{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T05:08:29Z","timestamp":1788930509551,"version":"build-2803163510"},"reference-count":45,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100007288","name":"Mahasarakham University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007288","id-type":"DOI","asserted-by":"publisher"}]},{"id":[{"id":"https:\/\/ror.org\/0453j3c58","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Bull semen cryopreservation is essential for preserving valuable genetics and supporting cattle breeding programs; however, freeze\u2013thaw stress compromises post-thaw sperm quality through oxidative damage and membrane injury. This study evaluated the effects of L-serine supplementation on post-thaw semen quality, susceptibility to induced lipid peroxidation, and antioxidant enzyme activity in bull semen. Semen collected from four Brahman bulls was cryopreserved in a Tris-based extender supplemented with 0, 2, 4, or 6 mM L-serine. Post-thaw total motility, progressive motility, viability, plasma membrane integrity, MDA concentration, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were assessed. Supplementation with 2 and 4 mM L-serine resulted in higher total motility and progressive motility, and greater plasma membrane integrity than the control and 6 mM treatments (p &lt; 0.05), while viability was higher with 4 mM than with the control and 6 mM treatments (p &lt; 0.05). MDA concentration following induced lipid peroxidation was lower in all L-serine-supplemented treatments than in the control (p &lt; 0.05). SOD activity was higher with 2 mM than with the control and 6 mM treatments (p &lt; 0.05), whereas GPx activity was highest with 2 mM, followed by 4 and 6 mM, with the lowest activity in the control (p &lt; 0.05). CAT activity was unaffected by treatment (p = 0.1192). Overall, supplementation with 2\u20134 mM L-serine improved selected post-thaw sperm quality parameters and was associated with reduced susceptibility to induced lipid peroxidation and changes in apparent extracellular antioxidant enzyme activities. These findings support the potential of L-serine as a functional additive to improve bull semen cryopreservation.<\/jats:p>","DOI":"10.3390\/ani16172809","type":"journal-article","created":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T13:53:28Z","timestamp":1788789208000},"page":"2809","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Influence of L-Serine on Sperm Quality, Malondialdehyde, and Antioxidant Enzyme Activity After Cryopreservation in Bull Semen"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3467-3134","authenticated-orcid":false,"given":"Ruthaiporn","family":"Ratchamak","sequence":"first","affiliation":[{"name":"Division of Animal Science, Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Maha Sarakham 44150, Thailand"},{"name":"The Network Center for Animal Breeding and Omics Research, Khon Kaen University, Khon Kaen 40002, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khomsan","family":"Buathalad","sequence":"additional","affiliation":[{"name":"Division of Animal Science, Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Maha Sarakham 44150, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maharach","family":"Matra","sequence":"additional","affiliation":[{"name":"Division of Animal Science, Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Maha Sarakham 44150, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4100-7570","authenticated-orcid":false,"given":"Eakapol","family":"Wangkahart","sequence":"additional","affiliation":[{"name":"Division of Fisheries, Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Maha Sarakham 44150, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2239-2300","authenticated-orcid":false,"given":"Wuttigrai","family":"Boonkum","sequence":"additional","affiliation":[{"name":"The Network Center for Animal Breeding and Omics Research, Khon Kaen University, Khon Kaen 40002, Thailand"},{"name":"Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3619-8680","authenticated-orcid":false,"given":"Vibuntita","family":"Chankitisakul","sequence":"additional","affiliation":[{"name":"The Network Center for Animal Breeding and Omics Research, Khon Kaen University, Khon Kaen 40002, Thailand"},{"name":"Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen 