Variation of the effects of melatonin on the motility and functionality of chilled ovine semen
DOI:
https://doi.org/10.70208/3007.8245.v6.n1.351Keywords:
Melatonin, cryopreservation, linearity, vitality, progressivenessAbstract
Melatonin is a potent indolaminic antioxidant that protects cell membrane integrity against oxidative stress and stabilizes mitochondrial functionality. To determine the functional dose of melatonin on chilled ram semen based on the variation of its effect on motility and sperm functionality was the objective of this study, evaluating various concentrations on Katahdin and Blackbelly ram semen at 5 °C. A logarithmic scale from 1/16 mM to 16 mM was used, with 8 replicates during 2, 4, and 24 hours. Results demonstrated that melatonin acts in a dose-dependent manner under a second-grade polynomial model as a function of log₂, reaching its maximum biological efficacy at 4 hours. During this period, optimal values were recorded for vitality (80%) with 1.46 mM, progressivity (40.20%) with doses near 16 mM, and linearity (38.06%) with 0.5 mM. The parabolic behavior of linearity warns of a biphasic effect, where excessive concentrations are counterproductive for cellular kinematics. It is concluded that a concentration of 1.5 mM represents the optimal balance to preserve the structure and displacement capacity of the ovine sperm in these breeds, being the suggested dose for subsequent cryopreservation.
References
Casao, A., Peña-Delgado, V., & Pérez-Pe, R. (2025). From spermatogenesis to fertilisation: the role of melatonin on ram spermatozoa. Domestic Animal Endocrinology, 91(106916), 106916. https://doi.org/10.1016/j.domaniend.2025.106916
Dai, G.-C., Meng, Y., Zhang, L.-K., Du, Y.-Q., Wen, F., Feng, T.-Y., & Hu, J.-H. (2019). Effect of addition of melatonin on liquid storage of ram semen at 4°C. Andrologia, 51(5), e13236. https://doi.org/10.1111/and.13236
Deng, S.-L., Sun, T.-C., Yu, K., Wang, Z.-P., Zhang, B.-L., Zhang, Y., Wang, X.-X., Lian, Z.-X., & Liu, Y.-X. (2017). Melatonin reduces oxidative damage and upregulates heat shock protein 90 expression in cryopreserved human semen. Free Radical Biology & Medicine, 113, 347–354. https://doi.org/10.1016/j.freeradbiomed.2017.10.342
Durá, A. (2023). Efecto del protocolo de refrigeración y de congelación sobre la calidad del semen criocongelado de machos cabríos. Gva.es. https://redivia.gva.es/bitstream/handle/20.500.11939/8947/2023_Dura_Efecto.pdf?sequence=1&isAllowed=y
Farfán-Alvarado, D., Alvarado-Alvarado, J., & Moscoso-Piedra, A. (2025). Determinación Del Efecto Post Congelación De Microdosis De Menaquinona-4 En Semen Ovino. Journal Scientific Investigar ISSN: 2588–0659. https://www.investigarmqr.com/ojs/index.php/mqr/article/view/2133/6225
Flores, C., Meléndez, C., Mendoza, C., Márquez, Y., & Vilanova, L. T. (2018). Vista de Efecto antioxidante de la melatonina durante la conservación de semen de cerdo. https://revistas.unne.edu.ar/index.php/vet/article/view/2780/2461
Hernández, A. (2024). CRIOCONSERVACIÓN DE SEMEN DE CAPRINO ADICIONADO CON MELATONINA COMO ANTIOXIDANTE [UNIVERSIDAD AUTÓNOMA DE PUEBLA]. https://repositorioinstitucional.buap.mx/server/api/core/bitstreams/15c33e24-91a5-4e24-adb1-173f583b8a91/content
Ji, K., Jinbo, Wei, Zhiwei, Fan, Mengkang, Zhu, Xin, Yuan, Sihuan, Zhang, Shuang, Li, Han, Xu, & Ling, Y. (2024). Preservative effects of curcumin on semen of Hu sheep. Animals. https://www.mdpi.com/2076-2615/14/6/947
Jiménez, S. M., Gascón, A. C., Pérez, J. Á. C., Blanco, M. T. M., & Pe, R. P. (2018). Acción protectora de la melatonina frente al daño por frío en espermatozoides ovinos. XLIII Congreso Nacional y XIX Congreso Internacional de la Sociedad Española de Ovinotecnia y Caprinotecnia (SEOC), 603–608.
