Ảnh hưởng của HEPES đến chất lượng tinh trùng heo pha loãng bằng dung dịch Beltsville Thawing Solution (BTS)
,
,
,
và
Article Sidebar
Full Text: 
PDF
Abstract
This study compared the effectiveness of two semen extenders: Beltsville Thawing Solution (BTS) and a modified BTS in which sodium bicarbonate was replaced with HEPES buffer (BTSH) during storage at 17°C for 96 h. The evaluated parameters included pH, osmotic pressure, sperm viability, membrane integrity assessed by the HOST, and sperm motility characteristics analyzed using a computer-assisted sperm analysis (CASA) system. The results demonstrated that BTSH maintained more stable pH and osmotic pressure, while significantly improving sperm motility and viability compared with BTS (p < 0.05), thereby reducing unfavorable fluctuations in the semen extenders throughout the storage period. After 96 hours of storage, BTSH achieved a viability of 73.67 ± 0.88% and total motility of 76.01 ± 1.79%, which were markedly higher than those obtained with BTS. Overall, the findings indicate that BTSH is the most suitable extender for boar semen storage at 17°C for up to 96 hours, effectively preserving overall sperm quality, motility, and plasma membrane stability throughout the storage duration.
Tóm tắt
Nghiên cứu được thực hiện nhằm so sánh hiệu quả của hai môi trường bảo quản tinh trùng heo gồm BTS (Beltsville Thawing Solution) và BTS được thay thế hệ đệm bicarbonate bằng HEPES (BTSH) trong quá trình bảo quản ở 17°C trong 96 giờ. Các chỉ tiêu được đánh giá gồm pH, áp suất thẩm thấu, tỷ lệ sống, tính toàn vẹn màng (HOST) và các chỉ số vận động tinh trùng được phân tích bằng hệ thống CASA. Kết quả cho thấy môi trường BTSH duy trì pH và áp suất thẩm thấu ổn định hơn, đồng thời cải thiện rõ rệt hoạt lực và tỷ lệ sống của tinh trùng so với BTS (p < 0,05), qua đó giúp hạn chế các biến động bất lợi của môi trường pha loãng trong suốt quá trình bảo quản. Sau 96 giờ bảo quản, BTSH đạt tỷ lệ sống 73,67 ± 0,88% và hoạt lực 76,01 ± 1,79%, vượt trội hơn so với BTS. Việc tổng hợp kết quả cho thấy BTSH là môi trường phù hợp nhất cho bảo quản tinh trùng heo ở 17°C đến 96 giờ, giúp duy trì chất lượng tổng thể, khả năng vận động và tính ổn định của màng tinh trùng trong suốt quá trình bảo quản.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Tài liệu tham khảo
Alquézar-Baeta, C., Gimeno-Martos, S., Miguel-Jiménez, S., Santolaria, P., Yániz, J., Palacín, I., Casao, A., Cebrián-Pérez, J. Á., Muiño-Blanco, T., & Pérez-Pé, R. (2019). OpenCASA: A new open-source and scalable tool for sperm quality analysis. PLOS Computational Biology, 15(1), e1006691. https://doi.org/10.1371/journal.pcbi.1006691
Dang-Nguyen, T. Q., Tich, N. K., Nguyen, B. X., Ozawa, M., Kikuchi, K., Manabe, N., Ratky, J., Kanai, Y., & Nagai, T. (2010). Introduction of various Vietnamese indigenous pig breeds and their conservation by using assisted reproductive techniques. The Journal of Reproduction and Development, 56(1), 31–35. https://doi.org/10.1262/jrd.09-165k
Buathalad, K., Koedkanmark, T., Boonkum, W., & Chankitisakul, V. (2025). Optimizing rooster semen preservation: Effect of oxygen exposure, sample rotation, and HEPES buffer supplementation. Animals, 15(16), 2391. https://doi.org/10.3390/ani15162391
Chanapiwat, P., & Kaeoket, K. (2016). The effect of adding antioxidants on quality of chilled boar semen. Thai Journal of Veterinary Medicine, 46(1), 83–90.
Dziekońska, A., Kinder, M., Fraser, L., Strzeżek, J., & Kordan, W. (2017). Metabolic activity of boar semen stored in different extenders supplemented with ostrich egg yolk lipoproteins. Journal of Veterinary Research, 61(1), 127–132. https://doi.org/10.1515/jvetres-2017-0016
Gadea, J. (2003). Semen extenders used in the artificial insemination of swine. Spanish Journal of Agricultural Research, 1(2), 17–27. https://doi.org/10.5424/sjar/2003012-17
Good, N. E., Winget, G. D., Winter, W., Connolly, T. N., Izawa, S., & Singh, R. M. (1966). Hydrogen ion buffers for biological research. Biochemistry, 5(2), 467–477. https://doi.org/10.1021/bi00866a011
Guthrie, H. D., & Welch, G. R. (2012). Effects of storage solutions on boar sperm motility, membrane integrity and mitochondrial function. Theriogenology, 78(2), 240–251.
