Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2022-0010
Animal Reproduction (AR)
ORIGINAL ARTICLE

Effect of different concentrations of Trolox® in association with docosahexaenoic acid on equine semen freezing

Cristiane Silva Aguiar; Celso Henrique Souza Costa Barros; William Morais Machado; Ivan Bezerra Allaman; Antônio de Oliveira Leite Filho; Larissa Pires Barbosa; Paola Pereira das Neves Snoeck

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Abstract

The aim of this study was to evaluate the association of different concentrations of Trolox® and the addition of a fixed concentration of DHA in the freezing of semen of Mangalarga Marchador stallions. To that end, 16 ejaculates were frozen in the following extenders: E1) BotuCrio® (BC; Control); E2) BC + 50 ngml-1 DHA + 30 µM Trolox® (BCDHA30T); E3) BC + 50 ngml-1 DHA + 40 µM Trolox® (BCDHA40T); E4) BC + 50 ngml-1 DHA + 50 µM Trolox® (BCDHA50T). All the tested extenders were similar in preserving different kinematic parameters, cell functional integrity, compacted DNA, and high and intermediate mitochondrial activity (P>0.05). However, sperm cryopreserved in BCDHA40T showed higher velocities than sperm frozen in the control extender (P<0.05). The 30 µM concentration of Trolox® was worse for sperm motility and the 50 µM concentration of Trolox® did not adequately preserve the structural integrity of the membranes in an extender containing DHA when compared to the BotuCrio® (P<0.05) extender. The use of Trolox® in freezing extenders containing DHA did not maximize the effect of BotuCrio®, except for in the case of sperm velocity parameters when at a concentration of 40 µM.

Keywords

antioxidant, fatty acids, lipid peroxidation, cryopreservation, stallion

References

Agarwal A, Said TM. Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update. 2003;9(4):331-45. http://dx.doi.org/10.1093/humupd/dmg027. PMid:12926527.

Aguiar CS, Barros CHSC, Machado WM, Allaman IB, Barbosa LP, Snoeck PPN. Efeito do ácido docosa-hexaenoico e do Trolox® no diluidor de refrigeração de sêmen de garanhões da raça Mangalarga Marchador. Arq Bras Med Vet Zootec. 2020;72(1):71-8. http://dx.doi.org/10.1590/1678-4162-10823.

Albertini R, Abuja PM. Pro-oxidant and antioxidant properties of Trolox C, analogue of vitamin E, in oxidation of low-density lipoprotein. Free Radic Res. 1999;30(3):181-8. http://dx.doi.org/10.1080/10715769900300201. PMid:10711788.

Alvarenga MA, Papa FO, Landim-Alvarenga FC, Medeiros ASL. Amides as cryoprotectants for freezing stallion semen: a review. Anim Reprod Sci. 2005;89(1-4):105-13. http://dx.doi.org/10.1016/j.anireprosci.2005.07.001. PMid:16099609.

Ball BA, Medina V, Gravance CG, Baumber J. Effect of antioxidants on preservation of motility,viability and acrosomal integrity of equine spermatozoa during storage at 5°C. Theriogenology. 2001;56(4):577-89. http://dx.doi.org/10.1016/S0093-691X(01)00590-8. PMid:11572439.

Bolle P, Evandri MG, Saso L. The controversial efficacy of vitamin E for human male infertility. Contraception. 2002;65(4):313-5. http://dx.doi.org/10.1016/S0010-7824(02)00277-9. PMid:12020785.

Breininger E, Beorlegui NB, O’Flaherty CM, Beconi MT. Alpha-tocopherol improves biochemical and dynamic parameters in cryopreserved boar semen. Theriogenology. 2005;63(8):2126-35. http://dx.doi.org/10.1016/j.theriogenology.2004.08.016. PMid:15826678.

Brigelius-Flohé R, Traber MG. Vitamin E: function and metabolism. FASEB J. 1999;13(10):1145-55. http://dx.doi.org/10.1096/fasebj.13.10.1145. PMid:10385606.

Brinsko SP, Varner DD, Love CC, Blanchard TL, Day BC, Wilson ME. Effect of feeding a DHA-enriched nutriceutical on the quality of fresh, cooled and frozen stallion semen. Theriogenology. 2005;63(5):1519-27. http://dx.doi.org/10.1016/j.theriogenology.2004.07.010. PMid:15725455.

Buege JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol. 1978;52:302-10. http://dx.doi.org/10.1016/S0076-6879(78)52032-6. PMid:672633.

