Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2019-0130
Animal Reproduction (AR)
Original Article

Improvement of in vitro fertilization by a tannin rich vegetal extract addition to frozen thawed boar sperm

Giovanna Galeati; Diego Bucci; Chiara Nerozzi; Beatrice Gadani; Carlo Tamanini; Beatrice Mislei; Marcella Spinaci

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Abstract

Abstract: Boar spermatozoa are very susceptible to cryopreservation injuries and, for this reason, pig remains one of the few species in which fresh semen is still preferred to thawed one for routine artificial insemination (AI). The present work evaluated the effect of supplementing boar sperm thawing medium with Silvafeed SP (SSP), a mixture of Chestnut and Quebracho wood extracts (60/40 w/w) rich in polyphenols (92.4% tannin content) on in vitro fertilization (IVF) and on the following sperm parameters: sperm motility (assessed by CASA), viability, acrosome integrity, mitochondrial function and lipid peroxidation (assessed by flow cytometry) and capacitation status (immunolocalization of tyrosine phosphorylated proteins). Thawed spermatozoa were incubated 1 h at 37°C in BTS without (CTR) or with (5, 10, 20 µg/mL) SSP. After incubation sperm suspension was divided in three aliquots: one was used for IVF trials, one for sperm analysis, and the last one was capacitated for 1 h at 39°C 5% CO2 in IVF medium. Sperm motility parameters, viability, acrosome integrity, mitochondrial functionality, lipid peroxidation and tyrosine phosphorylated protein immunolocalization, used as capacitation parameter, were not influenced by SSP. However, oocytes inseminated with thawed spermatozoa pretreated with all the different SSP concentrations presented a significant (P < 0.01) increase in penetration rate compared to CTR. In addition, 5 µg/mL SSP exerted a positive effect (P<0.05) on the total efficiency of fertilization. These results encourage the use of SSP in the thawing medium since post-thawing fertility is a limit for the large-scale use of boar frozen semen.

Keywords

pig, frozen semen, polyphenols, fertilization

References

Bailey JL, Lessard C, Jacques J, Brèque C, Dobrinski I, Zeng W, Galantino-Homer HL. Cryopreservation of boar semen and its future importance to the industry. Theriogenology. 2008;70(8):1251-9. http://dx.doi.org/10.1016/j.theriogenology.2008.06.014. PMid:18653225.

Brackett BG, Oliphant G. Capacitation of rabbit spermatozoa in vitro. Biol Reprod. 1975;12(2):260-74. http://dx.doi.org/10.1095/biolreprod12.2.260. PMid:1122333.

Bucci D, Galeati G, Tamanini C, Vallorani C, Rodriguez-Gil JE, Spinaci M. Effect of sex sorting on CTC staining, actin cytoskeleton and tyrosine phosphorylation in bull and boar spermatozoa. Theriogenology. 2012;77(6):1206-16. http://dx.doi.org/10.1016/j.theriogenology.2011.10.028. PMid:22192401.

Bucci D, Spinaci M, Yeste M, Mislei B, Gadani B, Prieto Martinez N, Love C, Mari G, Tamanini C, Galeati G. Combined effects of resveratrol and epigallocatechin-3-gallate on post thaw boar sperm and IVF parameters. Theriogenology. 2018;117:16-25. http://dx.doi.org/10.1016/j.theriogenology.2018.05.016. PMid:29807254.

Casas I, Sancho S, Briz M, Pinart E, Bussalleu E, Yeste M, Bonet S. Freezability prediction of boar ejaculates assessed by functional sperm parameters and sperm proteins. Theriogenology. 2009;72(7):930-48. http://dx.doi.org/10.1016/j.theriogenology.2009.07.001. PMid:19651432.

Didion BA, Braun GD, Duggan MV. Field fertility of frozen boar semen: a retrospective report comprising over 2600 AI services spanning a four year period. Anim Reprod Sci. 2013;137(3-4):189-96. http://dx.doi.org/10.1016/j.anireprosci.2013.01.001. PMid:23348011.

Funahashi H. Polyspermic penetration in porcine IVM-IVF systems. Reprod Fertil Dev. 2003;15(3):167-77. http://dx.doi.org/10.1071/RD02076. PMid:12921703.

Funahashi H, Cantley T, Day BN. Synchronization of meiosis in porcine oocytes by exposure to dibutyryl cyclic adenosine monophosphate improves developmental competence following in vitro fertilization. Biol Reprod. 1997;57(1):49-53. http://dx.doi.org/10.1095/biolreprod57.1.49. PMid:9209079.

Gadani B, Bucci D, Spinaci M, Tamanini C, Galeati G. Resveratrol and epigallocatechin-3-gallate addition to thawed boar sperm improves in vitro fertilization. Theriogenology. 2017;90:88-93. http://dx.doi.org/10.1016/j.theriogenology.2016.11.020. PMid:28166993.

Giaretta E, Estrada E, Bucci D, Spinaci M, Rodríguez-Gil JE, Yeste M. Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance. Theriogenology. 2015;83(3):399-407. http://dx.doi.org/10.1016/j.theriogenology.2014.10.002. PMid:25459422.

Guthrie HD, Welch GR. Effects of reactive oxygen species on sperm function. Theriogenology. 2012;78(8):1700-8. http://dx.doi.org/10.1016/j.theriogenology.2012.05.002. PMid:22704396.

Johnson LA, Weitze KF, Fiser P, Maxwell WM. Storage of boar semen. Anim Reprod Sci. 2000;62(1-3):143-72. http://dx.doi.org/10.1016/S0378-4320(00)00157-3. PMid:10924823.

