This dataset gathers information about the macroinvertatebrate samples and environmental variables collected on rivers of the Ebro River Basin (NE Iberian Peninsula), the second largest catchment in the Iberian Peninsula. The collection is composed of 1,776 sampling events carried out between 2005 and 2015 at more than 400 sampling sites. This dataset is part of a monitoring network set up by the Ebro Hydrographic Confederation, the official body entrusted with the care of the basin, to fulfill the requirements of the European Water Framework Directive. Biological indices based on the freshwater macroinvertebrate communities were used to evaluate the ecological status of the water bodies within the basin. Samples were qualitatively screened for all occurring taxa. Then, all individuals from all taxa in a quantitative subsample of each sample were counted. Biological indices were calculated to estimate water quality at each sampling site. All samples are kept at the Museum of Zoology of the University of Navarra.