Consumption of water polluted by faecal contaminants is responsible for 2 million deaths annually, most of which occur in developing countries without adequate sanitation. In tropical aquatic systems, streambeds can be reservoirs of persistent pathogenic bacteria and high rainfall can lead to contaminated soils entering streams and to the resuspension of sediment-bound microbes in the streambed. Here, we present a novel method using fallout radionuclides (Be-7 and Pb-210(xs)) to estimate the proportions of Escherichia coli, an indicator of faecal contamination, associated with recently eroded soil particles and with the resuspension of streambed sediments. We show that using these radionuclides and hydrograph separations we are able to characterize the proportion of particles originating from highly contaminated soils and that from the resuspension of particle-attached bacteria within the streambed. We also found that although overland flow represented just over one tenth of the total flood volume, it was responsible for more than two thirds of the downstream transfer of E. coli. We propose that data obtained using this method can be used to understand the dynamics of faecal indicator bacteria (FIB) in streams thereby providing information for adapted management plans that reduce the health risks to local populations.