This study investigated the occurrence of corticosteroid signaling disruptors in wastewaters and rivers in the Czech Republic and in Switzerland. 36 target compounds were detected using HPLC-MS/MS, with up to 6.4 mu g/L for azole antifungals that indirectly affect corticosteroid signaling. Glucocorticoid receptor (GR)-mediated activity was determined using the GR-CALUX bioassay with dexamethasone equivalent concentrations ranging from <LOD-2.6, 19-37, and 78-542 ng/L for river water, treated, and untreated wastewater, respectively. For most samples, the chemically predicted GR-mediated response was higher than that determined by the bioassay. Correspondingly, antiglucocorticoid activity was observed in some fractions. The fish plasma model (FPM), which predicts plasma concentrations, was applied to evaluate the potential of detected pharmaceuticals to cause receptor-mediated effects in fish. With one exception, medroxyprogesterone, the FPM applied to individual compounds predicted fish plasma concentrations to be below the level of human therapeutic plasma concentrations. To account for the activity of the sum of GR-active compounds, we introduce the "cortisol equivalents fish plasma concentration" approach, through which an increase in fish glucocorticoid plasma levels comparable to 0.9-83 ng/mL cortisol after exposure to the analyzed river waters was estimated. The results suggest that these chemicals may impact wild fish.