{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,28]],"date-time":"2026-03-28T03:54:50Z","timestamp":1774670090593,"version":"3.50.1"},"reference-count":61,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,7,8]],"date-time":"2023-07-08T00:00:00Z","timestamp":1688774400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["UIDP\/CVT\/00772\/2020"],"award-info":[{"award-number":["UIDP\/CVT\/00772\/2020"]}]},{"name":"Portuguese Foundation for Science and Technology (FCT)","award":["LA\/P\/0059\/2020"],"award-info":[{"award-number":["LA\/P\/0059\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>The present study aimed to evaluate the effects of melatonin (MLT), glutathione (GSH), and their combination on ram semen quality after thawing. During eight weekly sessions, semen from three Merino rams was pooled, diluted with an egg-yolk-based semen extender, and divided into four groups: control, 1 mM MLT, 5 mM GSH, and 1 mM MLT + 5 mM GSH. Diluted semen was evaluated before and after the freezing process. The supplementation of diluted semen with GSH at 5 mM had a deleterious effect on total motility progressive (TPM), curvilinear velocity (VCL), straight-line velocity (VSL), average-path velocity (VAP), linearity (LIN), and straightness (STR) and increased slow spermatozoa (%). MLT at 1 mM also had a negative effect on TPM, VSL, and STR in diluted semen. In thawed semen, 1 mM MLT increased the TPM compared with the control group. VSL was lower in the 5 mM GSH group than in the 1 mM MLT group. Additionally, the combination of both antioxidants attenuated the negative effect of 5 mM GSH on TPM, VSL, and BCF. These results indicate that 5 mM GSH impairs or does not improve sperm kinetic parameters in either diluted or thawed semen. They also suggest that MLT combined with GSH plays a protective role against these effects.<\/jats:p>","DOI":"10.3390\/vetsci10070446","type":"journal-article","created":{"date-parts":[[2023,7,10]],"date-time":"2023-07-10T00:54:23Z","timestamp":1688950463000},"page":"446","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Effect of In Vitro Addition of Melatonin and Glutathione on Seminal Parameters of Rams in Diluted Semen and after Thawing"],"prefix":"10.3390","volume":"10","author":[{"given":"Cristiana","family":"Carri\u00e7o","sequence":"first","affiliation":[{"name":"Department of Veterinary Sciences, Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), School of Agricultural and Veterinary Sciences, University of Tr\u00e1s-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal"}]},{"given":"Jo\u00e3o Pedro","family":"Barbas","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Genetic Resources of Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV), Quinta da Fonte Boa, 2005-048 Vale de Santar\u00e9m, Portugal"},{"name":"CIISA\u2014AL4AnimalS, Faculty of Veterinary Medicine, University of Lisbon, 1649-004 Lisboa, Portugal"}]},{"given":"Jorge","family":"Pimenta","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Genetic Resources of Instituto Nacional de Investiga\u00e7\u00e3o Agr\u00e1ria e Veterin\u00e1ria (INIAV), Quinta da Fonte Boa, 2005-048 Vale de Santar\u00e9m, Portugal"},{"name":"CIISA\u2014AL4AnimalS, Faculty of Veterinary Medicine, University of Lisbon, 1649-004 Lisboa, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4997-3933","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"Department of Veterinary Sciences, Animal and Veterinary Research Centre (CECAV), Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), School of Agricultural and Veterinary Sciences, University of Tr\u00e1s-os-Montes and Alto Douro (UTAD), 5000-801 Vila Real, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1079\/PAVSNNR201914052","article-title":"Recent Trends in Bovine Reproductive Biotechnologies","volume":"14","author":"Binyameen","year":"2019","journal-title":"CAB Rev."},{"key":"ref_2","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_3","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/S0378-4320(00)00099-3","article-title":"The Causes of Reduced Fertility with Cryopreserved Semen","volume":"60\u201361","author":"Watson","year":"2000","journal-title":"Anim. Reprod. Sci."