Animal Reproduction (AR)
https://animal-reproduction.org/article/doi/10.1590/1984-3143-AR2022-0068
Animal Reproduction (AR)
ORIGINAL ARTICLE

Post-thaw quality of ram sperm frozen with different concentrations of low-density lipoproteins associated with non-enzymatic antioxidants

Paola Pereira das Neves Snoeck; Diogo Ribeiro Câmara; Luís Cláudio de Oliveira Moura; Maíra Corona da Silva; Mariana Machado-Neves; Milton Rezende Teixeira-Neto; Marc Henry

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Abstract

The cryopreservation reduces ram sperm quality, decreasing the pregnancy rate of ewes inseminated with thawed sperm. Hence, we aimed to improve the post-thaw quality of ram sperm replacing egg yolk on Tris-Glucose extender with different concentrations of LDL (2 or 8%), associated with the addition of 10 mM non-enzymatic antioxidants (ascorbic acid, hydroxytoluene butylate, ascorbyl palmitate, and trehalose). Semen samples were collected from six rams, split into different treatments, and frozen. After thawing, kinematic (CASA), structural (propidium iodide and carboxyfluorescein diacetate) and functional (hypoosmotic test) sperm membrane integrity was assessed. Total motility, VCL, and LIN were also assessed in thawed samples during 3 h of incubation (38 °C). The results showed that hydroxytoluene butylate at 10 mM in Tris-Glucose extender with 8% LDL improved velocity parameters immediately post-thaw compared with Tris-Glucose egg yolk extender, as well as prevented the reduction of total motility and VCL after incubation. There was no benefit of adding ascorbic acid and trehalose. Moreover, for the first time, it was shown the motility impairment promoted by ascorbyl palmitate to ram sperm.

Supplementary Material

http://s3.amazonaws.com/host-client-assets/files/animreprod/1984-3143-ar-20-1-e20220068-Suppl.pdf

Keywords

ascorbyl palmitate, cryopreservation, hydroxytoluene butylate, sheep, spermatozoa

References

Agarwal A, Tadros H, Panicker A, Tvrdá E. Role of oxidants and antioxidants in male reproduction. In: Armstrong D, Stratton RD, editors. Oxidative stress and antioxidant protection. Hoboken: John Wiley & Sons, Inc; 2016. p. 221-52. http://dx.doi.org/10.1002/9781118832431.ch15

Alcay S, Toker MB, Gokce E, Ustuner B, Onder NT, Sagirkaya H, Nur Z, Soylu MK. Successful ram semen cryopreservation with lyophilized egg yolk-based extender. Cryobiology. 2015;71(2):329-33. http://dx.doi.org/10.1016/j.cryobiol.2015.08.008. PMid:26277039.

Amidi F, Pazhohan A, Shabani Nashtaei M, Khodarahmian M, Nekoonam S. The role of antioxidants in sperm freezing: a review. Cell Tissue Bank. 2016;17(4):745-56. http://dx.doi.org/10.1007/s10561-016-9566-5. PMid:27342905.

Azawi OI, Hussein EK. Effect of vitamins C or E supplementation to Tris diluent on the semen quality of Awassi rams preserved at 5 ˚C. Vet Res Forum. 2013;4(3):157-60. PMid:25653790.

Bag S, Joshi A, Naqvi SM, Mittal J. Effect of post-thaw incubation on sperm kinematics and acrosomal integrity of ram spermatozoa cryopreserved in medium-sized French straws. Theriogenology. 2004;62(3-4):415-24. http://dx.doi.org/10.1016/j.theriogenology.2003.10.018. PMid:15225998.

Balaro MFA, Fonseca JF, Oba E, Cardoso EC, Brandão FZ. Is the Santa Inês sheep a typical non-seasonal breeder in the Brazilian Southeast? Trop Anim Health Prod. 2014;46(8):1533-7. http://dx.doi.org/10.1007/s11250-014-0672-8. PMid:25252775.

Baldi E, Tamburrino L, Muratori M, Degl’Innocenti S, Marchiani S. Adverse effects of in vitro manipulation of spermatozoa. Anim Reprod Sci. 2020;220:106314. http://dx.doi.org/10.1016/j.anireprosci.2020.106314. PMid:32089373.

