1 2 3 4 5 6 7 8 9 10 0 2 4 6 8 C O Cu Intensity Energy (keV) Spec 1 Spec 2 Ag Eapp = 0V, tapp = 5 min 1 2 3 8 0 1 2 70 71 72 73 74 75 Ag S Cu Cu O Intensity (a.u) Energy (keV) C Low-power electrically-polarized nanoscale metallic surfaces composed of cupric salt and silver metal exhibited effective deactivation against a broad range of microbes. Six log reduction of five different bacteria – both gram-positive and gram-negative, was observed with electrically-polarized ENM devices. ENM devices were also effective against one viral particle and two medically-relevant fungi tested in the studies. In-situ hydrogen peroxide in micromolar concentration was produced through the oxygen reduction reaction mediated by cuprous ions. The electrochemical steady concentration of cuprous ions in molar concentration over ten days is key to a sustainable production of highly reactive hydroxyl radicals in the solution. These studies add a new toolbox to producing highly reactive oxygenated species by utilizing an external power battery to electrically-polarizing the nanoscale metallic films. 0 10 20 30 40 50 0.0 0.5 1.0 [AA]/[AA]o tapp (min) Control Ag, 0V Ag, 3V ENM, 0V ENM, 3V 350 400 450 500 550 600 0 100 200 300 non-ENM, Eapp= 0V 0 h 48 h 96 h 120 h Intensity (au) Wavelength (nm) 0 60 120 0 1 2 3 4 5 non-ENM 0V ENM 3V [OH.] nanomoles/cm2 tapp (hr) 0.0 0.1 5 10 15 0 8 16 24 32 ENM, 3V non-ENM, 0V Ag, 0V Ag, 3V [Cu(I)] M/cm2 tapp ( 1000 min) 𝐂𝐮+ Accelerated deactivation of broad-range microbes via in-situ generation of reactive oxygenated species M. Aswad Ali1a, Annie Y. Vargas-Lizarazo1a, Tyler Sons2, Michelle Garnett2, Jose Vargas2, Michael Olson3, Philip Jensik4, Scott Hamilton- Brehm2, Punit Kohli1 1Department of Chemistry and Biochemistry, Southern Illinois University, Carbondale, Illinois 62901, USA; 2Department of Microbiology, Southern Illinois University, Carbondale, Illinois 62901, USA; 3Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901, USA; 4Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901, USA; aEqual contributors Abstract Development of new strategies that are effective against a broad range of microbes is a crucial area of research because of dwindling new antibiotics and emergence of antibiotic-resistant microbes. Here, we report on implementing electrically-polarized nanoscale metallic coatings (ENM) that deactivate a broadband microbes including Gram-negative and Gram-positive bacteria with six log reduction, within five minutes of treatment. ENM devices were also effective in the deactivation of a virus and a fungus with high efficiency. Highly damaging in-situ reactive oxygenated species (ROS), including hydrogen peroxide and hydroxyl radicals, were produced by electrically-polarized metallic coatings. A sustainable micromolar ROS concentration was observed for about three days in ENM devices allowing effective antimicrobial surfaces for long-term usages. ENM represents a non-conventional tool providing a highly effective antimicrobial mode of deactivation through the production of highly damaging ROS to microbes. Fabrication of ENM Characterization Conclusion Acknowledgement Microbe Droplet on ENM Deactivation through ROS/Free radical mediated processes triggered by electrical polarization 2. Soaking in Cu2+ 3. Sputtering Ag O2, H2O 𝐞− 𝐂𝐮𝟐+ Battery ENM Method - Deactivation of Microbes by ENM Cu Ka1 & Ag La1 mix Results – Deactivation of Microbes 𝐂𝐮𝟐+/𝐂𝐮+couple mediates formation of ROS by ENM through 𝐂𝐮+diffusion into bulk SEM Mix Map and EDS Confirmation of the Species Present in ENM Water Static Contact Angle Measurements = 118o ± 8o AFM Determination of Thickness Ag Coating and Particle Size Distribution 20 µm 20 µm Non-homogeneity in the Junction of the Fibers-Two Electrode Equivalency Junction of overlapping fibers does not have Ag coating – Prevents shorting Spec 1 Spec