<jats:p>(1) Urbanization significantly influences the ecosystems of rivers in various ways, including the so-called loading effect of wastewater production. Benthic diatoms are used in ecological status assessments of waters. Beside species composition, traits can be used as indicators. We aimed to evaluate how the loading of the large city of Budapest manifests in the physico-chemical variables of the River Danube and what species composition and trait response this loading results in for the benthic diatom communities. (2) Weekly samplings were performed at points upstream and downstream of Budapest on both riverbanks. Samples were compared, based on general physical-chemical variables and the concentration of thirty-four elements, as well as species composition and seven traits of species of diatom communities. Ecological status was assessed using the Specific Pollution Sensitivity Index (IPS). (3) Only a few measured environmental variables showed differences between the sampling points, suggesting that the nutrient loading has significantly decreased due to the installation of several efficiently working wastewater treatment plants since the introduction of the European Union Water Framework Directive. In contrast, the species composition and traits of species showed the effect of land use. Benthic diatoms indicate the environmental changes caused by land use in the longer-term, while chemical measurements reflect instantaneous status.</jats:p>