40002, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,7]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ugur, M.R., Saber Abdelrahman, A., Evans, H.C., Gilmore, A.A., Hitit, M., Arifiantini, R.I., Purwantara, B., Kaya, A., and Memili, E. (2019). Advances in Cryopreservation of Bull Sperm. Front. Vet. Sci., 6.","DOI":"10.3389\/fvets.2019.00268"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Capela, L., Leites, I., Rom\u00e3o, R., Lopes-da-Costa, L., and Pereira, R.M.L.N. (2022). Impact of Heat Stress on Bovine Sperm Quality and Competence. Animals, 12.","DOI":"10.3390\/ani12080975"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.ijvsm.2017.11.001","article-title":"Evaluation of bull spermatozoa during and after cryopreservation: Structural and ultrastructural insights","volume":"6","author":"Khalil","year":"2017","journal-title":"Int. J. Vet. Sci. Med."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Castro, M., Leal, K., Pezo, F., and Contreras, M.J. (2025). Sperm Membrane: Molecular Implications and Strategies for Cryopreservation in Productive Species. Animals, 15.","DOI":"10.3390\/ani15121808"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"703","DOI":"10.21451\/1984-3143-AR2018-0041","article-title":"Impacts of Oxidative Stress on Bovine Sperm Function and Subsequent In Vitro Embryo Development","volume":"15","author":"Bollwein","year":"2018","journal-title":"Anim. Reprod."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ugur, M.R., Dinh, T., Hitit, M., Kaya, A., Topper, E., Didion, B., and Memili, E. (2020). Amino Acids of Seminal Plasma Associated with Freezability of Bull Sperm. Front. Cell Dev. Biol., 7.","DOI":"10.3389\/fcell.2019.00347"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Li, S., Ren, J., Zhang, W., Wang, B., Ma, Y., Su, L., Dai, Y., and Liu, G. (2023). Glutathione and Selenium Nanoparticles Have a Synergistic Protective Effect during Cryopreservation of Bull Semen. Front. Vet. Sci., 10.","DOI":"10.3389\/fvets.2023.1093274"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.anireprosci.2018.08.024","article-title":"Reduction of dissolved oxygen in semen extender with nitrogen gassing reduces oxidative stress and improves post-thaw semen quality of bulls","volume":"197","author":"Kumar","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_9","first-page":"71","article-title":"Status of Prooxidant and Antioxidant Systems in the Sperm and Seminal Plasma of Breeding Boars of Large White Breed and SS23 Synthetic Line","volume":"73","author":"Polishchuk","year":"2022","journal-title":"J. Physiol. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"10027","DOI":"10.3168\/jds.2024-24872","article-title":"Application of antioxidants in extender on bull sperm cryopreservation to reduce the male effect in dairy fertility","volume":"107","author":"Shepherd","year":"2024","journal-title":"J. Dairy Sci."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Gonzalez, M., Prashar, T., Connaughton, H., Barry, M., Robker, R., and Rose, R. (2022). Restoring Sperm Quality Post-Cryopreservation Using Mitochondrial-Targeted Compounds. Antioxidants, 11.","DOI":"10.3390\/antiox11091808"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"13","DOI":"10.17221\/104\/2020-CJAS","article-title":"Effect of Semen Extender Supplementation with Sericin on Post-Thaw Dairy Bull Sperm Quality and Lipid Peroxidation","volume":"66","author":"Yangngam","year":"2021","journal-title":"Czech J. Anim. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Authaida, S., Boonkum, W., and Chankitisakul, V. (2025). Enhancement of Semen Cryopreservation from Native Thai Bulls Through Moringa oleifera Leaf Extract Supplementation. Animals, 15.","DOI":"10.3390\/ani15030439"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Loetjettanarom, T., Authaida, S., Boonkum, W., and Chankitisakul, V. (2025). Effect of Kaempferia parviflora Supplementation in Semen Extenders on Post-Thaw Sperm Quality in Thai Native Bulls. Animals, 15.","DOI":"10.3390\/ani15070962"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2517","DOI":"10.14202\/vetworld.2022.2517-2524","article-title":"Effect of Supplementing L-Cysteine and Its Group Analogs on Frozen Semen Quality of Bulls: A Meta-Analysis","volume":"15","author":"Hayanti","year":"2022","journal-title":"Vet. World"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1700262","DOI":"10.1002\/mnfr.201700262","article-title":"Serine