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High sensitivity is essential for the application of bioelectrochemical system-based sensor (BES sensor) in water quality early-warning, where the electroactive biofilm is of vital importance as it delivers a responsive electric signal to toxic substances. This study artificially designed the morphology of a naturally formed biofilm by employing a serrated knife to scrape the biofilm and thus obtained a reduced thickness and roughness. Then it was further cut by half to halve the biomass. BES sensors equipped with control and processed biofilms were operated under constant anode potential (CAP) and tested at different Cu(II) concentrations to study their sensitivities. Results revealed that the scraped biofilms delivered much increased sensitivity towards Cu(II) shock, which was attributed to a reduced thickness as illustrated by macroscopic and microscopic morphology analysis. Another finding was that biomass per unit …