Vitellogenin, a yolk-precursor protein normally found only in the blood plasma of sexually mature female teleosts and other egg-laying vertebrates, was used as an indicator of exposure of male rainbow trout (Oncorhynchus mykiss) to exogenous estrogens. Vitellogenin concentrations were measured using a specific radioimmunoassay for trout. Cages containing adult male trout were placed at the points of discharge and at varying distances downstream of treated sewage effluent outfalls into the River Lea (UK) during the summer and winter months of 1992. After 3 weeks exposure at the majority of sites, fish held up to 15 km downstream of inputs showed an increase in plasma vitellogenin concentration, with statistically significant elevations up to 4.5 km downstream. A repeat survey in November 1992 below Harpenden sewage treatment works showed that the only two stations to give a significant response were 3 m and 1.6 km downstream of the outfall. This reduced effect compared to the first survey is thought to be due to dilution of both the influent sewage to the treatment works and of the river water itself by increased rainfall, the overall increase in dilution being approx. 36%. Trout were also placed in 15 raw water storage reservoirs in southeast England during the summer of 1993 for an exposure period of 6 weeks. No biologically significant increases in plasma vitellogenin concentration were observed in any of the reservoirs.