Summary The Okavango Delta of Botswana is a large arid-zone wetland comprising 20,000km2 of permanent and seasonal floodplains and over 100,000 islands. It has been shown that island groundwater can have very high dissolved arsenic (As) concentration, but the abiotic and biotic controls on As mobility are not well understood in this setting. At New Island, an island located in the seasonal swamp, dissolved As concentration increased from below detection limits in the surface water to 180μg/L in groundwater, present as As(III) species. We investigated the relative importance of hydrologic, geochemical, and geomicrobial processes, as well as influences of recent extreme flooding events, in mobilizing and sequestering As in the shallow groundwater system under this island. Our results suggest that evapotranspiration and through-flow conditions control the location of the high arsenic zone. A combination of processes is hypothesized to control elevated As in the concentration zone of New Island: high evapotranspiration rates concentrate As and other solutes, more alkaline pH leads to desorption of arsenic or dissolution of arsenic sulfides, and formation of thioarsenic complexes acts to keep arsenic in solution. Evidence from X-ray absorption near-edge structure spectroscopy (XANES) and sulfate reducing bacteria (SRB) measurements further suggests that SRBs influence arsenic sequestration as orpiment (As2S3). Although dissolved organic matter (DOM) was not significantly correlated to dissolved As in the groundwater, our results suggest that DOM may serve as an electron donor for sulfate reduction or other microbial reactions that influence redox state and As mobility. These results have important implications for water management in the region and in other large wetland environments. The processes evaluated in this study are also relevant for arsenic removal in subsurface constructed wetland systems that may exhibit rapidly changing processes over small spatial scales.