{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,3]],"date-time":"2026-11-03T17:30:28Z","timestamp":1793727028136,"version":"4.6.1"},"reference-count":60,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2021,1,23]],"date-time":"2021-01-23T00:00:00Z","timestamp":1611360000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100014180","name":"Junta de Castilla y Le\u00f3n","doi-asserted-by":"publisher","award":["LE253P18"],"award-info":[{"award-number":["LE253P18"]}],"id":[{"id":"10.13039\/501100014180","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100010198","name":"MINECO","doi-asserted-by":"publisher","award":["AGL2017-83098-R"],"award-info":[{"award-number":["AGL2017-83098-R"]}],"id":[{"id":"10.13039\/501100010198","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The optimization of sperm cryopreservation protocols in ram is a feasible tool to reinforce artificial insemination technologies considering the desirable application of sperm by vaginal\/cervical or transcervical deposition. Cryopreservation provokes different types of damage on spermatozoa and many of these detrimental effects are triggered by redox deregulation. For this reason, the antioxidant supplementation in sperm cryopreservation protocols to decrease reactive oxygen species (ROS) levels and to equilibrate redox status has been widely employed in different species. Despite this, more fertility trials are necessary to provide the definitive tool to ensure the antioxidant effectiveness on sperm quality. For this reason, in this work, we performed a multiparametric analysis of some previously tested antioxidants (crocin, GSH and Trolox) on ram sperm cryopreservation from field trials to sperm quality analyses focused on new strategies to measure redox balance. Attending to fertility trial, Trolox supplementation registered an improvement concerning to fertility (when we considered high fertility males) and multiple lambing frequency and other complementary and descriptive data related to lambing performance such as prolificacy and fecundity. This positive effect was more evident in multiple lambing frequency when we considered low fertility males than in global male analysis. In vitro analyses of sperm quality confirmed in vivo trials registering a positive effect on sperm viability and redox balance. In this study, we provided the definitive evidence that the role of trolox on redox balance maintenance has a direct effect on fertility parameters, such as prolificacy. The effectiveness of antioxidant treatments was tested, for the first time in ovine species, using an integrative and multiparametric approach combining in vivo and in vitro analyses and novel approaches, such as RedoxSYS. These types of strategies should be applied to improve sperm conservation methods and optimize AI technologies upgrading the correlation between in vitro and in vivo analyses.<\/jats:p>","DOI":"10.3390\/ani11020283","type":"journal-article","created":{"date-parts":[[2021,1,25]],"date-time":"2021-01-25T02:07:00Z","timestamp":1611540420000},"page":"283","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":41,"title":["Multiparametric Study of Antioxidant Effect on Ram Sperm Cryopreservation\u2014From Field Trials to Research Bench"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3655-6401","authenticated-orcid":false,"given":"Marta F.","family":"Riesco","sequence":"first","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Cell Biology, Department of Molecular Biology, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mercedes","family":"Alvarez","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7179-5742","authenticated-orcid":false,"given":"Luis","family":"Anel-Lopez","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Anatomy, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta","family":"Neila-Montero","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7901-228X","authenticated-orcid":false,"given":"Cristina","family":"Palacin-Martinez","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Montes-Garrido","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan Carlos","family":"Boixo","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2531-595X","authenticated-orcid":false,"given":"Paulino","family":"de Paz","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Cell Biology, Department of Molecular Biology, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Anel","sequence":"additional","affiliation":[{"name":"Assisted Reproduction Techniques Research Group (Itra-ULE), INDEGSAL, University of Le\u00f3n, 24071 Le\u00f3n, Spain"},{"name":"Animal Reproduction and Obstetrics, Department of Veterinary Medicine, Surgery and Anatomy, University of Le\u00f3n, 24071 Le\u00f3n, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1186\/s40104-019-0395-9","article-title":"Bovine sperm selection procedure prior to cryopreservation for improvement of post-thawed semen quality and fertility","volume":"10","author":"Nagata","year":"2019","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1111\/rda.13523","article-title":"Current challenges in sheep artificial insemination: A particular insight","volume":"54","author":"Alvarez","year":"2019","journal-title":"Reprod. Domest. Anim."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"803","DOI":"10.21451\/1984-3143-AR2018-0129","article-title":"Technical recommendations for artificial insemination in sheep","volume":"16","author":"Gibbons","year":"2019","journal-title":"Anim. Reprod."