##plugins.themes.bootstrap3.article.main##

Majid Alaeipour Mohammad Roostaei-Ali Mehr

Abstract

This study was conducted to evaluate the effect of replacing glycerol with erythritol on cryopreservation of ram spermatozoa. Semen samples (n=24) were collected from four rams in six times. In each session, the collected ejaculates (n=4) were pooled and split into 12 equal parts. The amount of 0.032 M glycerol (G32E0, equal to 3% glycerol), 0.016 M glycerol and 0.016 M erythritol (G16E16), 0.008 M glycerol and 0.024 M erythritol (G8E24), 0.032 M erythritol (G0E32), 0.054 M glycerol (G54E0, equal to 5% glycerol), 0.027 M glycerol and 0.027 M erythritol (G27E27), 0.013 M glycerol and 0.041 M erythritol (G13E41), 0.054 M erythritol (G0E54), 0.076 M glycerol (G76E0, equal to 7% glycerol ), 0.038 M glycerol and 0.038 M erythritol (G38E38), 0.019 M glycerol and 0.057 M erythritol (G19E57) and 0.076 M erythritol (G0E76) were added. The diluted samples were frozen using standard protocol. After thawing, the samples were incubated at 37°C for 6 h. Results showed that progressive sperm motility and acrosome integrity were higher in G13E41 (18.85 % and 27.41 %, respectively) than treatments that contained only glycerol at 6 h (p < 0.05). At the level of 0.032 and 0.054 M cryoprotectant, the highest of total sperm motility was observed in G8E24 (19.16 %) and G13E41 (18.85 %) at 6 h, respectively (p < 0.05). Therefore, the quality of frozen-thawed ram spermatozoa can be improved by using the mixture of 0.013 M glycerol plus 0.041 M erythritol or 0.008 M glycerol plus 0.024 M erythritol.