},{"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. Biobanking"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"241","DOI":"10.4103\/2305-0500.217317","article-title":"Melatonin as an Antioxidant Preserving Sperm from Domestic Animals","volume":"6","author":"Medrano","year":"2017","journal-title":"Asian Pac. J. Reprod."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e13577","DOI":"10.1111\/and.13577","article-title":"Reactive Oxygen Species in Male Reproduction: A Boon or a Bane?","volume":"53","author":"Baskaran","year":"2021","journal-title":"Andrologia"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Agarwal, A., Aitken, R.J., and Alvarez, J.G. (2012). Studies on Men\u2019s Health and Fertility, Humana Press. Oxidative Stress in Applied Basic Research and Clinical Practice.","DOI":"10.1007\/978-1-61779-776-7"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"107215","DOI":"10.1016\/j.anireprosci.2023.107215","article-title":"A Review of the Use of Antioxidants in Bovine Sperm Preparation Protocols","volume":"251","author":"Sapanidou","year":"2023","journal-title":"Anim. Reprod. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"797","DOI":"10.3906\/vet-1603-76","article-title":"Influence of Season on Seminal Antioxidant Enzymes in Karan Friesbulls under Tropical Climatic Conditions","volume":"40","author":"Soren","year":"2016","journal-title":"Turk. J. Vet. Anim. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"459","DOI":"10.2164\/jandrol.107.003459","article-title":"Effect of the Cryopreservation Process on the Activity and Immunolocalization of Antioxidant Enzymes in Ram Spermatozoa","volume":"29","author":"Marti","year":"2008","journal-title":"J. Androl."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"690","DOI":"10.1016\/j.theriogenology.2003.11.013","article-title":"Decrease in Glutathione Content in Boar Sperm after Cryopreservation","volume":"62","author":"Gadea","year":"2004","journal-title":"Theriogenology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e13236","DOI":"10.1111\/and.13236","article-title":"Effect of Addition of Melatonin on Liquid Storage of Ram Semen at 4 \u00b0C","volume":"51","author":"Dai","year":"2019","journal-title":"Andrologia"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.repbio.2013.01.174","article-title":"Supplementation of the Thawing Medium with Reduced Glutathione Improves Function of Frozen-Thawed Goat Spermatozoa","volume":"13","author":"Gadea","year":"2013","journal-title":"Reprod. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.cryobiol.2020.11.007","article-title":"Antioxidants and Their Effect on the Oxidative\/Nitrosative Stress of Frozen-Thawed Boar Sperm","volume":"98","author":"Pezo","year":"2021","journal-title":"Cryobiology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.smallrumres.2007.09.002","article-title":"Effect of Anti-Oxidants and Oxidative Stress Parameters on Ram Semen after the Freeze\u2013Thawing Process","volume":"75","author":"Bucak","year":"2008","journal-title":"Small Rumin. Res."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e13145","DOI":"10.1111\/jfbc.13145","article-title":"Understanding Oxidants and Antioxidants: Classical Team with New Players","volume":"44","author":"Ali","year":"2020","journal-title":"J. Food Biochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.cryobiol.2010.12.001","article-title":"Reduced Glutathione Content in Human Sperm Is Decreased after Cryopreservation: Effect of the Addition of Reduced Glutathione to the Freezing and Thawing Extenders","volume":"62","author":"Gadea","year":"2011","journal-title":"Cryobiology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"771440","DOI":"10.3389\/fvets.2021.771440","article-title":"Effect of Glutathione on Sperm Quality in Guanzhong Dairy Goat Sperm During Cryopreservation","volume":"8","author":"Zou","year":"2021","journal-title":"Front. Vet. