The correct definition of ecosystem needs is essential in order to guide policy and management strategies to optimize the increasing use of freshwater by human activities. Commonly, the assessment of the optimal or minimum flow rates needed to preserve ecosystem functionality has been done by habitat-based models that define a relationship between in-stream flow and habitat availability for various species of fish. We propose a new approach for the identification of optimal flows using the limiting factor approach and the evaluation of basic ecological relationships, considering the appropriate spatial scale for different organisms. We developed density-environment relationships for three different life stages of brown trout that show the limiting effects of hydromorphological variables at habitat scale. In our analyses, we found that the factors limiting the densities of trout were water velocity, substrate characteristics and refugia availability. For all the life stages, the selected models considered simultaneously two variables and implied that higher velocities provided a less suitable habitat, regardless of other physical characteristics and with different patterns. We used these relationships within habitat based models in order to select a range of flows that preserve most of the physical habitat for all the life stages. We also estimated the effect of varying discharge flows on macroinvertebrate biomass and used the obtained results to identify an optimal flow maximizing habitat and prey availability.