A total of 28 ejaculates collected from four Swamp buffalo bulls aged five to eight years maintained at the Network Project on Swamp Buffalo, College of Veterinary Science, Khanapara, Guwahati, Assam were used to study the leakage of sperm membrane protein, cholesterol, ALT and AST during cryopreservation of Swamp buffalo bull semen in Tris-egg yolk-citrate-glycerol extender. The mean levels of sperm membrane protein (SMP) in fresh, after equilibration and after freezing were 5.13±0.12, 3.95±0.10 and 3.83±0.11 mg/ 109 spermatozoa, respectively. The mean levels of cholesterol in fresh, after equilibration and after freezing were 21.95±0.44, 18.27±0.40 and 14.74±0.60 µg/ 108 spermatozoa, respectively. The mean levels of sperm membrane proteins and cholesterol were significantly (p<0.01) lower after equilibration and freezing. The mean AST activity was 12.63±1.14, 28.60±1.71 and 57.20±4.52 units/ 108 spermatozoa while the mean ALT activity was 0.89±0.11, 1.73±0.21 and 4.00±0.20 units/ 108 spermatozoa in fresh, after equilibration and freezing, respectively. The mean activity of AST and ALT was significantly (p<0.01) higher after equilibration and freezing. From the present study, it can be concluded that cryopreservation induces leakage of sperm membrane protein, cholesterol, ALT and AST of the swamp buffalo bull semen, which is an indicator of cryodamage.
Talukdar, D.J., Ahmed, K., Talukdar, P., Bora, P.P., 2025. Leakage of sperm membrane protein, cholesterol, ALT and AST during cryopreservation of swamp buffalo bull semen. Research Biotica 7(1), 11-15. DOI: 10.54083/ResBio/7.1.2025/11-15.
ALT, AST, Bull semen, Cholesterol, Sperm membrane protein, Swamp buffalo
Abdelnour, S.A., Khalil, W.A.,. Khalifa, N.E, Khalil, F.M.A., Hassan, M.A.E., 2024. L-Proline: A promising tool for boosting cryotolerance and fertilizing ability of cryopreserved sperm in animals. Animal Reproduction Science 263, 107429. DOI: https://doi.org/10.1016/j.anireprosci.2024.107429.
Arunkumar, R., Kumaresan, A., Sinha, M.K., Elango, K., King, J.P.E.S., Nag, P., Karuthadurai, T., Baithalu, R.K., Mohanty, T.K., Kumar, R., Datta, T.K., 2022. The cryopreservation process induces alterations in proteins associated with bull sperm quality: The equilibration process could be a probable critical control point. Frontiers in Endocrinology 13, 1064956. DOI: https://doi.org/10.3389/fendo.2022.1064956.
Chavda, B.P., Vala, K.B., Solanki, G.B., Prajapati, S.G., Singh, V.K., 2022. Caffeine supplementation in semen extender protects buffalo spermatozoa from cryodamage. Indian Journal of Veterinary Sciences and Biotechnology 18(4), 1-5. DOI: https://doi.org/10.48165/ijvsbt.18.4.01.
Cheema, R.S., Bansal, A.K., Bilaspuri, G.S., Gandotra, V.K., 2011.Correlation between the proteins and protein profile(s) of different regions of epididymis and their contents in goat buck. Animal Science Papers and Reports 29(1), 75-84.
De Leeuw, F.E., Chen, H.C., Colenbrander, B., Verkleij, A.J., 1990. Cold-induced ultrastructural changes in bull and boar sperm plasma membranes. Cryobiology 27(2), 171-183. DOI: https://doi.org/10.1016/0011-2240(90)90009-S.
Dhanju, C.K., Cheema, R.S., Kaur, S.P., 2001. Effect of freezing on proteins and protein profiles of sperm membrane extracts and seminal plasma of buffalo bulls. Asian-Australasian Journal of Animal Sciences 14(12), 1678-1682. DOI: https://doi.org/10.5713/ajas.2001.1678.
El-Harairy, M.A., Eid, L.N., Zeidan, A.E.B., El-Salaam, A.M.A., El-Kishk, M.A.M., 2011. Quality and fertility of the frozen-thawed bull semen as affected by the different cryoprotectants and glutathione levels. Journal of American Science 7(5), 791-801.
Foote, R.H., 1970. Fertility of bull semen at high extension rates in tris-buffered extenders. Journal Dairy Science 53(10), 1475-1477. DOI: https://doi.org/10.3168/jds.S0022-0302(70)86417-7.
Graham, E.F., Crabo, B.G., Schmel, M.K.L., 1974. Utilization of enzyme assay in developing techniques for freezing semen. In: Proceedings of VIIIth International Zootechnical Symposium, Milan, Italy. pp. 95-111. [c.f. Animal Breeding Abstract 42: 411].
Ibrahim, M.A., 2024. Bull sperm cryopreservation: An overview on the current status and future perspectives. The German Journal of Veterinary Research 4(1), 9-22. DOI: https://doi.org/10.51585/gjvr.2024.1.0071.
