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We injected 20 mL per testicle. We took 5 mL blood from all donkeys before injection. Castration was performed under general anesthesia 60 days later. Samples included blood and testicular tissue. Total motility (TM), progressive motility (PM), movementy features, DNA damage, morphology, viability, and plasma membrane functionality were evaluated. Hormone analyses, histomorphometric studies and oxidative stress indices including total antioxidant capacity (TAC), glutathione peroxidase (GPx), glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and NADP+\/NADPH were evaluated. Apoptosis, pyroptosis-related\n                      <jats:italic>Bax<\/jats:italic>\n                      ,\n                      <jats:italic>Caspase-1<\/jats:italic>\n                      ,\n                      <jats:italic>GSDMD<\/jats:italic>\n                      , and\n                      <jats:italic>Bcl-2<\/jats:italic>\n                      expression were also assessed.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      In HS and HM groups, testosterone, epididymal sperm count, motility, viability, and plasma membrane functionality dropped while sperm DNA damage increased. HS and HM groups had significantly lower histomorphometric parameters, TAC, GPx, SOD, GSH, and\n                      <jats:italic>Bcl-2<\/jats:italic>\n                      gene expression. MDA, NADP\n                      <jats:sup>+<\/jats:sup>\n                      \/NADPH,\n                      <jats:italic>Bax<\/jats:italic>\n                      ,\n                      <jats:italic>Caspase-1<\/jats:italic>\n                      , and\n                      <jats:italic>GSDMD<\/jats:italic>\n                      gene expression were substantially higher in the HS and HM groups than in the control group.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Toxic effects of hypertonic saline and mannitol on reproductive parameters were seen following, hence, they might be considered as a good chemical sterilizing treatment in donkeys.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12917-024-03915-1","type":"journal-article","created":{"date-parts":[[2024,3,11]],"date-time":"2024-03-11T03:01:37Z","timestamp":1710126097000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Effects of intratesticular injection of hypertonic mannitol and saline on the quality of donkey sperm, indicators of oxidative stress and testicular tissue pathology"],"prefix":"10.1186","volume":"20","author":[{"given":"Mohammadreza","family":"Baqerkhani","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Soleimanzadeh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rahim","family":"Mohammadi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,3,11]]},"reference":[{"key":"3915_CR1","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1186\/1746-6148-7-39","volume":"7","author":"K Jana","year":"2011","unstructured":"Jana K, Samanta PK. 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