Alpha-tocopherol improves frozen-thawed sperm quality by reducing hydrogen peroxide during cryopreservation of bull semen
Abstract
This study was conducted to investigate the effects of different levels of α-tocopherol in cryopreservation media on the bull sperm quality after thawing. Semen samples were collected from six Holstein bulls using artificial vagina twice a week. Semen samples were pooled to eliminate individual differences and then divided into four equal parts for freezing with extenders containing different concentrations of α-tocopherol according to experimental groups as follows: 0 (control), 1.2 mM (E1), 2.4 mM (E2) and 4.8 mM (E4). Motion characteristics, viability, plasma membrane functionality, lipid peroxidation and H2O2 status were determined after thawing. Results showed that malondialdehyde (MDA) concentration was significantly lower in E4 (6.1±0.6 nmol/mL) than E1 and E2 extenders (8.1±0.6 nmol/mL and 8±0.6 nmol/mL, respectively). Also, the lowest significant concentration of H2O2 was observed in E4 (3.2±0.13 nmol/mL) compared to E1 (4 ±0.1 nmol/mL), E2 (5.3± 0.1 nmol/mL) and control (6.7±0.1 nmol/mL). Moreover, E2 and E4 produced the highest significant motility (74.2±1.6%, 75.9±1.6%), viability (78.2±1.8%, 76.1±1.8%) and membrane functionality (73±1.6%, 70.5±1.6%) compared to other groups. It can be concluded that α-tocopherol at the concentration of 4.8 mM can be an efficient antioxidant additive in Bioxcell extender for cryopreservation of bull semen.
Downloads
References
Alvarez-Rodríguez M, Alvarez M, Anel-López L, Martínez-Rodríguez C, Martínez-Pastor F, Borragan S, Anel L, de Paz P, 2013. The antioxidant effects of soybean lecithin-or low-density lipoprotein-based extenders for the cryopreservation of brown-bear (Ursus arctos) spermatozoa. Reprod Fertil Dev 25: 1185-1193. https://doi.org/10.1071/RD12181
Amini MR, Kohram H, Shahaneh AZ, Zhandi M, Sharideh H, Nabi MM, 2015. The effects of different levels of vitamin E and vitamin C in modified Beltsville extender on rooster post-thawed sperm quality. Cell Tissue Bank 16: 587-592. https://doi.org/10.1007/s10561-015-9506-9
Anel-López L, Álvarez-Rodríguez M, García-Álvarez O, Álvarez M, Maroto-Morales A, Anel L, de Paz P, Garde JJ, Martínez-Pastor F, 2012. Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa. Anim Reprod Sci 135: 37-46. https://doi.org/10.1016/j.anireprosci.2012.09.001
Bansal AK, Bilaspuri GS, 2009. Antioxidant effect of vitamin E on motility, viability and lipid peroxidation of cattle spermatozoa under oxidative stress. Anim Sci Pap Rep 27: 5-14.
Bansal AK, Bilaspuri GS, 2011. Impacts of oxidative stress and antioxidants on semen functions. Vet Med In 2011: 1-7. https://doi.org/10.4061/2011/686137
Barbas JP, Mascarenhas RD, 2009. Cryopreservation of domestic animal sperm cells. Cell Tissue Bank 10: 49-62. https://doi.org/10.1007/s10561-008-9081-4
Breininger E, Beorlegui NB, O'Flaherty CM, Beconi MT, 2005. Alpha-tocopherol improves biochemical and dynamic parameters in cryopreserved boar semen. Theriogenology 63: 2126-2135. https://doi.org/10.1016/j.theriogenology.2004.08.016