Kadirvel, G., Kumar, S., Kumaresan, A., Kathiravan, P., 2009. Capacitation status of fresh and frozen-thawed buffalo spermatozoa in relation to cholesterol level, membrane fluidity and intracellular calcium. Animal Reproduction Science 116(3-4), 244-253. DOI: https://doi.org/10.1016/j.anireprosci.2009.02.003.
Kadirvel, G., Kathiravan, P., Kumar, S., 2011. Protein tyrosine phosphorylation and zona binding ability of in vitro capacitated and cryopreserved buffalo spermatozoa. Theriogenology 75(9), 1630-1639. DOI: https://doi.org/10.1016/j.theriogenology.2011.01.003.
Nagendrakumar, S.B., Kathiresan, D., 2009. Effect of type of freezing on the functional characteristics of frozen thawed bull spermatozoa. Indian Journal of Animal Reproduction 30(2), 13-18.
Lessard, C., Parent, S., Leclerc, P., Bailey, J.L., Sullivan, R., 2000. Cryopreservation alters the level of the bull sperm surface protein P25b. Journal of Andrology 21(5), 700-707. DOI: https://doi.org/10.1002/j.1939-4640.2000.tb02138.x.
Nath, K.C., Ahmed, K., Dutta, G., Barthakur, T., Borgohain, B.N., 1996. Semen quality and release of certain enzymes during the course of freezing bull spermatozoa. Indian Journal of Animal Reproduction 17(2), 130-131.
Pratap, N., Reddy, V.N.V., Sharma, P.A., Honnappa, T.G., Devraj, M., Krishnaswamy, A., Arrora, V.K., 1999. Estimation of transaminases (AST and ALT) in cryopreserved Murrah buffalo semen. Indian Journal of Animal Reproduction 20(2), 159-160.
Said, S., Maulana, T., Iskandar, H., Kaiin, E.M., Khaerunnisa, I., Putra, W.P.B., Hasan, F., Arifiantini, R.I., 2024. Sperm protein profile and their correlation with frozen semen quality of indigenous Indonesian buffalo bulls. Journal of Advanced Veterinary and Animal Research 11(4), 846-855. DOI: http://doi.org/10.5455/javar.2024.k836.
Singh, J., Pangaonkar, G.R., Biswas, R.K., Shrivastava, A.K., 1991. Studies on leakage of transaminases from buffalo spermatozoa in relation to deep freezing in certain extenders. Indian Journal of Animal Reproduction 12(2), 135-140.
Srivastava, N., Jerome, A., Srivastava, S.K., Ghosh, S.K., Amit, K., 2013a. Bovine seminal PDC-109 protein: An overview of biochemical and functional properties. Animal Reproduction Science 138(1-2), 1-13. DOI: https://doi.org/10.1016/j.anireprosci.2013.02.008.
Srivastava, N., Srivastava, S.K., Ghosh, S.K., Amit, K., Perumal, P., Jerome, A., 2013b. Acrosome membrane integrity and cryocapacitation are related to cholesterol content of bull spermatozoa. Asian Pacific Journal of Reproduction 2(2), 126-131. DOI: https://doi.org/10.1016/S2305-0500(13)60132-3.
Taha, T.A., Abdel-Gawad, E.I., Ayoub, M.A., 2000. Monthly variations in some reproductive parameters of Barki and Awassi rams throughout 1 year under subtropical conditions 2. Biochemical and enzymatic properties of seminal plasma. Animal Science 71(2), 325-332. DOI: https://doi.org/10.1017/S135772980005517X.
Talukdar, D.J., 2014. Morphological and functional characterization of in vitro capacitated and frozen thawed swamp buffalo spermatozoa. Ph.D. Thesis, Assam Agricultural University, Guwahati, India.
Talukdar, D.J., Ahmed, K., Talukdar, P., 2015a. Cryocapacitation and fertility of cryopreserved semen. International Journal of Livestock Research 5(6), 11-18.
Talukdar, D.J., Ahmed, K., Deori, S., Das, G.C., 2015b. Heparin-induced in vitro capacitation changes of swamp buffalo spermatozoa. Turkish Journal of Veterinary & Animal Sciences 39(5), 629-633. DOI: https://doi.org/10.3906/vet-1501-16.
Talukdar, D.J., Ahmed, K., Deka, B.C., Sinha, S., Deori, S., Das G.C., 2016. Cryo-capacitation changes during cryopreservation of swamp buffalo spermatozoa. The Indian Journal of Animal Sciences 86(4), 397-400. DOI: https://doi.org/10.56093/ijans.v86i4.57723.
Talukdar, D.J., Ahmed, K., Sinha, S., Deori, S., Das, G.C., Talukdar, P., 2017. Cryopreservation induces capacitation-like changes of the swamp buffalo spermatozoa. Buffalo Bulletin 36(1), 221-230.
Watson, P.F., 1995. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reproduction, Fertility and Development 7(4), 871-891. DOI: https://doi.org/10.1071/RD9950871.
