Effect of Antioxidant D-Aspartic Acid and Thawing Rate on the Freeze-Thawing Process of Ram Semen
الموضوعات :ح. دقیق کیا 1 , ص. وطنخواه 2
1 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
2 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
الکلمات المفتاحية: ram, cryopreservation, antioxidants, D-aspartic acid, thawing rate,
ملخص المقالة :
The study was conducted to determine the influence of D-aspartic acid (D-Asp) as antioxidant supplement and thawing rates on ram sperm motility, membrane integrity, abnormality, viability, mitochondria activity, malondialdehyde and antioxidant activities, and total antioxidant capacity after freezing-thawing process. Semen samples from five mature rams (3-4 years old) were diluted with extenders (1.5% soybean lecithin, 7% glycerol) containing no supplements (control) and D-Asp (5, 10, and 15 mg/L) and cryopreserved. Frozen straws were thawed at water bath temperatures at 37 ˚C for 30s and at 60 ˚C for 6s. Addition of 10 mg/L of D-Asp improved significantly progressive motility, average path velocity, and straight-line velocity percentages after freeze-thaw (P<0.05). Plasma membrane integrity, mitochondria activity, viability, total antioxidant capacity, and glutathione peroxidase were higher in group receiving 10 mg/L in comparison to other treatments (P<0.05). Meanwhile, total abnormality significantly decreased at this concentration (10 mg/L) in comparison to the level of 15 mg/L and control group (P<0.05). Our results revealed that malondialdehyde level was lower in group receiving 10 mg/L D-Asp compared to other treatments (P<0.05). There were no significant interactions between concentrations of D-Asp and thawing protocols for any semen samples. Also, significant improvement in sperm viability and mitochondria activity were observed at 37 ˚C to 30 s thawing method (P<0.05). In overall, results of the present study demonstrate that addition of D-Asp at level of 10 mg/L have beneficial effect on quality of post-thawed ram semen cryopreserved in an extender. Therefore, this antioxidant in the suitable dose can be recommended as an additional component of ram freezing extender. The 60 ˚C to 6s thawing procedure is not an appropriate replacement for 37 ˚C to 30 s.
Aamdal J. and Andersen K. (1968). Freezing of ram semen in straws. Pp. 977-980 in Int. Congr. Anim. Reprod. Artif. Insem., Paris, France.
Agarwal A. and Anandh Prabakaran S. (2005). Oxidative stress and antioxidants in male infertility: a difficult balance. Iranian J. Reprod. Med. 3, 1-8.
Agarwal A. and Said T.M. (2003). Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum. Reprod. Update. 9, 331-345.
Aitken R.J., Fisher H.M., Fulton N., Gomez E., Knox W., Lewis B. and Irvine S. (1997). Reactive oxygen species generation by human spermatozoa is induced by exogenous NADPH and inhibited by the flavoprotein inhibitors diphenylene iodonium and quinacrine. Mol. Reprod. Dev. 47, 468-482.
Alvarez J.G. and Storey B.T. (1992). Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation. J. Androl. 13, 232-241.
Andersen V.K., Aamdal J. and Fougner J.A. (1973). Intrauterine and deep cervical insemination with frozen semen in sheep. Reprod. Domest. Anim. 8, 113-118.
Anghel A., Zamfirescu S., Dragomir C., Nadolu D., Elena S. and Florica B. (2010). The effects of antioxidants on the cytological parameters of cryopreserved buck semen. Romanian Biotechnol. Lett. 15, 26-32.
Ansari M., Zhandi M., Kohram H., Zaghari M. and Sadeghi M. (2016). The effect of D-aspartic acid on sperm quality of broiler breeder roosters. Iranian J. Agric. Sci. 47, 165-174.
Ansari M., Zhandi M., Kohram H., Zaghari M., Sadeghi M. and Sharafi M. (2017). Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid. Theriogenology. 92, 69-74.