Brzezińska-Ślebodzińska E, Ślebodziński A, Pietras B, Wieczorek G, 1995. Antioxidant effect of vitamin E and glutathione on lipid peroxidation in boar semen plasma. Biol Trace Elem Res 47: 69-74. https://doi.org/10.1007/BF02790102
Bucak MN, Atessahin A, Varisli O, Yuce A, Tekin N, Akcay A, 2007. The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen-Microscopic and oxidative stress parameters after freeze-thawing process. Theriogenology 67: 1060-1067. https://doi.org/10.1016/j.theriogenology.2006.12.004
Bucak MN, Ateşşahin A, Yüce A, 2008. Effect of anti-oxidants and oxidative stress parameters on ram semen after the freeze–thawing process. Small Rumin Res 75: 128–134. https://doi.org/10.1016/j.smallrumres.2007.09.002
Büyükleblebici S, Tuncer PB, Bucak MN, Eken A, Sarıözkan S, Taşdemir U, Endirlik BÜ, 2014. Cryopreservation of bull sperm: Effects of extender supplemented with different cryoprotectants and antioxidants on sperm motility, antioxidant capacity and fertility results. Anim Reprod Sci 150: 77-83. https://doi.org/10.1016/j.anireprosci.2014.09.006
Cerolini S, Maldjian A, Surai P, Noble R, 2000. Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Anim Reprod Sci 58: 99-111. https://doi.org/10.1016/S0378-4320(99)00035-4
Dalvit GC, Cetica PD, Beconi MT, 1998. Effect of α-tocopherol and ascorbic acid on bovine in vitro fertilization. Theriogenology 49: 619-627. https://doi.org/10.1016/S0093-691X(98)00012-0
da Silva Maia M, Bicudo SD, Sicherle CC, Rodello L, Gallego ICS, 2010. Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants. Anim Reprod Sci 122: 118-123. https://doi.org/10.1016/j.anireprosci.2010.08.004
Davies MJ, Forni LG, Willson RL, 1988. Vitamin E analogue Trolox CEsr and pulse-radiolysis studies of free-radical reactions. Biochem J 255: 513-522.
Domínguez-Rebolledo ÁE, Fernández-Santos MR, Bisbal A, Ros-Santaella JL, Ramón M, Carmona M, Martínez-Pastor F, Garde JJ, 2010. Improving the effect of incubation and oxidative stress on thawed spermatozoa from red deer by using different antioxidant treatments. Reprod Fertil Dev 22: 856–870. https://doi.org/10.1071/RD09197
Donoghue A, Donoghue D, 1997. Effects of water-and lipid-soluble antioxidants on turkey sperm viability, membrane integrity, and motility during liquid storage. Poult Sci 76: 1440-1445. https://doi.org/10.1093/ps/76.10.1440
Emamverdi M, Zhandi M, Shahneh A, Sharafi M, Akhlaghi A, Motlagh M, Dadkhah F, Davachi N, 2014. Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders. Anim Prod Sci 55: 551-558. https://doi.org/10.1071/AN13215
Ernster L, Forsmark P, Nordenbrand K, 1992. The mode of action of lipid-soluble antioxidants in biological membranes. Relationship between the effects of ubiquinol and vitamin E as inhibitors of lipid peroxidation in submitochondrial particles. J Nut Sci Vita 38: 548-551. https://doi.org/10.3177/jnsv.38.Special_548
Esterbauer H, Cheeseman KH, 1990. Determination of aldehydic lipid peroxidation products: Malonaldehyde and 4-hydroxynonenal. Meth Enzym 186: 407-421. https://doi.org/10.1016/0076-6879(90)86134-H
Evans G, Maxwell WMC, Salamon S, 1987. Salamon's artificial insemination of sheep and goats. vol XI, 194 pp. Butterworths, Sydney; Boston.
