The cryoprotective effects of erythritol on frozen-thawed ram sperm

Document Type : Research Articles

Authors

Department of Animal Science, Faculty of Agricultural Sciences, University of Guilan, Rasht, Iran

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.

Keywords

Main Subjects


1. Sanchez-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, Tyihak 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.
CAPTCHA Image