{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T17:48:13Z","timestamp":1790876893593,"version":"4.1.0"},"reference-count":51,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T00:00:00Z","timestamp":1746748800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Madrid Institute for Rural, Agricultural, and Food Research and Development (IMIDRA)","award":["FP24-CONSERVANIMAL"],"award-info":[{"award-number":["FP24-CONSERVANIMAL"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>This study evaluated the impact of different processing techniques on microbial load and sperm quality in frozen\u2013thawed equine semen to identify alternatives to reduce the preventive use of antibiotics. Semen was obtained and processed under rigorous hygiene measures from ten stallions, using four protocols: Simple Centrifugation with antibiotics (S+) and Simple Centrifugation (S\u2212), Filtration (F\u2212) and Single-Layer Colloidal Centrifugation (C\u2212) in an antibiotic-free extender. Microbial load in different culture media, sperm viability and motility were assessed. Microbial load results were consistent across protocols, except in Columbia 5% Sheep Blood Agar media, where S\u2212 exhibited higher microbial load than S+ (p &lt; 0.05). However, F\u2212 and C\u2212 showed similar microbial loads to S+. No significant differences were observed in progressive motility, average path velocity, straight-line velocity or wobble parameters between protocols. Total motility and viability were significantly higher in S+ compared to other treatments (p &lt; 0.05). Thus, regardless of antibiotics, the proposed methods achieved results similar to the traditional antibiotic-inclusive protocol in terms of microbial load and the most relevant semen quality parameters. These findings suggest that the use of F\u2212 and C\u2212, combined with optimized hygiene measures, offers an effective alternative to reduce the prophylactic use of antibiotics in semen extenders.<\/jats:p>","DOI":"10.3390\/ani15101368","type":"journal-article","created":{"date-parts":[[2025,5,9]],"date-time":"2025-05-09T08:20:23Z","timestamp":1746778823000},"page":"1368","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Innovative Approaches to Avoid Antibiotic Use in Equine Semen Cryopreservation: Advancing Sustainable Reproductive Technologies"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-6595-8871","authenticated-orcid":false,"given":"Sonsoles Mercedes","family":"Zabala","sequence":"first","affiliation":[{"name":"Animal Medicine and Surgery Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"},{"name":"Animal Selection and Reproduction Center, Madrid Institute for Rural, Agricultural and Food Research and Development (IMIDRA), Ctra. Colmenar Viejo a Guadalix de la Sierra, km 1, Colmenar Viejo, 28770 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6713-5124","authenticated-orcid":false,"given":"Consuelo","family":"Serres","sequence":"additional","affiliation":[{"name":"Animal Medicine and Surgery Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Montero","sequence":"additional","affiliation":[{"name":"Animal Health Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6946-3527","authenticated-orcid":false,"given":"Francisco","family":"Crespo","sequence":"additional","affiliation":[{"name":"Animal Medicine and Surgery Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"},{"name":"Centro Militar de Cr\u00eda Caballar de \u00c1vila (CCFAA), C\/Arsenio Guti\u00e9rrez Palacios s\/n, 05005 \u00c1vila, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro Luis","family":"Lorenzo","sequence":"additional","affiliation":[{"name":"Physiology Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3404-2137","authenticated-orcid":false,"given":"Ver\u00f3nica","family":"P\u00e9rez-Aguilera","sequence":"additional","affiliation":[{"name":"Centro Militar de Cr\u00eda Caballar de \u00c1vila (CCFAA), C\/Arsenio Guti\u00e9rrez Palacios s\/n, 05005 \u00c1vila, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0307-1256","authenticated-orcid":false,"given":"Agust\u00edn","family":"Oliet","sequence":"additional","affiliation":[{"name":"Animal Selection and Reproduction Center, Madrid Institute for Rural, Agricultural and Food Research and Development (IMIDRA), Ctra. Colmenar Viejo a Guadalix de la Sierra, km 1, Colmenar Viejo, 28770 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Virginia","family":"Hij\u00f3n","sequence":"additional","affiliation":[{"name":"Animal Selection and Reproduction Center, Madrid Institute for Rural, Agricultural and Food Research and Development (IMIDRA), Ctra. Colmenar Viejo a Guadalix de la Sierra, km 1, Colmenar Viejo, 28770 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Santiago","family":"Moreno","sequence":"additional","affiliation":[{"name":"Animal Selection and Reproduction Center, Madrid Institute for Rural, Agricultural and Food Research and Development (IMIDRA), Ctra. Colmenar Viejo a Guadalix de la Sierra, km 1, Colmenar Viejo, 28770 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno","family":"Gonz\u00e1lez-Zorn","sequence":"additional","affiliation":[{"name":"Animal Health Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. Puerta de Hierro s\/n, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5238-3590","authenticated-orcid":false,"given":"Luna","family":"Guti\u00e9rrez-Cepeda","sequence":"additional","affiliation":[{"name":"Animal Medicine and Surgery Department, Veterinary Faculty, Complutense University of Madrid (UCM), Avda. 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