Sustainable management of dryland river systems is often complicated by extreme variability of precipitation in time and space, especially across large catchment areas. Understanding regional water quality changes in southern African dryland rivers and wetland systems is especially important because of their high subsistence value and provision of ecosystem services essential to both public and animal health. We quantified seasonal variation of Escherichia coli (E. coli) and Total Suspended Solids (TSS) in the Chobe River using spatiotemporal and geostatistical modeling of water quality time series data collected along a transect spanning a mosaic of protected, urban, and developing urban land use. We found significant relationships in the dry season between E. coli concentrations and protected land use (p = 0.0009), floodplain habitat (p = 0.016), and fecal counts from elephant (p = 0.017) and other wildlife (p = 0.001). Dry season fecal loading by both elephant (p = 0.029) and other wildlife (p = 0.006) was also an important predictor of early wet season E. coli concentrations. Locations of high E. coli concentrations likewise showed close spatial agreement with estimates of wildlife biomass derived from aerial survey data. In contrast to the dry season, wet season bacterial water quality patterns were associated only with TSS (p<0.0001), suggesting storm water and sediment runoff significantly influence E. coli loads. Our data suggest that wildlife populations, and elephants in particular, can significantly modify river water quality patterns. Loss of habitat and limitation of wildlife access to perennial rivers and floodplains in water-restricted regions may increase the impact of species on surface water resources. Our findings have important implications to land use planning in southern Africa’s dryland river ecosystems