{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T16:12:19Z","timestamp":1791994339359,"version":"4.4.0"},"reference-count":54,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T00:00:00Z","timestamp":1769904000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Cryopreservation of ram semen is an essential tool in assisted reproductive technology; however, oxidative stress generated during the freezing process may compromise sperm quality. This study evaluated the effects of Se and SeNPs on post-thaw sperm quality, PRDX5 expression, and oxidative status in cryopreserved ram semen. In this study, semen samples collected from five mature rams (three collections at 2-week intervals, yielding a total of 15 ejaculates) were frozen in liquid nitrogen using extenders supplemented with selenium (1 \u03bcg\/mL, S1; 10 \u03bcg\/mL, S2) or selenium nanoparticles (SeNPs; 1 \u03bcg\/mL, N1; 2 \u03bcg\/mL, N2) alongside a nonsupplemented control extender. Post-thaw sperm quality was evaluated using computer-assisted sperm analysis (CASA) for motility, kinematic parameters, viability, membrane integrity (HOST) assays, chromatin condensation assessment, and morphological analysis. Total oxidant status (TOS) measurements and PRDX5 gene expression analysis were performed separately. Low-dose SeNPs (1 \u00b5g\/mL) significantly improved total motility (55.73 \u00b1 19.01%), progressive motility (25.05 \u00b1 15.34%), viability (57.27 \u00b1 19.30%), HOST-positive spermatozoa (50.87 \u00b1 18.91%), and morphologically normal spermatozoa (88.27 \u00b1 4.10%) compared with the control and high-dose sodium selenite groups (p &lt; 0.05). Chromatin condensation abnormalities were lowest in the SeNP-treated group. S1 and N2 also improved motility and morphology compared with the control; however, the increases were numerically smaller than those observed in the N1 group. In contrast, S2 supplementation showed limited benefit, with values that were similar to those of the control. Morphologically normal spermatozoa were highest in N1, followed by S1 and N2, while S2 and the control exhibited the lowest values (p &lt; 0.05). In contrast, no significant differences were detected in TOS or PRDX5 gene expression among the experimental groups (p &gt; 0.05). These findings indicate that low-dose SeNPs enhance post-thaw sperm functional integrity and cryotolerance without inducing measurable changes in bulk oxidative markers or gene transcription.<\/jats:p>","DOI":"10.3390\/ani16030457","type":"journal-article","created":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T14:12:46Z","timestamp":1770041566000},"page":"457","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Effects of Selenium Nanoparticles and Sodium Selenite Supplementation on Cryopreserved Ram Sperm Quality, Oxidative Status, and PRDX5 Gene Expression"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0666-5359","authenticated-orcid":false,"given":"Cumali","family":"Kaya","sequence":"first","affiliation":[{"name":"Department of Animal Reproduction and Artificial Insemination, University of Ondokuz Mayis, 55200 Samsun, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7904-0459","authenticated-orcid":false,"given":"Cansu","family":"Can","sequence":"additional","affiliation":[{"name":"Department of Medical Biology, Faculty of Medicine, University of Ondokuz Mayis, 55200 Samsun, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5728-1478","authenticated-orcid":false,"given":"Burcu","family":"Esin","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction and Artificial Insemination, University of Ondokuz Mayis, 55200 Samsun, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0230-8968","authenticated-orcid":false,"given":"Emre","family":"D\u00fcnder","sequence":"additional","affiliation":[{"name":"Department of Statistics, Faculty of Science, University of Ondokuz Mayis, 55200 Samsun, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0754-6116","authenticated-orcid":false,"given":"Mesut","family":"\u00c7evik","sequence":"additional","affiliation":[{"name":"Department of Animal Reproduction and Artificial Insemination, University of Ondokuz Mayis, 55200 Samsun, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9160-5026","authenticated-orcid":false,"given":"Melih","family":"Akar","sequence":"additional","affiliation":[{"name":"Department of Production Animal Medicine, Faculty of Veterinary Medicine, University of Helsinki, 00014 Helsinki, Finland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.5534\/wjmh.2014.32.1.1","article-title":"Effect of Oxidative Stress on Male Reproduction","volume":"32","author":"Agarwal","year":"2014","journal-title":"World