Forouzanfar M, Sharafi M, Hosseini SM, Ostadhosseini S, Hajian M, Hosseini L, Abedi P, Nili N, Rahmani HR, Nasr-Esfahani MH, 2010. In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology 73: 480-487. https://doi.org/10.1016/j.theriogenology.2009.10.005
Khan R, Rahman Z-u, Javed I, Muhammad F, 2012. Effect of vitamins, probiotics and protein on semen traits in post-molt male broiler breeders. Anim Reprod Sci 135: 85-90. https://doi.org/10.1016/j.anireprosci.2012.09.005
Majid A, Qureshi MS, Khan RU, 2015. In vivo adverse effects of alpha-tocopherol on the semen quality of male bucks. J Anim Phys Anim Nut 99: 841-846. https://doi.org/10.1111/jpn.12284
Martínez-Páramo S, Diogo P, Dinis M, Herráez M, Sarasquete C, Cabrita E, 2012. Incorporation of ascorbic acid and α-tocopherol to the extender media to enhance antioxidant system of cryopreserved sea bass sperm. Theriogenology 77: 1129-1136. https://doi.org/10.1016/j.theriogenology.2011.10.017
Masoudi R, Sharafi M, Shahneh AZ, Towhidi A, Esmaeili V, Shahverdi A, Davachi, ND, 2016a. Fertility and flow cytometry study of frozen-thawed sperm in cryopreservation medium supplemented with soybean lecithin. Cryobiology 73: 69-72. https://doi.org/10.1016/j.cryobiol.2016.05.010
Masoudi R, Sharafi M, Shahneh AZ, Towhidi A, Zhandi M, Esmaeili V, Shahverdi A, 2016b. Effect of dietary fish oil supplementation on ram semen freeze ability and fertility using soybean lecithin and egg yolk-based extenders. Theriogenology 86: 1583-1588. https://doi.org/10.1016/j.theriogenology.2016.05.018
Masoudi R, Shahneh AZ, Towhidi A, Kohram H, Sharif AA, Sharafi M, 2017. Fertility response evaluation of artificial insemination methods in sheep with fresh and frozen-thawed semen. Cryobiology 73: 77-80. https://doi.org/10.1016/j.cryobiol.2016.11.012
Mata-Campuzano M, Álvarez-Rodríguez M, Tamayo-Canul J, López-Urueña E, de Paz P, Anel L, Martínez-Pastor F, Álvarez A, 2014. Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants: Effect of temperature, extender and storage time. Anim Reprod Sci 151: 137-147. https://doi.org/10.1016/j.anireprosci.2014.10.006
Najafi A, Zhandi M, Towhidi A, Sharafi M, Sharif AA, Motlagh MK, Martinez-Pastor F, 2013. Trehalose and glycerol have a dose-dependent synergistic effect on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender. Cryobiology 66: 275-282. https://doi.org/10.1016/j.cryobiol.2013.03.002
Najafi A, Kia HD, Mohammadi H, Najafi MH, Zanganeh Z, Sharafi M, Martinez-Pastor F, Adeldust H, 2014a. Different concentrations of cysteamine and ergothioneine improve microscopic and oxidative parameters in ram semen frozen with a soybean lecithin extender. Cryobiology 69: 68-73. https://doi.org/10.1016/j.cryobiol.2014.05.004
Najafi A, Najafi M, Zanganeh Z, Sharafi M, Martinez‐Pastor F, Adeldust H, 2014b. Cryopreservation of ram semen in extenders containing soybean lecithin as cryoprotectant and hyaluronic acid as antioxidant. Reprod Domest Anim 49: 934-940. https://doi.org/10.1111/rda.12405
Nasiri AH, Towhidi A, Zeinoaldini S, 2012. Combined effect of DHA and α‐tocopherol supplementation during bull semen cryopreservation on sperm characteristics and fatty acid composition. Andrologia 44: 550-555. https://doi.org/10.1111/j.1439-0272.2011.01225.x
NRC, 2001. Nutrient requirements of dairy cattle, 7th rev vers. National Research Council, 408 pp. https://doi.org/10.17226/9825
O'Flaherty C, Beconi M, Beorlegui N, 1997. Effect of natural antioxidants, superoxide dismutase and hydrogen peroxide on capacitation of frozen‐thawed bull sperm. Andrologia 29: 269-275. https://doi.org/10.1111/j.1439-0272.1997.tb00481.x
Purdy P, 2006. A review on goat sperm cryopreservation. Small Rumin Res 63: 215-225. https://doi.org/10.1016/j.smallrumres.2005.02.015
Revell S, Mrode R, 1994. An osmotic resistance test for bovine semen. Anim Reprod Sci 36: 77-86. https://doi.org/10.1016/0378-4320(94)90055-8
Sarıözkan S, Bucak MN, Tuncer PB, Büyükleblebici S, Eken A, Akay C, 2015. Influence of fetuin and hyaluronan on the post-thaw quality and fertilizing ability of Holstein bull semen. Cryobiology 71: 119-124. https://doi.org/10.1016/j.cryobiol.2015.04.011
Shahverdi A, Sharafi M, Gourabi H, Amiri Yekta A, Esmaeili V, Sharbatoghli M, Janzamin E, Hajnasrollahi M, Mostafayi F, 2015. Fertility and flow cytometric evaluations of frozen-thawed rooster semen in cryopreservation medium containing low-density lipoprotein. Theriogenology 1: 78-85 https://doi.org/10.1016/j.theriogenology.2014.07.044
Silva SV, Soares AT, Batista AM, Almeida FC, Nunes JF, Peixoto CA, Guerra MMP, 2013. Vitamin E (Trolox) addition to Tris-egg yolk extender preserves ram spermatozoon structure and kinematics after cryopreservation. Anim Reprod Sci 137: 37-44. https://doi.org/10.1016/j.anireprosci.2012.12.002
Suleiman SA, Ali ME, Zaki ZM, El-Malik EM, Nasr MA, 1996. Lipid peroxidation and human sperm motility: protective role of vitamin E. Andrology 17: 530-537.