Burton GW, Joyce A, Ingold KU. Is vitamin E the only lipid-soluble, chain-breaking antioxidant in human blood plasma and erythrocyte membranes? Arch Biochem Biophys. 1983;221(1):281-90. http://dx.doi.org/10.1016/0003-9861(83)90145-5. PMid:6830261.

Cao G, Alessio HM, Cutler RG. Oxygen-radical absorbance capacity assay for antioxidants. Free Radic Biol Med. 1993;14(3):303-11. http://dx.doi.org/10.1016/0891-5849(93)90027-R. PMid:8458588.

Cao G, Cutler RG. High concentrations of antioxidants may not improve defense against oxidative stress. Arch Gerontol Geriatr. 1993;17(3):189-201. http://dx.doi.org/10.1016/0167-4943(93)90050-R. PMid:15374318.

Colégio Brasileiro de Reprodução Animal – CBRA. Manual for andrological examination and semen evaluation of animal semen. 3rd ed. Belo Horizonte: CBRA; 2013.

Connor WE, Lin DS, Wolf DP, Alexander M. Uneven distribution of desmosterol and docosahexaenoic acid in the heads and tails of monkey sperm. J Lipid Res. 1998;39(7):1404-11. http://dx.doi.org/10.1016/S0022-2275(20)32521-9. PMid:9684743.

Costa JMS, Souza WL, Moraes EA, Lima DIB, Magalhães LMV, Coelho VG. Efeito da adição dos antioxidantes trolox c e ácido ascórbico no sêmen fresco de carneiros sobre a atividade mitocondrial de espermatozoides descongelados. Ciênc Vet Tróp. 2015;18:333-6.

Domosławska A, Zdunczyk S, Franczyk M, Kankofer M, Janowski T. Selenium and vitamin E supplementation enhances the antioxidant status of spermatozoa and improves semen qualityin male dogs with lowered fertility. Andrologia. 2018;50(6):e13023. http://dx.doi.org/10.1111/and.13023. PMid:29744899.

Evans HC, Dinh TTN, Ugur MR, Hitit M, Sajeev D, Kaya A, Topper E, Nicodemus MC, Smith GD, Memili E. Lipidomic markers of sperm cryotolerance in cattle. Sci Rep. 2020;10(1):20192. http://dx.doi.org/10.1038/s41598-020-77089-9. PMid:33214639.

Evenson D, Jost L. Current Protocols in Cytometry. Hoboken: John Wiley & Sons, Inc.; 2001. Unit 7.13, Chapter 7, Sperm chromatin structure assay for fertility assessment: current protocols in cytometry. http://dx.doi.org/10.1002/0471142956.cy0713s13.

Fraser L. Cryopreservation of stallion semen using different amides.tructural damage to nuclear DNA in mammalian spermatozoa: its evaluation techniques and relationship with male infertility. Pol J Vet Sci. 2004;7:311-21.

García BM, Fernández LG, Ferrusola CO, Rodríguez AM, Bolaños JMG, Martinez HR, Tapia JA, Morcuende D, Peña FJ. Fatty acids and plasmalogens of the phospholipids of the sperm membranes and their relation with the post-thaw quality of stallion spermatozoa. Theriogenology. 2011;75(5):811-8. http://dx.doi.org/10.1016/j.theriogenology.2010.10.021. PMid:21144567.

Gholami H, Chamani M, Towhidi A, Fazeli MH. Effect of feeding a docosahexaenoic acid-enriched nutriceutical on the quality of fresh and frozen-thawed semen in Holstein bulls. Theriogenology. 2010;74(9):1548-58. http://dx.doi.org/10.1016/j.theriogenology.2010.06.025. PMid:20708237.

Gomes GM, Jacob JCF, Medeiros ASL, Papa FO, Alvarenga MA. Improvement of stallion spermatozoa preservation with alternative cryoprotectants for the Mangalarga Marchador breed. Theriogenology. 2002;58(2-4):277-9. http://dx.doi.org/10.1016/S0093-691X(02)00899-3.

Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. Oxford: Oxford University Press; 2015. http://dx.doi.org/10.1093/acprof:oso/9780198717478.001.0001.

Hammerstedt RH, Graham JK, Nolan JP. Cryopreservation of mammalian sperm: what we ask them to survive. J Androl. 1990;11(1):73-88. PMid:2179184.

Harris MA, Baumgard LH, Arns MJ, Webel SK. Stallion spermatozoa membrane phospholipid dynamics following dietary n-3 supplementation. Anim Reprod Sci. 2005;89(1-4):234-7. PMid:16265723.

