Diluents are substances added to the semen to increase the semen volume, reduce sperm density, and maintain sperm viability. This study aimed to compare the effects of various semen diluents on the quality of frozen semen of Limousin bull. Three mature Limousin bulls were used as semen sources. The semen was collected using an artificial vagina and then evaluated for its quality. Sperms with motility less than 70% and abnormality greater than 20% were excluded from this study. After semen evaluation, each ejaculate was individually divided into four equal tubes and diluted with skimmed milk-egg yolk (SMEY), Tris–egg yolk (TEY), Tris–egg yolk–skimmed milk (TEYSM), or Andromed®. Diluted semen was loaded into a mini straw and then equilibrated at 5 °C for 4 h. Following equilibration, the straws were frozen. The percentages of sperm motility, viability, intact membrane, and sperm DNA integrity were evaluated after freezing. Furthermore, malondialdehyde (MDA) and aspartate aminotransferase (AST) enzyme concentrations were assessed after thawing. The sperm motility with TEY, TEYSM, and Andromed® did not differ significantly but was higher than with SMEY. Sperm viability, membrane intactness, and sperm DNA integrity did not differ among treatments. By contrast, the MDA levels and concentration of AST in the Andromed® diluent semen showed the lowest values compared with SMEY semen (p<0.05) but did not differ from those of TEY and TEYSM semen. To conclude, TEY, TEYSM, and Andromed® diluents performed equally well, and although they showed better results than SMEY, sperm diluted in SMEY are still suitable for artificial insemination.
Al-Badry, K. I., Q. M. Aboud, S. J. Zalzala, F. F. Ibrahim, & W. Y. Lateef. 2018. Effect of trehalose and month of collection on DNA fragmentation in Holstein bull semen during dilution, cooling and thawing. J. Vet. Res. 22:237-246.
Andersen, J. M., P. O. Ronning, H. Herning, S. D. Bekken, T. B. Haugen, & O. Witczak. 2016. Fatty acid composition of spermatozoa is associated with BMI and with semen quality. Andrology 4:857-865. https://doi.org/10.1111/andr.12227
Anghel, A. & Z. Stela. 2010. Role of antioxidant additives in the protection of the cryopreserved semen against free radicals. Rom. Biotechnol. Lett. 15:33-41.
Arifiantini, R. I. 2012. Teknik Koleksi dan Evaluasi Semen pada Hewan. IPB Press, Bogor, Indonesia.
Arif, A. A., T. Maulana, E. M. Kaiin, B. Purwantara, R. I. Arifiantini, & E. Memili. 2020. Comparative analysis of various step-dilution techniques on the quality of frozen Limousin bull semen. Vet. World. 13:2422-2428. https://doi.org/10.14202/vetworld.2020.2422-2428
Asadpour, R., R. Jafari, & H. Tayefi-Nasrabadi. 2012. The effect of antioxidant supplementation in semen extenders on semen quality and lipid peroxidation of chilled bull spermatozoa. Iran J. Vet. Res.13:246-249.
Ax, R., M. Dally, B. A. Didion, R. W. Lenz, C. C. Love, D. D. Varner, B. Hafez, & M. E. Bellin. 2000. Semen Evaluation. In: Hafez B, Hafez ESE. Reproduction in Farm Animals 7th (Ed). Philadelphia (US): Lippincott Williams and Wilkins. pp 365-389.
Badan Standardisasi Nasional. 2017. SNI Semen Beku-Bagian 1: Sapi. BSN, Jakarta.
