No abstract

Recent research has proven the ability of cold atmospheric plasma (CAP) for assuring food safety. A more flexible and transportable alternative is the use of plasma activated liquids (PAL), which are also known to have antimicrobial properties. However, within the context of food safety, little is known on its potential regarding decontamination. This research therefore focusses on identifying the impact of (i) the microbial species and its cell type (planktonic cells or biofilms), (ii) the CAP settings (i.e., gas composition and generation time) and (iii) PAL related factors (treatment time and PAL age) on the technologies efficacy. Cell densities were monitored using the plate counting technique for which the results were analyzed by means of predictive inactivation models. Moreover, the pH and the concentrations of long-lived species (i.e., hydrogen peroxide, nitrite, and nitrate) were measured to characterize the PAL solutions. The results indicated that although the type of pathogen impacted the efficacy of the treatment, mainly the cell mode had an important effect. The presence of oxygen in the operating gas ensured the generation of PAL solutions with a higher antimicrobial activity. Moreover, to ensure a good microbial inactivation, PAL generation times needed to be sufficiently long. Both CAP related factors resulted in a higher amount of long-lived species, enhancing the inactivation. For 30 min. PAL generation using O 2 , this resulted in log reductions up to 3.9 for biofilms or 5.8 for planktonic cells. However, loss of the PAL activity for stored solutions, together with the frequent appearance of a tailing phase in the inactivation kinetics, hinted at the importance of the short-lived species generated. Different factors, related to (i) the pathogen and its cell mode, (ii) the CAP settings and (iii) PAL related factors, proved to impact the antimicrobial efficacy of the solutions and should be considered with respect to future applications of the PAL technology.

The developed model biofilms can be applied as a standard to study biofilms (in different research fields) and their subsequent inactivation by different methods. In addition, the results of this study could be used to control biofilm formation (e.g. by setting a maximum allowed surface temperature).

The biofilm mode of growth protects bacterial cells against currently applied disinfection methods for abiotic (food) contact surfaces. Therefore, innovative methods, such as Cold Atmospheric Plasma (CAP), should be investigated for biofilm inactivation. However, more knowledge is required concerning the influence of the biofilm age on the inactivation efficacy in order to comment on a possible application of CAP in the (food) processing industry. L. monocytogenes and S. Typhimurium biofilms with five different ages (i.e., 1, 2, 3, 7, and 10 days) were developed. For the untreated biofilms, the total biofilm mass and the cell density were determined. To investigate the biofilm resistance towards CAP treatment, biofilms with different ages were treated for 10 min and the remaining cell density was determined. Finally, for the one-day old reference biofilms and the most resistant biofilm age, complete inactivation curves were developed to examine the influence of the biofilm age on the inactivation kinetics. For L. monocytogenes, an increased biofilm age resulted in (i) an increased biomass, (ii) a decreased cell density prior to CAP treatment, and (iii) an increased resistance towards CAP treatment. For S. Typhimurium, similar results were obtained, except for the biomass, which was here independent of the biofilm age.