Background: Dairy cow manure applied to pastures is a significant potential source of estrogenic contamination in nearby streams. One possible pathway is through infiltration via preferential flow to drainage pipes, particularly after heavy rainfall events. In a period of 73 days in the spring of 2010, a drainage catchment in a cattle pasture in the Swiss lowlands was closely monitored. Manure was applied three times during the study, and part of the catchment was also subjected to grazing. During five field campaigns, water samples from two sampling locations were taken for 4-24 h in consecutive sampling intervals. 17β-estradiol equivalents (EEQ) were determined with the yeast estrogen screen (YES) and the ER-CALUX assay. Some water chemistry parameters, pH, conductivity, oxygen content and soil moisture tension were also monitored. Results: Washout of estrogenic activity was highest during or right after heavy rainfall events, shortly after manure spreading, when peak values of >10 ng/l EEQ were found in several samples. However, in two field campaigns, high EEQ values were also found 14 and 28 days, after the last manure application, in one case during a dry weather period. This indicates that estrogenic compounds are more stable in natural soils than what is expected from data gathered in lab studies. Conclusions: Streams in agricultural areas with a high proportion of drained land may be subject to numerous peaks of EEQ during the course of the year. This may have a negative effect on aquatic organisms, namely fish embryos, living in these streams.