Highly sensitive standardized toxicity biosensors for rapid water quality warning

[Display omitted] •A layered design strategy is effective to standardize biotoxicity sensors.•The sensitivity of microbial electrochemical sensors was improved by 2.3 times.•The highly repeatable performance decreased coefficient of variation by 27 times.•SEAB also exhibited a better tolerance to low concentration of formaldehyde. Microbial electrochemical sensor (MES) using electroactive biofilm (EAB) as the sensing element represents a broad-spectrum technology for early warning of biotoxicity of water samples. However, its commercial application is impeded by limited sensitivity and repeatability. Here, we proposed a layered standardized EAB (SEAB) with enriched Geobacter anodireducens SD-1 in the inner layer and self-matched outer layer. The SEAB sensors showed a 2.3 times higher sensitivity than conventional EAB acclimated directly from wastewater (WEAB). A highly repeatable response sensitivity was concentrated at 0.011 ± 0.0006 A/m2/ppm in 4 replicated batches of SEAB sensors (R2 > 0.95), highlighting their potential for reliable toxicity monitoring in practical applications. In contrast, the sensing performance of all WEAB sensors was unpredictable. SEAB also exhibited a better tolerance towards low concentration of formaldehyde, with only a 4 % loss in viability. Our findings improved the sensitivity and reproducibility of standardized MES for toxicity early warning.