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To this end, in Experiment 1 SP was added to a sperm model without previous contact with this substance (ram epididymal sperm) at the beginning or the end of a 48-hour preservation protocol at 5\u00a0\u00b0C (<jats:italic>n<\/jats:italic>\u2009=\u200913). Sperm motility and kinetic parameters and sperm functionality in terms of sperm viability, apoptosis, mitochondrial activity and reacted acrosomes were assessed after 6\u00a0h of storage at 15\u00a0\u00b0C (standard liquid preservation method) and 24 and 48\u00a0h at 5\u00a0\u00b0C. Extended sperm showed better results after 48\u00a0h when stored in the absence than in the presence of SP in most sperm quality parameters. Moreover, the final SP supplementation of this experimental group resulted in the highest sperm motility and kinetic parameters, viability and mitochondrial activity. These results suggested that initial SP deprivation could be beneficial in a medium-term ram sperm preservation protocol in liquid form, as well as a final supplementation. Therefore, we conducted Experiment 2 to evaluate the effect of SP removal from freshly ejaculated ram semen under the same storage conditions as in Experiment 1 (<jats:italic>n<\/jats:italic>\u2009=\u200912). Surprisingly, SP withdrawal impaired sperm functionality, leading to increased apoptosis and decreased mitochondrial activity after 24 and 48\u00a0h at 5\u00a0\u00b0C. Conversely, SP supplementation at the end of the preservation protocol of the ejaculate processed as usual had a positive effect on sperm quality and fertility. To summarize, SP absence was beneficial for a medium-term preservation protocol (up to 48\u00a0h at 5\u00a0\u00b0C) of ram epididymal sperm, but the same preservation protocol for ram ejaculated sperm revealed a possible failure of the SP removal method in avoiding the sperm-SP interaction effect. Meanwhile, SP supplementation of ram semen at the end of the preservation protocol increased in vitro sperm quality and fertility after artificial insemination.<\/jats:p>","DOI":"10.1186\/s12917-024-04214-5","type":"journal-article","created":{"date-parts":[[2024,8,12]],"date-time":"2024-08-12T02:02:29Z","timestamp":1723428149000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Seminal plasma removal for medium-term preservation of ram sperm at 5\u00a0\u00b0C"],"prefix":"10.1186","volume":"20","author":[{"given":"Marta","family":"Neila-Montero","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mercedes","family":"Alvarez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marta F.","family":"Riesco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Soriano-\u00dabeda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Montes-Garrido","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristina","family":"Palacin-Martinez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulino","family":"de Paz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Anel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis","family":"Anel-Lopez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,8,12]]},"reference":[{"key":"4214_CR1","doi-asserted-by":"publisher","first-page":"1235","DOI":"10.1016\/j.theriogenology.2004.07.001","volume":"63","author":"L Anel","year":"2005","unstructured":"Anel L, Kaabi M, Abroug B, Alvarez M, Anel E, Boixo JC, et al. 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