Microorganisms are predominantly organized in biofilms, where cells live in dense communities and are more resistant to external stresses compared to their planktonic counterparts. With in vitro experiments, the susceptibility of Candida albicans biofilms to a non-thermal plasma treatment (plasma source kINPen®09), in terms of growth, survival, and cell viability was investigated. Behind that background, the C. albicans strain SC5314 (ATCC® MYA-2876™) was plasma treated for different time periods (30 s, 60 s, 120 s, 180 s, 300 s). The results of the experiments embracing colony forming units, fluorescence LIVE/DEAD™ assays, and XTT assays and revealed a negative influence of the plasma treatment on the proliferation ability, vitality, and the metabolism of C. albicans biofilms, respectively. Morphological analysis of plasma treated biofilms using atomic force microscopy supported the indications for lethal plasma effects concomitant with membrane disruptions and the loss of intracellular fluid. Controversial to other publications, fluorescence- and confocal laser scanning-microscopic inspection of plasma treated biofilms indicated that an inactivation of cells mainly appeared on the bottom side of the biofilms. If this inactivation leads to a detachment of the biofilms from the overgrown surface, it might offer completely new approaches in the plasma treatment of biofilms. Because of its biochemical-mechanical mode of action, resistance of microbial cells against plasma are unknown at this state of research. Importance Microbial communities are an increasing problem in medicine but also in industry. Thus, an efficient and rapid removal of biofilms is becoming increasingly important. With the aid of the kINPen®09, a radiofrequency plasma jet (RFPJ), decisive new findings on the effects of plasma on C. albicans biofilms could be obtained. This work was able to show that the inactivation of biofilms mainly takes place on the bottom side, which in turn offers new possibilities for the removal of biofilms by other strategies, e.g. mechanical treatment.This results demonstrated that non-thermal atmospheric pressure plasma is well-suited for biofilm decontamination.

Summary: The susceptibility of Candida albicans biofilms to a non‐thermal plasma treatment has been investigated in terms of growth, survival and cell viability by a series of in vitro experiments. For different time periods, the C. albicans strain SC5314 was treated with a microwave‐induced plasma torch (MiniMIP). The MiniMIP treatment had a strong effect (reduction factor (RF) = 2.97 after 50 s treatment) at a distance of 3 cm between the nozzle and the superior regions of the biofilms. In addition, a viability reduction of 77% after a 20 s plasma treatment and a metabolism reduction of 90% after a 40 s plasma treatment time were observed for C. albicans . After such a treatment, the biofilms revealed an altered morphology of their cells by atomic force microscopy (AFM). Additionally, fluorescence microscopy and confocal laser scanning microscopy (CLSM) analyses of plasma‐treated biofilms showed that an inactivation of cells mainly appeared on the bottom side of the biofilms. Thus, the plasma inactivation of the overgrown surface reveals a new possibility to combat biofilms.

For the decontamination of surfaces in the food production industry, plasma-generated compounds such as plasma-treated water or plasma-processed air offer many promising possibilities for future applications. Therefore, the antimicrobial effect of water treated with microwave-induced plasma (MidiPLexc) on Pseudomonas fluorescens biofilms was investigated. A total of 10 mL deionized water was treated with the MidiPLexc plasma source for 100, 300 and 900 s (pretreatment time) and the bacterial biofilms were exposed to the plasma-treated water for 1, 3 and 5 min (post-treatment time). To investigate the influence of plasma-treated water on P. fluorescens biofilms, microbiological assays (colony-forming units, fluorescence and XTT assay) and imaging techniques (fluorescence microscopy, confocal laser scanning microscopy, and atomic force microscopy) were used. The colony-forming units showed a maximum reduction of 6 log₁₀ by using 300 s pretreated plasma water for 5 min. Additionally, a maximum reduction of 81% for the viability of the cells and a 92% reduction in the metabolic activity of the cells were achieved by using 900 s pretreated plasma water for 5 min. The microscopic images showed evident microbial inactivation within the biofilm even at the shortest pretreatment (100 s) and post-treatment (1 min) times. Moreover, reduction of the biofilm thickness and increased cluster formation within the biofilm was detected. Morphologically, the fusion of cell walls into a uniform dense cell mass was detectable. The findings correlated with a decrease in the pH value of the plasma-treated water, which forms the basis for the chemically active components of plasma-treated water and its antimicrobial effects. These results provide valuable insights into the mechanisms of inactivation of biofilms by plasma-generated compounds such as plasma-treated water and thus allow for further parameter adjustment for applications in food industry.