Hackerova, L., Pilsova, A., Pilsova, Z., Zelenkova, N., Tymich Hegrova, P., Klusackova, B., Chmelikova, E., Sedmikova, M., Simonik, O., & Postlerova, P. (2025). Boar sperm motility assessment using computer-assisted sperm analysis: Current practices, limitations, and methodological challenges. Animals, 15(3), 305. https://doi.org/10.3390/ani15030305
Harrison, R. A., Ashworth, P. J., & Miller, N. G. (1996). Bicarbonate/CO₂, an effector of capacitation, induces a rapid and reversible change in the lipid architecture of boar sperm plasma membranes. Molecular Reproduction and Development, 45(3), 378–391. https://doi.org/10.1002/(SICI)1098-2795(199611)45:3<378::AID-MRD16>3.0.CO;2-V
Henning, H., Nguyen, Q. T., Wallner, U., & Waberski, D. (2022). Temperature limits for storage of extended boar semen from the perspective of the sperm's energy status. Frontiers in Veterinary Science, 9, 953021. https://doi.org/10.3389/fvets.2022.953021
Johnson, L. A., Weitze, K. F., Fiser, P., & Maxwell, W. M. C. (2000). Storage of boar semen. Animal Reproduction Science, 62(1–3), 143–172. https://doi.org/10.1016/S0378-4320(00)00157-3
Keeratikunakorn, K., Chanapiwat, P., Aunpad, R., Ngamwongsatit, N., & Kaeoket, K. (2024). Effect of antimicrobial peptide BiF2_5K7K on contaminated bacteria isolated from boar semen and semen qualities during preservation and subsequent fertility test on pig farm. Antibiotics, 13(7), 579. https://doi.org/10.3390/antibiotics13070579
Kondracki, S., Wysokińska, A., Mańkowska, M., & Iwanina, M. (2017). The use of eosin–nigrosin staining for the evaluation of boar semen quality. Annals of Warsaw University of Life Sciences – SGGW Animal Science, 56(1), 39–46.
Kwon, W. S., Rahman, M. S., Lee, J. S., Yoon, S. J., Park, Y. J., & Pang, M. G. (2017). Improvement of sperm functions by the addition of HEPES in boar semen extender. Reproduction in Domestic Animals, 52(2), 213–220. https://doi.org/10.1111/rda.12902
Ho, T. K., Vo, C. T., Lam, T. N., Nguyen, T. K. K., & Nguyen, T. T. (2023). Effect of different extender on extended boar semen quality in the long-term storage. In Proceedings of the International Food Animal Conference 2023 (pp. 28–29). Can Tho University.
Murase, T., Imaeda, N., Yamada, H., Takasu, M., Taguchi, K., & Katoh, T. (2010). Dilution of boar ejaculates with BTS containing HEPES in place of bicarbonate immediately after ejaculation can reduce the increased inducibility of the acrosome reaction. Journal of Reproduction and Development, 56(3), 309–314. https://doi.org/10.1262/jrd.09-157N
Osinowote, A., Oguejiofor, C., & Olatunbosun, O. (1982). Hypo-osmotic swelling test for assessing boar sperm membrane integrity. Journal of Reproductive Fertility, 66(2), 283–288.
Park, Y. J., Shin, D. H., Pang, W. K., Ryu, D. Y., Rahman, M. S., Adegoke, E. O., & Pang, M. G. (2021). Short-term storage of semen samples in acidic extender increases the proportion of females in pigs. BMC Veterinary Research, 17(1), 362. https://doi.org/10.1186/s12917-021-03078-3
Revell, S. G., & Mrode, R. A. (1994). An osmotic resistance test for bovine semen. Animal Reproduction Science, 36(1–2), 77–86. https://doi.org/10.1016/0378-4320(94)90055-8
Rigau, T., Piedrafita, J., Reverter, A., Canal, M., & Rodríguez-Gil, J. E. (1996). The rate of L-lactate production: A feasible parameter for the fresh diluted boar semen quality analysis. Animal Reproduction Science, 43(2–3), 161–172. https://doi.org/10.1016/0378-4320(96)01496-0
Rivas, C. U., Ayala, M. E., & Aragón, A. (2022). Effect of various pH levels on the sperm kinematic parameters of boars. South African Journal of Animal Science, 52(5), 693–704. https://doi.org/10.4314/sajas.v52i5.13
Trentin, J. M., Centeno, L. A. M., de Souza Balestrin, T., Minela, T., Zanatta, G. M., Pessoa, G. A., & Rubin, M. I. B. (2018). Sodium bicarbonate and HEPES as buffer components for cooling the semen of pony stallions. Semina: Ciências Agrárias, 39(2), 631–642. https://doi.org/10.5433/1679-0359.2018v39n2p631
Will, M. A., Clark, N. A., & Swain, J. E. (2011). Biological pH buffers in IVF: Help or hindrance to success. Journal of Assisted Reproduction and Genetics, 28(8), 711–724. https://doi.org/10.1007/s10815-011-9582-0
Matabane, M. B., Thomas, R., Netshirovha, T. R., Tsatsimpe, M., Ng’ambi, J. W., Nephawe, K. A., & Nedambale, T. L. (2017). Relationship between sperm plasma membrane integrity and morphology and fertility following artificial insemination. South African Journal of Animal Science, 47(1), 102-106. https://doi.org/10.4314/sajas.v47i1.15