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Methods: To address this challenge, this study developed an ISKNV \u0394orf037l vaccine strain, where the orf037l gene was knocked out. Infection assays conducted at 28 \u00b0C showed that the knocking out the orf037l gene decreased the virulence of ISKNV and reduced lethality against mandarin fish by 26.7% compared to wild-type ISKNV. To further diminish residual virulence, the effect of low-temperature (22 \u00b0C) immersion immunization was evaluated. Results: The results indicate that low temperature significantly diminished the virulence of the \u0394orf037l vaccine strain, elevating the survival rate of mandarin fish to 90%. Furthermore, the vaccine strain effectively triggered the expression of crucial immune-related genes, such as IFN-h, IL-1, I\u03baB, Mx, TNF-\u03b1, and Viperin, while inducing the production of specific neutralizing antibodies. Low-temperature immersion with \u0394orf037l achieved a high relative percentage of survival of 92.6% (n = 30) in mandarin fish, suggesting the potential of \u0394orf037l as a promising immersion vaccine candidate. Conclusions: These findings contribute to advancing fish immersion vaccine development and demonstrate the importance and broad applicability of temperature optimization strategies in vaccine development. Our work carries profound implications for both the theoretical understanding and practical application in aquaculture disease control.<\/jats:p>","DOI":"10.3390\/vaccines12101170","type":"journal-article","created":{"date-parts":[[2024,10,14]],"date-time":"2024-10-14T12:44:31Z","timestamp":1728909871000},"page":"1170","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Evaluation of a Low-Temperature Immersion Immunization Strategy for the Infectious Spleen and Kidney Necrosis Virus orf037l Gene-Deleted Attenuated Vaccine"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3282-9641","authenticated-orcid":false,"given":"Weiqiang","family":"Pan","sequence":"first","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiajie","family":"Fu","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruoyun","family":"Zeng","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingcong","family":"Liang","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanlin","family":"You","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhipeng","family":"Zhan","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhoutao","family":"Lu","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shaoping","family":"Weng","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3223-6176","authenticated-orcid":false,"given":"Changjun","family":"Guo","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2460-7061","authenticated-orcid":false,"given":"Jianguo","family":"He","sequence":"additional","affiliation":[{"name":"School of Marine Sciences, State Key Laboratory for Biocontrol & Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Guangdong Province Key Laboratory of Aquatic Economic Animals\/Guangdong Provincial Observation and Research Station for Marine Ranching of the Lingdingyang Bay, Sun Yat-sen University, 135 Xingang Road West, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/S0044-8486(01)00639-1","article-title":"Experimental Transmission, Pathogenicity and Physical\u2013Chemical Properties of Infectious Spleen and Kidney Necrosis Virus (Isknv)","volume":"204","author":"He","year":"2002","journal-title":"Aquaculture"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1007\/s00705-023-05797-4","article-title":"Changes to Virus Taxonomy and the ICTV Statutes Ratified by the International Committee on Taxonomy of Viruses (2023)","volume":"168","author":"Zerbini","year":"2023","journal-title":"Arch. 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