The aim of this study was to investigate the effect of diet extruder conditions, such as feed-loading rate and screw speed, on growth performance and biochemical responses in olive flounder Paralichthys olivaceus. Over 8 wks, we used four identical diets (triplicated per treatment) with differing ratios of feed-loading rate (kg/h):screw speed (rpm/min) in a laboratory-scaled twin-screw extruder of 50:640, 80:640, 120:640, and 80:400, designated as EP1, EP2, EP3, and EP4, respectively. Screw speed impacted the buoyancy of experimental diets. Diets produced at a screw speed of 640 rpm/min floated for > 24 hrs, whereas those produced at a speed of 400 rpm/min sank between 10 s and 5 min. Fish that were fed EP1 and EP4 diets grew significantly faster than those fed EP2 and EP3 diets. Fish fed EP1 diets ate and gained weight most efficiently among treatments, a result that is likely to be related to feed-loading rate, i.e., ingredients extruded at a low feed-loading rate may have more time to cook in the pre-conditioner of the extruder. A cooked diet may be easier to digest in fish. Fish fed EP4 diets also showed significant weight gain, as compared to those fed EP2 and EP3 diets. However, we found no differences among treatments in proximate compositions of dorsal muscle, liver, and viscera of fish. Our results suggest that extruder conditions, may influence feed quality, impacting feed efficiency and growth of fish.