{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T12:54:45Z","timestamp":1791204885423,"version":"4.1.0"},"reference-count":54,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"vor","delay-in-days":27,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T00:00:00Z","timestamp":1788220800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Medicine &amp;amp; Sci"],"published-print":{"date-parts":[[2026,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Supplementation of semen extender with natural antioxidants represents a promising strategy to mitigate cryopreservation\u2010induced damage in ram spermatozoa.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objectives<\/jats:title>\n                    <jats:p>\n                      This study evaluated the dose\u2010dependent effects of\n                      <jats:italic>Spirulina platensis<\/jats:italic>\n                      extract (0\u201070 \u00b5g\/mL) supplemented to Tris\u2010based extenders on post\u2010thaw sperm quality in Lori rams.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Semen was collected twice weekly over a four\u2010week period (eight replicates) from four mature rams using an artificial vagina. On each collection day, ejaculates meeting the acceptable criteria were pooled and extended in a Tris\u2010based diluent containing 0, 10, 20, 30, 40, 50, 60, or 70 \u00b5g\/mL of\n                      <jats:italic>Spirulina<\/jats:italic>\n                      extract, designated as Spir\u20100 through Spir\u201070, respectively. All samples were equilibrated at 5\u00b0C for 3 h, then cryopreserved in liquid nitrogen and subsequently thawed in a 37\u00b0C water bath for 30\u00a0s. Post\u2010thaw sperm quality was comprehensively assessed for motility (using CASA), viability, plasma membrane integrity, acrosome morphology, mitochondrial membrane potential (\u0394\u03a8m), DNA denaturation (via AO), and oxidative biomarkers, including malondialdehyde (MDA), hydrogen peroxide (H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      ), and antioxidant enzyme activities (superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GPx]). Data were analysed using a completely randomised design with one\u2010way ANOVA, followed by Tukey's HSD post\u2010hoc test, with statistical significance set at\n                      <jats:italic>P<\/jats:italic>\n                      \u00a0&lt;\u00a00.05.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Supplementation at the concentration of 50 \u00b5g\/mL\n                      <jats:italic>Spirulina<\/jats:italic>\n                      extract (Spir\u201050) yielded the highest post\u2010thaw total (TM; 50.23%) and progressive motility (PM; 33.0%), as compared to the control (TM; 32.30%; PM; 18.8%). Moreover, this concentration significantly enhanced viability (59.16%), plasma membrane functionality (56.95%), and the proportion of spermatozoa exhibiting high mitochondrial membrane potential (hMMP; 54.5%). In addition, Spir\u201050 markedly increased glutathione peroxidase (GPx) activity (35.49 vs. 23.73 mU\/mL in control) and superoxide dismutase (SOD) activity (213.07 vs. 190.74 U\/mL in control), along with total antioxidant capacity (TAC; 4.53 \u00b5mol\/L vs. 3.34 \u00b5mol\/L in control). Conversely, this treatment substantially reduced malondialdehyde (MDA; 2.60 nmol\/mL vs. 7.69 nmol\/mL in control), DNA denaturation (2.87% vs. 6.67% in control), and hydrogen peroxide levels (H\n                      <jats:sub>2<\/jats:sub>\n                      O\n                      <jats:sub>2<\/jats:sub>\n                      ; 16.4% vs. 27.7% in control).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Supplementation with the concentration of 50 \u00b5g\/mL\n                      <jats:italic>Spirulina<\/jats:italic>\n                      extract exhibited the most effective cryoprotective effects among the tested concentrations by improving sperm motility, viability, membrane and DNA integrity, mitochondrial function, and endogenous antioxidant defences, while attenuating lipid peroxidation and oxidative stress. These findings support the incorporation of\n                      <jats:italic>Spirulina<\/jats:italic>\n                      extract in Tris\u2010based extenders to improve post\u2010thaw ram sperm quality in ram semen cryopreservation protocols.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/vms3.71186","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T17:09:21Z","timestamp":1790615361000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effects of Supplementing Semen Extenders With\n                    <i>Spirulina platensis<\/i>\n                    Extract on Post\u2010Thaw Ram Sperm Quality"],"prefix":"10.1002","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-5435-8992","authenticated-orcid":false,"given":"Kimia","family":"Jaberi","sequence":"first","affiliation":[{"name":"Department of Animal Science Faculty of 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