Taking into account that the relationship between metabolic substrate and the characteristics of the diet is essential for understanding the mechanisms associated with the use of the components of a certain food substance, the aim of this study was to examine the metabolic substrate used by Penaeus setiferus,P. schmitti, P. duorurum and P. notialis postlarvae PL 35–40 (35–40 days after metamorphosis) fed purified diets with different levels of proteins (40, 50, 60 and 65%). Oxygen consumption and ammonia excretion were measured in animals in intermoult which had been acclimatized to each diet over a minimum of 5 days. Measurements were taken on fasted (24 h) and fed animals. Oxygen consumption of fasting animals increased with a rise in the proteins of the diet in the four species. In the fed animals, this tendency was observed in P. setiferus and P. notialis, while in P. schmitti and P. duorarum oxygen consumption was a weak relationship, decreasing or increasing with a diet with 50% protein. Both in fasting and feeding animals, the ammonia excretion increased with the increase in proteins in the diet. The O : N atomic ratios of the four species decreased (between 30 and 3), according to increase in dietetic protein : energy ratio, (10.4 to 17.0 mg protein/kcal). The highest value of O : N ratio was observed in P. setiferus (40% of protein requirement, O : N = 24 to 34) and lowest in P. schmitti (60% of protein requirement, O : N = 4 to 11), with intermediate values in P. duorarum and P. notialis (50% protein requirement, O : N = 8 to 14 and 13 to 19, respectively). These results support previous research that showed that the omnivorous-herbivorous species, like P. setiferus, use protein-lipids normally as energy source in contrast to omnivorous-carnivorous species such as P. duorarum which use protein preferentially as a source of energy.