Numerous studies have identified the issue of road surface runoff as a source of contamination into waterways but the impact of vehicular wash-off is less well understood. A ford crossing provides a pathway for vehicle-derived contaminants emanating from both road surface runoff and vehicular wash-off into a river system. Twyford Lane Ford (Ford 1) and Birchgrove Lane Ford (Ford 2), located ca. 600 m apart on a tributary of the River Ouse in Sussex (UK), were the focus of this study. A combination of biomonitoring (assessment of benthic macroinvertebrates) and chemical assessments of water and sediments has been undertaken to determine any detrimental impacts, such as a lack in biodiversity, resulting from the ford crossings. Sediment concentrations of chromium (Cr3+), lead (Pb) and zinc (Zn) were generally elevated at Ford 1, attenuating at sampling points between the fords to then peak at Ford 2. However, sediment particle size was seen to have an influence on elemental concentrations. In general, an increase in elemental concentrations was associated with a higher percentage of fine-grained sediments (≤63 μm). Elevated concentrations of Zn and magnesium (Mg) were identified within water samples taken during a precipitation event following a prolonged dry period. The biomonitoring results found reduced BMWP (Biological Monitoring Working Party) scores at positions close to the ford crossings, and where the stream was in proximity to the roadside. Sensitive Ephemeroptera were largely absent at sampling points closest to the fords, which is likely to be associated with elevated Zn. The results suggest that careful consideration should be applied when selecting crossing points over sensitive waters.