Evaluation of Centrifuged Egg Yolk Extender with L-Methionine for Improving Cryosurvival of Kankrej Bull Spermatozoa
DOI:
https://doi.org/10.48165/ijvsbt.2026.22.06.26Keywords:
Centrifuged egg yolk extender, Enzymatic parameters, Kankrej bull, L-methionine, Oxidative stress, Semen cryopreservation, Sperm qualityAbstract
The present study was undertaken to evaluate the effect of a centrifuged Tris-fructose egg yolk glycerol (TFEYG) extender supplemented with L-methionine on post-dilution and post-thaw semen quality, oxidative stress, and enzymatic activity in Kankrej bull semen. Eighteen ejaculates collected from three healthy, sexually mature Kankrej bulls were processed using three extender formulations: conventional TFEYG (Group I), centrifuged TFEYG (Group II), and centrifuged TFEYG supplemented with 2.5 mM L-methionine (Group III). Semen quality was assessed in terms of individual sperm motility, viability, abnormalities, hypo-osmotic swelling test (HOST)-reacted sperm, malondialdehyde (MDA), superoxide dismutase (SOD), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) at the post-dilution and post-thaw stages. Cryopreservation reduced semen quality in all treatment groups; however, semen extended with centrifuged TFEYG, particularly when supplemented with L-methionine, retained significantly (p
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Akalin, P. P., Baspinar, N., Coyan, K., Bucak, M. N., Ataman, M. B., Ömür, A. D., & Aci̇baeva, B. (2015). Effects of extenders containing methionine and trehalose on ram sperm acrosome integrity and some biochemical parameters after freeze-thawing. Journal of Bahri Dagdas Animal Research, 3(1), 7–15.
Akhter, S., Ansari, M. S., Andrabi, S. M. H., Ullah, N., & Qayyum, M. (2008). Effect of antibiotics in extender on bacterial and spermatozoal quality of cooled buffalo (Bubalus bubalis) bull semen. Reproduction in Domestic Animals, 43(3), 272–278.
Alam, K., Srivastava, S., Verma, S. K., Verma, A. K., Singh, B., Kumar, R., & Singh, N. (2025). Methionine supplementation improves freezability of Jamnapari buck semen. Journal of Environmental Biology, 46(3), 329–335.
Bilodeau, J. F., Blanchette, S., Cormier, N., & Sirard, M. A. (2002). Reactive oxygen species-mediated loss of bovine sperm motility in egg yolk Tris extender: Protection by pyruvate, metal chelators and bovine liver or oviductal fluid catalase. Theriogenology, 57(3), 1105–1122.
Bucak, M. N., Tuncer, P. B., Sarıözkan, S., Başpınar, N., Taşpınar, M., Çoyan, K., & Öztuna, D. (2010). Effects of antioxidants on post-thawed bovine sperm and oxidative stress parameters: Antioxidants protect DNA integrity against cryodamage. Cryobiology, 61(3), 248–253.
Chaudhari, P. J., Dhami, A. J., Chaudhari, D. V., & Parmar, S. C. (2017). Freezability of cattle and buffalo semen and association of fresh and frozen-thawed sperm quality parameters. International Journal of Current Microbiology and Applied Sciences, 6(12), 1445–1454.
Chaudhary, A. S., Rathod, B. S., Makawana, G. S., Kapadiya, P. S., & Chaudhary, R. K. (2026). Evaluation of alternatives to conventional egg yolk extender for cryopreservation of Kankrej bull semen. The Indian Journal of Veterinary Sciences and Biotechnology, 22(6), 145–149.
Çoyan, K., Başpınar, N., Bucak, M. N., Akalın, P. P., Ataman, M. B., Ömür, A. D., & Sarıözkan, S. (2010). Influence of methionine and dithioerythritol on sperm motility, lipid peroxidation and antioxidant capacities during liquid storage of ram semen. Research in Veterinary Science, 89(3), 426–431.
Dheerib, G. S., Banana, H. J. H., & Al-Sarray, M. A. R. (2020). Effect of adding lyophilized low-density lipoproteins (L-LDL), methionine and their combinations on semen quality of Holstein bulls after cryopreservation. Systematic Reviews in Pharmacy, 11(11), 1813–1817.
El-Battawy, K. A., & El-Nattat, W. S. (2018). Cryopreservation of buck semen with emphasis on its chilling and its characteristics after some amino acids supplementation. Current Science International, 7(4), 731–736.