Plasma-treated water (PTW) possess anti-microbial potential against Pseudomonas fluorescence, which is observable for both suspended cells and cells organized in biofilms. Against that background, the chemical composition of PTW tends to focus. Various analytical techniques have been applied for analyses, which reveal various traceable reactive oxygen and nitrogen compounds (RONS). Based on these findings, it is our aim to generate a PTW analog (anPTW), which has been compared in its anti-microbial efficiency with freshly generated PTW. Additionally, a solution of every traceable compound of PTW has been mixed according to their PTW concentration. As references, we treated suspended cells and mature biofilms of P. fluorescence with PTW that originates from a microwave-driven plasma source. The anti-microbial efficiency of all solutions has been tested based on a combination of a proliferation, an XTT, and a live–dead assay. The outcomes of the test proved an anti-microbial power of PTW that suggests more active ingredients than the traceable compounds HNO₃, HNO₂, and H₂O₂ or the combined mixture of the analog.

Sustainable and microbiologically secure foodstuff production lines are of increasing scientific interest and are in the focus of recent research programs. Additionally, they are of great importance for the production industry due to the prevention of food-borne illnesses caused by pathogens such as Salmonella sp., Listeria monocytogenes, or Escherichia coli. These pathogens are responsible for production losses, loss of customer acceptance, and severe food-borne illnesses. A pathogenic threat is frequently combated with sanitizing steps of the production lines. For conveyor band cleaning, this study compares the cleaning abilities of nitric acid (HNO₃) and plasma treated water (PTW), which have been sprayed via a commercially available nozzle on two different polymeric surfaces (polysiloxane and polyurethane). Additionally, the cleaning agents HNO₃ and PTW have been characterized through their pH and their conductivity. These findings have been underpinned by experiments that focus on a possible influence of nozzle abrasion, such as brass and stainless-steel nanoparticles, on the antimicrobial potential of PTW and HNO₃. Adversely acting effects like an enhanced abrasion of conveyer band materials due to PTW or HNO₃ treatment have been checked by using light microscopic micrographs and topographic scans in high-resolution mode. Based on the presented results of the experiments, the suitability of an in-place sanitation step in foodstuff production lines has been demonstrated on a laboratory scale.

The control of the pathogenic load on foodstuffs is a key element in food safety. Particularly, seafood such as cold-smoked salmon is threatened by pathogens such as Salmonella sp. or Listeria monocytogenes. Despite strict existing hygiene procedures, the production industry constantly demands novel, reliable methods for microbial decontamination. Against that background, a microwave plasma-based decontamination technique via plasma-processed air (PPA) is presented. Thereby, the samples undergo two treatment steps, a pre-treatment step where PPA is produced when compressed air flows over a plasma torch, and a post-treatment step where the PPA acts on the samples. This publication embraces experiments that compare the total viable count (tvc) of bacteria found on PPA-treated raw (rs) and cold-smoked salmon (css) samples and their references. The tvc over the storage time is evaluated using a logistic growth model that reveals a PPA sensitivity for raw salmon (rs). A shelf-life prolongation of two days is determined. When cold-smoked salmon (css) is PPA-treated, the treatment reveals no further impact. When PPA-treated raw salmon (rs) is compared with PPA-untreated cold-smoked salmon (css), the PPA treatment appears as reliable as the cold-smoking process and retards the growth of cultivable bacteria in the same manner. The experiments are flanked by quality measurements such as color and texture measurements before and after the PPA treatment. Salmon samples, which undergo an overtreatment, solely show light changes such as a whitish surface flocculation. A relatively mild treatment as applied in the storage experiments has no further detected impact on the fish matrix.