This study evaluated the bio-economic assessment, growth performance, and nutrient utilization of non-conventional protein sources as substitutes for traditional feed ingredients in Nile tilapia (Oreochromis niloticus) diets. Four test diets were formulateded with various combinations of non-conventional protein sources, including agricultural by-products, rendered animal meals, and insect components, detailed in Table 4. These diets were formulated using the novel FUTA aquafeed software and administered to tilapia over a period of 56 days. Biological parameters such as weight gain, feed efficiency ratio (FER), protein efficiency ratio (PER), and survival rate were compared with the control diet containing the conventional fishmeal. Economic analysis, based on feed formulation costs and production yields, was also conducted. The results presented indicated that significantly higher (P>0.05) mean weight gain, FER, PER and better FCR was recorded in fish fed with a diet comprising 30% poultry viscera meal and 10% Mopane worm meal (Diet C), in comparison to the control and diets containing Palm kernel cake (Diet A) and blood meal (P<0.05). However, the control diet recorded the best profitability per unit biomass produced. These findings highlighted the potential for the widespread adoption of specific non-conventional feed ingredients, including blood meal, poultry intestine, mopane worms, water fern, palm kernel meal, and wheat meal, to decrease feed costs without compromising tilapia growth.