Athurupana R., Ioki S. and Funahashi H. (2015). Rapid thawing and stabilizing procedure improve postthaw survival and in vitro penetrability of boar spermatozoa cryopreserved with a glycerol-free trehalose-based extender. Theriogenology. 84, 940-947.
Bansal A.K. and Bilaspuri G. (2011). Impacts of oxidative stress and antioxidants on semen functions. Vet. Med. Int. 32, 1-7.
Bilodeau J.F., Blanchette S., Cormier N. and Sirard M.A. (2002). Reactive oxygen species-mediated loss of bovine sperm motility in egg yolk Tris extender: Protection by pyruvate, metal chelators and bovine liver or oviductal fluid catalase. Theriogenology. 57, 1105-1122.
Borah B.K., Deka B.C., Biswas R.K., Chakravarty P., Deori S., Sinha S. and Ahmed K. (2015). Effect of thawing methods on frozen semen quality of yak (Poephagus grunniens) bulls. Vet. World. 8, 831-834.
Bucak M.N., Ateşşahin A. and 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.
Budai C. (2014). The protective effect of antioxidants on liquid and frozen stored ram semen. Sci. Pap. Anim. Sci. Biotechnol. 47, 46-52.
Chandrashekar K.N. and Muralidhara (2010). D-aspartic acid induced oxidative stress and mitochondrial dysfunctions in testis of prepubertal rats. Amino Acids. 38, 817-827.
Correa J.R., Rodriguez M.C., Patterson D.J. and Zavos P.M. (1996). Thawing and processing of cryopreserved bovine spermatozoa at various temperatures and their effects on sperm viability, osmotic shock and sperm membrane functional integrity. Theriogenology. 46, 413-420.
Çoyan K., Başpınar N., Bucak M.N. and Akalın P.P. (2011). Effects of cysteine and ergothioneine on post-thawed Merino ram sperm and biochemical parameters. Cryobiology. 63, 1-6.
D'Aniello A. and Giuditta A. (1977). Identification of D-aspartic acid in the brain of Octopus vulgaris lam. J. Neurochem. 29, 1053-1057.
D’Aniello G., Grieco N., Di Filippo M.A., Cappiello F., Topo E., D'Aniello E. and Ronsini S. (2007). Reproductive implication of D-aspartic acid in human pre-ovulatory follicular fluid. Hum. Reprod. 22, 3178-3183.
D’Aniello G., D’Aniello G., Ronsini S., Notari T., Grieco N., Infante V., D’Angelo N., Mascia F., Maddalena Di Fiore M., Fisher G. and D’Aniello A. (2012). D-aspartate, a key element for the improvement of sperm quality. Adv. Sex. Med. 2, 47-53.
Deka B. and Rao A. (1987). Effect of extenders and thawing methods on post thawing preservation of goat semen. Indian Vet. J. 64, 591-594.
Dhami A.J. and Sahni K.L. (1993). Evaluation of different cooling rates, equilibration periods and diluents for effects on deep-freezing, enzyme leakage and fertility of taurine bull spermatozoa. Theriogenology. 40, 1269-1280.
Di Fiore M.M., Santillo A. and Chieffi Baccari G. (2014). Current knowledge of D-aspartate in glandular tissues. Amino Acids. 46, 1805-1818.
Dodaran H.V., Zhandi M., Sharafi M., Nejati-Amiri E., Nejati-Javaremi A., Mohammadi-Sangcheshmeh A., Shehab-El-Deen M.A. and Shakeri M. (2015). Effect of ethanol induced mild stress on post-thawed bull sperm quality. Cryobiology. 71, 12-17.
Eriksson B.M. and Rodriguez-Martinez H. (2000). Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in FlatPacks and Maxi-straws. Anim. Reprod. Sci. 63, 205-220.
Esterbauer H. and Cheeseman K.H. (1990). Determination of aldehydic lipid peroxidation products: Malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 186, 407-421.
