Generation of Plasma-Activated Fluids for Successful Disinfection of Pseudomonas aeruginosa in Liquid Environments and Determination of Microbial Damage

Hummert, Mareike; Leenders, Paul; Mellmann, Alexander; Becker, Karsten; Kuczius, Thorsten

Abstract

Abstract The application of the non-thermal atmospheric pressure plasma technology is a promising tool for microbial inactivation. During the activation process, many reactive substances and radicals arise associated with physicochemical changes in the fluid and massive pH drop. In this study, we analyzed and optimized plasma activation settings and conditions of water and liquids to obtain inactivation of the waterborne microorganism Pseudomonas aeruginosa in a liquid environment. The minimal electrical output was 60 Watt with 20 min activation time followed by 30 min contact time with 108 cells/mL. Using higher electrical power (>90 W) with a Lab Unit generating plasma-activated water, a shorter activation time (<10 min) was sufficient for bacterial inactivation. The organic and inorganic composition of the activated liquid with different mineral salt concentrations is of utmost importance for the yield of reactive species during the plasma activation process and consequently for the antimicrobial effect. Plasma-activated fluids with high organic and inorganic contents demonstrated lower inactivation efficiencies than low loaded fluids; yet antimicrobial efficacy could be achieved by increasing the electrical power and activation time. For sufficient inactivation of bacterial suspensions, at least half a volume unit of plasma-activated water had to be added after appropriately optimized activation. Further dilutions reduced the antimicrobial effect. PAW lost activity after being left standing for a prolonged time after activation, so for maximizing the antimicrobial effect a direct use after activation is recommendable. Bacterial inactivation was shown by the absence of colony forming units on culture media and, at the molecular level, damage to the membrane and inactivation of enzymes were observed. Plasma-activated fluids demonstrated a high potential in applications as microbiological disinfectant in liquids.