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    Reduce and refine: Plasma treated water vs conventional disinfectants for conveyor-belt cleaning in sustainable food-production lines
    (Melville, NY : American Inst. of Physics, 2021) Weihe, Thomas; Schnabel, Uta; Winter, Hauke; Möller, Timon; Stachowiak, Jörg; Neumann, Sabine; Schlüter, Oliver; Ehlbeck, Jörg
    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.
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    Characterization of antimicrobial effects of Plasma-Treated Water (PTW) produced by Microwave-Induced Plasma (MidiPLexc) on pseudomonas fluorescens biofilms
    (Basel : MDPI, 2020) Handorf, Oliver; Pauker, Viktoria Isabella; Schnabel, Uta; Weihe, Thomas; Freund, Eric; Bekeschus, Sander; Riedel, Katharina; Ehlbeck, Jörg
    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. © 2020 by the authors.
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    The combination of plasma-processed air (PPA) and plasma-treated water (PTW) causes synergistic inactivation of Candida albicans SC5314
    (Basel : MDPI, 2020) Schnabel, Uta; Yarova, Kateryna; Zessin, Björn; Stachowiak, Jörg; Ehlbeck, Jörg
    Microwave-induced plasma was used for the generation of plasma-processed air (PPA) and plasma-treated water (PTW). By this way, the plasma was able to functionalize the compressed air and the used water to antimicrobial effective agents. Their fungicidal effects by single and combined application were investigated on Candida albicans strain SC5314. The monoculture of C. albicans was cultivated on specimens with polymeric surface structures (PE-stripes). The additive as well as the synergistic fungicidal potential of PPA and PTW was investigated by different process windows of plasma exposure time (5-50 s) and sample treatment time with PPA/PTW (1-5 min). For a single PTW or PPA treatment, an increase in the reduction factor with the indicated treatment time was observed (maximum reduction factor of 1.1 and 1.6, respectively). In comparison, the combined application of PTW and then PPA resulted in antagonistic, additive and synergistic effects, depending on the combination. An application of the synergistically acting processes of PTW for cleaning and PPA for drying can be an innovative alternative to the sanitary processes currently used in production plants. © 2020 by the authors.
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    Plasma-Treated Water Affects Listeria monocytogenes Vitality and Biofilm Structure
    (Lausanne : Frontiers Media, 2021) Handorf, Oliver; Pauker, Viktoria Isabella; Weihe, Thomas; Schäfer, Jan; Freund, Eric; Schnabel, Uta; Bekeschus, Sander; Riedel, Katharina; Ehlbeck, Jörg
    Background: Plasma-generated compounds (PGCs) such as plasma-processed air (PPA) or plasma-treated water (PTW) offer an increasingly important alternative for the control of microorganisms in hard-to-reach areas found in several industrial applications including the food industry. To this end, we studied the antimicrobial capacity of PTW on the vitality and biofilm formation of Listeria monocytogenes, a common foodborne pathogen. Results: Using a microwave plasma (MidiPLexc), 10 ml of deionized water was treated for 100, 300, and 900 s (pre-treatment time), after which the bacterial biofilm was exposed to the PTW for 1, 3, and 5 min (post-treatment time) for each pre-treatment time, separately. Colony-forming units (CFU) were significantly reduced by 4.7 log10 ± 0.29 log10, as well as the metabolic activity decreased by 47.9 ± 9.47% and the cell vitality by 69.5 ± 2.1%, compared to the control biofilms. LIVE/DEAD staining and fluorescence microscopy showed a positive correlation between treatment and incubation times, as well as reduction in vitality. Atomic force microscopy (AFM) indicated changes in the structure quality of the bacterial biofilm. Conclusion: These results indicate a promising antimicrobial impact of plasma-treated water on Listeria monocytogenes, which may lead to more targeted applications of plasma decontamination in the food industry in the future.
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    The Effect of Plasma Treated Water Unit Processes on the Food Quality Characteristics of Fresh-Cut Endive
    (Lausanne : Frontiers Media, 2021-1-27) Schnabel, Uta; Handorf, Oliver; Winter, Hauke; Weihe, Thomas; Weit, Christoph; Schäfer, Jan; Stachowiak, Jörg; Boehm, Daniela; Below, Harald; Bourke, Paula; Ehlbeck, Jörg
    This study evaluated the impact of a defined plasma treated water (PTW) when applied to various stages within fresh-cut endive processing. The quality characteristic responses were investigated to establish the impact of the PTW unit processes and where PTW may be optimally applied in a model process line to retain or improve produce quality. Different stages of application of PTW within the washing process were investigated and compared to tap water and chlorine dioxide. Fresh-cut endive (Cichorium endivia L.) samples were analyzed for retention of food quality characteristics. Measurements included color, texture, and nitrate quantification. Effects on tissue surface and cell organelles were observed through scanning electron and atomic force microscopy. Overall, the endive quality characteristics were retained by incorporating PTW in the washing process. Furthermore, promising results for color and texture characteristics were observed, which were supported by the microscopic assays of the vegetal tissue. While ion chromatography detected high concentrations of nitrite and nitrate in PTW, these did not affect the nitrate concentration of the lettuce tissue post-processing and were below the concentrations within EU regulations. These results provide a pathway to scale up the industrial application of PTW to improve and retain quality characteristic retention of fresh leafy products, whilst also harnessing the plasma functionalized water as a process intervention for reducing microbial load at multiple points, whether on the food surface, within the process water or on food-processing surfaces.