Fiser P.S., Fairfull R.W., Hansen C., Panich P.L., Shrestha J.N. and Underhill L. (1993). The effect of warming velocity on motility and acrosomal integrity of boar sperm as influenced by the rate of freezing and glycerol level. Mol. Reprod. Dev. 34, 190-195.
Froman D. (2003). Deduction of a model for sperm storage in the oviduct of the domestic fowl (Gallus domesticus). Biol. Reprod. 69, 248-253.
Froman D.P., Wardell J.C. and Feltmann A.J. (2006). Sperm mobility: deduction of a model explaining phenotypic variation in roosters (Gallus domesticus). Biol. Reprod. 74, 487-491.
Giulini S., Sblendorio V., Xella S., La Marca A., Palmieri B. and Volpe A. (2009). Seminal plasma total antioxidant capacity and semen parameters in patients with varicocele. Reprod. Biomed. Online. 18, 617-621.
Graham J.K., Kunze E. and Hammerstedt R.H. (1990). Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry. Biol. Reprod. 43, 55-64.
Gualtieri R., Boni R., Tosti E., Zagami M. and Talevi R. (2005). Intracellular calcium and protein tyrosine phosphorylation during the release of bovine sperm adhering to the fallopian tube epithelium in vitro. Reproduction. 129, 51-60.
Hashemi A., Farhoomand P., Pirmohammadi R., Nayebpor M. and Razzaghzadeh S. (2007). Effect of extender and thawing methods on post thawing preservation. J. Anim. Vet. Adv. 6, 1337-1339.
Jones R. and Mann T. (1977). Toxicity of exogenous fatty acid peroxides towards spermatozoa. J. Reprod. Fertil. 50, 255-260.
Khosrowbeygi A., Zargham N. and Farzadi L. (2008). Fatty acid composition in normozoospermic, asthenozoopermic, asthenoteratozoospermic and oligoasthenoteratozoospermic ejaculates. Int. J. Reprod. Biomed. 6, 39-43.
Lenzi A., Picardo M., Gandini L. and Dondero F. (1996). Lipids of the sperm plasma membrane: from polyunsaturated fatty acids considered as markers of sperm function to possible scavenger therapy. Hum. Reprod. Update. 2, 246-256.
Macchia G., Topo E., Mangano N., D'Aniello E. and Boni R. (2010). DL-aspartic acid administration improves semen quality in rabbit bucks. Anim. Reprod. Sci. 118, 337-343.
Mahat R.K., Kumar S., Arora M., Bhale D.V., Mehta R. and Batra J. (2015). Role of oxidative stress and antioxidants in male infertility. Int. J. Health Sci. Res. 5, 324-333.
Maia Mda S., Bicudo S.D., Sicherle C.C., Rodello L. and Gallego I.C. (2010). Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants. Anim. Reprod. Sci. 122, 118-123.
Malo C., Gil L., Gonzalez N., Martínez F., Cano R., de Blas I. and Espinosa E. (2010). Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: comparison between cysteine and rosemary (Rosmarinus officinalis). Cryobiology. 61, 142-147.
Marshall C.E. (1984). Considerations for cryopreservation of semen. Zoo Biol. 3, 343-356.
Maxwell W.M. and Salamon S. (1993). Liquid storage of ram semen: A review. Reprod. Fertil. Dev. 5, 613-638.
Mazur P. (1984). Freezing of living cells: Mechanisms and implications. American J. Physiol. Cell Physiol. 247, 125-142.
Mazur P. (1985). Basic Concepts in Freezing Cells. Oak Ridge National Lab., Oak Ridge, Tennessee, USA.
Mehdipour M., Daghigh Kia H., Najafi A., Vaseghi Dodaran H. and García-Álvarez O. (2016). Effect of green tea (Camellia sinensis) extract and pre-freezing equilibration time on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender. Cryobiology. 73, 297-303.
