Efficiency of Long-term Storage at Chilling Temperatures (4 ºC) of Lyophilized Tris Egg Yolk Extender on Frozen Bovine Semen

Main Article Content

Napak Rungroekrit
Jatuporn Kajaysri
Chullachat Chapanya

Abstract

The lyophilized extender was proved that it had efficiency to be cryoprotectant for spermatozoa in frozen bovine semen. But its stability after long term storage was in question. This study also was assigned to evaluate the long-term storage (3 months) of lyophilized Tris egg yolk extender at 4 0C by analysis the quality of bovine semen after dilution, equilibration and freezing and compare their parameters of semen in fresh Tris egg yolk extender. Eight Holstein bulls was semen collected that each batch of collection was divided equally into two groups. Control group (n=8) and Treatment group (n=8) were diluted with fresh extender and fresh lyophilized extender of Tris egg yolk, respectively. Optimal sperm concentration was adjusted as 80x106/mL. The semen diluents were further processed for cooling and freezing, after that it would be stored at -196 °C for 24 hours. The sperm motilities of both groups were evaluated in terms of sperm motility, progressive sperm motility, rapid sperm motility using computer assisted semen analysis (CASA) and sperm plasma membrane integrity by hypoosmotic swelling test (HOST). The semen diluents of both groups were analyzed three points such as after semen dilution, equilibration and thawing after freezing. The experiments were repeated for three more times. They were repeated of semen quality analysis that the semen in lyophilized extender after storage at 4 0C in vacuum bottles for 1 month, 2 and 3 months of treatment group were analyzed and compared to the semen in fresh extender of control group. The results showed, the percentages of sperm motility, progressive sperm motility, rapid sperm motility and HOST positive after dilution, equilibration and freezing of semen in fresh extender of control group were not significantly different from those parameters of the semen in fresh, 1-month, 2-months and 3 months storages of lyophilized extenders of treatment group (P>0.05) that the all values of spermatozoa in treatment groups were accepted for good cryoprotectant of these lyophilized extenders in frozen semen. In conclusion, the long-term storage (3 months) of lyophilized Tris egg yolk extender still had good efficiency to preserve the frozen bovine semen quality.

Downloads

Download data is not yet available.

Article Details

Section
Research Article

References

Alcay, S., E. Gokce, M. B. Toker, N. T. Onder, B. Ustuner, E. Uzabacı, Z. Gul and S. Cavus. 2016. Freeze-dried egg yolk based extenders containing various antioxidants improve post-thawing quality and incubation resilience of goat spermatozoa. Cryobiology. 72: 269-273.
Alcay, S. , M. B. Toker, E. Gokce, B. Ustuner, N. T. Onder, H. Sagirkaya, Z. Nur and M. K. Soylu. 2015. Successful ram semen cryopreservation with lyophilized egg yolk-based extender. Cryobiology. 71: 329-333.
Barszcz, K., D. Wiesetek, M. Wasowicz and M. Kupczynska. 2012. Bull semen collection and analysis for artificial insemination. J. Agri. Sci. 4: 1-10.
Chaudhary, P. J., A. J. N. Dhami, D. V. Chaudhari and S. C. Parmar. 2017. Freezability of cattle and buffalo semen and association of fresh and frozen-thawed sperm quality parameters. Int. J. Curr. Microbiol. App. Sci. 6: 1445-1454.
Correa, J. R. and P. M. Zavos. 1994. The hypoosmotic swelling test: Its employment as an assay to evaluate the functional integrity of the frozen-thawed bovine sperm membrane. Theriogenology. 42: 351-360.
Edmonson, A. J., I. J. Lean, L. D. Weaver, T. Farver and G. Webster. 1989. A body condition scoring chart of Holstein dairy cows. J. Dairy Sci. 72: 68-78.
Forouzanfar, M., M. Sharafi, S. M. Hosseini, S. Ostadhosseini, M. Hajian, L. Hosseini, P. Abedi, N. Nili, H. R. Rahmani and M. H. Nasr-Esfahani. 2010. In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology. 73: 480-487.
Gaidhani, K. A., M. Harwalkar, D. Bhambere and P. S. Nirgude. 2015. Lyophilization/freezing drying: A review. World J. Phar. Res. 4: 516-543.
Hu, J. H., Q. W. Li, L. S. Zan, Z. L. Jiang, J. H. An, L .Q. Wang and Y. H. Jia. 2010. The cryoprotective effect of low density lipoproteins in extenders on bull spermatozoa following freezing-thawing. Anim. Reprod. Sci. 117: 11-17.
Jeyendran, R. S., H. H. Vander-Ven, M. Perez-Pelaez, B. G. Crabo and L. J. D. Zanevld. 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characters. J. Repord. Fertil. 70: 219-228.
Kajaysri, J., U. Rungroekrit, S. Ruenphet and C. Chapanya. 2017. Efficiency of lyophilized Tris egg yolk extender on bull semen cryopreservation. Proc. SAADC Anim. Sci. Cong. 323-325.
Kumar, S., J. D. Millar and P. F. Watson. 2003. The effect of cooling rate on survival of cryopreserved bull, ram and boar spermatozoa: A comparison of two controlled-rate cooling machines. Cryobiology. 46: 246-253.
Moustacas, V. S., F. G. Zaffalon, M. A. Lagares, A. M. Loaiza-Eccheverri, F. C. Varago, M. M. Neves, L. G. Heneine, R. P. Arruda and M. Henry. 2011. Natural, but not lyophilized, low density lypoproteins were an acceptable alternative to egg yolk for cryopreservation of ram semen. Theriogenology. 75: 300-307.
Rasul, Z., M. Anzar, S. Jalali and N. Ahmad. 2000. Effect of buffering system on post-thaw motion characteristics, plasma membrane integrity and acrosome morphology of buffalo spermatozoa. Anim. Reprod. Sci. 59: 31-41.
Sundararaman, M. N., J. Kalatharan and K. Thilak Pon Jawahar. 2012. Computer assisted semen analysis for quantification of motion characteristics of bull sperm during cryopreservation cycle. Vet. World. 5: 723-726.
Wall, R. J. and R. H. Foote. 1999. Fertility of bull sperm frozen and stored in clarified egg yolk-Tris-glycerol extender. J. Dairy Sci. 82: 817-821.