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This study examined the effects of dietary silkworm pupae powder (SWP) supplementation on the growth performance, muscle fatty acid composition, and intestinal function in mandarin fish (Siniperca chuatsi) during a 60-day feeding trial. A total of 360 fish (initial body weight 52.44 ± 0.06 g) were distributed randomly into three groups with three replicates per group and 40 fish per replicate. The three groups were fed either the basal diet, supplementation with 1.0 % fermented SWP (T1), or supplementation with 1.0 % fermented induced SWP (T2, induced by Beauveria bassiana). Results showed that fish fed T1 had significantly higher (P < 0.05) condition factor, C16:0, C20:4n-6, C20:5 n3, C22:6n3 C22:5n6, C22:5n-3, and n-3/n-6 levels, but lower (P < 0.05) fat body ratio, C18:1, C18:2n-6 and crude lipid levels than those fish fed basal diet. All fish had similar (P > 0.05) weight gain rate, specific growth rate, feed conversion rate, feed intake, and hepatosomatic index. Fish fed T2 had higher (P < 0.05) trypsin activity than those fish fed basal diet. Compared with the basal diet, fish fed T1 or T2 had higher (P < 0.05) villus length, muscular layer thickness, glutathione peroxidase, alkaline phosphatase, and lysozyme activities, but lower (P < 0.05) triglyceride and total cholesterol levels. Comparative transcriptomic analysis indicated that T1 or T2 enhanced immunity and lipid metabolism ability, as manifested in the up-regulated expression of il12rb1, scdb, tlr2, and FABP7, and the down-regulated expression of EBP and hsd17b7. These findings suggest that dietary supplementation with fermented SWP could enhance the health and quality of mandarin fish, potentially offering a valuable strategy for improving aquaculture productivity.