Abstract Ecological studies on food webs have considerably increased in recent decades, especially in aquatic communities. Because Chironomidae family are highly specious, occurring in almost all aquatic habitats is considered organisms-key to initiate studies on ecological relationships and trophic webs. We tested the hypothesis that the diversity of the morphospecies diet reflects differences on both the food items available among habitats and the preferences of larval feeding. We analyzed the gut content of the seven most abundant Chironomidae morphospecies of the different habitats from the Upper Paraná River. We categorized the food items found into algae, fungal spores, fragments of plants, algae and animal fragments and sponge spicules. We observed the algae predominance in the gut content of morphospecies from lakes. Considering the different regions from each lake, we registered the highest food abundance in the littoral regions in relation to the central regions. From the variety of feeding habits (number of item kinds), we classified Chironomus strenzkei, Tanytarsus sp.1, Procladius sp.1 as generalist morphospecies. We found a nested pattern between food items and Chironomidae morphospecies, where some items were common to all taxa (e.g., Bacillariophyceae algae, especially), while others were found in specific morphospecies (e.g., animals fragments found in Procladius sp.1). The algae represented the most percentage of gut contents of Chironomidae larvae. This was especially true for the individuals from littoral regions, which is probably due to the major densities of algae associated to macrophytes, which are abundant in these regions. Therefore, the feeding behavior of these morphospecies was generalist and not selective, depending only of the available resources.