Harrison RA, Vickers SE. Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. J Reprod Fertil. 1990;88(1):343-52. http://dx.doi.org/10.1530/jrf.0.0880343. PMid:1690300.

Hatamoto LK, Baptista CA So, Nichi M, Barnabe VH, Barnabe RC, Cortada CNM. Effects of dexamethasone treatment (to mimic stress) and vitamin E oral supplementation on the spermiogram and on seminal plasma spontaneous lipid peroxidation and antioxidant enzyme activities in dogs. Theriogenology. 2006;66(6-7):1610-4. http://dx.doi.org/10.1016/j.theriogenology.2006.02.012. PMid:16581116.

Hishikawa D, Valentine WJ, Iizuka-Hishikawa Y, Shindou H, Shimizu T. Metabolism and functions of docosahexaenoic acid-containing membrane glycerophospholipids. FEBS Lett. 2017;591(18):2730-44. http://dx.doi.org/10.1002/1873-3468.12825. PMid:28833063.

Hrudka F. Cytochemical and ultracytochemical demonstration of cytochrome c oxidase in spermatozoa and dynamics of its changes accompanying ageing or induced by stress. Int J Androl. 1987;10(6):809-28. http://dx.doi.org/10.1111/j.1365-2605.1987.tb00385.x. PMid:2828243.

Iizuka-Hishikawa Y, Hishikawa D, Sasaki J, Takubo K, Goto M, Nagata K, Nakanishi H, Shindou H, Okamura T, Ito C, Toshimori K, Sasaki T, Shimizu T. Lysophosphatidic acid acyltransferase 3 tunes the membrane status of germ cells by incorporating docosahexaenoic acid during spermatogenesis. J Biol Chem. 2017;292(29):12065-76. http://dx.doi.org/10.1074/jbc.M117.791277. PMid:28578315.

Jonathan M SC, Wildelfrancys LS, Elenice AM, Dailli I BL, Laiane MVM, Vitória GC. Efeito da adição dos antioxidantes trolox c e ácido ascórbico no sêmen fresco de carneiros sobre a atividade mitocondrial de espermatozoides descongelados. Cienc Vet Trop. 2015;18:333-6.

Kaeoket K, Sang-urai P, Thamniyom A, Chanapiwat P, Techakumphu M. Effect of docosahexaenoic acid on quality of cryopreserved boar semen in different breeds. Reprod Domest Anim. 2010;45(3):458-63. http://dx.doi.org/10.1111/j.1439-0531.2008.01239.x. PMid:19090818.

Kao SH, Chao HT, Chen HW, Hwang TIS, Liao TL, Wei YH. Increase of oxidative stress in human sperm with lower motility. Fertil Steril. 2008;89(5):1183-90. http://dx.doi.org/10.1016/j.fertnstert.2007.05.029. PMid:17669405.

Kasimanickam R, Pelzer KD, Kasimanickam V, Swecker WS, Thatcher CD. Association of classical semen parameters, sperm DNA fragmentation index, lipid peroxidation and antioxidant enzymatic activity of semen in ram-lambs. Theriogenology. 2006;65(7):1407-21. http://dx.doi.org/10.1016/j.theriogenology.2005.05.056. PMid:16188307.

Lenzi A, Gandini L, Maresca V, Rago R, Sgro P, Dondero F, Picardo M. Fatty acid composition of spermatozoa and immature germ cells. Mol Hum Reprod. 2000;6(3):226-31. http://dx.doi.org/10.1093/molehr/6.3.226. PMid:10694269.

Maia MS, Bicudo SD. Radicais livres, antioxidantes e função espermática em mamíferos: uma revisão. Rev Bras Reprod Anim. 2009;33:183-93.

Martínez-Soto JC, Landeras J, Gadea J. Spermatozoa and seminal plasma fatty acids as predictors of cryopreservation success. Andrology. 2013;1(3):365-75. http://dx.doi.org/10.1111/j.2047-2927.2012.00040.x. PMid:23596043.

McCarthy MJ, Meyers SA. Antioxidant treatment in the absence of exogenous lipids and proteins protects rhesus macaque sperm from cryopreservation- induced cell membrane damage. Theriogenology. 2011;76(1):168-76. http://dx.doi.org/10.1016/j.theriogenology.2011.01.029. PMid:21458048.

Melo MIV, Henry M. Teste hiposmótico na avaliação do sêmen equino. Arq Bras Med Vet Zootec. 1999;51:71-8.

