title: Cold Plasma Inactivation of Bacterial Biofilms and Reduction of Quorum Sensing Regulated Virulence Factors.
abstract: The main objectives of this work were to investigate the effect of atmospheric cold plasma (ACP) against a range of microbial biofilms commonly implicated in foodborne and healthcare associated human infections and against P. aeruginosa quorum sensing (QS)-regulated virulence factors, such as pyocyanin, elastase (Las B) and biofilm formation capacity post-ACP treatment. The effect of processing factors, namely treatment time and mode of plasma exposure on antimicrobial activity of ACP were also examined. Antibiofilm activity was assessed for E. coli, L. monocytogenes and S. aureus in terms of reduction of culturability and retention of metabolic activity using colony count and XTT assays, respectively. All samples were treated 'inpack' using sealed polypropylene containers with a high voltage dielectric barrier discharge ACP generated at 80 kV for 0, 60, 120 and 300 s and a post treatment storage time of 24 h. According to colony counts, ACP treatment for 60 s reduced populations of E. coli to undetectable levels, whereas 300 s was necessary to significantly reduce populations of L. monocytogenes and S. aureus biofilms. The results obtained from XTT assay indicated possible induction of viable but non culturable state of bacteria. With respect to P. aeruginosa QS-related virulence factors, the production of pyocyanin was significantly inhibited after short treatment times, but reduction of elastase was notable only after 300 s and no reduction in actual biofilm formation was achieved post-ACP treatment. Importantly, reduction of virulence factors was associated with reduction of the cytotoxic effects of the bacterial supernatant on CHO-K1 cells, regardless of mode and duration of treatment. The results of this study point to ACP technology as an effective strategy for inactivation of established biofilms and may play an important role in attenuation of virulence of pathogenic bacteria. Further investigation is warranted to propose direct evidence for the inhibition of QS and mechanisms by which this may occur.
authors.affiliation: Plasma Research Group, School of Food Science and Environmental Health, Dublin Institute of Technology, Dublin 1, Ireland.
authors.forename: Dana
authors.initials: D
authors.lastname: Ziuzina
authors.affiliation: Plasma Research Group, School of Food Science and Environmental Health, Dublin Institute of Technology, Dublin 1, Ireland.
authors.forename: Daniela
authors.initials: D
authors.lastname: Boehm
authors.affiliation: Plasma Research Group, School of Food Science and Environmental Health, Dublin Institute of Technology, Dublin 1, Ireland.
authors.forename: Sonal
authors.initials: S
authors.lastname: Patil
authors.affiliation: School of Chemical Engineering, University of New South Wales, Sydney, Australia.
authors.forename: P J
authors.initials: PJ
authors.lastname: Cullen
authors.affiliation: Plasma Research Group, School of Food Science and Environmental Health, Dublin Institute of Technology, Dublin 1, Ireland.
authors.forename: Paula
authors.initials: P
authors.lastname: Bourke
mesh_terms: D000818:Animals; D001419:Bacteria / Q000472:pathogenicity* / Q000648:ultrastructure; D018441:Biofilms; D016466:CHO Cells; D003080:Cold Temperature; D006224:Cricetinae; D003412:Cricetulus; D008855:Microscopy, Electron, Scanning; D058626:Plasma Gases; D053038:Quorum Sensing; D037521:Virulence Factors
chemical_list: D058626:Plasma Gases; D037521:Virulence Factors