A study was conducted to determine the possible synergistic effects between dietary rutin (a bioflavonoid) and vitamin C, and to evaluate their antioxidant effects in fingerling channel catfish. Purified casein/gelatin diets containing two levels of rutin (0 and 1000 mg/kg diet) and three levels of L-ascorbic acid (0, 1500 and 3000 mg/kg diet) in a factorial arrangement were fed to fingerling channel catfish for 16 weeks. Fish fed the diets without supplemental vitamin C showed deformed spinal columns, external hemorrhages and fin erosion after 10 to 12 weeks. Also these fish had significantly (p < 0.05) depressed body weight gain, feed efficiency, hematocrit, hepatosomatic index (% liver weight), as well as reduced liver, fillet and plasma vitamin concentrations after 16 weeks. Liver, fillet and plasma vitamin C concentrations were correlated with dietary vitamin C levels. Forced oxidation of fillet samples significantly (p < 0.05) increased 2-thiobarbituric acid (TBA) values of fillets from fish fed diets without vitamin C and rutin. However, results from the present study indicated only limited synergistic effects of dietary rutin on vitamin C nutrition of channel catfish.