Ball BA. Oxidative stress, osmotic stress and apoptosis: impacts on sperm function and preservation in the horse. Anim Reprod Sci. 2008;107(3-4):257-67. http://dx.doi.org/10.1016/j.anireprosci.2008.04.014. PMid:18524506.

Bast A, Haenen GRMM, Doelman CJA. Oxidants and antioxidants: state of the art. Am J Med. 1991;91(3C, Suppl 3):S2-13. http://dx.doi.org/10.1016/0002-9343(91)90278-6. PMid:1928207.

Belala R, Briand-Amirat L, Martinot A, Thorin C, Michaud S, Desherces S, Youngs CR, Bencharif D. A comparison of liquid and lyophilized egg yolk plasma to low density lipoproteins for freezing of canine spermatozoa. Reprod Domest Anim. 2019;54(8):1131-8. http://dx.doi.org/10.1111/rda.13476. PMid:31145501.

Bergeron A, Manjunath P. New insights towards understanding the mechanisms of sperm protection by egg yolk and milk. Mol Reprod Dev. 2006;73(10):1338-44. http://dx.doi.org/10.1002/mrd.20565. PMid:16868925.

Bergeron A, Villemure M, Lazure C, Manjunath P. Isolation and characterization of the major proteins of ram seminal plasma. Mol Reprod Dev. 2005;71(4):461-70. http://dx.doi.org/10.1002/mrd.20310. PMid:15892046.

Brouwers JFHM, Gadella BM. In situ detection and localization of lipid peroxidation in individual bovine sperm cells. Free Radic Biol Med. 2003;35(11):1382-91. http://dx.doi.org/10.1016/j.freeradbiomed.2003.08.010. PMid:14642386.

Caritá AC, Fonseca-Santos B, Shultz JD, Michniak-Kohn B, Chorilli M, Leonardi GR. Vitamin C: one compound, several uses. Advances for delivery, efficiency and stability. Nanomedicine. 2020;24:102117. http://dx.doi.org/10.1016/j.nano.2019.102117. PMid:31676375.

Cirit Ü, Bağış H, Demir K, Agca C, Pabuccuoğlu S, Varışlı Ö, Clifford-Rathert C, Agca Y. Comparison of cryoprotective effects of iodixanol, trehalose and cysteamine on ram semen. Anim Reprod Sci. 2013;139(1-4):38-44. http://dx.doi.org/10.1016/j.anireprosci.2013.03.010. PMid:23602486.

Denisov IG, Makris TM, Sligar SG, Schlichting I. Structure and chemistry of cytochrome P450. Chem Rev. 2005;105(6):2253-77. http://dx.doi.org/10.1021/cr0307143. PMid:15941214.

Dresser GK, Wacher V, Wong S, Wong HT, Bailey DG. Evaluation of peppermint oil and ascorbyl palmitate as inhibitors of cytochrome P4503A4 activity in vitro and in vivo. Clin Pharmacol Ther. 2002;72(3):247-55. http://dx.doi.org/10.1067/mcp.2002.126409. PMid:12235445.

El-Hamid ISA, Khalifa MA, Shedeed HA, Rateb SA. Cryosurvival of ram spermatozoa after supplementing the diluent with L-ascorbic acid or α-tocopherol. Int J Vet Sci. 2018;7:197-204.

Fang Y, Zhong R. Effects of oxidative stress on spermatozoa and male infertility. In: Das K, Das S, Biradari MS, Bobbarala V, Tata SS, editors. Free radical medicine and biology. London: IntechOpen; 2020. http://dx.doi.org/10.5772/intechopen.86585.

Farshad A, Khalili B, Jafaroghli M. Effects of butylated hydroxytoluene on freezability of ram spermatozoa. Asian-Australas J Anim Sci. 2010;23(10):1276-81. http://dx.doi.org/10.5713/ajas.2010.90590.

Freitas AP, dos Santos GFF, Fernandes AR, Mendonça GG, de Paz CCP, Vercesi Filho AE, El Faro L. Effect of thermal stress on basic seminal characteristics of Gyr bulls. Int J Biometeorol. 2020;64(10):1649-56. http://dx.doi.org/10.1007/s00484-020-01945-6. PMid:32504143.

