Effect of Different Freezing Rates on Post-Thaw Semen Quality in Yak

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B. K. D. Borah
B. C. Deka
R. K. Biswas
S. Deori
P. Chakravarty
K. Ahmed
S. Sinha


A study was conducted on yak semen to evaluate the effect of different freezing rates on the quality of frozen semen. The semen was frozen following three freezing rates viz., Freezing rate I (@ 5ºC/minute from +4oC to -10oC, @ 40ºC/minute from -10ºC to -100ºC and @ 20ºC/minute from -100ºC to -140ºC), II (@ 4ºC/minute from +4ºC to -12ºC, @ 40ºC/minute from -12ºC to -40ºC and @ 50ºC/minute from -40ºC to -140ºC and III (@ 5oC/minute from +4ºC to -10ºC, @50ºC/minute from -10ºC to -100ºC and @ 20ºC/minute from -100oC to -140ºC). The sperm  motility, live sperm, HOST-reacted sperm, total incidence of acrosomal changes and  extracellular release of Alanine aminotransferase and Aspartate aminotransferase of yak semen extended in Tris extender differed significantly (P <0.01) between stages of processing and freezing but did not differ significantly between freezing rates and due to stage × freezing rate interaction. Analysis of variance revealed that in frozen semen, a freezing rate significantly (P<0.05) influenced the percentage of live sperm with Mitochondrial Membrane Potential (MMP) but did not significantly influence the percentage of sperm with DNA damage. The percentage of live sperm with MMP in frozen semen was significantly higher for freezing rate III than for freezing rate II but the difference between freezing rates I and III was not significant.

Freezing rate, frozen semen, quality, yak

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How to Cite
Borah, B. K. D., Deka, B. C., Biswas, R. K., Deori, S., Chakravarty, P., Ahmed, K., & Sinha, S. (2019). Effect of Different Freezing Rates on Post-Thaw Semen Quality in Yak. Current Journal of Applied Science and Technology, 35(3), 1-6. https://doi.org/10.9734/cjast/2019/v35i330179
Original Research Article


Deori S. The effect of freezing on cryosurvival of yak semen. SAARC J Agri. 2017;15:215-218.

Clulow JR, Mansfield LJ, Morris LHA, Evans G, Maxwell WMC. A comparison between freezing methods for the cryopreservation of stallion spermatozoa. Anim Reprod Sci. 2008;108:298–308.

Mazur P. Basic concepts in freezing cells. In: Johnson LA, Larsson K. editors. Deep Freezing of Boar Semen. Swedish University of Agricultural Sciences, Uppsala; 1985.

Watson PF. Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reprod Fertil Dev. 1995;7:871–891.

Nur Z, Zik B, Ustuner B, Tutuncu S, Sagirkaya H, Ozguden CG, et al. Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniv Vet Fak Derg. 2011;58:267-272.

Sarma JP, Sinha S, Biswas RK, Deka BC, Sarmah BC, Gogoi T, et al. Effect of freezing rate on quality of cryopreserved goat spermatozoa using a programmable freezer. IOSR-JAVS. 2015;8:67-69.

Dalal J, Kumar A, Kumar P, Honparkhe M, Malik VS, Singhal S, et al. Improvement in cryosurvival of buffalo bull (Bubalus bubalis) sperm by altering freezing rate within critical temperature range. Asian Pacific J Reprod. 2018;7:72-78.

Ahmed MU. Effect of freezing rate and thawing method on quality of frozen Swamp buffalo semen. MVSc thesis, Assam Agricultural University, Khanapara, Guwahati, India; 2011.

Ustuner B, Nur Z, Alcay S, Toker MB, Sagirkaya H, Soylu MK. Effect of freezing rate on goat sperm morphology and DNA integrity. Turkish J Vet Anim Sci. 2015;39: 110-114.

Pugliesi G, Furst R, Carvalho GR. Impact of using a fast-freezing technique and different thawing protocols on viability and fertility of frozen equine spermatozoa. Andrologia. 2014;46:1055-1062.

Garner DL, Thomas CL, Joerg HW, Dejarnette JM, Marshall CE. Fluorometric assessment of mitochondrial function and viability in cryopreserved bovine spermatozoa. Biol Reprod. 1997;57:1401-1406.

Shahani SK, Revell SG, Argo CM, Murray RD. Effect of some respiratory and glycolytic inhibitors on mitochondrial functionality in bovine semen. The Open Androl J. 2012;4:12-20.

Rastegarnia A, Shahverdi A, Topraggaleh TR, Ebrahimi B. Shafipour V. Effect of different thawing rates on post-thaw viability, kinetic parameters and chromatin structure of buffalo (Bubalus bubalis) spermatozoa. Cell J. 2013;14:306-313.

Woelders H. Fundamental and recent development in cryopreservation of bull and boar semen. Vet Quart. 1997;19:135-138.

Pickett BW, Berndtson WE. Principles and techniques of frozen spermatozoa. In: Salisbury GW, VanDemark NL, Lodge JR. editors. Physiology of Reproduction and Artificial Insemination of Cattle. W.H. Freeman and Company, San Francisco; 1978.

Byrne GP, Lonergan P, Wade M, Duffy P, Donovan A, Hanrahan JP, et al. Effect of freezing rate of ram spermatozoa on subsequent fertility in vivo and in vitro. Anim Reprod Sci. 2000;62:265-75.

Anel L, Paz P, Alvarez M, Chamorro CA, Boixo JC, Manso A, et al. Field and in vitro assay of three methods for freezing ram semen. Theriogenology. 2003;60:1293-1308.

Barbas JP, Mascarenhas RD. Cryopreservation of domestic animal sperm cells. Cell Tissue Bank. 2009;10: 49-62.

Eriksson BM, Rodriguez-Martinez H. Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa. Anim Reprod Sci. 2000;63:205-220.