As the global population grows, there is an increasing need to produce food; aquaculture can help meet some of this growing demand. The majority of protein for aquafeeds is fish meal. But due to growing costs and potential scarcity of fish meal supplies in coming years, there is an impending need to find alternative protein sources. DDGS, a coproduct of the fuel ethanol industry, may be suitable for this purpose. The objective of this study was to examine the effect of extrusion processing conditions on two feed blends containing DDGS and other ingredients. Based on previous research, two optimized aquafeeds were formulated for Nile Tilapia. The first blend contained 15% DDGS, 58% soybean meal, 21.0% corn flour, 2.4% mono calcium phosphate, 0.75% vitamin and mineral mix, and 2.85% soybean oil. The second contained 20% DDGS, 44.2% soybean meal, 25.0% corn flour, 8% Menhaden fish meal, 0.8% vitamin and mineral mix, and 2% soybean oil. The blends were processed in a pilot-scale twinscrew extruder at speeds between 360 and 475 rpm, using a 3.175-mm circular die. Extruder parameters, including moisture content after the conditioner and at the die, as well as mass flow rate, were measured to quantify the extruder behavior during processing. The resulting extrudates were subjected to extensive physical property analysis, which included moisture content, water activity, color (Hunter L, a, and b), unit density, bulk density, pellet durability index (PDI), water stability, and floatability. All process settings produced viable extrudates. Extrudate bulk density ranged from 0.35 to 0.41 g/cm3; PDI ranged from 87.42 to 94.35%, all extrudates were stable in water and floated for more than 30 min. This study highlights the importance of experimentally quantifying the effects of feed ingredients and process variables when developing aquafeeds with novel materials