Characteristics and Kinematics of Fresh Belgian Blue Semen and Its Resistance to Frozen–Thaw Using Different Extenders
Abstract
Extenders are used to preserve semen quality during storage. The Belgian Blue is a cattle breed known for its high dressing percentage; however, male reproductive performance is relatively low due to small testes size. This study aimed to analyze the characteristics and kinematics of fresh and frozen Belgian Blue semen. Fresh semen samples were collected from four Belgian Blue bulls, and their quality was assessed macroscopically, microscopically, and kinematically using portable computer-assisted sperm analysis (CASA) immediately after collection. Semen samples with progressive sperm motility ≥70% were diluted with the following extenders: Tris–egg yolk (TEY) and the commercial extenders AndroMed®, BoviFree®, and Steridyl®. CASA results showed that bulls differed significantly in ejaculate volume, color, consistency, sperm kinematics, including velocity curvilinear (VCL), velocity straight-line (VSL), and velocity average path (VAP), and sperm motility; however, none of these variables differed among frozen–thawed semen samples treated with the different extenders. In contrast, total motility, slow motility, and immotile sperm in frozen–thawed semen differed significantly between bulls. In conclusion, the characteristics of fresh Belgian Blue semen vary between individuals. Moreover, TEY, AndroMed®, BoviFree®, and Steridyl® showed comparable protective capabilities for frozen–thawed semen.
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References
Aminurrahman, A., Priyanto, R., & Jakaria, J. (2021). Evaluasi ukuran-ukuran tubuh pada sapi Belgian Blue, peranakan ongole dan silangannya. Journal Agripet, 21(1): 49-54. https://doi.org/10.17969/agripet.v21i1.17684 DOI: https://doi.org/10.17969/agripet.v21i1.17684
Arif, A. A., Maulana, T., Kaiin, E. M., Purwantara, B., & Arifiantini, R. I. (2022). The quality of frozen semen of Limousin bull in various semen diluents. Tropical Animal Science Journal, 45(3), 284-290. https://doi.org/10.5398/tasj.2022.45.3.284 DOI: https://doi.org/10.5398/tasj.2022.45.3.284
Arifiantini, R. I., Karja, N. W. K., Susilawati, T., Said, S., & Mahendra, H. C. (2024). Manual produksi semen beku sapi di Indonesia. IPB Press.
Badan Standarisasi Nasional (BSN). (2024). SNI 4869-1 Semen beku - Bagian 1: Sapi. https://www.scribd.com/document/967157579/rsni3-4869-1-2024-1
Baharun, A., Arifiantini, R. I., & Yusuf, T. L. (2017). Freezing capability of Pasundan bull sperm using tris-egg yolk, tris-soy, and AndroMed® diluents. Indonesian Journal of Veterinary Sciences, 11(1), 45-49. https://doi.org/10.21157/j.ked.hewan.v11i1.5810 DOI: https://doi.org/10.21157/j.ked.hewan.v11i1.5810
Baharun, A., Pertiwi, P. P. P., Rahmi, A., Rachmadanti, N. C., Handarini, R., Iskandar, H., Samsudewa, D., Maulana, T., Said, S., Darussalam, I., Nurcholis, N., & Arifiantini, R. I. (2025). Frozen semen quality of Pasundan bulls with different individual variations. Open Veterinary Journal, 15(1), 244–251. https://doi.org/10.5455/OVJ.2025.v15.i1.22 DOI: https://doi.org/10.5455/OVJ.2025.v15.i1.22
Bahmid, N. A., Karja, N. W. K., & Arifiantini, R. I. (2023). The quality of frozen Friesian Holstein semen after long-term storage. Tropical Animal Science Journal, 46(1), 13-19. https://doi.org/10.5398/tasj.2023.46.1.13 DOI: https://doi.org/10.5398/tasj.2023.46.1.13
