Streambed sediment is commonly analyzed to assess occurrence of hydrophobic pesticides and risks to aquatic communities. However, stream biofilms also have the potential to accumulate pesticides and may be consumed by aquatic organisms. To better characterize risks to aquatic life, the U.S. Geological Survey Regional Stream Quality Assessment measured 93 current-use and 3 legacy pesticides in bed sediment and biofilm from 54 small streams in California across a range of land-use settings. On average, 4 times as many current-use pesticides were detected in biofilm at a site (median of 2) as in sediment (median of 0.5). Of 31 current-use pesticides detected, 20 were detected more frequently in biofilm than sediment and 10 equally frequently. Pyrethroids as a class were the most potentially toxic to benthic invertebrates, and of the nine pyrethroids detected, seven occurred more frequently in biofilm than sediment. We constructive General Additive Models to investigate relations between pesticides and six metrics of benthic community structure. Pesticides in biofilm improved fit in four of the six models, and pesticides in sediment improved fit in two. The results indicate that sampling of stream biofilms can complement bed-sediment sampling by identifying more current-use pesticides present and better estimating ecological risks.