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    Plasma-treated air and water-assessment of synergistic antimicrobial effects for sanitation of food processing surfaces and environment
    (Basel : MDPI, 2019) Schnabel, Uta; Handorf, Oliver; Yarova, Kateryna; Zessin, Björn; Zechlin, Susann; Sydow, Diana; Zellmer, Elke; Stachowiak, Jörg; Andrasch, Mathias; Below, Harald; Ehlbeck, Jörg
    The synergistic antimicrobial effects of plasma-processed air (PPA) and plasma-treated water (PTW), which are indirectly generated by a microwave-induced non-atmospheric pressure plasma, were investigated with the aid of proliferation assays. For this purpose, microorganisms (Listeria monocytogenes, Escherichia coli, Pectobacterium carotovorum, sporulated Bacillus atrophaeus) were cultivated as monocultures on specimens with polymeric surface structures. Both the distinct and synergistic antimicrobial potential of PPA and PTW were governed by the plasma-on time (5–50 s) and the treatment time of the specimens with PPA/PTW (1–5 min). In single PTW treatment of the bacteria, an elevation of the reduction factor with increasing treatment time could be observed (e.g., reduction factor of 2.4 to 3.0 for P. carotovorum). In comparison, the combination of PTW and subsequent PPA treatment leads to synergistic effects that are clearly not induced by longer treatment times. These findings have been valid for all bacteria (L. monocytogenes > P. carotovorum = E. coli). Controversially, the effect is reversed for endospores of B. atrophaeus. With pure PPA treatment, a strong inactivation at 50 s plasma-on time is detectable, whereas single PTW treatment shows no effect even with increasing treatment parameters. The use of synergistic effects of PTW for cleaning and PPA for drying shows a clear alternative for currently used sanitation methods in production plants. Highlights: Non-thermal atmospheric pressure microwave plasma source used indirect in two different modes—gaseous and liquid; Measurement of short and long-living nitrite and nitrate in corrosive gas PPA (plasma-processed air) and complex liquid PTW (plasma-treated water); Application of PTW and PPA in single and combined use for biological decontamination of different microorganisms.
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    Optimizing the application of plasma functionalised water (PFW) for microbial safety in fresh-cut endive processing
    (New York, NY [u.a.] : Elsevier Science, 2021) Schnabel, Uta; Balazinski, Martina; Wagner, Robert; Stachowiak, Jörg; Boehm, Daniela; Andrasch, Mathias; Bourke, Paula; Ehlbeck, Jörg
    The microbiological profiles and responses of native microflora of endive were investigated using a model process line, to establish where a defined PFW should be optimally applied to retain or improve produce microbiological quality. The PFW processes were compared with tap water and ClO2. The antimicrobial efficacy of PFW was quantified by determining the reduction in microbial load, the microbial viability and vitality. Depending on the stage of application of PFW, up to 5 log10-cycles reduction was achieved, accompanied by a reduction of metabolic activity, but not necessarily with a decrease in metabolic vitality. Multiple application (3-step-PFW-application) was more effective than single application (1-step-PFW-application) and PFW showed stronger antimicrobial effect in pre-cleaned endive. High concentrations of nitrite (315 mg l−1) and nitrate (472 mg l−1) in PFW were the main factors for the antimicrobial efficacy of PFW against bacteria. Furthermore, H2O2 and an acidic pH supported the mechanism of action against the endive microflora. These results identify the pathway to scale up successful industrial application of PFW targeting microbiological quality and safety of fresh leafy products.Industrial relevance The safety, quality and shelf life of freshly cut vegetables, e.g. lettuce, are strongly influenced by the microbial load. In addition, the hygienic design of production line, and a good handling/ production practice are indispensable. This study shows that the application of PFW, as a promising non-thermal sanitation technology, enables the inactivation of native microbial contamination on fresh-cut endive depending on the process stage of application. It further describes the impact of PFW on the metabolic activity and metabolic vitality of the lettuce-associated microflora. For higher acceptance, the mechanism of action of PFW was assumed based on previous chemical analyses and compared to the industrial standard of ClO2. The results contribute to the understanding and product-specificity of PFW-induced effects on safety, quality and shelf life of fresh cut lettuce and could be a basis for a possible industrial implementation and complement of common technologies.
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    Microbial Control of Raw and Cold-Smoked Atlantic Salmon (Salmo salar) through a Microwave Plasma Treatment
    (Basel : MDPI, 2022) Weihe, Thomas; Wagner, Robert; Schnabel, Uta; Andrasch, Mathias; Su, Yukun; Stachowiak, Jörg; Noll, Heinz Jörg; Ehlbeck, Jörg
    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.
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    Antimicrobial effects of microwave-induced plasma torch (MiniMIP) treatment on Candida albicans biofilms
    (Oxford : Wiley-Blackwell, 2019) Handorf, Oliver; Schnabel, Uta; Bösel, André; Weihe, Thomas; Bekeschus, Sander; Graf, Alexander Christian; Riedel, Katharina; Ehlbeck, Jörg
    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. © 2019 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.