viability. Research has explored various methods and additives to optimize semen handling and storage, focusing on factors such as sperm motility, concentration, and the effects of cryoprotectants. These studies provide valuable insights into the challenges and potential solutions for enhancing the reproductive success of honey (Tsvetkov et al., 2024). The studies have shown epididymal epithelium, the lining of the epididymis, plays a significant role in the processing of sperm (Rodriguez-Martinez et al., 1990). Studies have also been conducted to understand the impact of various factors on sperm maturation in the epididymis of rams (Wegener et al., 2014; Martínez-Fresneda et al., 2019). For instance, research on the human epididymis has focused on the consequences of vasectomy on the epididymal transcriptome, which is the complete set of RNA transcripts produced by the epididymis (Martin-DeLeon, 2006). DOI: https://dx.doi.org/10.54203/scil.2025.wvj13 PII: S232245682500013-15