Two experiments were conducted to assess the nutritional value and potential use of mesquite bean (Prosopis juliflora) meal (MBM) to substitute corn in extruded diets for Nile tilapia (Oreochromis niloticus). Digestible energy and the content of protein, soluble carbohydrates and amino acids were measured for 160 Nile tilapia juveniles (average weight 25.58 ± 5.52 g), allocated to eight treatment groups (four fed the experimental diet, and four fed a reference diet). To determine the inclusion potential of MBM, 384 fish (average weight 21.85 ± 1.01 g) were used and allocated to 24 tanks (1,000 L capacity). The experimental design was completely randomized with six levels of substitution (0%, 20%, 40%, 60%, 80% and 100%), and four repetitions. After the feeding experiment, tissue samples (blood, liver, muscle and intestine) were used to analyse metabolic and haematological variables. MBM produced apparent digestibility coefficients (ADC) similar to those of traditional diet ingredients: 81.85% dry matter (DM), 83.02% crude energy, 88.57% crude protein, 82.88% lysine and 82.57% methionine.Differences were observed in the digestibility of soluble carbohydrates (p < 0.05), with MBM having higher levels of starch (72.19%) and sucrose (88.99%). A 20% substitution with MBM produced a decrease in zootechnical performance (p < 0.05). Enzymatic activity of amylase and sucrose seemed to be induced in response to the proportion of their respective substrate in the diets. We observed a reduction in the efficiency of dietary protein utilization, with reduced levels of plasma amino acids (p < 0.05), owing to the increase in the enzymatic activity of muscular alanine and aspartate transaminases (ALT and AST) and the deaminase glutamate dehydrogenase in the hepatic and muscular tissues. Haematological variables did not indicate any adverse effects of MBM on the health status or survival of the fish (100%). Therefore, we conclude that MBM may be a viable alternative to corn meal in the diet, enabling substitution of up to 20%. K E Y W O R D Sdigestion, fish, haematology, metabolism, performance Abbreviations: ALT, alanine aminotransferase; AST, aspartate aminotransferase; CM, corn meal; GDH, glutamate dehydrogenase; MBM, Mesquite bean meal.