Bustani, G. S., & Baiee, F. H. (2021). Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders. Veterinary World, 14(5), 1220-1233. https://doi.org/10.14202/vetworld.2021.1220-1233 DOI: https://doi.org/10.14202/vetworld.2021.1220-1233
Castro, M., Leal, M., Pezo, F., & Contreras, M. J. (2025). Sperm membrane: Molecular implications and strategies for cryopreservation in productive species. Animals, 15(12), 1808. https://doi.org/10.3390/ani15121808 DOI: https://doi.org/10.3390/ani15121808
Chowdhury, S., Das, S., Gupta, T., Sana, D., & Bose, S. (2014). Evaluation of frozen semen by acrosomal integrity and sperm concentration: Two vital quality parameters of male fertility in bovines. Exploratory Animal and Medical Research, 4(1), 101-107. https://www.researchgate.net/publication/301147419
Contri, A., Gloria, A., Robbe, D., Valorz, C., Wegher, L., & Carluccio, A. (2013). Kinematic study on the effect of pH on bull sperm function. Animal Reproduction Science, 136(4), 252-259. https://doi.org/10.1016/j.anireprosci.2012.11.008 DOI: https://doi.org/10.1016/j.anireprosci.2012.11.008
Dey, S., Brothag, C., & Vijayaraghavan, S. (2019). Signaling enzymes required for sperm maturation and fertilization in mammals. Frontiers in Cell and Developmental Biology, 7, 341. https://doi.org/10.3389/fcell.2019.00341 DOI: https://doi.org/10.3389/fcell.2019.00341
Fiems, L. O., De Campeneere, S., Van Caelenbergh, W., De Boever, J. L., & Vanacker, J. M. (2003). Carcass and meat quality in double-muscled Belgian Blue bulls and cows. Meat Science, 63(3), 345-352. https://doi.org/10.1016/S0309-1740(02)00092-X DOI: https://doi.org/10.1016/S0309-1740(02)00092-X
Gunawan, M., Said, S., Karja, N. W. K., Arifiantini, R. I., Setiadi, M., A., & Iskandar, H. (2025). Development and evaluation of soy lecithin-derived nanoliposomes as a plant-based alternative to egg-yolk extender for Ongole-grade bull semen cryopreservation. Veterinary World, 18(11), 3433-3446. https://doi.org/10.14202/vetworld.2025.3433-3446 DOI: https://doi.org/10.14202/vetworld.2025.3433-3446
Hoesni, F., Afzalani, A., Fitriani, Nurhayati, N., Raguati, R., & Firmansyah, F. (2024). Semen quality of Bali bulls fed supplement of mungbean sprouts and mineral mix. International Journal of Agricultural Technology, 20(6), 2315-2326. https://li04.tci-thaijo.org/index.php/IJAT/article/view/5595
Hoflack, G., Opsomer, G., Van Soom, A., Maes, D., de Kruif, A., & Duchateau, L. (2006). Comparison of sperm quality of Belgian Blue and Holstein Friesian bulls. Theriogenology, 66(8), 1834-1846. https://doi.org/10.1016/j.theriogenology.2006.05.007 DOI: https://doi.org/10.1016/j.theriogenology.2006.05.007
Kathiravan, P., Kalatharan, J., Karthikeya, G., Rengarajan, K., & Kadirvel, G. (2011). Objective sperm motion analysis to assess dairy bull fertility using computer‐aided system - A review. Reproduction in Domestic Animals, 46(1), 165-172. https://doi.org/10.1111/j.1439-0531.2010.01603.x DOI: https://doi.org/10.1111/j.1439-0531.2010.01603.x
Keady S. M., Waters S. M., Hamill R. M., Dunne P. G., Keane M. G., Richardson, R. I, Kenny, D. A., & Moloney, A. P. (2017). Compensatory growth in crossbred Aberdeen Angus and Belgian Blue steers: Effects on the colour, shear force and sensory characteristics of longissimus muscle. Meat Science, 125(3), 128-136. https://doi.org/10.1016/j.meatsci.2016.11.020 DOI: https://doi.org/10.1016/j.meatsci.2016.11.020
