{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T22:55:58Z","timestamp":1777416958923,"version":"3.51.4"},"reference-count":71,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2025,7,27]],"date-time":"2025-07-27T00:00:00Z","timestamp":1753574400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31972996"],"award-info":[{"award-number":["31972996"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["Z2022152"],"award-info":[{"award-number":["Z2022152"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Third Livestock and Poultry Genetic Resources Census, Conservation, and Development and Utilization in Guangxi","award":["31972996"],"award-info":[{"award-number":["31972996"]}]},{"name":"Third Livestock and Poultry Genetic Resources Census, Conservation, and Development and Utilization in Guangxi","award":["Z2022152"],"award-info":[{"award-number":["Z2022152"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Semen cryopreservation is associated with sperm vulnerability to oxidative stress and ice crystal-induced damage, adversely affecting in vitro fertilization (IVF) success. This study aimed to investigate the effects of freezing diluent supplemented with antioxidant limonin (Lim), myo-inositol (MYO), and the ice crystal formation inhibitor L-proline (LP) through sperm motility, morphological integrity, and antioxidant capacity. The Lim (150 mM), MYO (90 mM), and LP (100 mM) significantly ameliorated the quality of post-thaw sperm in Debao boar, and combined treatment of these agents significantly enhanced sperm motility, structural integrity, and antioxidant capacity compared with individual agents (p &lt; 0.05). Notably, the combined use of these agents reduced glycerol concentration in the freezing diluent from 3% to 2%. Meanwhile, the integrity of the sperm plasma membrane, acrosome membrane, and mitochondrial membrane potential was significantly improved (p &lt; 0.05), and the result of IVF revealed the total cell count of the blastocysts was also greater in the 2% glycerol group (p &lt; 0.05). In conclusion, the newly developed freezing diluent for semen, by adding Lim (150 mM), MYO (90 mM), and LP (100 mM), can enhance the quality of frozen\u2013thawed Debao boar sperm and reduce the concentration of glycerol from 3% to 2% as high concentrations of glycerol can impair the quality of thawed sperm and affect in vitro fertilization outcomes. In conclusion, the improved dilution solution formulated demonstrated efficacy in enhancing the quality of porcine spermatozoa following cryopreservation and subsequent thawing.<\/jats:p>","DOI":"10.3390\/ani15152204","type":"journal-article","created":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T07:57:24Z","timestamp":1753689444000},"page":"2204","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The Effects of Limonin, Myo-Inositol, and L-Proline on the Cryopreservation of Debao Boar Semen"],"prefix":"10.3390","volume":"15","author":[{"given":"Qianhui","family":"Feng","sequence":"first","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanyan","family":"Yang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bing","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wen","family":"Shi","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yizhen","family":"Fang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunrong","family":"Xu","sequence":"additional","affiliation":[{"name":"Guangxi Work Station of Livestock & Poultry Breed Improvement, Nanning 530001, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhuxin","family":"Deng","sequence":"additional","affiliation":[{"name":"Guangxi Work Station of Livestock & Poultry Breed Improvement, Nanning 530001, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanyou","family":"Feng","sequence":"additional","affiliation":[{"name":"School of Environmental and Life Sciences, Nanning Normal University, Nanning 530023, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8735-5925","authenticated-orcid":false,"given":"Deshun","family":"Shi","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3640450","DOI":"10.1155\/2020\/3640450","article-title":"Fatty Acid Profile and Metabolism Are Related to Human Sperm Parameters and Are Relevant in Idiopathic Infertility and Varicocele","volume":"2020","author":"Collodel","year":"2020","journal-title":"Mediat. 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