Advanced SearchSearch Tips
The Cryoprotective Effect on Frozen-thawed Boar Semen of Egg Yolk Low Density Lipoproteins
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
The Cryoprotective Effect on Frozen-thawed Boar Semen of Egg Yolk Low Density Lipoproteins
Hu, Jian-hong; Li, Qing-Wang; Li, Gang; Chen, Xiao-Yu; Hai-Yang, Hai-Yang; Zhang, Shu-Shan; Wang, Li-Qiang;
  PDF(new window)
In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.
Boar Semen;Cryopreservation;Yolk LDL;Sperm Motility;Acrosome Integrity;Plasma Membrane Integrity;
 Cited by
Avaliação de diferentes crioprotetores intra e extracelulares na criopreservação de sêmen de touros, Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 2013, 65, 2, 415  crossref(new windwow)
Cryoprotective effects of low-density lipoproteins, trehalose and soybean lecithin on murine spermatogonial stem cells, Zygote, 2014, 22, 02, 158  crossref(new windwow)
) spermatozoa, Reproduction in Domestic Animals, 2016, 51, 5, 708  crossref(new windwow)
Andersson, M., T. Hellman, B. G. Holmstrom and L. Jokinen. 1992. Computerized and subjective assessments of post- thaw motility of semen from Finnish Ayrshire Al bulls in relation to nonreturn rates. Acta. Vet. Scand. 33(l):89-93

Bogart, R. and D. Mayer. 1950. The effects of egg yolk on the various physical and chemical factors detrimental to spermatozoa viability. J. Anim. Sci. 9:143

Check, J. H., B. Shanis, A. Bollendorf, H. Adelson and E. Breen. 1989. Semen characteristics and infertility in ageing. Arch. Androl. 23:275-277 crossref(new window)

Cheon, Y. M., H. K. Kim, C. B. Yang, Y. J. Yi and C. S. Park.2002. Effect of season influencing semen characteristics, frozenthawed sperm viability and testosterone concentration in Duroc boars. Asian-Aust. J. Anim. Sci. 15(4):500-503

Clarke, R. N. and L. A. Johnson. 1987. Effect of liquid storage and cryopreservation of boar spermatozoa on acrosomal integrity and the penetration of zona-free hamster ova in vitro. Gamete. Res. 16:193-204 crossref(new window)

Comizzoli, P., R. Mauget and P. Mermillod. 2001. Assessment of in vitro fertility or deer spermatozoa by heterologous IVF with zona-free bovine oocytes. Theriogenol. 56:261-274 crossref(new window)

Cook, W. H. and W. G. Martin. 1969. Egg lipoproteins. In: (Ed. E. Tria and A. M. Scanu). Structural and functional aspects of lipoproteins in living systems. New York: Academic Press. 579-615

Correa, J. R. and P. M. Zavos. 1994. The Hypoosmotic swelling test: it's employment as an assay to evaluate the functional integrity of the frozen-thawed bovine sperm membrane. Theriogenol. 42:351-360 crossref(new window)

Cotirtens, J. L. and J. M. Rety. 2001. Numerical simulation for freezing and thawing mammalian spermatozoa Evaluation of cell injuries at different depth in bags or straws during all steps of the technique. Genet. Sel. Evol. 33(Suppl l):85-104

Demianowicz, W. and J. Strezek. 1996. The effect of lipoprotein fraction of egg yolk on some of the biological properties of boar spermatozoa during storage of the semen in liquid state. Reprod. Dom. Anim. 31:279-280 crossref(new window)

Evans, R. C., D. H. Bauer, S. L. Bandemer, S. B. Vaghelf and C. J. Flegal. 1973. Structure of egg yolk very low density lipoprotein. Polydispersity of the very low density lipoprotein and the role of lipovitellenin in the structure. Arch. Biochem. Biophys.154:493-500 crossref(new window)

Fazeli, A., W. J. Hage, F. P. Cheng, W. F. Voorhout, A. Marks, M. M. Bevers and B. Colenbrander. 1997. Acrosome-intact boar spermatozoa initiate binding to the homologous zona pellucida in vitro. Biol. Reprod. 56:430-438 crossref(new window)

Garcia-Lopez, Z., M. Ollero, T. Muino-Blanco and J. A. Cebrian- Perez. 1996. A dextran swim-up procedure for separation of highly motile and viable ram spermatozoa from semen plasma. Theriogenol. 46:141-151 crossref(new window)

Gillmore, J. A., J. Liu, A. T. Peter and J. K. Critser. 1998. Determination of plasma membrane characteristics of boar spermatozoa and their relevance to cryopreservation. Biol. Reprod. 58:28-36 crossref(new window)

Graham, J. K. and R. H. Foote. 1987. Effect of several lipids fatty acyl chain length and degree of unsaturation on the motility of bull spermatozoa after cold shock and freezing. Cryobiol. 24:42-52 crossref(new window)

Hammadeh, M. E., T. Georg, P. Rosenbaum and W. Shmidt. 2001. Association between freezing agent and acrosome damage of human spermatozoa from subnormal and normal semen. Andrologia. 33:331-336 crossref(new window)

Hossain, A. M., B. Rizk, S. Barik, C. Huff, I. H. Thorneycroft. 1998. Time course of hypo-osmotic swellings of human spermatozoa: evidence of ordered transition between swelling subtypes. Hum. Reprod.13:1578-1583 crossref(new window)

Jeyendran, R. S., Van, Der, Ven, H. H., M. Perez-Pelaez, B. G. Grabo and L. J. D. Zaneveld. 1984. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J. Reprod. Fertil. 70:219-225