El-Hawary, A. F., El-Gohary, E. S. H., Gabr, A. A., El-Kishk, M. A. M., & El-Malky, O. M. (2017). Frozen-thawed bull spermatozoa quality and fertility rate added with seleno-zinc methionine. 1—Effect of zinc methionine and seleno-methionine on freezability and fertility of bull semen. Journal of the Egypt. Veterinary Medical Association, 17(2), 259–275.
Fernandez-Santos, M. R., Esteso, M. C., Montoro, V., Soler, A. J., & Garde, J. J. (2006). Cryopreservation of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa: Effects of egg yolk, glycerol and cooling rate. Theriogenology, 66(8), 1931–1942.
Gloria, A., Contri, A., Wegher, L., Vignola, G., Dellamaria, D., & Carluccio, A. (2014). The effects of antibiotic additions to extenders on fresh and frozen-thawed bull semen. Animal Reproduction Science, 150(1–2), 15–23.
Jeyendran, R. S., Van der Ven, H. H., Perez-Pelaez, M., Crabo, B. G., & Zaneveld, L. J. D. (1984). Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Reproduction, 70(1), 219–228.
Kapadiya, P. S., Nakhashi, H. C., Chauhan, P. M., Sutaria, T. V., & Suthar, B. N. (2018). Efficacy of melatonin as an antioxidant for cryopreservation of Kankrej bull semen. Ruminant Science, 7(1), 89–92.
Kim, G., Weiss, S. J., & Levine, R. L. (2014). Methionine oxidation and reduction in proteins. Biochimica et Biophysica Acta, 1840(2), 901–905.
Moussa, M., Martinet, V., Trimeche, A., Tainturier, D., & Anton, M. (2002). Low density lipoproteins extracted from hen egg yolk by an easy method: Cryoprotective effect on frozen-thawed bull semen. Theriogenology, 57(6), 1695–1706.
Nair, S. J., Brar, A. S., Ahuja, C. S., Sangha, S. P. S., & Chaudhary, K. C. (2006). A comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Animal Reproduction Science, 96(2), 21–29.
Nouri, H., Towhidi, A., Zhandi, M., & Sadeghi, R. (2013). The effects of centrifuged egg yolk used with INRA plus soybean lecithin extender on semen quality to freeze miniature Caspian horse semen. Journal of Equine Veterinary Science, 33(12), 1050–1053.
Prasad, S., & Kumar, S. (2022). Improving animal production, productivity and economic gain to the farmers through increasing coverage of artificial insemination in India. The Indian Journal of Animal Reproduction, 43(1), 1–7.
Said, S., Agung, P. P., Putra, W. P. B., & Kaiin, E. M. (2020). The role of biotechnology in animal reproduction. IOP Conference Series: Earth and Environmental Science, 462(1), 012035.
Sariozkan, S., Bucak, M. N., Tuncer, P. B., Buyukleblebici, S., & Canturk, F. (2014). Influence of various antioxidants added to TCM-199 on post-thaw bovine sperm parameters, DNA integrity and fertilizing ability. Cryobiology, 68(1), 129–133.
Singh, P., Agarwal, S., Singh, H., Singh, S., Verma, P. K., Butt, M. S., & Sharma, U. (2020). Effects of ascorbic acid as antioxidant semen additive in cryopreservation of crossbred cattle bull semen. International Journal of Current Microbiology and Applied Sciences, 9(7), 3089–3099.
Verma, A. K., Kumar, R., Srivastava, S., Alam, K., Kumar, R., Sakhahari, D., & Verma, S. (2023). Methionine as a semen additive to improve Murrah buffalo (Bubalus bubalis) semen cryopreservation. The Indian Journal of Veterinary Science and Biotechnology, 19(6), 39–43.
Wafa, W. M., El-Nagar, H. A., Hussein, Y. S., & Saeed, A. M. (2021). Effect of different antioxidant sources added to buffalo semen extender during cryopreservation on freezability and fertility of buffalo spermatozoa. Journal of Animal Health & Production, 9(3), 222–228.
Wall, R. J., & Foote, R. H. (1999). Fertility of bull sperm frozen and stored in clarified egg yolk-Tris-glycerol extender. Journal of Dairy Science, 82(4), 817–821.
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