A 3 (protein levels, 380, 460 and 520 g kg–1 diet) × 2 (lipid levels, 65 and 140 g kg–1 diet) factorial experiment with three replicates was conducted. Weight gain, feed efficiency and daily feed intake were not significantly affected by dietary protein level, but were by dietary lipid level. Weight gains of fish fed 65 g lipid kg–1 diet were significantly, or slightly, higher than for 140 g lipid kg–1 diet at all protein levels. Daily protein intake was significantly affected by both dietary protein and lipid levels (P < 0.002). Daily lipid intake was not significantly affected by dietary protein level, but was by dietary lipid level (P < 0.001). Protein efficiency ratio was significantly affected by dietary protein level (P < 0.02), but not by dietary lipid level. Protein efficiency ratio tended to improve with the decrease of dietary protein level at the same lipid level. Moisture, protein and lipid contents of whole fish were significantly affected by dietary lipid level (P < 0.01). Increased dietary lipid did not improve growth or feed efficiency, but increased body fat deposition. It was concluded that the optimum dietary protein and lipid level for growth of juvenile ayu may be 380 and 65 g kg–1 diet, respectively, when fish were fed to satiety three times daily in seawater.