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1089\/bio.2018.0113","article-title":"Spermatozoa Cryopreservation: State of Art and Future in Small Ruminants","volume":"17","author":"Lv","year":"2019","journal-title":"Biopreserv. Biobank."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3199","DOI":"10.1210\/jc.2007-2616","article-title":"Significance of Mitochondrial Reactive Oxygen Species in the Generation of Oxidative Stress in Spermatozoa","volume":"93","author":"Koppers","year":"2008","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Martin, H.D., Bragado, M.J., Batista, A.R., Oliveira, P.F., and Alves, M.G. (2019). Antioxidants and male fertility: From molecular studies to clinical evidence. Antioxidants, 8.","DOI":"10.3390\/antiox8040089"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Len, J.S., Koh, W.S.D., and Tan, S.X. (2019). The roles of reactive oxygen species and antioxidants in cryopreservation. Biosci. Rep., 39.","DOI":"10.1042\/BSR20191601"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Pe\u00f1a, F.J., Flaherty, C., Ortiz, R.J.M., Mart\u00edn, C.F.E., Gaitskell, P.G.L., Gil, M.C., Ferrusola, C.O., Flaherty, C., Ortiz, R.J.M., and Mart\u00edn, C.F.E. (2019). Redox regulation and oxidative stress: The particular case of the stallion spermatozoa. Antioxidants, 8.","DOI":"10.3390\/antiox8110567"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1002\/(SICI)1098-2795(200003)55:3<326::AID-MRD11>3.0.CO;2-A","article-title":"Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: Implications for sperm lipoperoxidative damage","volume":"55","author":"Ollero","year":"2000","journal-title":"Mol. Reprod. Dev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"4","DOI":"10.4103\/jhrs.JHRS_150_18","article-title":"Role of Oxidative Stress in Male Infertility: An Updated Review","volume":"12","author":"Alahmar","year":"2019","journal-title":"J. Hum. Reprod. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1518","DOI":"10.1016\/j.fertnstert.2014.10.020","article-title":"Male infertility testing: Reactive oxygen species and antioxidant capacity","volume":"102","author":"Ko","year":"2014","journal-title":"Fertil. Steril."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/S0015-0282(02)04948-8","article-title":"Role of reactive oxygen species in the pathophysiology of human reproduction","volume":"79","author":"Agarwal","year":"2003","journal-title":"Fertil. Steril."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/S1383-5718(01)00300-X","article-title":"Oxidative stress associated DNA damage in testis of mice: Induction of abnormal sperms and effects on fertility","volume":"513","author":"Rajesh","year":"2001","journal-title":"Mutat. Res.Genet. Toxicol. Environ. Mutagen."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"R189","DOI":"10.1530\/REP-19-0452","article-title":"Impact of oxidative stress on male and female germ cells: Implications for fertility","volume":"159","author":"Aitken","year":"2020","journal-title":"Reproduction"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1080\/2090598X.2019.1599624","article-title":"Oxidative stress and sperm function: A systematic review on evaluation and management","volume":"17","author":"Dutta","year":"2019","journal-title":"Arab J. Urol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1590\/S1677-55382007000500002","article-title":"Clinical relevance of oxidative stress and sperm chromation damage in male infertility: An evidence based analysis","volume":"33","author":"Cocuzza","year":"2007","journal-title":"Int. Braz. J. Urol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.anireprosci.2018.03.019","article-title":"Supplementation of ram semen extender to improve seminal quality and fertility rate","volume":"192","author":"Allai","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Agarwal, A., Durairajanayagam, D., and Plessis, S.S. (2014). Utility of antioxidants during assisted reproductive techniques: An evidence based review. Reprod. Biol. Endocrinol., 12.","DOI":"10.1186\/1477-7827-12-112"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1007\/s10561-016-9566-5","article-title":"The role of antioxidants in sperm freezing: A review","volume":"17","author":"Amidi","year":"2016","journal-title":"Cell Tissue Bank."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.anireprosci.2012.05.009","article-title":"Application of antioxidants and centrifugation for cryopreservation of boar spermatozoa","volume":"132","author":"Zhang","year":"2012","journal-title":"Anim. Reprod. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Peris, F.P., Soler, A.J., Iniesta, C.M., Mart\u00edn, M.A., S\u00e1nchez, A.I., Medina, C.D.A., Fern\u00e1ndez, S.M.R., Garc\u00eda, \u00c1.O., Maroto, M.A., and Montoro, V. (2020). Sperm Cryodamage in Ruminants: Understanding the Molecular Changes Induced by the Cryopreservation Process to Optimize Sperm Quality. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21082781"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.anireprosci.2015.09.012","article-title":"Reduced glutathione addition improves both the kinematics and physiological quality of post-thawed red deer sperm","volume":"162","author":"Alvarez","year":"2015","journal-title":"Anim. Reprod. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1111\/rda.12694","article-title":"The Effect of Oxidative Stress on Thawed Bulk-Sorted Red Deer Sperm","volume":"51","author":"Parrilla","year":"2016","journal-title":"Reprod. Domest. Anim."