Influence of Attached Bacteria and Biofilm on Double-layer Capacitance During Biofilm Monitoring by Electrochemical Impedance Spectroscopy

Development of an effective strategy for biofilm control in water-related system has become a matter of significant concern nowadays. Electrochemical monitoring, especially electrochemical impedance spectroscopy (EIS), is one of the efficient approaches to dealing with biofilm-related issues. However, currently used EIS methods without a redox probe intend to detect all effects generated from media components, bacteria, and bacterial metabolites, which used to make the signals from the attached bacteria and biofilm weakened. In this study, we tried improved EIS measurement to monitor bacterial adhesion and biofilm maturation using a double-layer capacitance. In this improved method, we minimized background signal by subtracting the interference of electrolyte caused by bacterial metabolism. Pseudomonas aeruginosa PA14 wild type and wspF mutant that form the biofilm of distinct nature were used for the model strains to test our method. During bacterial adhesion and biofilm maturation, EIS data were collected and equivalent circuit analysis was carried out to obtain constant phase element (CPE) values representing double-layer capacitance. Since the influence by the bacterial growth-related culture media condition was eliminated by adopting fresh electrolyte at the measurement, the contribution of attached bacteria and biofilm was exclusively measured. As a result, the bacterial adhesion at the early stage of biofilm development was specifically monitored from reduction in double-layer capacitance. Particularly, the plateau in double-layer capacitance appeared upon biofilm maturation, indicating that biofilm maturation could be expected beyond this point. In conclusion, this study found that measurement of double-layer capacitance based on EIS could provide a monitoring parameter suggesting bacterial adhesion and the initiation point of biofilm maturation.