ABSTRACT The stable carbon isotopic variability and the contributions of autochthonous (sediment microbial biomass, phytoplankton, and periphyton) and allochthonous (soil microbial biomass) sources available to the detritivorous fish Prochilodus lineatus were investigated in three environments of the floodplain of the Upper Paraná River. The isotopic composition of carbon sources and fish varied significantly among the studied environments. The autochthonous resources, represented by the phytoplankton, were the most assimilated by the species, followed by periphyton and sediment microbial biomass. The species used the sources differently in each environment. This study suggests that the inherent characteristics of this area, as well as the size of the watershed, the dry season, anthropogenic actions, and phytoplankton productivity, favor the use of autochthonous resources by the species studied. Therefore, studies in the floodplain should employ an eco-hydrological approach that quantifies the magnitude of energy subsidies, as well as an access route to consumers, knowledge about the selectivity of detritivorous species and the effects of different land uses.