As flood events are expected to become more frequent due to climate change, investigating how overland flow exports terrestrial nutrients, carbon and living organisms into aquatic systems is essential for understanding both soil and stream ecosystem status. Here we assessed how dissolved organic carbon (DOC), total suspended sediments (TSS), and stream bacterial diversity responded to stream discharge and overland flow during stormflow in a tropical catchment. A higher humification index and a decreasing ratio of allochthonous to autochthonous DOC indicated that DOC from soils was exported to stream during the flood. The δ 13 C and δ 15 N of particulate matter was indicative of a source in the cultivated areas of the upper catchment and of subsurface soils (stream banks and gullies) in the downstream section. Bacterial richness of particle-attached (PA) and the free-living (FL) fractions increased with the flood progression in the upstream section. Moreover, the community structure of the PA fraction in the stream was more similar to that of overland flow than was the FL fraction. This suggests that the soil PA bacterial community was washed-out with overland flow during the flood recession. The relative contribution of sources and the composition of TSS, rather than hydrological regime, significantly drove the composition of bacterial community. In conclusion, our results emphasize that overland flow during a flood event strongly influences the structure of stream bacterial communities further underlining the biological connectivity between terrestrial runoff and stream flow.