Kaeoket K, Chanapiwat P, Tummaruk P, Techakumphu M. Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen. Asian J Androl. 2010;12(5):760-5. http://dx.doi.org/10.1038/aja.2010.48. PMid:20601963.

Knox RV. The fertility of frozen boar sperm when used for artificial insemination. Reprod Domest Anim. 2015;50(Suppl 2):90-7. http://dx.doi.org/10.1111/rda.12552. PMid:26174925.

Knox RV. Artificial insemination in pigs today. Theriogenology. 2016;85(1):83-93. http://dx.doi.org/10.1016/j.theriogenology.2015.07.009. PMid:26253434.

Koleckar V, Kubikova K, Rehakova Z, Kuca K, Jun D, Jahodar L, Opletal L. Condensed and hydrolysable tannins as antioxidants influencing the health. Mini Rev Med Chem. 2008;8(5):436-47. http://dx.doi.org/10.2174/138955708784223486. PMid:18473933.

Küntzel A. Die quantitative Gerbstoffanalyse nach dem Filterverfahren. Das Leder. 1954;2:28-31.

Lee JA, Spidlen J, Boyce K, Cai J, Crosbie N, Dalphin M, Furlong J, Gasparetto M, Goldberg M, Goralczyk EM, Hyun B, Jansen K, Kollmann T, Kong M, Leif R, McWeeney S, Moloshok TD, Moore W, Nolan G, Nolan J, Nikolich-Zugich J, Parrish D, Purcell B, Qian Y, Selvaraj B, Smith C, Tchuvatkina O, Wertheimer A, Wilkinson P, Wilson C, Wood J, Zigon R, Scheuermann RH, Brinkman RR. International Society for Advancement of Cytometry Data Standards Task Force, Scheuermann RH, Brinkman RR. MIFlowCyt: the Minimum Information about a Flow Cytometry Experiment. Cytometry A. 2008;73(10):926-30. http://dx.doi.org/10.1002/cyto.a.20623. PMid:18752282.

Peña FJ, Saravia F, Núñez-Martínez I, Johannisson A, Wallgren M, Rodriguez Martinez H. Do different portions of the boar ejaculate vary in their ability to sustain cryopreservation? Anim Reprod Sci. 2006;93(1-2):101-13. http://dx.doi.org/10.1016/j.anireprosci.2005.06.028. PMid:16084673.

Petrunkina AM, Waberski D, Bollwein H, Sieme H. Identifying non-sperm particles during flow cytometric physiological assessment: a simple approach. Theriogenology. 2010;73(7):995-1000. http://dx.doi.org/10.1016/j.theriogenology.2009.12.006. PMid:20171719.

Petters RM, Wells KD. Culture of pig embryos. J Reprod Fertil Suppl. 1993;48:61-73. PMid:8145215.

Roca J, Gil MA, Hernandez M, Parrilla I, Vazquez JM, Martinez EA. Survival and fertility of boar spermatozoa after freeze-thawing in extender supplemented with butylated hydroxytoluene. J Androl. 2004;25(3):397-405. http://dx.doi.org/10.1002/j.1939-4640.2004.tb02806.x. PMid:15064318.

Rooke JA, Shao CC, Speake BK. Effects of feeding tuna oil on the lipid composition of pig spermatozoa and in vitro characteristics of semen. Reproduction. 2001;121(2):315-22. http://dx.doi.org/10.1530/rep.0.1210315. PMid:11226056.

Spinaci M, Muccilli V, Bucci D, Cardullo N, Gadani B, Tringali C, Tamanini C, Galeati G. Biological effects of polyphenol-rich extract and fractions from an oenological oak-derived tannin on in vitro swine sperm capacitation and fertilizing ability. Theriogenology. 2018;108:284-90. http://dx.doi.org/10.1016/j.theriogenology.2017.12.015. PMid:29277068.

Tokeshi I, Yoshimoto T, Muto N, Nakamura S, Ashizawa K, Nakada T, Tatemoto H. Antihyaluronidase action of ellagic acid effectively prevents polyspermy as a result of suppression of the acrosome reaction induced by sperm-zona interaction during in vitro fertilization of porcine oocytes. J Reprod Dev. 2007;53(4):755-64. http://dx.doi.org/10.1262/jrd.18173. PMid:17389774.

Weng XG, Cai MM, Zhang YT, Liu Y, Gao ZL, Song J, Liu ZH. Effect of Astragalus polysaccharide addition to thawed boar sperm on in vitro fertilization and embryo development. Theriogenology. 2018;121:21-6. http://dx.doi.org/10.1016/j.theriogenology.2018.07.030. PMid:30125824.

White IG. Lipids and calcium-uptake of sperm in relation to cold shock and preservation- a review. Reprod Fertil Dev. 1993;5(6):639-58. http://dx.doi.org/10.1071/RD9930639. PMid:9627725.

Yeste M. Recent advances in boar sperm crypreservation: state of the art and current perspectives. Reprod Domest Anim. 2015;70(Suppl 2):51-9.

Yeste M. Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs. Theriogenology. 2016;85(1):47-64. http://dx.doi.org/10.1016/j.theriogenology.2015.09.047. PMid:26506124.

Yeste M, Rodríguez-Gil IE, Bonet S. Artificial insemination with frozen-thawed boar sperm. Mol Reprod Dev. 2017;84(9):802-13. http://dx.doi.org/10.1002/mrd.22840. PMid:28608609.
 

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