Alleviates Oxidative Stress via Supporting Glutathione Synthesis and Methionine Cycle in Mice","volume":"61","author":"Zhou","year":"2017","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/j.tem.2023.03.001","article-title":"Serine Signaling Governs Metabolic Homeostasis and Health","volume":"34","author":"He","year":"2023","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1185","DOI":"10.1016\/j.psj.2019.10.040","article-title":"Different Concentrations of Cysteamine, Ergothioneine, and Serine Modulate Quality and Fertilizing Ability of Cryopreserved Chicken Sperm","volume":"99","author":"Thananurak","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Chankitisakul, V., Saiyamanon, H., Boonkum, W., Wangkahart, E., and Ratchamak, R. (2025). Dose-Dependent Effects of L-Serine Supplementation on Boar Sperm Quality During Chilled and Cryopreserved Storage. Animals, 15.","DOI":"10.3390\/ani15182670"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.anireprosci.2018.05.012","article-title":"Evaluation of cooling and freezing systems of bovine semen","volume":"195","author":"Dias","year":"2018","journal-title":"Anim. Reprod. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1111\/rda.13121","article-title":"Investigation of in vitro measurable sperm attributes and their influence on electroejaculated bull semen with a fixed-time artificial insemination protocol in Australian Bos indicus cattle","volume":"53","author":"Satake","year":"2018","journal-title":"Reprod. Domest. Anim."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1016\/j.theriogenology.2016.10.008","article-title":"Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility","volume":"89","author":"Campanholi","year":"2017","journal-title":"Theriogenology"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Montoya-Monsalve, G., S\u00e1nchez-Calabuig, M.-J., Blanco-Murcia, J., Elvira, L., Guti\u00e9rrez-Ad\u00e1n, A., Ramos-Ibeas, P., and Campanile, G. (2021). Impact of Overuse and Sexually Transmitted Infections on Seminal Parameters of Extensively Managed Bulls. Animals, 11.","DOI":"10.3390\/ani11030827"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Elkhawagah, A.R., Ricci, A., Bertero, A., Poletto, M.L., Nervo, T., Donato, G.G., Vincenti, L., and Martino, N.A. (2024). Supplementation with MitoTEMPO before cryopreservation improves sperm quality and fertility potential of Piedmontese beef bull semen. Front. Vet. Sci., 11.","DOI":"10.3389\/fvets.2024.1376057"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ratchamak, R., Authaida, S., Koedkanmark, T., Boonkum, W., Semaming, Y., and Chankitisakul, V. (2023). Supplementation of Freezing Medium with Ginseng Improves Rooster Sperm Quality and Fertility Relative to Free Radicals and Antioxidant Enzymes. Animals, 13.","DOI":"10.3390\/ani13162660"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e12970","DOI":"10.1111\/and.12970","article-title":"Study on Correlation of Sperm Quality Parameters with Antioxidant and Oxidant Status of Buffalo Bull Semen During Various Stages of Cryopreservation","volume":"50","author":"Lone","year":"2018","journal-title":"Andrologia"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1016\/j.cryobiol.2010.09.009","article-title":"The Effect of Cysteine and Glutathione on Sperm and Oxidative Stress Parameters of Post-Thawed Bull Semen","volume":"61","author":"Tuncer","year":"2010","journal-title":"Cryobiology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1007\/s10561-013-9412-y","article-title":"Effects of in Combination Antioxidant Supplementation on Microscopic and Oxidative Parameters of Freeze-Thaw Bull Sperm","volume":"15","author":"Ashrafi","year":"2014","journal-title":"Cell Tissue Bank."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"98","DOI":"10.5424\/sjar\/2014121-4486","article-title":"Effect of Rosemary (Rosmarinus officinalis) Extracts and Glutathione Antioxidants on Bull Semen Quality after Cryopreservation","volume":"12","author":"Hoseinkhani","year":"2014","journal-title":"Span. J. Agric. