The responses of cytochrome P4501A1 in the liver, the appearance of fluorescent metabolites in bile, and genotoxic damage in erythrocytes were studied in Oncorhynchus mykiss exposed under laboratory conditions to sediments taken from the Biobio River, central Chile. Samples were taken at four sampling sites in the Biobio River, following a pollution gradient from the discharge, where Stations 2 and 3 are impacted by a petrochemical industrial discharge effluent. Chemical analysis indicates polycyclic aromatic hydrocarbon (PAH) presence in a range of 2000 to 7000 ng g(-1) dry weight in sediments from Stations 2 and 3, respectively. Both the CYP1A1 activity and the bile metabolites of O. mykiss exposed to the PAH-contaminated sediments present statistically significant differences with respect to the other stations. Additionally, the comet assay revealed notable genotoxic damage in trout erythrocytes from Stations 2 and 3, an effect that was not observed at the other stations.