Article Details

Keywords

Cryopreservation, Polyol, Glycerol, Ram sperm

References
1. Sánchez-Partida LG, Windsor DP, Eppleston J, Setchell BP, Maxwell WM. Fertility and its relationship to motility characteristics of spermatozoa in ewes after cervical, transcervical, and intrauterine insemination with frozen-thawed ram semen. J. Androl. 1999; 20(2): 280–288.
2. Roostaei-Ali Mehr M, Parisoush P. Effect of different levels of silymarin and caproic acid on storage of ram semen in liquid form. Reprod. Domest. Anim. 2016; 51(4): 569–574.
3. Salamon S, Maxwell WMC. Storage of ram semen. Anim. Reprod. Sci. 2000; 62(1-3): 77–111.
4. Abdelhakeam AA, Graham EF, Vazquez IA, Chaloner KM. Studies on the absence of glycerol in unfrozen and frozen ram semen. Development of an extender for freezing: effects of osmotic pressure, egg yolk levels, type of sugars and the method of dilution. Cryobiology 1991; 28(1): 43–49.
5. Abdelhakeam AA, Graham EF, Vazquez IA. Studies on the absence of glycerol in unfrozen and frozen ram semen: Fertility trials and the effect of dilution methods on freezing ram semen in the absence of glycerol. Cryobiology 1991; 28(1): 36–42.
6. Slavik T. Effect of glycerol on the penetrating ability of fresh ram spermatozoa with zona free hamster eggs. J. Reprod. Fertil. 1987; 79(1): 99–103.
7. Colas G. Effect of initial freezing temperature, addition of glycerol and dilution of the survival and fertilizing ability of deep-frozen ram semen. J. Reprod. Fertil. 1975; 42(2): 277–285.
8. Morrier A, Castonguay F, Bailey J. Glycerol addition and conservation of fresh and cryopreserved ram spermatozoa. Can. J. Anim. Sci. 2002; 82(3): 347–356.
9. Utsumi K, Hochi S, Iritani A. Cryoprotective effect of polyols rat embryos during two-step freezing. Cryobiology 1992; 29(3): 332–41.
10. Til HP, Juper CF, Faile HE, Bruyntjes JP, Bar A. Subchronic feeding studies with erythritol in rats and mice. Regul. Toxicol. Pharm. 1996; 24(2): 221–231
11. Bernet WO, Borzelleca JF, Flamm G, Munro IC. Erythritol: a review of biological and toxicological studies. Regul. Toxicol. Pharm. 1996; 24(2): 191–197.
12. Maxwell WMC, Watson PF. Recent progress in the preservation of ram semen. Anim. Reprod. Sci. 1996; 42(1-4): 55–65.
13. Lovelock JE. The protective action of neutral solutes against haemolysis by freezing and thawing. Biochem. J. 1954; 56(2): 265–270.
14. Miyamoto H, Ishibashi T. Survival of mouse embryos after freezing and thawing in the presence of erythritol. J. Exp. Zool. B. Mol. Dev. Evol. 1981; 216(2): 337–40.
15. Molinia FC, Evans G, Maxwell WMC. Effect of polyols on the post-thawing motility of pellet-frozen ram spermatozoa. Theriogenology 1994; 42(1): 15–23.
16. Kim, SW, Park, CH, Kim, HJ, Yoon, JH, Hwang, YJ, Kim, D. Antioxidant effect of Erythritol on boar spermatozoa during cryopreservation. Dev. Biol. 2011; 356(1): 195.
17. Willadsen SM, Polge CL, Rowson E, Rowson A, Moor RM. Deep freezing of sheep embryos. J. Reprod. Fertil. 1976; 46(1): 151–154.
18. Drevius LO. Permeability coefficients of bull spermatozoa for water and polyhydric alcohols. Experi. Cell Res. 1971; 69(1): 212–6.
19. Chung YS, Lee M. Genotoxicity assessment of erythritol by using short-term assay. Toxi. Res. 2013; 29(4): 249–255.
20. Szende B, Tyihák E. Effect of formaldehyde on cell proliferation and death. Cell Biol. Int. 2010; 34(12): 1273–1282.
21. De Leeuw FE, De Leeuw AM, Den Daas JH, Colenbrander B, Verkleij AJ. Effects of various cryoprotective agents and membrane-stabilizing compounds on bull sperm membrane integrity after cooling and freezing. Cryobiology 1993; 30(1): 32–44.
22. Daily WA, Higgens CE. Preservation and storage of microorganisms in the gas phase of liquid nitrogen. Cryobiology 1973; 10 (5): 364–367.
23. Motamedi-Mojdehi R, Roostaei-Ali Mehr M, Rajabi-Toustani R. Effect of different levels of glycerol and cholesterol-loaded cyclodextrin on cryosurvival of ram spermatozoa. Reprod. Domest. Anim. 2014; 49(1): 65–70.
24. Redway KF, Lapage SP. Effect of carbohydrates and related compounds on the long–term preservation of freez-dried Bacteria. Cryobiology 1974; 11(1): 73–79.
25. Neild DM, Gadella BM, Chaves MG, Miragaya MH, Colenbrander B, Aguero A. Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation. Theriogenology 2003; 59(8): 1693–1705.
26. Soylu MK, Nur Z, Ustuner B, Dogan I, Sagirkaya H, Gunay U, Ak K. Effects of various cryoprotective agents and extender osmolality on post-thaw ram semen. B. Vet. I. Pulawy 2007; 51: 241–6.
27. Rashba-step J, Eugene S, Nicholas J, Turro A, Cederbaum I. Oxidation of glycerol to formaldehyde by microsomes: Are glycerol radicals produced in the reaction pathway? Biochemistry 1994; 33(32): 9504–9510.
28. Chi Y, Zhang X, Cai J, Liu FY, Xing GG, Wan Y. Formaldehyde increases intracellular calcium concentration in primary cultured hippocampal neurons partly through NMDA receptors and T-type calcium channels. Neurosci. Bull. 2012; 28(6): 715–722.
29. Gillan L, Evans G, Maxwell WMC. Capacitation status and fertility of frozen-thawed ram spermatozoa. J. Reprod. Fertil. Develop. 1997; 9(5): 481–487.
30. Roostaei-Ali Mehr M, Mousavi M, Ghadamyari M. Effect of seminal plasma proteins on membrane cholesterol efflux of ram epididymal spermatozoa. Small Rumin. Res. 2015; 129: 88–91.
How to Cite
AlaeipourM., & Roostaei-Ali MehrM. (2020). The cryoprotective effects of erythritol on frozen-thawed ram sperm. Iranian Journal of Veterinary Science and Technology, 11(2), 1-10. https://doi.org/10.22067/veterinary.v11i2.82921
Section
Original Articles