Galarza DA, López‐Sebastián A, Santiago‐Moreno J. Supplementing a skimmed milk-egg yolk‐based extender with L‐carnitine helps maintain the motility, membrane integrity and fertilizing capacity of chilled ram sperm. Reprod Domest Anim. 2020;55(7):805-13. http://dx.doi.org/10.1111/rda.13687. PMid:32311171.

Gibb Z, Griffin RA, Aitken RJ, De Iuliis GN. Functions and effects of reactive oxygen species in male fertility. Anim Reprod Sci. 2020;220:106456. http://dx.doi.org/10.1016/j.anireprosci.2020.106456. PMid:32381272.

Graham JK, Hammerstedt RH. Differential effects of butylated hydroxytoluene analogs on bull sperm subjected to cold-induced membrane stress. Cryobiology. 1992;29(1):106-17. http://dx.doi.org/10.1016/0011-2240(92)90010-Y. PMid:1318820.

Hammerstedt RH, Amann RP, Rucinsky T, Morse PD Jr, Lepock J, Snipes W, Keith AD. Use of spin labels and electron spin resonance spectroscopy to characterize membranes of bovine sperm: effect of butylated hydroxytoluene and cold shock. Biol Reprod. 1976;14(4):381-97. http://dx.doi.org/10.1095/biolreprod14.4.381. PMid:179626.

Hanada H. Disruption of vitamin E and butylated hydroxytoluene antioxidant function in response to paraquat-induced chromosomal damage in cultured anuran leukocytes. Biol Med (Aligarh). 2014;07(01):222. http://dx.doi.org/10.4172/0974-8369.1000222.

Harrison RAP, 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.

Hu JH, Jiang ZL, Lv RK, Li QW, Zhang SS, Zan LS, Li YK, Li X. The advantages of low-density lipoproteins in the cryopreservation of bull semen. Cryobiology. 2011;62(1):83-7. http://dx.doi.org/10.1016/j.cryobiol.2010.12.007. PMid:21199643.

Hu JH, Li QW, Zan LS, Jiang ZL, An JH, Wang LQ, Jia YH. The cryoprotective effect of low-density lipoproteins in extenders on bull spermatozoa following freezing-thawing. Anim Reprod Sci. 2010;117(1-2):11-7. http://dx.doi.org/10.1016/j.anireprosci.2009.04.001. PMid:19586728.

Landaeta-Hernández AJ, Gil-Araujo MA, Ungerfeld R, Owen Rae D, Urdaneta-Moyer A, Parra-Núñez A, Kaske M, Bollwein H, Chenoweth PJ. Effect of season and genotype on values for bull semen variables under tropical conditions. Anim Reprod Sci. 2020;221:106592. http://dx.doi.org/10.1016/j.anireprosci.2020.106592. PMid:32927186.

Loaiza-Echeverri AM, Cruz BC, Snoeck PPN, Moura LCO, Neves BP, Neves MM, Heneine LGD, Henry M. Low density lipoproteins added to an extender frozen or lyophilized are evenly efficient in cryoprotecting ovine sperm cells than when 16% whole egg yolk was added. Semina: Ciênc Agrár. 2015;36(3):1335-46. http://dx.doi.org/10.5433/1679-0359.2015v36n3p1315.

Marti E, Marti JI, Muiño-Blanco T, Cebrián-Pérez JA. Effect of the cryopreservation process on the activity and immunolocalization of antioxidant enzymes in ram spermatozoa. J Androl. 2008;29(4):459-67. http://dx.doi.org/10.2164/jandrol.107.003459. PMid:18296478.

Martínez-Fresneda L, O’Brien E, Velázquez R, Toledano-Díaz A, Martínez-Cáceres CM, Tesfaye D, Schellander K, García-Vázquez FA, Santiago-Moreno J. Seasonal variation in sperm freezability associated with changes in testicular germinal epithelium in domestic (Ovis aries) and wild (Ovis musimon) sheep. Reprod Fertil Dev. 2019;31(10):1545-57. http://dx.doi.org/10.1071/RD18511. PMid:31182186.

