Juvenile Paralichthys olivaceus (mean length 19.8±2.6 cm, mean weight 97.8±15.8 g) were exposed for 96 hours to different nitrate concentrations of 0, 62.5, 125, 250, 500, 1,000, and 1,500 mg L-1 in biofloc and 0, 62.5, 125, 250, 500, and 1,000 mg L-1 in seawater. Median lethal concentration values (LC50, the concentration at which 50% of mortality occurred after 96 hours of exposure) of nitrate to P. olivaceus in biofloc and seawater were 1,226 and 597 mg NO3 L-1 (P<0.05), respectively, revealing a higher toxicity of nitrate to P. olivaceus in seawater than in biofloc. In hematological parameters, hematocrit level in P. olivaceus exposed to nitrate was significantly increased only at a concentration of 1,000 mg L-1 in biofloc and at concentrations higher than 250 mg L-1 in seawater, but no significant changes in hemoglobin were found in biofloc and seawater. In plasma parameters, aspartate aminotransferase (AST) and alanine aminotransminase (ALT) were significantly increased by nitrate exposure in biofloc and seawater, but no significant changes in alkaline phosphatase (ALP) were found in biofloc and seawater. Results of this study indicate that nitrate exposure to P. olivaceus have a lethal toxic effect and alter hematological and plasma constituents of flatfish P. olivaceus. Given relatively lower toxicity of nitrate in biofloc than in seawater, the use of biofloc in aquaculture may reduce potential toxic effect caused by nitrate in feces and feed residue.