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Twenty\u2010four ejaculations were collected from four Simmental bulls using the artificial vagina method. Ejaculates exhibiting over 70% motility were pooled to eliminate individual variations and allocated into five experimental groups: antibiotic\u2010free control group, antibiotic group, LA, BL and LR groups, respectively. Three probiotic strains,\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Lactobacillus rhamnosus<\/jats:italic>\n                    <\/jats:styled-content>\n                    (LR),\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Lactobacillus acidophilus<\/jats:italic>\n                    <\/jats:styled-content>\n                    (LA) and\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Bifidobacterium longum<\/jats:italic>\n                    <\/jats:styled-content>\n                    (BL), were incorporated into a Tris egg yolk\u2010based semen extender at a concentration of 10\n                    <jats:sup>9<\/jats:sup>\n                    \u2009CFU\/mL. Following dilution, samples were equilibrated and subsequently stored in liquid nitrogen. Post\u2010thaw evaluations were conducted for sperm motility and kinematic parameters using CASA, hypo\u2010osmotic swelling test, fluorescent staining for chromatin integrity, acrosomal integrity and mitochondrial membrane potential, as well as oxidative stress biomarkers. Additionally, apoptosis and metabolism related proteins were quantified by ELISA. The results exhibited that probiotic treated groups had significantly higher total and progressive motility as compared with controls (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001). In comparison with the control group, BL supplementation showed significantly reduced lipid peroxidation and increased antioxidant enzyme activities, along with lowered TGF\u2010\u03b2 and Caspase\u20103 levels, indicating attenuated oxidative stress and apoptotic signalling (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001). In contrast with the control group, LR supplementation resulted in elevated MDA and apoptosis\u2010related markers, suggesting strain specific pro\u2010oxidative effects (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.001). There were no significant differences observed in STAR or ATP synthase levels across the treatments. Altogether, the results exhibited that strains of probiotics, particularly BL, have the potential to serve as an alternative to antibiotics in bovine semen cryopreservation. Nevertheless, strain\u2010dependent responses and limitations, especially with respect to DNA integrity, require further investigation.\n                  <\/jats:p>","DOI":"10.1111\/rda.70197","type":"journal-article","created":{"date-parts":[[2026,3,19]],"date-time":"2026-03-19T09:42:19Z","timestamp":1773913339000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Probiotic Supplementation as an Alternative to Antibiotics in Bovine Sperm Cryopreservation: Effects on Motility, Oxidative Stress and Apoptosis"],"prefix":"10.1111","volume":"61","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2976-4368","authenticated-orcid":false,"given":"Ali Do\u011fan","family":"\u00d6m\u00fcr","sequence":"first","affiliation":[{"name":"Department of Reproduction and Artificial Insemination, Faculty of Veterinary Medicine Atat\u00fcrk University  Erzurum 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