Melo MIV, Henry M, Beker ARCL. Hypoosmotic test to predict viability of equine chilled semen in different extenders. Arq Bras Med Vet Zootec. 2005;57(6):757-63. http://dx.doi.org/10.1590/S0102-09352005000600009.

Memon AA, Wahid H, Rosnina Y, Goh YM, Ebrahimi M, Nadia FM, Audrey G. Effect of butylated hydroxytoluene on cryopreservation of Boer goat semen in Tris egg yolk extender. Anim Reprod Sci. 2011;129(1-2):44-9. http://dx.doi.org/10.1016/j.anireprosci.2011.10.004. PMid:22024366.

Merino O, Aguagüiña WE, Esponda P, Risopatrón J, Isachenko E, Isachenko V, Sánchez R. Protective effect of butylated hydroxytoluene on sperm function in human spermatozoa cryopreserved by vitrification technique. Andrologia. 2015;47(2):186-93. http://dx.doi.org/10.1111/and.12246. PMid:24612426.

Morris GJ, Faszer K, Green JE, Draper D, Grout BWW, Fonseca F. Rapidly cooled horse spermatozoa: loss of viability is due to osmotic imbalance during thawing, not intracellular ice formation. Theriogenology. 2007;68(5):804-12. http://dx.doi.org/10.1016/j.theriogenology.2007.06.009. PMid:17645937.

Moustacas VS, Zaffalon FG, Lagares MA, Loaiza-Eccheverri AM, Varago FC, Neves MM, Heneine LGD, Arruda RP, Henry M. Natural, but not lyophilized, low density lypoproteins were an acceptable alternative to egg yolk for cryopreservation of ram semen. Theriogenology. 2011;75(2):300-7. http://dx.doi.org/10.1016/j.theriogenology.2010.08.016. PMid:20934209.

Naijian HR, Kohram H, Shahneh AZ, Sharafi M, Bucak MN. Effects of different concentrations of BHT on microscopic and oxidative parameters of Mahabadi goat semen following the freeze-thaw process. Cryobiology. 2013;66(2):151-5. http://dx.doi.org/10.1016/j.cryobiol.2012.12.010. PMid:23313785.

Najafi A, Zhandi M, Towhidi A, Sharafi M, Akbari Sharif A, Khodaei Motlagh M, Martinez-Pastor F. 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. 2013;66(3):275-82. http://dx.doi.org/10.1016/j.cryobiol.2013.03.002. PMid:23500077.

Neves MM, Heneine LGD, Henry M. Cryoprotection effectiveness of low concentrations of natural and lyophilized LDL (low density lipoproteins) on canine spermatozoa. Arq Bras Med Vet Zootec. 2014;66(3):769-77. http://dx.doi.org/10.1590/1678-41626184.

Öztürk AE, Bodu M, Bucak MN, Ağır V, Özcan A, Keskin N, İli P, Topraggaleh TR, Sidal H, Başpınar N, Dursun Ş. The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters. Cryobiology. 2020;95:157-63. http://dx.doi.org/10.1016/j.cryobiol.2020.03.008. PMid:32259524.

Palomo MJ, García W, Tabarez A. Effect of seminal plasma and butylated hydroxytoluene (BHT) concentration on ram sperm freezability. Small Rumin Res. 2017;153:66-70. http://dx.doi.org/10.1016/j.smallrumres.2017.05.010.

Peris-Frau P, Soler AJ, Iniesta-Cuerda M, Martín-Maestro A, Sánchez-Ajofrín I, Medina-Chávez DA, Fernández-Santos MR, García-Álvarez O, Maroto-Morales A, Montoro V, Garde JJ. Sperm cryodamage in ruminants: understanding the molecular changes induced by the cryopreservation process to optimize sperm quality. Int J Mol Sci. 2020;21(8):2781. http://dx.doi.org/10.3390/ijms21082781. PMid:32316334.

Pini T, Farmer K, Druart X, Teixeira-Gomes AP, Tsikis G, Labas V, Leahy T, Graaf SP. Binder of sperm proteins protect ram spermatozoa from freeze-thaw damage. Cryobiology. 2018;82:78-87. http://dx.doi.org/10.1016/j.cryobiol.2018.04.005. PMid:29653077.

