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Ruminant livestock are particularly susceptible\u00a0to Lm, \u00a0thus effective strategies are needed\u00a0for controlling ovine listeriosis. In this study, we developed two inactivated vaccines and evaluated their efficacy against Lm infection in murine and ovine models. We inactivated the Lm serotype 4h XYSN strain and adjuvanted it with water-in-oil ISA 61 VG (61 VG-AIV) or aluminum (Al-AIV). Pathological observations confirmed the safety of both vaccines in mice and sheep. The immunological assays demonstrated that, compared with the Al-AIV, the 61\u00a0VG-AIV induced higher levels of Lm-specific antibodies and proinflammatory cytokines, suggesting that the ISA\u00a061\u00a0VG adjuvant has superior immunostimulatory effects compared with the alum adjuvant. 61\u00a0VG-AIV elicited greater immunoprotection than Al-AIV (83.4% vs. 50%) against serotype 4h Lm strain challenge in mice. Additionally, 61 VG-AIV afforded cross-protection against challenges with serotypes 1\/2a, 1\/2b, and 4b Lm strains. Importantly, high immunoprotection in sheep was conferred by the 61 VG-AIV group (83.4%). Taken together, our findings demonstrate that the ISA 61 VG adjuvant contributes to enhancing the humoral and cellular immune responses of inactivated Lm, and 61 VG-AIV is a promising vaccine candidate for the prevention and control of animal listeriosis. This research lays a solid foundation for its application in veterinary medicine.<\/jats:p>","DOI":"10.1186\/s13567-025-01483-2","type":"journal-article","created":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T11:26:52Z","timestamp":1742470012000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The inactivated and\u00a0ISA 61 VG adjuvanted vaccine enhances protection against cross-serotype Listeria monocytogenes"],"prefix":"10.1186","volume":"56","author":[{"given":"Fanzeng","family":"Meng","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ye","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chao","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tianxiang","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yao","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zegang","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Yao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin\u2019an","family":"Jiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2429-5715","authenticated-orcid":false,"given":"Yuelan","family":"Yin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,3,20]]},"reference":[{"key":"1483_CR1","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1016\/j.siny.2009.01.006","volume":"14","author":"KM Posfay-Barbe","year":"2009","unstructured":"Posfay-Barbe KM, Wald ER (2009) Listeriosis. 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The experiments conducted on animals adhered to internationally recognized guidelines for biomedical research involving animals. The animals were housed and handled in animal biosafety facilities by individuals who possessed the necessary qualifications to carry out experimental procedures. All animals in the challenge experiment were closely monitored after infection and euthanized when recovery was deemed impossible.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"The authors declare that they have no competing interests.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"60"}}