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1111\/and.12236","article-title":"Antioxidative Effects of Cysteamine, Hyaluronan and Fetuin on Post-Thaw Semen Quality, DNA Integrity and Oxidative Stress Parameters in the Brown Swiss Bull","volume":"47","author":"Tuncer","year":"2015","journal-title":"Andrologia"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e0401","DOI":"10.5424\/sjar\/2017151-9761","article-title":"Alpha-Tocopherol Improves Frozen-Thawed Sperm Quality by Reducing Hydrogen Peroxide during Cryopreservation of Bull Semen","volume":"15","author":"Motemani","year":"2017","journal-title":"Span. J. Agric. Res."},{"key":"ref_32","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_33","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1083\/jcb.201604085","article-title":"The importance of serine metabolism in cancer","volume":"214","author":"Mattaini","year":"2016","journal-title":"J. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1016\/j.theriogenology.2012.05.002","article-title":"Effects of reactive oxygen species on sperm function","volume":"78","author":"Guthrie","year":"2012","journal-title":"Theriogenology"},{"key":"ref_35","first-page":"622","article-title":"Polyunsaturated Fatty Acids in Male Ruminant Reproduction\u2014A Review","volume":"30","author":"Malla","year":"2017","journal-title":"Asian-Australas. J. Anim. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wang, Y., Fu, X., and Li, H. (2025). Mechanisms of oxidative stress-induced sperm dysfunction. Front. Endocrinol., 16.","DOI":"10.3389\/fendo.2025.1520835"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"385","DOI":"10.5713\/ab.22.0325","article-title":"Role of antioxidants in fertility preservation of sperm\u2014A narrative review","volume":"36","author":"Qamar","year":"2023","journal-title":"Anim. Biosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"14804","DOI":"10.1074\/jbc.M117.788901","article-title":"Hydrogen peroxide produced by superoxide dismutase SOD-2 activates sperm in Caenorhabditis elegans","volume":"292","author":"Sakamoto","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1002\/(SICI)1098-2795(200003)55:3<282::AID-MRD6>3.0.CO;2-7","article-title":"Levels of antioxidant defenses are decreased in bovine spermatozoa after a cycle of freezing and thawing","volume":"55","author":"Bilodeau","year":"2000","journal-title":"Mol. Reprod. Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.anireprosci.2015.02.006","article-title":"Inter- and intra-individual variability of total antioxidant capacity of bovine seminal plasma and relationships with sperm quality before and after cryopreservation","volume":"155","author":"Calisici","year":"2015","journal-title":"Anim. Reprod. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"102648","DOI":"10.1016\/j.psj.2023.102648","article-title":"Solid Storage Supplemented with Serine of Rooster Semen Enhances Higher Sperm Quality and Fertility Potential during Storage at 5 \u00b0C for up to 120 h","volume":"102","author":"Kheawkanha","year":"2023","journal-title":"Poult. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Valencia, J., and Javier Henao, F. (2018). Biological Signals of Sperm Membrane Resistance to Cryoinjury in Boars. Cryopreservation Biotechnology in Biomedical and Biological Sciences, IntechOpen.","DOI":"10.5772\/intechopen.80824"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"106884","DOI":"10.1016\/j.anireprosci.2021.106884","article-title":"\u201cFine feathers make fine birds\u201d\u2014The mammalian sperm plasma membrane lipid composition and effects on assisted reproduction","volume":"246","author":"Gautier","year":"2022","journal-title":"Anim. Reprod. Sci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.cmet.2016.08.009","article-title":"One-carbon metabolism in health and disease","volume":"25","author":"Ducker","year":"2017","journal-title":"Cell Metab."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/j.plipres.2008.01.003","article-title":"Roles of l-serine and sphingolipid synthesis in brain development and neuronal survival","volume":"47","author":"Hirabayashi","year":"2008","journal-title":"Prog. Lipid Res."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2809\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T04:29:40Z","timestamp":1788928180000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/17\/2809"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,7]]},"references-count":45,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ani16172809"],"URL":"https:\/\/doi.org\/10.3390\/ani16172809","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,7]]}}}