Rostami B, Ebrahimi D, Sadeghipanah H, Masoumi R, Shahir MH. Effects of supplementation of tris-egg yolk extender with different sugars and antioxidants on freezability of ram semen. Cryobiology. 2020;92:62-6. http://dx.doi.org/10.1016/j.cryobiol.2019.10.198. PMid:31669434.

Said L, Saad A, Carreau S. Differential expression of mRNA aromatase in ejaculated spermatozoa from infertile men in relation to either asthenozoospermia or teratozoospermia. Andrologia. 2014;46(2):136-46. http://dx.doi.org/10.1111/and.12058. PMid:23252370.

Sieme H, Oldenhof H, Wolkers WF. Sperm membrane behaviour during cooling and cryopreservation. Reprod Domest Anim. 2015;50(Suppl 3):20-6. http://dx.doi.org/10.1111/rda.12594. PMid:26382025.

Silva MC, Moura LCO, Melo MIV, Mambrini JVM, Neves MM, Henry MRJM, Snoeck PPN. Prolonged post cooling but not pre-cooling equilibrium length improves the viability of ram sperm cryopreserved in an extender containing low-density lipoproteins. Small Rumin Res. 2014;119(1-3):88-95. http://dx.doi.org/10.1016/j.smallrumres.2014.02.006.

Snoeck PPN, Moura LCO, Clemente CAA, Loaiza-Echeverri AM, Neves MM, Allaman IB, Henry M. Effect of catalase, superoxide dismutase and reduced glutathione in LDL extender on ovine cryopreserved sperm viability. Semina: Ciênc Agrár. 2015;36(4):2593-601. http://dx.doi.org/10.5433/1679-0359.2015v36n4p2593.

Sönmez M, Demirci E. The effect of ascorbic acid on the freezability of ram semen diluted with extenders containing different proportions of glycerole. Turk J Vet Anim Sci. 2004;28:893-9.

St-Pierre NR. Design and analysis of pen studies in the animal sciences. J Dairy Sci. 2007;90(Suppl 1):E87-99. http://dx.doi.org/10.3168/jds.2006-612. PMid:17517755.

Tonieto RA, Goularte KL, Gastal GDA, Schiavon RS, Deschamps JC, Lucia T Jr. Cryoprotectant effect of trehalose and low-density lipoprotein in extenders for frozen ram semen. Small Rumin Res. 2010;93(2-3):206-9. http://dx.doi.org/10.1016/j.smallrumres.2010.05.003.

Tremellen K. Oxidative stress and male infertility: a clinical perspective. Hum Reprod Update. 2008;14(3):243-58. http://dx.doi.org/10.1007/978-1-61779-776-7_16. PMid:18281241.

Varela E, Rojas M, Restrepo G. Membrane stability and mitochondrial activity of bovine sperm frozen with low-density lipoproteins and trehalose. Reprod Domest Anim. 2020;55(2):146-53. http://dx.doi.org/10.1111/rda.13599. PMid:31802545.

Vera-Munoz O, Amirat-Briand L, Bencharif D, Anton M, Desherces S, Shmitt E, Thorin C, Tainturier D. Effect of low-density lipoproteins, spermatozoa concentration and glycerol on functional and motility parameters of bull spermatozoa during storage at 4 °C. Asian J Androl. 2011;13(2):281-6. http://dx.doi.org/10.1038/aja.2010.84. PMid:21113174.

Watson PF, Anderson WJ. Influence of butylated hydroxytoluene (BHT) on the viability of ram spermatozoa undergoing cold shock. J Reprod Fertil. 1983;69(1):229-35. http://dx.doi.org/10.1530/jrf.0.0690229. PMid:6887138.

Yu L, Yang X, Ma B, Ying H, Shang X, He B, Zhang Q. Abnormal arachidonic acid metabolic network may reduce sperm motility via P38 MAPK. Open Biol. 2019;9(4):180091. http://dx.doi.org/10.1098/rsob.180091. PMid:31014201.

Zobel M. Toxicological evaluation of some food additives including anticaking agents, antimicrobials, antioxidants, emulsifiers and thickening agents. Mol Nutr Food Res. 1976;20(6):681-2.
 


Submitted date:
10/09/2022

Accepted date:
03/13/2023

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