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"106139","DOI":"10.1016\/j.smallrumres.2020.106139","article-title":"Effects of Reduced Glutathione on Ram Sperm Parameters, Antioxidant Status, Mitochondrial Activity and the Abundance of Hexose Transporters during Liquid Storage at 5 \u00b0C","volume":"189","author":"Shi","year":"2020","journal-title":"Small Rumin. Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Y\u00e1nez-Ortiz, I., Catal\u00e1n, J., Delgado-Berm\u00fadez, A., Carluccio, A., Mir\u00f3, J., and Yeste, M. (2021). Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm. Vet. Sci., 8.","DOI":"10.3390\/vetsci8120302"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1016\/j.jevs.2013.05.001","article-title":"Addition of Glutathione to an Extender for Frozen Equine Semen","volume":"33","author":"Wolf","year":"2013","journal-title":"J. Equine Vet. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.cryobiol.2016.02.001","article-title":"Effect of Reduced Glutathione (GSH) in Canine Sperm Cryopreservation: In Vitro and in Vivo Evaluation","volume":"72","author":"Lucio","year":"2016","journal-title":"Cryobiology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/j.theriogenology.2014.10.002","article-title":"Combining Reduced Glutathione and Ascorbic Acid Has Supplementary Beneficial Effects on Boar Sperm Cryotolerance","volume":"83","author":"Giaretta","year":"2015","journal-title":"Theriogenology"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.anireprosci.2017.05.006","article-title":"Effect of Reduced Glutathione Supplementation in Semen Extender on Tyrosine Phosphorylation and Apoptosis like Changes in Frozen Thawed Hariana Bull Spermatozoa","volume":"182","author":"Shah","year":"2017","journal-title":"Anim. Reprod. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1111\/jpi.12360","article-title":"Melatonin as an Antioxidant: Under Promises but over Delivers","volume":"61","author":"Reiter","year":"2016","journal-title":"J. Pineal Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Minucci, S., and Venditti, M. (2022). New Insight on the In Vitro Effects of Melatonin in Preserving Human Sperm Quality. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23095128"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1111\/rda.13851","article-title":"Melatonin Supplementation Improved Cryopreserved Thai Swamp Buffalo Semen","volume":"56","author":"Inyawilert","year":"2021","journal-title":"Reprod. Domest. Anim."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.cryobiol.2019.05.001","article-title":"Antioxidative Effect of Melatonin on Cryopreserved Chicken Semen","volume":"89","author":"Appiah","year":"2019","journal-title":"Cryobiology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1111\/rda.13853","article-title":"Melatonin Can Improve Viability and Functional Integrity of Cooled and Frozen\/Thawed Rabbit Spermatozoa","volume":"56","author":"Fadl","year":"2021","journal-title":"Reprod. Domest. Anim."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1530\/RAF-20-0022","article-title":"Melatonin Added to Freezing Diluent Improves Canine (Bulldog) Sperm Cryosurvival","volume":"1","author":"Carbajal","year":"2020","journal-title":"Reprod. Fertil."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"87","DOI":"10.5653\/cerm.2022.05267","article-title":"Melatonin and Selenium Supplementation in Extenders Improves the Post-Thaw Quality Parameters of Rat Sperm","volume":"49","author":"Shahandeh","year":"2022","journal-title":"Clin. Exp. Reprod. Med."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"990","DOI":"10.1210\/er.2018-00084","article-title":"Melatonin as a Hormone: New Physiological and Clinical Insights","volume":"39","author":"Amaral","year":"2018","journal-title":"Endocr. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1046\/j.1600-079X.2003.00092.x","article-title":"Regulation of Antioxidant Enzymes: A Significant Role for Melatonin","volume":"36","author":"Rodriguez","year":"2004","journal-title":"J. Pineal Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"106850","DOI":"10.1016\/j.anireprosci.2021.106850","article-title":"Use of Melatonin in Sperm Cryopreservation of Farm Animals: A Brief Review","volume":"233","author":"Ofosu","year":"2021","journal-title":"Anim. Reprod. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Manuela Dr\u0103goi, C., and Crengu\u0163a Nicolae, A. (2018). Melatonin-Molecular Biology, Clinical and Pharmaceutical Approaches, IntechOpen.","DOI":"10.5772\/intechopen.74993"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Barbas, J.P., Pimenta, J., Baptista, M.C., Marques, C.C., Pereira, R.M.L.N., Carolino, N., and Sim\u00f5es, J. (2023). Ram Semen Cryopreservation for Portuguese Native Breeds: Season and Breed Effects on Semen Quality Variation. Animals, 13.","DOI":"10.3390\/ani13040579"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1017\/S0021859600024874","article-title":"Subjective Assessment of Body Fat in Live Sheep","volume":"72","author":"Russel","year":"1969","journal-title":"J. Agric. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Fernandes, M., Hern\u00e1ndez, P.R., Sim\u00f5es, J., and Barbas, J.P. (2021). Effects of Three Semen Extenders, Breeding Season Month and Freezing\u2013Thawing Cycle on Spermatozoa Preservation of Portuguese Merino Sheep. Animals, 11.","DOI":"10.3390\/ani11092619"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1111\/j.1600-079X.2010.00843.x","article-title":"Melatonin Protects Ram Spermatozoa from Cryopreservation Injuries in a Dose-Dependent Manner: Melatonin Mitigates Cryopreservation Injuries of Ram Spermatozoa","volume":"50","author":"Succu","year":"2011","journal-title":"J. Pineal Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e14304","DOI":"10.1111\/and.14304","article-title":"Melatonin and Canthaxanthin Enhances Sperm Viability and Protect Ram Spermatozoa from Oxidative Stress during Liquid Storage at 4 \u00b0C","volume":"54","author":"Bhalothia","year":"2022","journal-title":"Andrologia"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1007\/s11250-022-03135-5","article-title":"Shielding Effect of Melatonin Improves Seminal Quality and Oxidative Stress Indices during Chilled Storage of Ram Semen","volume":"54","author":"Kumar","year":"2022","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1111\/j.1439-0531.2011.01758.x","article-title":"In Vitro and In Vivo Evaluation of Ram Sperm Frozen in Tris Egg-Yolk and Supplemented with Superoxide Dismutase and Reduced Glutathione","volume":"46","author":"Silva","year":"2011","journal-title":"Reprod. Dom. Anim."},{"key":"ref_43","unstructured":"(2023, June 23). Proiser: The Global Solution in Automatic Semen Analysis. Available online: https:\/\/proiser.com\/home\/."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"106904","DOI":"10.1016\/j.anireprosci.2021.106904","article-title":"Advances in Sperm Cryopreservation in Farm Animals: Cattle, Horse, Pig and Sheep","volume":"246","author":"Yeste","year":"2022","journal-title":"Anim. Reprod. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.theriogenology.2013.09.004","article-title":"Computer-Assisted Sperm Analysis (CASA): Capabilities and Potential Developments","volume":"81","author":"Amann","year":"2014","journal-title":"Theriogenology"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1002\/vms3.224","article-title":"Effect of Melatonin on Bovine Sperm Characteristics and Ultrastructure Changes Following Cryopreservation","volume":"6","author":"ChaithraShree","year":"2020","journal-title":"Vet. Med. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.cryobiol.2016.12.008","article-title":"Use of Antioxidants Reduce Lipid Peroxidation and Improve Quality of Crossbred Ram Sperm during Its Cryopreservation","volume":"74","author":"Banday","year":"2017","journal-title":"Cryobiology"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.smallrumres.2012.12.015","article-title":"Effect of Glutathione in Soybean Lecithin-Based Semen Extender on Goat Semen Quality after Freeze-Thawing","volume":"112","author":"Salmani","year":"2013","journal-title":"Small Rumin. Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.smallrumres.2016.01.009","article-title":"Effects of Low Molecular Weight Cryoprotectants on the Post-Thaw Ram Sperm Quality and Fertilizing Ability","volume":"136","author":"Alcay","year":"2016","journal-title":"Small Rumin. Res."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1089\/bio.2020.0056","article-title":"Cryopreservation Effects on Ram Sperm Ultrastructure","volume":"18","author":"Keskin","year":"2020","journal-title":"Biopreserv. Biobanking"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"e1569","DOI":"10.1002\/wsbm.1569","article-title":"Energy Metabolism in Mammalian Sperm Motility","volume":"14","author":"Amaral","year":"2022","journal-title":"WIREs Mech. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Van De Hoek, M., Rickard, J.P., and De Graaf, S.P. (2022). Motility Assessment of Ram Spermatozoa. Biology, 11.","DOI":"10.3390\/biology11121715"},{"key":"ref_53","first-page":"235","article-title":"Effect of Exogenous Glutathione in Extender on the Freezability of Nili-Ravi Buffalo (Bubalus Bubalis) Bull Spermatozoa","volume":"28","author":"Ansari","year":"2010","journal-title":"Anim. Sci. Pap. Rep."},{"key":"ref_54","first-page":"6670","article-title":"Protective Effect of Melatonin on Sperm Motility Parameters on Liquid Storage of Ram Semen at 5 \u00b0C","volume":"10","author":"Ashrafi","year":"2011","journal-title":"Afr. J. Biotechnol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"106516","DOI":"10.1016\/j.domaniend.2020.106516","article-title":"Melatonin Improves the Motility and DNA Integrity of Frozen-Thawed Ram Spermatozoa Likely via Suppression of Mitochondrial Superoxide Production","volume":"74","author":"Pool","year":"2021","journal-title":"Domest. Anim. Endocrinol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"24","DOI":"10.58292\/ct.v13.9359","article-title":"Andrology Laboratory Review: Evaluation of Sperm Motility","volume":"13","author":"Peter","year":"2021","journal-title":"Clin. Theriogenol."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Riesco, M.F., Alvarez, M., Anel-Lopez, L., Neila-Montero, M., Palacin-Martinez, C., Montes-Garrido, R., Boixo, J.C., De Paz, P., and Anel, L. (2021). Multiparametric Study of Antioxidant Effect on Ram Sperm Cryopreservation\u2014From Field Trials to Research Bench. Animals, 11.","DOI":"10.3390\/ani11020283"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.theriogenology.2022.11.035","article-title":"The Effects of Melatonin, Glutathione and Vitamin E on Semen Cryopreservation of Mediterranean Buffalo","volume":"197","author":"Luo","year":"2023","journal-title":"Theriogenology"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.reprotox.2018.02.005","article-title":"Melatonin Attenuates \u0394 9-Tetrahydrocannabinol-Induced Reduction in Rat Sperm Motility and Kinematics In-Vitro","volume":"77","author":"Alagbonsi","year":"2018","journal-title":"Reprod. Toxicol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1016\/j.anireprosci.2009.05.014","article-title":"Osmotic Stress Stimulates Generation of Superoxide Anion by Spermatozoa in Horses","volume":"117","author":"Burnaugh","year":"2010","journal-title":"Anim. Reprod. Sci."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Va\u0161\u00ed\u010dek, J., Bal\u00e1\u017ei, A., Svoradov\u00e1, A., Vozaf, J., Duj\u00ed\u010dkov\u00e1, L., Makarevich, A.V., Bauer, M., and Chrenek, P. (2022). Comprehensive Flow-Cytometric Quality Assessment of Ram Sperm Intended for Gene Banking Using Standard and Novel Fertility Biomarkers. IJMS, 23.","DOI":"10.3390\/ijms23115920"}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/10\/7\/446\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:08:54Z","timestamp":1760126934000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/10\/7\/446"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,8]]},"references-count":61,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,7]]}},"alternative-id":["vetsci10070446"],"URL":"https:\/\/doi.org\/10.3390\/vetsci10070446","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,8]]}}}