Mitre R, Cheminade C, Allaume P, Legrand P, Legrand AB. Oral intake of shark liver oil modifies lipid composition and improves motility and velocity of boar sperm. Theriogenology. 2004;62(8):1557-66. http://dx.doi.org/10.1016/j.theriogenology.2004.02.004. PMid:15451263.

Murphy C, English AM, Holden SA, Fair S. Cholesterol-loaded-cyclodextrins improve the post-thaw quality of stallion sperm. Anim Reprod Sci. 2014;145(3-4):123-9. http://dx.doi.org/10.1016/j.anireprosci.2014.01.013. PMid:24583046.

Nasiri AH, Towhidi A, Zeinoaldini S. Combined effect of DHA and alpha-tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition. Andrologia. 2012;44(Suppl 1):550-5. http://dx.doi.org/10.1111/j.1439-0272.2011.01225.x. PMid:21951061.

Naves CS, Beletti ME, Duarte MB, Vieira RC. Avaliação da cromatina espermática em equinos com azul de toluidina e acridine orange. Biosci J. 2004;20:117-24.

Nguyen LN, Ma D, Shui G, Wong P, Cazenave-Gassiot A, Zhang X, Wenk MR, Goh ELK, Silver DL. Mfsd2a is a transporter for the essential omega-3 fatty acid docosahexaenoic acid. Nature. 2014;509(7501):503-6. http://dx.doi.org/10.1038/nature13241. PMid:24828044.

Nichi M. Efeito do tratamento com antioxidantes e ácidos graxos poli-insaturados em amostras espermáticas epididimárias de touros [dissertation]. São Paulo: Universidade de São Paulo; 2009. Portuguese.

Nichi M. Sistemas de proteção enzimática e níveis de peroxidação espontânea dos lipídios seminais de touros zebuínos e taurinos criados a campo na região de Dourados, MS [thesis]. São Paulo: Universidade de São Paulo; 2003. Portuguese.

Peña F, Martínez HR, Tapia J, Ferrusola C, Fernández LG, García BM. Mitochondria in mammalian sperm physiology and pathology: a review. Reprod Domest Anim. 2009;44(2):345-9. http://dx.doi.org/10.1111/j.1439-0531.2008.01211.x. PMid:19144010.

Rice DS, Calandria JM, Gordon WC, Jun B, Zhou Y, Gelfman CM, Li S, Jin M, Knott EJ, Chang B, Abuin A, Issa T, Potter D, Platt KA, Bazan NG. Adiponectin receptor 1 conserves docosahexaenoic acid and promotes photoreceptor cell survival. Nat Commun. 2015;6(1):6228. http://dx.doi.org/10.1038/ncomms7228. PMid:25736573.

Roqueta-Rivera M, Stroud CK, Haschek WM, Akare SJ, Segre M, Brush RS, Agbaga MP, Anderson RE, Hess RA, Nakamura MT. Docosahexaenoic acid supplementation fully restores fertility and spermatogenesis in male delta-6 desaturase-null mice. J Lipid Res. 2010;51(2):360-7. http://dx.doi.org/10.1194/jlr.M001180. PMid:19690334.

Sabeti P, Pourmasumi S, Rahiminia T, Akyash F, Talebi AR. Etiologies of sperm oxidative stress. Int J Reprod Biomed. 2016;14(4):231-40. http://dx.doi.org/10.29252/ijrm.14.4.231. PMid:27351024.

Safarinejad MR, Hosseini SY, Dadkhah F, Asgari MA. Relationship of omega-3 and omega-6 fatty acids with semen characteristics, and anti-oxidant status of seminal plasma: a comparison between fertile and infertile men. Clin Nutr. 2010;29(1):100-5. http://dx.doi.org/10.1016/j.clnu.2009.07.008. PMid:19666200.

Sarlós P, Molnár A, Kókai M, Gábor G, Rátky J. Comparative evaluation of the effect of antioxidants in the conservation of ram semen. Acta Vet Hung. 2002;50(2):235-45. http://dx.doi.org/10.1556/avet.50.2002.2.13. PMid:12113179.

Shindou H, Koso H, Sasaki J, Nakanishi H, Sagara H, Nakagawa KM, Takahashi Y, Hishikawa D, Iizuka-Hishikawa Y, Tokumasu F, Noguchi H, Watanabe S, Sasaki T, Shimizu T. Docosahexaenoic acid preserves visual function by maintaining correct disc morphology in retinal photoreceptor cells. J Biol Chem. 2017;292(29):12054-64. http://dx.doi.org/10.1074/jbc.M117.790568. PMid:28578316.