Kudratullah, Arifiantini, R. I., Said, S., Yuliani, E., & Purwantara, B. (2024). Semen characteristics, freezability, and application of motility-based protein markers (proAKAP4) in assessing the suitability of superior Bali bulls (Bos sondaicus) at the Regional AI Center. Reproduction and Breeding, 4, 279-286. https://doi.org/10.1016/j.repbre.2024.09.005 DOI: https://doi.org/10.1016/j.repbre.2024.09.005
Meyermans, R. Janssens, S., Coussé, A. Gorssen, W., Hubin, X., Mayeres, P., Veulemans W., Claerebout, E., Charlier, C., & Buys, N. (2022). Myostatin mutation causing double muscling could affect increased psoroptic mange sensitivity in dual purpose Belgian Blue cattle. Animal, 16 (3), 100460. https://doi.org/10.1016/j.animal.2022.100460 DOI: https://doi.org/10.1016/j.animal.2022.100460
Morrel, J. M., Valeanu, A. S., Lundeheim, N., & Johannisson, A. (2018). Sperm quality in frozen beef and dairy bull semen. Acta Veterinaria Scandinavica, 60(1), 41. https://doi.org/10.1186/s13028-018-0396-2 DOI: https://doi.org/10.1186/s13028-018-0396-2
Nsairat, H., Khater, D., Sayed, U., Odeh, F., Al Bawab, A., & Alshaer, W. (2022). Liposomes: Structure, composition, types, and clinical applications. Heliyon, 8(5), e09394. https://doi.org/10.1016/j.heliyon.2022.e09394 DOI: https://doi.org/10.1016/j.heliyon.2022.e09394
Oliveira, L. Z., Arruda, R. P. de, Andrade, A. F. C. de, Celeghini, E. C. C., Reeb, P. D., Martins, J. P. N., Santos, R. M. dos, Beletti, M. E., Peres, R. F. G., Monteiro, F. M., & Hossepian de Lima, V. F. M. (2013). Assessment of in vitro sperm characteristics and their importance in the prediction of conception rate in a bovine timed-AI program. Animal Reproduction Science, 137(3-4), 145-155. https://doi.org/10.1016/j.anireprosci.2013.01.010 DOI: https://doi.org/10.1016/j.anireprosci.2013.01.010
Pappa, A. Z., da Silva, H. M., Valadão, L., & da Silva, F. (2019). Effect of fructose on thawed bull semen’s viability obtained by post-mortem collection. Biomedical Journal of Scientific & Technical Research, 19(3), 14319-14323. https://doi.org/10.26717/BJSTR.2019.19.003300 DOI: https://doi.org/10.26717/BJSTR.2019.19.003300
Pardede, B. P., Agil, M., Yudi, Y., & Supriatna, I. (2020). Relationship of frozen-thawed semen quality with the fertility rate after being distributed in the Brahman Cross Breeding Program. Veterinary World, 13(12), 2649-2657. https://doi.org/10.14202/vetworld.2020.2649-2657 DOI: https://doi.org/10.14202/vetworld.2020.2649-2657
Pini, T., Leahy, T., & de Graaf, S.P. (2018). Sublethal sperm freezing damage: Manifestations and solutions. Theriogenology, 118, 172-181. https://doi.org/10.1016/j.theriogenology.2018.06.006 DOI: https://doi.org/10.1016/j.theriogenology.2018.06.006
Rahman, M. B., Vandaele, L., Rijsselaere, T., Maes, D., Hoogewijs, M., Frijters, A., Noordman, J., Granados, A., Dernelle, E., Shamsuddin, M., Parrish, J. J., & Van Soom, A. (2011). Scrotal insulation and its relationship to abnormal morphology, chromatin protamination and nuclear shape of spermatozoa in Holstein-Friesian and Belgian Blue bulls. Theriogenology, 76(7), 1246-1257. https://doi.org/10.1016/j.theriogenology.2011.05.031 DOI: https://doi.org/10.1016/j.theriogenology.2011.05.031
Ramírez-Vasquez, R. R. A., Cano, A., Hozbor, F. A., & Cesari, A. (2019). Cryopreservation and egg yolk extender components modify the interaction between seminal plasma proteins and the sperm surface. Theriogenology, 140, 153–163. https://doi.org/10.1016/j.theriogenology.2019.08.025 DOI: https://doi.org/10.1016/j.theriogenology.2019.08.025