J. Mens Health"},{"key":"ref_2","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_3","doi-asserted-by":"crossref","unstructured":"Berean, D.I., Bogdan, L.M., and Cimpean, R. (2024). Advancements in Understanding and Enhancing Antioxidant-Mediated Sperm Cryopreservation in Small Ruminants: Challenges and Perspectives. Antioxidants, 13.","DOI":"10.3390\/antiox13060624"},{"key":"ref_4","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_5","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_6","doi-asserted-by":"crossref","first-page":"14661","DOI":"10.1038\/s41598-020-71015-9","article-title":"Protein Signatures of Seminal Plasma from Bulls with Contrasting Frozen-Thawed Sperm Viability","volume":"10","author":"Gomes","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"247","DOI":"10.17221\/33\/2017-CJAS","article-title":"Curcumin Offers Antioxidant Protection to Cryopreserved Bovine Semen","volume":"63","author":"Mackovich","year":"2018","journal-title":"Czech J. Anim. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1080\/09712119.2020.1816549","article-title":"The Effects of Different Levels of Nano Selenium on the Quality of Frozen-Thawed Sperm in Ram","volume":"48","author":"Nateq","year":"2020","journal-title":"J. Appl. Anim. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1007\/s11259-024-10302-6","article-title":"Effects of Organic and Inorganic Selenium Supplementation on Frozen-Thawed Ram Semen at Two Cooling Periods","volume":"48","author":"Rodrigues","year":"2024","journal-title":"Vet. Res. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.anireprosci.2014.01.009","article-title":"Role of Selenium in Male Reproduction\u2014A Review","volume":"146","author":"Ahsan","year":"2014","journal-title":"Anim. Reprod. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e14652","DOI":"10.1111\/rda.14652","article-title":"Investigation of the Protective Effects of Different Forms of Selenium in Freezing Dog Semen: Comparison of Nanoparticle Selenium and Sodium Selenite","volume":"59","author":"Esin","year":"2024","journal-title":"Reprod. Domest. Anim."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Shahin, M.A., Khalil, W.A., Saadeldin, I.M., Swelum, A.A.-A., and El-Harairy, M.A. (2020). Comparison between the Effects of Adding Vitamins, Trace Elements, and Nanoparticles to Shotor Extender on the Cryopreservation of Dromedary Camel Epididymal Spermatozoa. Animals, 10.","DOI":"10.3390\/ani10010078"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8759","DOI":"10.1074\/jbc.M610078200","article-title":"Extracellular Production of Hydrogen Selenide Accounts for Thiol-Assisted Toxicity of Selenite against Saccharomyces cerevisiae","volume":"282","author":"Tarze","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"12360","DOI":"10.1080\/10408398.2022.2101093","article-title":"Selenium Nanoparticles: Enhanced Nutrition and Beyond","volume":"63","author":"Chen","year":"2023","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Abdelnour, S.A., Hassan, M.A.E., Mohammed, A.K., Alhimaidi, A.R., Al-Gabri, N., Al-Khaldi, K.O., and Swelum, A.A. (2020). The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm. Animals, 10.","DOI":"10.3390\/ani10091508"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1678","DOI":"10.1016\/j.psj.2019.12.006","article-title":"Effect of Astaxanthin Nanoparticles in Protecting the Post-Thawing Quality of Rooster Sperm Challenged by Cadmium Administration","volume":"99","author":"Najafi","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1016\/j.jinorgbio.2007.06.021","article-title":"Size Effect of Elemental Selenium Nanoparticles (Nano-Se) at Supranutritional Levels on Selenium Accumulation and Glutathione S-Transferase Activity","volume":"101","author":"Peng","year":"2007","journal-title":"J. Inorg. Biochem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.theriogenology.2018.12.017","article-title":"Impact of Selenium Nano-Particles in Semen Extender on Bull Sperm Quality after Cryopreservation","volume":"126","author":"Khalil","year":"2019","journal-title":"Theriogenology"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e12869","DOI":"10.1111\/and.12869","article-title":"Effect of in Vitro Selenium Supplementation on Sperm Quality in Asthenoteratozoospermic Men","volume":"50","author":"Ghafarizadeh","year":"2018","journal-title":"Andrologia"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1186\/s40104-021-00548-4","article-title":"Expression, Structure and Function Analysis of the Sperm-Oocyte Fusion Genes Juno and Izumo1 in Sheep (Ovis aries)","volume":"12","author":"Hu","year":"2021","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"100","DOI":"10.1016\/j.anireprosci.2016.09.011","article-title":"Effect of Vitamin E and Selenium Nanoparticles on Post-Thaw Variables and Oxidative Status of Rooster Semen","volume":"174","author":"Safa","year":"2016","journal-title":"Anim. Reprod. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1089\/ars.2005.7.768","article-title":"Peroxiredoxins, Oxidative Stress, and Cell Proliferation","volume":"7","author":"Immenschuh","year":"2005","journal-title":"Antioxid. Redox Signal."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1089\/ars.2023.0287","article-title":"Structural and Functional Diversity of the Peroxiredoxin 6 Enzyme Family","volume":"40","author":"Rahaman","year":"2024","journal-title":"Antioxid. Redox Signal."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1089\/ars.2010.3584","article-title":"Peroxiredoxin 5: Structure, Mechanism, and Function of the Mammalian Atypical 2-Cys Peroxiredoxin","volume":"15","author":"Knoops","year":"2011","journal-title":"Antioxid. Redox Signal."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/2051-4190-24-4","article-title":"Peroxiredoxins: Hidden Players in the Antioxidant Defence of Human Spermatozoa","volume":"24","year":"2014","journal-title":"Basic Clin. Androl."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1089\/ars.2019.7985","article-title":"Antioxidant Enzymes and Male Fertility: Lessons from Knockout Models","volume":"32","author":"Scarlata","year":"2020","journal-title":"Antioxid. Redox Signal."},{"key":"ref_27","first-page":"263","article-title":"Effects of in Vitro Selenium Addition to the Semen Extender on the Spermatozoa Characteristics before and after Freezing in Water Buffaloes (Bubalus bubalis)","volume":"3","author":"Dorostkar","year":"2012","journal-title":"Vet. Res. Forum"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1111\/rda.12903","article-title":"Effect of Selenium Supplementation on Semen Characteristics of Brazil\u2019s Ram","volume":"52","author":"Piagentini","year":"2017","journal-title":"Reprod. Domest. Anim."},{"key":"ref_29","first-page":"249","article-title":"In Vitro Supplementation of Nano Selenium Minimizes Freeze-Thaw Induced Damage to Ram Spermatozoa","volume":"8","author":"Hozyen","year":"2019","journal-title":"Int. J. Vet. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.theriogenology.2003.10.016","article-title":"Comparison between the Toluidine Blue Stain and the Feulgen Reaction for Evaluation of Rabbit Sperm Chromatin Condensation and Their Relationship with Sperm Morphology","volume":"62","author":"Beletti","year":"2004","journal-title":"Theriogenology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_32","unstructured":"Team, R.C. (2025, November 08). R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria. Available online: http:\/\/www.R-project.org\/."},{"key":"ref_33","unstructured":"Wei, T., and Simko, V.R. (Package \u201cCorrplot\u201d: Visualization of a Correlation Matrix, 2021). Package \u201cCorrplot\u201d: Visualization of a Correlation Matrix, Package Corrplot for R Software."},{"key":"ref_34","unstructured":"Revelle, W.R. (2017). Psych: Procedures for Personality and Psychological Research, Northwestern University."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1002\/j.1939-4640.2000.tb03420.x","article-title":"The Effect of Reactive Oxygen Species on Equine Sperm Motility, Viability, Acrosomal Integrity, Mitochondrial Membrane Potential, and Membrane Lipid Peroxidation","volume":"21","author":"Baumber","year":"2000","journal-title":"J. Androl."},{"key":"ref_36","first-page":"317","article-title":"Effect of in Vitro Selenium Addition to the Semen Extender on the Spermatozoa Characteristics before and after Freezing in Kundhi Buffalo Bull and in Vivo Fertility Rate","volume":"51","author":"Jamali","year":"2019","journal-title":"Pak. J. Zo\u00f6l."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"e20200102","DOI":"10.1590\/0103-8478cr20200102","article-title":"Selenium Supplementation Prevents DNA Damage in Ram Spermatozoa","volume":"51","author":"Moya","year":"2020","journal-title":"Ci\u00eancia Rural."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Mohammed Ali, I.A., AL-Ahmed, H.I., and Ben Ahmed, A. (2023). Evaluation of Green Synthesis (Withania somnifera) of Selenium Nanoparticles to Reduce Sperm DNA Fragmentation Diabetic Mice Induced with Streptozotocin. Appl. Sci., 13.","DOI":"10.3390\/app13020728"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"878","DOI":"10.1002\/mrd.20686","article-title":"Impact of Cryopreservation and Reactive Oxygen Species on DNA Integrity, Lipid Peroxidation, and Functional Parameters in Ram Sperm","volume":"74","author":"Peris","year":"2007","journal-title":"Mol. Reprod. Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.smallrumres.2009.11.015","article-title":"The Effect of Antioxidants on Post-Thawed Angora Goat (Capra hircus ancryrensis) Sperm Parameters, Lipid Peroxidation and Antioxidant Activities","volume":"89","author":"Bucak","year":"2010","journal-title":"Small Rumin. Res."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Abah, K.O., Ligocka-Kowalczyk, Z., Itodo, J.I., Ameh, G., Partyka, A., and Nizanski, W. (2025). Association between Sperm DNA Fragmentation and Fertility Parameters in Farm Animals: A Systematic Review and Meta-Analysis. BMC Vet. Res., 21.","DOI":"10.1186\/s12917-025-04652-9"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.smallrumres.2010.10.011","article-title":"Lipid Peroxidation and Generation of Hydrogen Peroxide in Frozen-Thawed Ram Semen Supplemented with Catalase or Trolox","volume":"95","author":"Sicherle","year":"2011","journal-title":"Small Rumin. Res."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Ribeiro, J.C., Braga, P.C., Martins, A.D., Silva, B.M., Alves, M.G., and Oliveira, P.F. (2021). Antioxidants Present in Reproductive Tract Fluids and Their Relevance for Fertility. Antioxidants, 10.","DOI":"10.3390\/antiox10091441"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.anireprosci.2012.08.027","article-title":"Proteomic Analysis of Boar Spermatozoa and Quantity Changes of Superoxide Dismutase 1, Glutathione Peroxidase, and Peroxiredoxin 5 during Epididymal Maturation","volume":"135","author":"Park","year":"2012","journal-title":"Anim. Reprod. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"20241516","DOI":"10.1098\/rspb.2024.1516","article-title":"Silent Cells? Potential for Context-Dependent Gene Expression in Mature Sperm","volume":"292","author":"Lymbery","year":"2025","journal-title":"Proc. R. Soc. B Biol. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Horky, P., Urbankova, L., Bano, I., Kopec, T., Nevrkla, P., Pribilova, M., Baholet, D., Chilala, P., Slama, P., and Skalickova, S. (2023). Selenium Nanoparticles as Potential Antioxidants to Improve Semen Quality in Boars. Animals, 13.","DOI":"10.3390\/ani13152460"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.mce.2005.12.026","article-title":"Oxidative Stress, Sperm Survival and Fertility Control","volume":"250","author":"Aitken","year":"2006","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"686137","DOI":"10.4061\/2011\/686137","article-title":"Impacts of Oxidative Stress and Antioxidants on Semen Functions","volume":"2011","author":"Bansal","year":"2011","journal-title":"Vet. Med. Int."},{"key":"ref_49","first-page":"157","article-title":"Effect of Cryopreservation on Lipid Composition and Antioxidant Enzyme Activity of Seabass (Lates calcarifer) Sperm","volume":"15","author":"Klaiwattana","year":"2016","journal-title":"Iran. J. Fish. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Peris-Frau, P., Soler, A.J., Iniesta-Cuerda, M., Mart\u00edn-Maestro, A., S\u00e1nchez-Ajofr\u00edn, I., Medina-Ch\u00e1vez, D.A., Fern\u00e1ndez-Santos, M.R., Garc\u00eda-\u00c1lvarez, O., Maroto-Morales, 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_51","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1089\/ars.2010.3624","article-title":"Structure-Based Insights into the Catalytic Power and Conformational Dexterity of Peroxiredoxins","volume":"15","author":"Hall","year":"2011","journal-title":"Antioxid. Redox Signal."},{"key":"ref_52","first-page":"626374","article-title":"The Enzymatic Antioxidant System of Human Spermatozoa","volume":"2014","year":"2014","journal-title":"Adv. Androl."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"127296","DOI":"10.1016\/j.jtemb.2023.127296","article-title":"Zinc Oxide and Selenium Nanoparticles Can Improve Semen Quality and Heat Shock Protein Expression in Cryopreserved Goat (Capra hircus) Spermatozoa","volume":"80","author":"Abedin","year":"2023","journal-title":"J. Trace Elem. Med. Biol."},{"key":"ref_54","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"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/3\/457\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T14:35:44Z","timestamp":1770042944000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/3\/457"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,1]]},"references-count":54,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["ani16030457"],"URL":"https:\/\/doi.org\/10.3390\/ani16030457","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,1]]}}}