Biofilms are sessile microbial communities living at interfaces, in which the extracellular matrix is responsible for the mechanical stability and adhesion to surfaces. Transition from planktonic to attached cells is a key step in the biofilm formation process. Monitoring of this transition is needed to prevent contamination of biomedical devices and mitigate microbially influenced corrosion. Under anoxic local conditions and in the presence of an exogenous redox mediator, biofilms can divert part of the electron flow associated with catabolism to electrodes maintained at a defined potential. Extracellular electron transfer (EET) follows upon the attachment of planktonic cells to the surface. Modification of the electrode increases bacterial attachment, thus allowing early bioelectrochemical detection of the biofilm. Here, we report a Ni electrode micropillar array to detect early attachment of Escherichia coli biofilm through mediated EET in potentiostat‐controlled electrochemical cells. The biofilm‐surface interaction is studied by using electrochemical impedance spectroscopy (EIS) at different potentials and temperatures over 24 h. The analysis of the EIS signature highlights the effect of temperature on mediated EET in biofilms. These results demonstrate that micropillared electrodes allow earlier biofilm detection than flat electrodes, which is relevant to biofilm sensing and investigation of microbially influenced corrosion in drinking water systems and biomedical devices.

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Innovative applications like energy production from wastewater via microbial fuel cells, production of high-added value products, biosensors for bacteria detection in food, environment, and clinical samples, as well as understanding microbially influenced corrosion can be achieved through bioelectrochemical systems (BES). BES are powered by the extracellular electron transfer (EET) from bacteria into electrodes. Bacteria accumulate at interfaces to form biofilms. When they grow on electrodes, biofilms are termed electrochemically active biofilms (EAB). While biofilm formation and electrochemical activity are thought to be affected by the material and the microstructured of the interface, this interaction is not well-understood for early bacterial biofilms. This dissertation aims to investigate early bacterial biofilms at electrode surfaces, through the fabrication and testing of ordered array microstructured electrodes with different topography and materials. The goals of this research were to understand (i) the EET from bacteria to the electrode, (ii) the role of the metal surface in the transfer mechanism, and (iii) the effect of electrode microstructure on early biofilm formation. The first study involved three different gold microstructured electrodes fabricated with photolithography. The charge production showed that micropillar structures enhanced the charge transfer up to 22% higher than without any microstructure. The characterization of the electrode surface using electrochemical impedance spectroscopy (EIS) showed a higher conductivity and lower impedance for the microstructured electrodes after early biofilm growth at 24 h. Microscope analysis of the electrodes showed that Escherichia coli biofilm formation ensued at the base of the pillar microstructures. Confocal laser scanning microscopy images revealed that 41% of the cells on the electrode were alive, composing an early biofilm of 400 nm thickness. Further, the combination of the microstructures, electrochemical analysis, and imaging shed light on the EET at the biofilm/gold electrode interface. The second study employed nickel microstructured electrodes analog to the gold electrodes used in the first study, to assess the effect of a different material on early biofilm formation. Microstructured electrodes showed a minimal enhancement on the Abstract ii charge transferred (3%) for the chronocoulometry test, although the surface area was increased by 3 and 6% for small and large pillars, respectively. The cell/electrode interface was monitored through EIS at 0, 8, and 24 after cell inoculum at three different temperatures (23, 30, and 37 °C). The results showed a drop of the impedance during the first hours of the experimental runs, followed by a constant increase of the impedance with time. EIS analysis was carried on a range of electric potentials, temperatures and stirring conditions, to gain further information on the electrochemical signature at the cell/electrode interface. The results showed that non-stirring conditions favored reproducible ...