Riwu, R. M. J., Arifiantini, R. I., & Karja, N. W. K. (2023). Comparison of different lecithin diluents for cryopreservation of Toraya buffalo semen. Tropical Animal Science Journal, 46(4), 396-402. https://doi.org/10.5398/tasj.2023.46.4.396 DOI: https://doi.org/10.5398/tasj.2023.46.4.396
Rotimi, D. E., Iyobhebhe, M., Oluwayemi, E. T., Olajide, O. P., Akinsanola, B. A., Evbuomwan, I. O., Asaleye, R. M., & Ojo, O. A. (2024). Energy metabolism and spermatogenesis. Heliyon, 10(19), e38591. https://doi.org/10.1016/j.heliyon.2024.e38591 DOI: https://doi.org/10.1016/j.heliyon.2024.e38591
Safa, F., Yekti, A. P. A., Utami, P., Syah, H. A., Febrianto, N., Rachmawati, A., Yuswati, A., Amaliya, A., Sulistyowati, D., & Susilawati, T. (2025). Sperm quality, kinematics, membrane integrity, and DNA fragmentation of frozen sexed semen in Holstein-Friesian bulls. Tropical Animal Science Journal, 48(3), 199-210. https://doi.org/10.5398/tasj.2025.48.3.199 DOI: https://doi.org/10.5398/tasj.2025.48.3.199
Santos, D., Monteiro, M.J., Voss, H.P., Komora, N., Teixeira, P., & Pintado, M. (2021). The most important attributes of beef sensory quality and production variables that can affect it: A review. Livestock Science, 250(August 2021), 104573. https://doi.org/10.1016/j.livsci.2021.104573 DOI: https://doi.org/10.1016/j.livsci.2021.104573
Sukirman, I., Sukmawati, E., Rasad, S.D., & Solihati, N. (2019). The influence of breed and type of extender on the quality of bull semen. Animal Reproduction, 21(2), 64-70. https://doi.org/10.20884/1.jap.2019.21.2.641 DOI: https://doi.org/10.20884/1.jap.2019.21.2.641
Taaffe, P., O’Meara, C. M., Stiavnicka, M., Byrne, C. J., Eivers, B., Lonergan, P., & Fair, S. (2022). Increasing the frequency of ejaculate collection in young dairy bulls increases semen production and field fertility. Theriogenology, 1(182), 45-52. https://doi.org/10.1016/j.theriogenology.2022.01.030 DOI: https://doi.org/10.1016/j.theriogenology.2022.01.030
Tethool, A. N., Ciptad, G., Wahjuningsih, S., & Susilawati, T. (2022). Deterioration of frozen semen of Bali cattle after cooling at 5°C. World’s Veterinary Journal, 12(4), 395-404. https://doi.org/10.54203/scil.2022.wvj50 DOI: https://doi.org/10.54203/scil.2022.wvj50
Vince, S., Zaja, I. Z., Zamardzija, M., Balic, I. M., Vilic, M., Duricic, D., Valpotic, H., Markovic, F., & Milinkovic-Tur, S. (2018). Age-related differences of semen quality, seminal plasma, and spermatozoa antioxidative and oxidative stress variables in bulls during cold and warm periods of the year. Animal, 12(3), 559-568. https://doi.org/10.1017/S1751731117001811 DOI: https://doi.org/10.1017/S1751731117001811
Xu, Z., Yan, Q., Zhang, K., Lei, Y., Zhou, C., Ren, T., Gao, N., Wen, F., & Li, X. (2025). Mitochondrial regulation of spermatozoa function: Metabolism, oxidative stress and therapeutic insights. Animals, 15(15), 2246. https://doi.org/10.3390/ani15152246 DOI: https://doi.org/10.3390/ani15152246
Yendraliza, Y., & Rahman, Y. G. (2023). Cryopreservation of Simmental cattle semen with egg yolk from different avian species and level glycerol of different in tris diluent. Livestock and Animal Research, 21(3), 147-152. https://doi.org/10.20961/lar.v21i3.61255 DOI: https://doi.org/10.20961/lar.v21i3.61255
Zhao, F., Li, R., Liu, Y., & Chen, H. (2023). Perspectives on lecithin from egg yolk: Extraction, physicochemical properties, modification, and applications. Frontiers in Nutrition, 9, 1082671. https://doi.org/10.3389/fnut.2022.1082671 DOI: https://doi.org/10.3389/fnut.2022.1082671
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