Distillers dried grains with solubles (DDGS), a feed coproduct from the fuel ethanol industry, has been shown to be a viable potential alternative protein source for aquaculture feeds. To investigate this, three isocaloric (3.5kcal/g) ingredient blends containing 20, 30, and 40% DDGS, with a net protein adjusted to 28% (wet basis, wb), were prepared for use as Nile tilapia feed. Extrusion processing was then conducted using three DDGS contents (20, 30, and 40%, wb), three moisture contents (15, 20, and 25%, wb), three barrel temperature gradients (90–100–100°C, 90–130–130°C, and 90–160–160°C), and five screw speeds (80, 100, 120, 140, and 160rpm) using a single screw laboratory extruder. Several processing parameters, including mass flow rate, net torque required, specific mechanical energy consumption, apparent viscosity, and temperature and pressure of the dough inside the barrel and die, were measured to quantify the extrusion behavior of the DDGS-based blends. For all blends, as the temperature profile increased, mass flow rate exhibited a slight decrease, die pressure decreased, and apparent viscosity exhibited a slight decrease as well. Likewise, the net torque requirement, specific mechanical energy consumption, and apparent viscosity decreased as screw speed increased, but mass flow rate increased. Additionally, as moisture content increased, die pressure decreased. At higher temperatures in the barrel and die, the viscosity of the dough was lower, leading to lower torque and specific mechanical energy requirements. Increasing the DDGS content, on the other hand, resulted in a higher mass flow rate and decreased pressure inside the die. As demonstrated in this study, the selection of suitable temperature and moisture content levels are critical for processing DDGS-based ingredient blends.