Abstract
Efficient wastewater treatment monitoring is vital for addressing water scarcity. Microbial fuel cells (MFCs) have emerged as real-time biosensors for biochemical oxygen demand (BOD) in urban wastewater. Discrepancies in signal generation may arise due to changes in the composition and metabolism of mixed-culture electroactive biofilms stemming from different wastewater compositions. In this study, 3D-printed MFC-based biosensors were employed to assess the BOD of sterile complex artificial wastewater and untreated urban wastewater. Alterations in the microbial composition of the anode were evaluated using 16S rRNA sequencing and metagenomics analysis. Results show that MFC-based biosensors can be effectively recalibrated for diverse types of wastewater, maintaining consistent sensitivity (0.64 ± 0.10 mA L mg
−1
m
−2
with synthetic wastewater and 0.78 ± 0.13 mA L mg
−1
m
−2
with urban wastewater) and limit of detection (49 ± 8 mg L
−1
for synthetic wastewater and 44 ± 7 mg L
−1
for urban wastewater). Crucially, pre-sterilization, conductivity adjustments, and nitrogen purging of wastewater are not required before its introduction into the biosensor. However, the presence of native aerobic microorganisms in the wastewater might affect the current output. Metagenomics and taxonomic analyses revealed that the alterations in biofilm composition are predominantly in response to the varied chemical and microbiological compositions of different substrates. Despite variations in anodic biofilm composition, the MFC-based biosensor maintains a relative error comparable to the standard BOD test. This highlights the resilience and flexibility of the biosensor when directly used with a variety of wastewater types before full biofilm adjustment.