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Agarwal, A., Selvam, M.K.P., Samanta, L., Vij, S.C., Parekh, N., Sabanegh, E., Tadros, N.N., Arafa, M., and Sharma, R. (2019). Effect of antioxidant supplementation on the sperm proteome of idiopathic infertile men. Antioxidants, 8.","DOI":"10.3390\/antiox8100488"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1016\/j.theriogenology.2005.04.009","article-title":"Relationship between in vitro fertilisation of ewe oocytes and the fertility of ewes following cervical artificial insemination with frozen-thawed ram semen","volume":"64","author":"Meara","year":"2005","journal-title":"Theriogenology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.rbmo.2016.10.008","article-title":"Diagnostic application of oxidation-reduction potential assay for measurement of oxidative stress: Clinical utility in male factor infertility","volume":"34","author":"Agarwal","year":"2017","journal-title":"Reprod. Biomed. Online"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.theriogenology.2014.10.018","article-title":"Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender","volume":"83","author":"Campuzano","year":"2015","journal-title":"Theriogenology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1293","DOI":"10.1016\/S0093-691X(03)00140-7","article-title":"Field and in vitro assay of three methods for freezing ram semen","volume":"60","author":"Anel","year":"2003","journal-title":"Theriogenology"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Riesco, F.M., Lopez, A.L., Montero, N.M., Martinez, P.C., Garrido, M.R., Alvarez, M., Paz, P., and Anel, L. (2020). ProAKAP4 as novel molecular marker of sperm quality in ram: An integrative study in fresh, cooled and cryopreserved sperm. Biomolecules, 10.","DOI":"10.3390\/biom10071046"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1007\/s11250-016-1000-2","article-title":"Lambing rate and prolificacy in inseminated hair sheep treated with bovine somatotropin","volume":"48","author":"Flores","year":"2016","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Li, F., Feng, X., Yang, H., Zhu, A., Pang, J., Han, L., Zhang, T., Yao, X., and Wang, F. (2017). Genome-wide analysis of DNA Methylation profiles on sheep ovaries associated with prolificacy using whole-genome Bisulfite sequencing. BMC Genom., 18.","DOI":"10.1186\/s12864-017-4068-9"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2369","DOI":"10.1093\/humrep\/15.11.2369","article-title":"Offspring sex ratio of subfertile men and men with abnormal sperm characteristics","volume":"15","author":"Jacobsen","year":"2000","journal-title":"Hum. Reprod."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1002\/mrd.20759","article-title":"Scrotal heat stress effects on sperm viability, sperm DNA integrity, and the offspring sex ratio in mice","volume":"75","author":"Crespo","year":"2008","journal-title":"Mol. Reprod. Dev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1002\/j.1939-4640.2002.tb02324.x","article-title":"Oxidative stress and male infertility: From research bench to clinical practice","volume":"23","author":"Saleh","year":"2002","journal-title":"J. Androl."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.cryobiol.2017.10.001","article-title":"Impact of supplementation of semen extender with antioxidants on the quality of chilled or cryopreserved Arabian stallion spermatozoa","volume":"79","author":"Ghallab","year":"2017","journal-title":"Cryobiology"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.theriogenology.2016.11.020","article-title":"Resveratrol and Epigallocatechin-3-gallate addition to thawed boar sperm improves in vitro fertilization","volume":"90","author":"Gadani","year":"2017","journal-title":"Theriogenology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.anireprosci.2016.01.014","article-title":"Effect on post-cryopreserved semen characteristics of Holstein bulls of adding combinations of vitamin C and either catalase or reduced glutathione to Tris extender","volume":"167","author":"Eidan","year":"2016","journal-title":"Anim. Reprod. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.cryobiol.2017.03.002","article-title":"Antioxidant effect of quercetin in an extender containing DMA or glycerol on freezing capacity of goat semen","volume":"75","author":"Jamadi","year":"2017","journal-title":"Cryobiology"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.anireprosci.2012.09.001","article-title":"Reduced glutathione and Trolox (vitamin E) as extender supplements in cryopreservation of red deer epididymal spermatozoa","volume":"135","author":"Morales","year":"2012","journal-title":"Anim. Reprod. Sci."},{"key":"ref_40","first-page":"374","article-title":"Human semen quality and the secondary sex ratio","volume":"18","author":"Bae","year":"2016","journal-title":"Asian J. Androl."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1038\/aja.2012.109","article-title":"Sperm speed is associated with sex bias of siblings in a human population","volume":"15","author":"Mossman","year":"2013","journal-title":"Asian J. Androl."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1080\/01485010500301982","article-title":"A follow-up expanded study of the correlation of sperm velocity in seminal plasma and offspring gender","volume":"52","author":"Maligaya","year":"2006","journal-title":"Arch. Androl."