Baharun, A., R. I. Arifiantini, & T. L. Yusuf. 2017. Freezing capability of pasundan bull sperm using tris-egg yolk, tris-soy, and andromed® diluents. J. Ked. Hew. 11:45- 49. https://doi.org/10.21157/j.ked.hewan.v11i1.5810
Bailee, F. H., H. Wahid, Y. Rosina, O. Ariff, & N. Yimer. 2018. Sperm DNA impairment in the bull: causes, influences on reproduction and evaluations. Cryobiology 41:63–80. https://doi.org/10.1016/j.cryobiol.2017.12.006
Best, B. P. 2015. Cryoprotectant toxicity: Facts, issues, and questions. Rejuvenation Res. 18:422-436. https://doi.org/10.1089/rej.2014.1656
Borah, B. K. D., B. C. Deka, R. K. Biswas, P. Chakravarty, S. Deori, S. Sinha, & K. Ahmed. 2015. Effect of thawing methods on frozen semen quality of yak (Poephagus grunniens L.) bulls. Vet. World. 8:831-834. https://doi.org/10.14202/vetworld.2015.831-834
Coppock, R. W. 2019. Nutraceuticals in Genitourinary Maladies. In: Gupta, R.C., A. Srivastava, & R. Lall. (editor). Nutraceuticals in Veterinary Medicine. Springer Nature Switzerland AG, Switzerland. pp. 489-514. https://doi.org/10.1007/978-3-030-04624-8_33
Choudhary, K. K., K. M. Kavya, A. Jerome, & R. K. Sharma. 2016. Advances in reproductive biotechnologies, Vet. World. 9:388-395. https://doi.org/10.14202/vetworld.2016.388-395
Durrant, B. S. 2009. The importance and potential of artificial insemination in CANDES (companion animals, non-domestic, endangered species). Theriogenology 71:113-322. https://doi.org/10.1016/j.theriogenology.2008.09.004
Ehala-Aleksejev, K., & M. Punab. 2016. Serum hepatic enzyme activity in relation to semen quality and serum reproductive hormone levels among Estonian fertile men. Andrology 4:152-159. https://doi.org/10.1111/andr.12136
El-Harairy, M. A., W. A. Khalil, E. I. Khalifa, & A. A. Saber. 2018. Effect of propolis ethanolic extract supplementation to ram semen extenders on sperm characteristics, lipid peroxidation and some enzymatic activities in seminal plasma in chilled semen. Journal of Animal and Poultry Production 9:235–243. https://doi.org/10.21608/jappmu.2018.41098
El-Nagar, H. A. 2017. Effect of some semen extenders as a natural source of antioxidants on quality of frozen Friesian semen. Journal of Animal and Poultry Production 8:391-397. https://doi.org/10.21608/jappmu.2017.46014
Feradis, M. P. 2010. Reproductive Biotechnology in Farm Animals. Alphabeta, Bandung.
Herrera, J. A., G. Calderón, A. Guzmána, A. K. Vargasa, A. Ávalosa, & A. M. Rosalesa. 2017. Evaluation of two diluents for the storage of fresh and cryopreserved semen of Harris hawk (Parabuteo unicinctus). Austral J. Vet. Sci. 1:39-43. https://doi.org/10.4067/S0719-81322017000100039
Inanc, M. E., C. Beste, K. Tekin, H. Alemdar, & A. Daskin. 2018. The combination of CASA kinetic parameters and fluorescein staining as a fertility tool in cryopreserved bull semen. Turk. J. Vet. Anim. Sci. 42:452-458. https://doi.org/10.3906/vet-1801-83
Jakaria, G. G., K. Puja, R. I. Arifiantini, S. Herliantien, B. Bakar, A. Hutasoit, S. H. M. Herwiyati, E. Hajirin, Jamarizal, Y. Maulana, H. Chakra, M. Zuljisman, F. Deflaizar, T. U. Abrianto, & S. Hidayati. 2018. Roadmap Swasembada Pejantan Unggul 2018-2022. Direktorat Perbibitan dan Produksi Ternak, Direktorat Jenderal Peternakan dan Kesehatan Hewan, Kementerian Pertanian, Jakarta (ID).
Jha, P. J., M. G. S. Alam, A. A. Mansur, N. Naher, T. Islam, M. U. Bhuiyan, & F. Y. Bari. 2019. Cryopreservation of Bangladeshi ram semen using different diluents and manual freezing techniques. Cryobiology 89:35–41. https://doi.org/10.1016/j.cryobiol.2019.06.001
Kucuk, N. 2018. Sperm DNA and detection of DNA fragmentations in sperm. Turk. J. Urol. 44:1-5. https://doi.org/10.5152/tud.2018.49321
Michos, I. A., G. Basioura, C. M. Boscos, & I. A. Tsakmakidis. 2013. Proper use and impact of ‘Computer Assisted Semen Analysis’ technique on semen evaluation of farm animals. Journal of the Hellenic Veterinary Medical Society 64:267-274. https://doi.org/10.12681/jhvms.15506
Morrell, J. M. 2011. Artificial Insemination: Current and Future Trends. https://www.intechopen.com/chapters/16096.
