The Salinas River watershed along the central coast of California, U.S.A., supports rapidly growing urban areas and intensive agricultural operations. The river drains to an estuarine National Wildlife Refuge and a National Marine Sanctuary. The occurrence, spatial patterns, sources and causes of aquatic toxicity in the watershed were investigated by sampling four sites in the main river and four sites in representative tributaries during 15 surveys between September 1998 and January 2000. In 96 hr toxicity tests, significant Ceriodaphnia dubia mortality was observed in 11% of the main river samples, 87% of the samples from a channel draining an urban/agricultural watershed, 13% of the samples from channels conveying agricultural tile drain runoff, and in 100% of the samples from a channel conveying agricultural surface furrow runoff. In six of nine toxicity identification evaluations (TIEs), the organophosphate pesticides diazinon and/or chlorpyrifos were implicated as causes of observed toxicity, and these compounds were the most probable causes of toxicity in two of the other three TIEs. Every sample collected in the watershed that exhibited greater than 50% C. dubia mortality (n = 31) had sufficient diazinon and/or chlorpyrifos concentrations to account for the observed effects. Results are interpreted with respect to potential effects on other ecologically important species.