AimsIn biosand filters (BSF), treatment is largely driven by the development of a biolayer (schmutzdecke) which establishes itself during the startup phase. In this study, the effect of changing influent total organic carbon (TOC) loading on the removal efficiency of Vibrio cholerae in laboratory-operated BSFs was quantified. Methods and ResultsBSFs were charged with high, medium or low TOC influents and removal efficacy and schmutzdecke composition was monitored over 2months. The highest V. cholerae removal efficiencies were observed in the BSF receiving the lowest TOC. Schmutzdecke composition was found to be influenced by influent TOC, in terms of microbial community structure and amount of extracellular polymeric substance (EPS). ConclusionsPhysical/chemical attachment was shown to be important during startup. The BSF receiving influent water with lower TOC had a higher attachment coefficient than the BSF receiving high TOC water, suggesting more physical/chemical treatment in the lower TOC BSF. The high TOC BSF had more EPS than did the biofilm from the low-TOC BSF, suggesting that schmutzdecke effects may be more significant at high TOC. Significance and Impact of the StudyOverall, this study confirms that influent water characteristics will affect BSF treatment efficacy of V. cholerae especially during the startup phase.