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"294","DOI":"10.1002\/j.1939-4640.1997.tb01922.x","article-title":"Effects of heat stress on mouse testicular cells and sperm chromatin structure","volume":"18","author":"Sailer","year":"1997","journal-title":"J. Androl."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3692","DOI":"10.1016\/j.cub.2019.09.031","article-title":"Differential Sperm Motility Mediates the Sex Ratio Drive Shaping Mouse Sex Chromosome Evolution","volume":"29","author":"Rathje","year":"2019","journal-title":"Curr. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Santolaria, P., Palacin, I., and Yaniz, J. (2011). Management Factors Affecting Fertility in Sheep. Artificial Insemination in Farm Animals, InTech.","DOI":"10.5772\/18013"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.anireprosci.2013.04.002","article-title":"A comparative study of sperm morphometric subpopulations in cattle, goat, sheep and pigs using a computer-assisted fluorescence method (CASMA-F)","volume":"139","author":"Fiel","year":"2013","journal-title":"Anim. Reprod. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.1016\/S0093-691X(03)00174-2","article-title":"Comparison of methods to evaluate the plasmalemma of bovine sperm and their relationship with in vitro fertilization rate","volume":"60","author":"Brito","year":"2003","journal-title":"Theriogenology"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Plaza, D.M., Mu\u00f1oz, M.P., Tapia, J.A., Ortega, F.C., Silva, B.C.C., Pe\u00f1a, F.J., Davila, M.P., Mu\u00f1oz, P.M., and Tapia, J.A. (2015). Inhibition of mitochondrial complex I leads to decreased motility and membrane integrity related to increased hydrogen peroxide and reduced ATP production, while the inhibition of glycolysis has less impact on sperm motility. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0138777"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.aju.2017.11.008","article-title":"Laboratory assessment of oxidative stress in semen","volume":"16","author":"Agarwal","year":"2018","journal-title":"Arab J. Urol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1177\/1756287216652779","article-title":"Oxidation-reduction potential of semen: What is its role in the treatment of male infertility?","volume":"8","author":"Agarwal","year":"2016","journal-title":"Ther. Adv. Urol."},{"key":"ref_51","first-page":"287","article-title":"Not every sperm is sacred; a perspective on male infertility","volume":"24","author":"Aitken","year":"2018","journal-title":"Mol. Hum. Reprod."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.3168\/jds.S0022-0302(98)75739-X","article-title":"The Relationship between the Number of Spermatozoa Inseminated and the Reproductive Efficiency of Individual Dairy Bulls","volume":"81","author":"Daas","year":"1998","journal-title":"J. Dairy Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1002\/j.1939-4640.2001.tb02219.x","article-title":"Relationship of bull fertility to sperm nuclear shape","volume":"22","author":"Ostermeier","year":"2001","journal-title":"J. Androl."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1016\/j.theriogenology.2008.04.012","article-title":"Sperm morphology: Its relevance to compensable and uncompensable traits in semen","volume":"70","author":"Saacke","year":"2008","journal-title":"Theriogenology"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.anireprosci.2014.02.005","article-title":"In vitro assessment of sperm quality from rams of high and low field fertility","volume":"146","author":"Fiel","year":"2014","journal-title":"Anim. Reprod. Sci."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"31","DOI":"10.4103\/1008-682X.122203","article-title":"Oxidative stress and male reproductive health","volume":"16","author":"Aitken","year":"2014","journal-title":"Asian J. Androl."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.anireprosci.2010.08.004","article-title":"Lipid peroxidation and generation of hydrogen peroxide in frozen-thawed ram semen cryopreserved in extenders with antioxidants","volume":"122","author":"Maia","year":"2010","journal-title":"Anim. Reprod. Sci."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.cryobiol.2016.02.008","article-title":"Effect of Trolox on sperm quality in normozospermia and oligozospermia during cryopreservation","volume":"72","author":"Nekoonam","year":"2016","journal-title":"Cryobiology"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.anireprosci.2012.12.002","article-title":"Vitamin E (Trolox) addition to Tris-egg yolk extender preserves ram spermatozoon structure and kinematics after cryopreservation","volume":"137","author":"Silva","year":"2013","journal-title":"Anim. Reprod. Sci."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Zhu, Z., Fan, X., Lv, Y., Zhang, N., Fan, C., Zhang, P., and Zeng, W. (2015). Vitamin E analogue improves rabbit sperm quality during the process of cryopreservation through its antioxidative action. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0145383"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/11\/2\/283\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:14:25Z","timestamp":1760159665000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/11\/2\/283"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,23]]},"references-count":60,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2021,2]]}},"alternative-id":["ani11020283"],"URL":"https:\/\/doi.org\/10.3390\/ani11020283","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,23]]}}}