Nair S.J., Brar A.S., Ahuja C.S., Sangha S.P. and Chaudhary K.C. (2006). A comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Anim. Reprod. Sci. 96, 21-29.
Najafi A., Najafi M.H., Zanganeh Z., Sharafi M., Martinez-Pastor F. and Adeldust H. (2014). Cryopreservation of ram semen in extenders containing soybean lecithin as cryoprotectant and hyaluronic acid as antioxidant. Reprod. Domest. Anim. 49, 934-940.
Nicolae D., Stela Z., Dragomir C. and Hortanse A.A. (2014). Effect of thawing time and temperature variation on the quality of frozen-thawed ram semen. Romanian Biotechnol. Lett. 19, 8935-8940.
Ogbuewu I.P., Aladi N.O., Etuk I.F., Opara M.N., Uchegbu M.C., Okoli I.C. and Iloeje M.U. (2010). Relevance of oxygen free radicals and antioxidants in sperm. Res. J. Vet. Sci. 3, 138-164.
Partyka A., Łukaszewicz E. and Niżański W. (2012). Effect of cryopreservation on sperm parameters, lipid peroxidation and antioxidant enzymes activity in fowl semen. Theriogenology. 77, 1497-1504.
Perry E.J. (1955). Artificial Insemination of Farm Animals. Rutgers University Press, New Brunswick.
Pontbriand D., Howard J.G., Schiewe M.C., Stuart L.D. and Wildt D.E. (1989). Effect of cryoprotective diluent and method of freeze-thawing on survival and acrosomal integrity of ram spermatozoa. Cryobiology. 26, 341-354.
Purdy P.H. (2006). A review on goat sperm cryopreservation. Small Rumin. Res. 63, 215-225.
Salamon S. and Maxwell W.M.C. (1995). Frozen storage of ram semen I. Processing, freezing, thawing and fertility after cervical insemination. Anim. Reprod. Sci. 37, 185-249.
SAS Institute. (2001). SAS®/STAT Software, Release 9.1. SAS Institute, Inc., Cary, NC. USA.
Shi Y.C., Shang X.J., Wang X.L. and Huang Y.F. (2006). Correlation of total antioxidant capacity in seminal plasma with sperm motility of infertile men. Natl. J. Androl. 12, 703-705.
Sies H. (1997). Oxidative stress: Oxidants and antioxidants. Exp. Physiol. 82, 291-295.
Talevi R., Barbato V., Fiorentino I., Braun S., Longobardi S. and Gualtieri R. (2013). Protective effects of in vitro treatment with zinc, d-aspartate and coenzyme Q10 on human sperm motility, lipid peroxidation and DNA fragmentation. Reprod. Biol. Endocrinol. 11, 81-91.
Topo E., Soricelli A., D'Aniello A., Ronsini S. and D'Aniello G. (2009). The role and molecular mechanism of D-aspartic acid in the release and synthesis of LH and testosterone in humans and rats. Reprod. Biol. Endocrinol. 7, 120-127.
Vazquez J.M., Martinez E.A., Martinez P., Garcia-Artiga C. and Roca J. (1997). Hypoosmotic swelling of boar spermatozoa compared to other methods for analysing the sperm membrane. Theriogenology. 47, 913-922.
Woolley D.M. and Richardson D.W. (1978). Ultrastructural injury to human spermatozoa after freezing and thawing. J. Reprod. Fertil. 53, 389-394.
Yue H.X., Li F.P., Jiang M., Lin L. and Xu K.H. (2005). Influence of cryoprotectant with glycerol and freezing-thawing procedure on the motility of human sperm. Natl. J. Androl. 11, 204-206.
Zarghami N.A. and Khosrow beygi A. (2004). Evaluation of lipid peroxidation as an indirect measure of oxidative stress in seminal plasma. Int. J. Reprod. Biomed. 2, 34-39.