Surai P, Kostjuk I, Wishart G, Macpherson A, Speake B, Noble R, Ionov I, Kutz E, 1998. Effect of vitamin E and selenium supplementation of cockerel diets on glutathione peroxidase activity and lipid peroxidation susceptibility in sperm, testes, and liver. Biol Trace Elem Res 64: 119-132. https://doi.org/10.1007/BF02783329
Towhidi A, Parks JE, 2012. Effect of n-3 fatty acids and α-tocopherol on post-thaw parameters and fatty acid composition of bovine sperm. J Assist Reprod Genetic 29: 1051-1056. https://doi.org/10.1007/s10815-012-9834-7
Tuncer PB, Bucak MN, Sarıözkan S, Sakin F, Yeni D, Çiğerci İH, Ateşşahin A, Avdatek F, Gündoğan M, Büyükleblebici O, 2010. The effect of raffinose and methionine on frozen/thawed Angora buck (Capra hircus ancryrensis) semen quality, lipid peroxidation and antioxidant enzyme activities. Cryobiology 61: 89-93. https://doi.org/10.1016/j.cryobiol.2010.05.005
Tuncer PB, Sariozkan S, Bucak MN, Ulutas PA, Akalin PP, Buyukleblebici S, Canturk F, 2011. Effect of glutamine and sugars after bull sperm cryopreservation. Theriogenology 75: 1459-1465. https://doi.org/10.1016/j.theriogenology.2010.12.006
Watson P, 1976. The protection of ram and bull sperm by the low density lipoprotein fraction of egg yolk during storage at 5 C and deep-freezing. J Therm Biol 1: 137-141. https://doi.org/10.1016/0306-4565(76)90003-6
Watson P, 2000. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci 60: 481-492. https://doi.org/10.1016/S0378-4320(00)00099-3
Yousef MI, 2010. Vitamin E modulates reproductive toxicity of pyrethroid lambda-cyhalothrin in male rabbits. Food Chem Toxico 48: 1152-1159. https://doi.org/10.1016/j.fct.2010.02.002
Zanganeh Z, Zhandi M, Zare-Shahneh A, Najafi A, Nabi MM, Mohammadi-Sangcheshmeh A, 2013. Does rosemary aqueous extract improve buck semen cryopreservation? Small Rumin Res 114: 120-125. https://doi.org/10.1016/j.smallrumres.2013.05.015
Zhandi M, Sharafi M, 2015. Negative effect of combined cysteine and glutathione in soy lecithin-based extender on post-thawed ram spermatozoa. Cell Tissue Bank 16: 443-448. https://doi.org/10.1007/s10561-014-9488-z
© CSIC. Manuscripts published in both the print and online versions of this journal are the property of the Consejo Superior de Investigaciones Científicas, and quoting this source is a requirement for any partial or full reproduction.
All contents of this electronic edition, except where otherwise noted, are distributed under a Creative Commons Attribution 4.0 International (CC BY 4.0) licence. You may read the basic information and the legal text of the licence. The indication of the CC BY 4.0 licence must be expressly stated in this way when necessary.
Self-archiving in repositories, personal webpages or similar, of any version other than the final version of the work produced by the publisher, is not allowed.