Sicherle CC, Maia MS, Bicudo SD, Green RE, Souza DB, Azevedo HC. The effect of Trolox addition to egg-yolk–Tris extender on the motility and membrane integrity of frozen-thawed ram spermatozoa. Nottingham: Nottingham niversity Press; 2006. Reproduction in domestic ruminants VI.

Sicherle CC, Maia MS, Bicudo SD, Rodello L, Azevedo HC. Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen supplemented with catalase or Trolox. Small Rumin Res. 2011;95(2-3):144-9. http://dx.doi.org/10.1016/j.smallrumres.2010.10.011.

Sieme H, Oldenhof H, Wolkers WF. Mode of action of cryoprotectants for sperm preservation. Anim Reprod Sci. 2016;169:2-5. http://dx.doi.org/10.1016/j.anireprosci.2016.02.004. PMid:26936658.

Sikka SC. Role of oxidative stress and antioxidants in andrology and assisted reproductive technology. J Androl. 2004;25(1):5-18. http://dx.doi.org/10.1002/j.1939-4640.2004.tb02751.x. PMid:14662779.

Silva KMG, Gamboa SC, Rodrigues AS, Santos JR, Guerra MMP. Adição de piruvato de sódio e trolox ao diluidor utilizado para congelação de sêmen de garanhões férteis e subférteis. Cienc Rural. 2008;38(8):2271-7. http://dx.doi.org/10.1590/S0103-84782008000800028.

The R Project for Statistical Computing [software]. Vienna: The R Project for Statistical Computing; 2016 [cited 2022 Oct 27]. Available from: https://www.R-project.org/.

Towhidi A, Parks JE. Effect of n-3 fatty acids and α-tocopherol on post-thaw parameters and fatty acid composition of bovine sperm. J Assist Reprod Genet. 2012;29(10):1051-6. http://dx.doi.org/10.1007/s10815-012-9834-7. PMid:22869241.

Towhidi A, Zeinoaldini S, Ardebili R, Davachi ND, Nasiri AH. Combined n-3 fatty acids and α-Tocopherol supplementation improved the ovine sperm cryosurvival. Iran J Biotechnol. 2013;11(4):238-43. http://dx.doi.org/10.5812/ijb.14469.

Upreti GC, Jensen K, Oliver JE, Duganzich DM, Munday R, Smith JF. Motility of ram spermatozoa during storage in a chemically-defined diluent containing antioxidants. Anim Reprod Sci. 1997;48(2-4):269-78. http://dx.doi.org/10.1016/S0378-4320(97)00054-7. PMid:9452879.

Venables WN, Ripley BD. Modern applied statistics with S. Berlin: Springer; 2002. http://dx.doi.org/10.1007/978-0-387-21706-2.

Verstegen J, Iguer-Ouada M, Onclin K. Computer assisted semen analyzers in andrology research and veterinary practice. Theriogenology. 2002;57(1):149-79. http://dx.doi.org/10.1016/S0093-691X(01)00664-1. PMid:11775967.

Wassall SR, Stillwell W. Polyunsaturated fatty acid-cholesterol interactions: domain formation in membranes. Biochim Biophys Acta. 2009;1788(1):24-32. http://dx.doi.org/10.1016/j.bbamem.2008.10.011. PMid:19014904.

Waterhouse KE, Hofmo PO, Tverdal A, Miller RR Jr. Within and between breed differences in freezing tolerance and plasma membrane fatty acid composition of boar sperm. Reproduction. 2006;131(5):887-94. http://dx.doi.org/10.1530/rep.1.01049. PMid:16672353.

Wood PL, Scoggin K, Ball BA, Troedsson MH, Squires EL. Lipidomics of equine sperm and seminal plasma: identification of amphiphilic (O-acyl)-ω-hydroxy-fatty acids. Theriogenology. 2016;86(5):1212-21. http://dx.doi.org/10.1016/j.theriogenology.2016.04.012. PMid:27180330.

Wu T-W, Hashimoto N, Au J-X, Wu J, Mickle DAG, Carey D. Trolox protects rat hepatocytes against oxyradical damage and the ischemic rat liver from reperfusion injury. Hepatology. 1991;13(3):575-80. http://dx.doi.org/10.1002/hep.1840130328. PMid:1999327.
 


Submitted date:
01/21/2022

Accepted date:
10/27/2022

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