Layered Design of a Highly Repeatable Electroactive Biofilm for a Standardized Biochemical Oxygen Demand Sensor

Microbial electrochemical sensors are promising to monitor bioavailable organics in real environments, but their application is restricted by the unpredictable performance of the electroactive biofilm (EAB), which is randomly acclimated from environmental microflora. With a long-term stable EAB as a template, we successfully designed EAB (DEAB) by the sequential growth of Geobacter anodireducens and automatched microbes, achieving a reproducible high current than those naturally acclimated from wastewater (NEAB). Pre-inoculation of planktonic aerobes as oxygen bioscavengers was necessary to ensure the colonization of Geobacter in the inner layer, and the abundant Geobacter (50%) in DEAB guaranteed 4 times higher current density with a 15-fold smaller variation among 20 replicates than those of NEAB. The sensor constructed with DEAB exhibited a shorter measuring time and a precise biochemical oxygen demand (BOD) measurement with acetate, real domestic wastewater, and supernatant of anaerobic digestion. Here, we for the first time proposed an applicable strategy to standardize EABs for BOD sensors, which is also crucial to ensure a stable performance of all bioelectrochemical technologies.