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An 8‐week feeding trial was conducted to evaluate the effect of replacement of dietary fishmeal with cottonseed meal (CSM) on the growth performance and health status of juvenile grass carp, Ctenopharyngodon idellus (6.67 g). Six isonitrogenous and isoenergetic diets were formulated with different CSM levels (0, 121, 241, 362, 482 and 603 g/kg diet). The first limiting amino acid was methionine, which decreased with an increase in dietary CSM from 361 to 538 g/kg. When the dietary CSM content was lower than 241 g/kg, the limiting amino acid was histidine. Thermal growth coefficient (TGC) and feed efficiency first increased and then decreased (p < 0.05), which was consistent with the change in the limiting amino acid. The feeding rate increased with a rise in the dietary CSM level (p < 0.05). The activity of digestive enzymes except trypsin decreased significantly (p < 0.05), serum aminotransferase activity and triglycerides increased, serum cholesterol decreased (p < 0.05). When dietary CSM was raised to 362 g/kg, intestinal villi and hepatopancreas tissue were injured. When the dietary CSM was 603 g/kg, alternative complement pathway haemolytic activity was inhibited (p < 0.05). The gossypol level in dorsal muscles from each treatment was below the detection limit. Based on the quadratic regression analysis of the TGC, the optimal dietary CSM level was 175.9 g/kg, and the maximum could be up to 351.8 g/kg and still result in the same TGC as that of control group. [ABSTRACT FROM AUTHOR]