Lema Guamán, M. F., Maldonado Cornejo, M. E., Alvarado Alvarado, J. C., & Moscoso Piedra, A. L. (2025). Evaluación del Efecto Combinado de la menaquiona-4 y L-Carnitina sobre la Crioconservación de Semen Ovino. ASCE MAGAZINE, 4(3), 1906–1926. https://doi.org/10.70577/asce/1906.1926/2025
Makris, A., Alevra, A. I., Exadactylos, A., Papadopoulos, S., & Griveas, I. (2023). The Role of Melatonin to Ameliorate Oxidative Stress in Sperm Cells. nternational Journal of Molecular Sciences, 24(20), 15056. https://doi.org/10.3390/ijms242015056
Manchev, S., Stefanov, R., Anev, G., Tzvetkova, P., & Maksimović, N. (2014). Effect of melatonin on the reproductive characteristics of rams of north east Bulgarian fine fleece breed. Comptes Rendus de l’Académie Bulgare Des Sciences. https://r.istocar.bg.ac.rs/handle/123456789/432
Martín-Hidalgo, D., Barón, F. J., Bragado, M. J., Carmona, P., Robina, A., García-Marín, L. J., & Gil, M. C. (2011). The effect of melatonin on the quality of extended boar semen after long-term storage at 17 °C. Theriogenology, 75(8), 1550–1560. https://doi.org/10.1016/j.theriogenology.2010.12.021
Motta, N. C., Egger, R. C., Monteiro, K. S., Vogel de Oliveira, A., & Solis Murgas, L. D. (2022). Effects of melatonin supplementation on the quality of cryopreserved sperm in the neotropical fish Prochilodus lineatus. Theriogenology, 179, 14–21. https://doi.org/10.1016/j.theriogenology.2021.11.012
Nallella, K. P., Sharma, R. K., Allamaneni, S. S. R., Aziz, N., & Agarwal, A. (2004). Cryopreservation of human spermatozoa: comparison of two cryopreservation methods and three cryoprotectants. Fertility and Sterility, 82(4), 913–918. https://doi.org/10.1016/j.fertnstert.2004.02.126
Paucar-Quito, E., Alvarado-Alvarado, J., Moscoso-Piedra, A., & Maldonado-Cornejo, M. (2025). Evaluación de tres diluidores comerciales sobre el semen ovino post-congelación. Polo del conocimiento. https://polodelconocimiento.com/ojs/index.php/es/article/view/8829/pdf
Şavkay, O. L., Yalçın, M. E., & Tavşanoğlu, V. (2020). Sperm motility analysis system implemented on a hybrid architecture to produce an intelligent analyzer. Informatics in Medicine Unlocked, 19(100324), 100324. https://doi.org/10.1016/j.imu.2020.100324
Shahat, A. M., Thundathil, J. C., & Kastelic, J. P. (2022). Melatonin improves testicular hemodynamics and sperm quality in rams subjected to mild testicular heat stress. Theriogenology, 188, 163–169. https://doi.org/10.1016/j.theriogenology.2022.05.029
Shayestehyekta, M., Mohammadi, T., Soltani, L., & PooyanMehr, M. (2022). Effect of different concentrations of melatonin on ram epididymal spermatozoa recovered post-mortem under oxidative stress conditions and storage at 4°C. Zuchthygiene [Reproduction in Domestic Animals], 57(12), 1520–1528. https://doi.org/10.1111/rda.14228
Sunder, M., & Leslie, S. W. (2025). Semen analysis. En StatPearls. StatPearls Publishing.
Tisalema-Shaca, M. O., Mira-Naranjo, J. M., Valle-Baldeón, S., & Llivi-Marcatoma, J. (2024). Caracterización sociocultural y económica de producción de ovinos en comunidades indígenas, Tungurahua - Ecuador. Telos, 26(3), 975–992. https://doi.org/10.36390/telos263.12
Tiwari, S., Dewry, R. K., Srivastava, R., Nath, S., & Mohanty, T. K. (2022). Targeted antioxidant delivery modulates mitochondrial functions, ameliorates oxidative stress and preserve sperm quality during cryopreservation. Theriogenology, 179, 22–31. https://doi.org/10.1016/j.theriogenology.2021.11.013
Valdez-Pinargote, G., Moscoso-Piedra, A., & Maldonado-Cornejo, M. (2024). Efecto de la concentración de la l-carnitina en congelación del semen ovino. Journal Scientific Investigar ISSN: 2588–0659. https://doi.org/10.56048/MQR20225.8.4.2024.7324-7341
Widyastuti, R., Prastowo, S., Jaswandi, J., Lubis, A., Setiawan, R., Ridlo, M. R., & Boediono, A. (2024). Effect of melatonin supplementation on sperm quality parameters and expression of antioxidant genes during cold storage of buck semen extenders. Veterinary World, 17(4), 863–870. https://doi.org/10.14202/vetworld.2024.863-870
Yadav, D. K., Kumar, A., Gupta, S., Sharma, P., Kumar, G., Sachan, V., Yadav, B., Yadav, S., Saxena, A., & Swain, D. K. (2023). Antioxidant additive melatonin in tris-based egg yolk extender improves post-thaw sperm attributes in Hariana bull. Animal Reproduction Science, 251(107214), 107214. https://doi.org/10.1016/j.anireprosci.2023.107214
Zhang, L., Sun, Y., Jiang, C., Sohail, T., Sun, X., Wang, J., & Li, Y. (2024). Damage to mitochondria during the cryopreservation, causing ROS leakage, leading to oxidative stress and decreased quality of ram sperm. Zuchthygiene [Reproduction in Domestic Animals], 59(10), e14737. https://doi.org/10.1111/rda.14737
