{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T15:09:31Z","timestamp":1790003371430,"version":"4.0.1"},"reference-count":69,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T00:00:00Z","timestamp":1789776000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2022YFD1300900"],"award-info":[{"award-number":["2022YFD1300900"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Mulberry leaf is a promising plant feed resource for aquaculture, yet high inclusion limits its application due to anti-nutritional factors. Exogenous enzyme supplementation can improve the utilization efficiency of bioactive compounds in mulberry leaf-containing diets. This study aimed to evaluate the dose-dependent effects of mulberry leaf powder supplemented with a multi-enzyme premix (MLE) on growth, intestinal health, antioxidant and immune responses, as well as intestinal microbiota and metabolome in juvenile Acipenser schrenckii. Four diets containing 0%, 2%, 4% and 6% MLE were formulated for a 10-week feeding trial. Moderate MLE supplementation maintained normal growth without significantly improving growth performance relative to the control, and was associated with reduced serum triglyceride, total cholesterol and glucose, as well as improved intestinal morphology and digestive enzyme activity. Appropriate MLE supplementation was associated with changes in the Nrf2\/Keap1 signaling pathway, enhanced antioxidant capacity, and upregulated anti-inflammatory gene il10. Multi-omics analysis revealed that MLE induced taxon-specific changes in the intestinal microbiota and was associated with altered metabolic profiles, enriching beneficial taxa including Priestia, Bacillus_P and Cetobacterium_A. These changes suggest a potential modulation of the microbiota\u2013metabolite\u2013immunity axis that may help sustain intestinal homeostasis. Nevertheless, the beneficial effects declined at the 6% inclusion level. Collectively, MLE was associated with dose-dependent protective effects on the intestinal barrier that may involve the microbiota\u2013metabolite\u2013immunity axis and the Nrf2\/Keap1 pathway; the optimal supplementary range is preliminarily estimated at 3.5\u20134.5% for juvenile Acipenser schrenckii, although this estimate is based on four dietary levels and requires validation in future dose\u2013response studies.<\/jats:p>","DOI":"10.3390\/foods15183319","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T14:12:09Z","timestamp":1789999929000},"page":"3319","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Mulberry Leaf Powder Supplemented with a Multi-Enzyme Premix Affects the Intestinal Microbiota\u2013Metabolite\u2013Immunity Axis and Nrf2\/Keap1 Signaling in Juvenile Acipenser schrenckii: A Dose-Dependent Evaluation"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4784-0907","authenticated-orcid":false,"given":"Wanwan","family":"Zhu","sequence":"first","affiliation":[{"name":"Department of Grassland Science, College of Animal Science, Guizhou University, Guiyang 550000, China"},{"name":"Beijing Tianyuanyugang Farm Co., Ltd. (Beijing Amur Sturgeon Elite Breeding Base), Beijing 101400, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"}]},{"given":"Nan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Grassland Science, College of Animal Science, Guizhou University, Guiyang 550000, China"},{"name":"Beijing Tianyuanyugang Farm Co., Ltd. (Beijing Amur Sturgeon Elite Breeding Base), Beijing 101400, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8072-0467","authenticated-orcid":false,"given":"Xuekai","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Grassland Science and Technology, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7133-1558","authenticated-orcid":false,"given":"Yi","family":"Xiong","sequence":"additional","affiliation":[{"name":"Frontier Technology Research Institute of China Agricultural University in Shenzhen, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"given":"Fuyu","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Grassland Science, College of Animal Science, Guizhou University, Guiyang 550000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Yongsheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Livestock and Poultry Multi-Omics of MARA, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1023\/A:1007370213924","article-title":"An overview of Acipenseriforms","volume":"48","author":"Bemis","year":"2002","journal-title":"Environ. 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