Mughal, D. H., A. Ijaz, M. S. Yousaf, F. Wadood, U. Farooq, S. A. Mahmood, & A. Riaz. 2018. Effect of osmotic pressure on spermatozoa characteristics of cryopreserved buffalo bull (Bubalus bubalis) semen. J. Appl. Anim. Res. 46:274-277. https://doi.org/10.1080/09712119.2017.1295971
Nagata, M. B., J. Egashira, N. Katafuchi, K. Endo, K. Ogata, K. Yamanaka, T. Yamanouchi, H. Matsuda, Y. Hashiyada, & K. Yamashita. 2019. Bovine sperm selection procedure prior to cryopreservation for improvement of post thawed semen quality and fertility. J. Anim. Sci. Biotechnol. 10:91. https://doi.org/10.1186/s40104-019-0395-9
Nagy, A., T. Polichronopoulos, A. Gáspárdy, L. Solti, & S. Cseh. 2015. Correlation between bull fertility and sperm cell velocity parameters generated by computer-assisted semen analysis. Acta Vet. Hung. 63:370–381. https://doi.org/10.1556/004.2015.035
Pena, A. & C. Linde-Forsberg. 2000. Effect of equex, one or two step dilution and two freezing and thawing rates on post-thaw survival of dog spermatozoa. Theriogenology 54:859-875. https://doi.org/10.1016/S0093-691X(00)00397-6
Raheja, N., S. Choudhary, S. Grewal, N. Sharma, & N. Kumar. 2018. A review on semen extenders and additives used in cattle and buffalo bull semen preservation. J. Entomol. Zool. Stud. 6:239-245.
Said, S., F. Afiati, & T. Maulana. 2015. Study on changes of sperm head morphometry and DNA integrity of freeze-dried bovine spermatozoa. J. Indones. Trop. Anim. Agric. 40:145-152. https://doi.org/10.14710/jitaa.40.3.145-152
Santoso S, H. Herdis, R. I. Arifiantini, A. Gunawan, & C. Sumantri. 2021. Characteristics and potential production of frozen semen of pasundan bull. Trop. Anim. Sci. J. 44:24-31. https://doi.org/10.5398/tasj.2021.44.1.24
Sharafi, M., M. Zhandi, M. Shahverdi, & M. Shakeri. 2015. Beneficial effect of nitric oxide-induced mild oxidative stress on post-thawed bull semen quality. Int J. Fert Ster. 9:230-237.
Sorongbe, T. A., J. O. Daramola, O. M Onagbesan, A. V. Jegede, & E.O. Adekunle. 2019. Soybean milk extender improves sperm functional and oxidative stress parameters of goat sperm during slow and rapid freezing. Agricultura Tropica et Subtropica 52:89-96. https://doi.org/10.2478/ats-2019-0010
Sukmawati, E., R. I. Arifiantini, & B. Purwantara. 2014. Freezing capacity of sperm on various type of superior bulls. Jurnal Ilmu Ternak dan Veteriner 19:168-175. https://doi.org/10.14334/jitv.v19i3.1079
Susilawati, T. 2013. Pedoman Inseminasi Buatan pada Ternak. UB Press, Malang, Indonesia.
Ugur, M.R., A. S. Abdelrahman, H. C. Evans, A. A. Gilmore, M. Hitit, R. I. Arifiantini, B. Purwantara, A. Kaya, & E. Meili. 2019. Advances in cryopreservation of bull sperm. Review article. Front. Vet. Sci. 6:268. https://doi.org/10.3389/fvets.2019.00268
Veerabramhaiah, K., A. S. Rao, V. H. Rao, K. V. Naidu, & S. T. V. Rao. 2011. Effect of tris and biociphos-plus extenders on the extracellular enzyme release of phosphatases and transferases in punganur bull semen. J. Adv. Vet. Anim. Res. 1:61–64.
Waheed, M. M., E. M. Gouda, & T. A. A. Khalifa. 2013. Impact of seminal plasma superoxide dismutase and glutathione peroxidase on cryopreserved buffalo spermatozoa. Anim. Reprod. Sci 142:126-30. https://doi.org/10.1016/j.anireprosci.2013.09.008
Vishwanath, R., & P. Shannon. 2000. Storage of bovine semen in liquid and frozen state. Anim. Reprod. Sci. 62:23-53. https://doi.org/10.1016/S0378-4320(00)00153-6
Widjaya, N. 2011. The effect of skimmed milk with Tris egg yolk diluent on the survival of bull spermatozoa at storage temperature of 5 ºC. Sains Peternakan 9:72-76. https://doi.org/10.20961/sainspet.v9i2.4796
Zega, I., S. Ilyas, & S. Hutahaean. 2015. Spermatozoa quality of Limousin in a two-stetm extender diluent with duck egg yolk supplementation during storage in the refrigerator. Jurnal Biosains 1:66-72. https://doi.org/10.24114/jbio.v1i3.2925
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Authors submitting manuscripts should understand and agree that copyright of manuscripts of the article shall be assigned/transferred to Tropical Animal Science Journal. The statement to release the copyright to Tropical Animal Science Journal is stated in Form A. This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License (CC BY-SA) where Authors and Readers can copy and redistribute the material in any medium or format, as well as remix, transform, and build upon the material for any purpose, but they must give appropriate credit (cite to the article or content), provide a link to the license, and indicate if changes were made. If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original.