Non-buffered plasma-activated liquids such as water and saline have shown bactericidal effects. In the present study, we investigated the anti-bacterial efficacy and stability of plasma-activated water (PAW) and plasma-activated saline (PAS), generated using a high voltage dielectric barrier discharge system. This study compares the potential of non-buffered plasma-activated liquids (PAL) for the inactivation of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) after storage of the solutions at five different temperatures for a storage time up to 18 months after their generation. The temperatures used were room temperature, 4 °C, −16 °C, −80 °C, −150 °C. Both PAW and PAS achieved 6 log reduction for both bacteria on the first day of their generation after 60 min contact time and they retained these effects after 18 months when stored at the lowest temperatures. Chemical analysis of PAL showed that plasma caused a drop in pH, generation of reactive oxygen species and nitrates, whereas no nitrites are detected in the system used. The concentrations of chemical species were affected by the storage at different temperatures and a thermocouple probe was used to investigate the freezing behaviour of the PAL.

Retention of plasma reactive species to liquids is a promising new technology, an alterna-5-11 and also as a new eco-friendly 12 13 Building an understanding of a

The spread of multidrug-resistant bacteria poses a significant threat to human health. Plasma activated liquids (PAL) could be a promising alternative for microbial decontamination, where different PAL can possess diverse antimicrobial efficacies and cytotoxic profiles, depending on the range and concentration of their reactive chemical species. In this research, the biological activity of plasma activated water (PAW) on different biological targets including both microbiological and mammalian cells was investigated in vitro. The aim was to further an understanding of the specific role of distinct plasma reactive species, which is required to tailor plasma activated liquids for use in applications where high antimicrobial activity is required without adversely affecting the biology of eukaryotic cells. PAW was generated by glow and spark discharges, which provide selective generation of hydrogen peroxide, nitrite and nitrate in the liquid. The PAW made by either spark or glow discharges showed similar antimicrobial efficacy and stability of activity, despite the very different reactive oxygen species (ROS) and reactive nitrogen species profiles (RNS). However, different trends were observed for cytotoxic activities and effects on enzyme function, which were translated through the selective chemical species generation. These findings indicate very distinct mechanisms of action which may be exploited when tailoring plasma activated liquids to various applications. A remarkable stability to heat and pressure was noted for PAW generated with this set up, which broadens the application potential. These features also suggest that post plasma modifications and post generation stability can be harnessed as a further means of modulating the chemistry, activity and mode of delivery of plasma functionalised liquids. Overall, these results further understanding on how PAL generation may be tuned to provide candidate disinfectant agents for biomedical application or for bio-decontamination in diverse areas.

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Plasma activated liquids have demonstrated antimicrobial effects and receive increasing attention due to the potential to strengthen the armoury of novel approaches against antibiotic resistant bacteria. However, the antibacterial activity and cytotoxic effects of these solutions need to be understood and balanced before exposure to humans. In this study, the antibacterial effects of plasma activated saline (PAS) were tested against Gram negative and positive bacteria, and HaCaT keratinocytes were used for cytotoxicity studies. For the first time, a co-culture model between these bacteria and eukaryotic cells under the influence of PAS has been described. Exposure of saline to plasma resulted in high concentrations of nitrate, hydrogen peroxide and a reduction of pH. PAS caused high antibacterial effects in the co-culture model, accompanied by high cytotoxic effects to the monolayer of mammalian cells. We present evidence and provide a deeper understanding for the hypothesis that upon treatment with PAS, chemical species generated in the liquid mediate high antimicrobial effects in the co-culture setup as well as mitochondrial depolarization and glutathione depletion in HaCaT cells and cell lysis due to acidic pH. In conclusion, PAS retains strong antibacterial effects in a co-culture model, which may have unintended negative biological effects on mammalian cells.