Kumar, A., J. Singh, A. S. Nanda and G. R. Pangaonkar. 2004. Quality and fertility of post thaw sephadex filtered bull semen. Asian-Aust. J. Anim. Sci. 17(6):755-759

Lamia Amirata, Daniel Tainturiera, Laetitia Jeanneaua, Chantal Thorina, Olivier Gerard, Jean Luc Courtensc and Marc Antond. 2004. Bull semen in vitro fertility after cryopreservation using egg yolk LDL: a comparison with optidyl, a commercial egg yolk extender. Theriogenol. 61:895-907 crossref(new window)

Larson, J. L. and D. J. Miller. 1999. Simple histochemical stain for acrosomes on sperm from several species. Mol. Reprod. Dev. 52:445-449 crossref(new window)

Mladenovic, I., S. Micic and O. Genbacev. 1995. Hypo-osmotic swelling test for quality control of spermatozoa prepared for assisted reproduction. Arch. Androl. 34:163-169 crossref(new window)

Moussa, M., V. Martinet, A. Trimeche, D. Tainturier and M. Anton. 2002. Low density lipoproteins extracted from hen egg yolk by an easy method: cryoprotective effect on frozen-thawed bull semen. Theriogenol. 57:1695-1706 crossref(new window)

Obando, H., T. Tamayo and C. Alvaro. 1984. Evaluation of some factors affecting swine spermatozoa during freezing. In: Proceedings of the Tenth International Congresson Animal Reproduction AI. Urbana. 2:193

Pace, M. M. and E. F. Graham. 1974. Components in egg yolk which protect bovine spermatozoa during freezing. J. Anim. Sci. 39:1144-1149

Polge, C., S. Salamon and I. Wilmut. 1970. Fertilizing capacity of frozen boar semen following surgical insemination. Vet. Rec. 87:424-428

Revell, S. G. and R. A. Mrode. 1994. An osmotic resistance test for bovine semen. Anim. Reprod. Sci. 36:77-86 crossref(new window)

Rodriguez-Martinez, H., B. Larsson, B. R. Zhang and L. Soderquist. 1997. In vitro assessment of viability and fertilizing capacity of bull spermatozoa. J. Reprod. Dev. 43:1- 11

Rota, A., N. Penzo, L. Vincenti and R. Mantovani. 2000. Hypoosmotic swelling (HOS) as a screening assay for testing in vitro fertility of bovine spermatozoa. Theriogenol. 53:1415- 1420 crossref(new window)

Roth, T. L., R. B. Weiss, J. L. Buff, D. E. Wildt and L. M. Bush. 1998. Heterologous in vitro fertilization and sperm capacitation in an endangered African antelope the scimitarhorned oryx (Oryx dammah). Biol. Reprod. 58:475-482 crossref(new window)

Stalhammar, E. M., L. Janson and J. Philipsson. 1994. The impact of sperm motility on non return rate in preselected dairy bulls. Reprod. Nutr. Dev. 34(l):37-45 crossref(new window)

Trimeche, A., M. Anton, B. Renard, G. Gandemer and D. Tainturier. 1996. Quail egg yolk: A novel cryoprotectant for the freeze preservation of Poitou jackass sperm. Cryobiol. 34:385- 393 crossref(new window)

Tsutsui, T. 1988. Functional properties of heat-treated egg yolk low density lipoprotein. J. Food Sci. 53:1103-1106 crossref(new window)

Vazquez, J. M., E. Martinez, P. Garcia-Artigac and J. Roca. 1997. Hypoosmotlc swelling of boar spermatozoa compared to other methods for analyzing the sperm membrane. Theriogenol. 47:913-922 crossref(new window)

Vazquez, J. M., E. Martinez, J. Roca, P. Coy and L. M. Pastor. 1993. Acrosome reaction of boar spermatozoa in homologous in vitro fertilization. Mol. Reprod. Dev. 36:84-88 crossref(new window)

Visser, D. and S. Salamon. 1974. Effect of composition of trisbased diluent on stirvival of boar spermatozoa following deepfreezing. Aust. J. Biol. Sci. 27:485-497

Watson, P. F. and C. A. Martin. 1975. The influence of some fractions of egg yolk on the survival of ram spermatozoa at 5$^{\circ}C$. Aust. J. Biol. Sci. 28:145-152

Yanagimachi, R. 1994. Mammalian fertilization. In: (Ed. E. Knobil and J. D. Neil). The physiology of reproduction. New York: Raven Press. 189-317

Yi, Y. J., H. J. Ko, S. H. Lee, C. B. Yang, D. S. Son, H. K. Kim and C. S. Park. 2004a. In vitro fertilization and development of pig oocytes inseminated with boar sperm by different sperm washing media after thawing of the frozen straws. Asian-Aust. J. Anim. Sci. 17(2):164-167

Yi, Y. J., S. H. Lee and C. S. Park. 2004b. Comparison of semen characteristics, frozen-thawed sperm viability, testosterone concentration and embryo development between Yorkshire boar A and B. Asian-Aust. J. Anim. Sci. 17(5):612-616

Zhou, J. B., K. Z. Yue, M. J. Luo, Z. L. Chang, H. Liang, Z. Y. Wang and J. H. Tan. 2004. Effect of extenders and temperatures on sperm viability and fertilizing capacity of Harbin white boar semen during long-term liquid storage. Asian-Aust. J. Anim. Sci. 17(11):1501-1508