Thomas Weihe

Non-thermal decontamination methods offer energy-efficient, lower-temperature alternatives to conventional heat treatments with potentially milder effects on nutritional and sensory quality. This critical review synthesizes evidence for ultraviolet (UV), electron-beam irradiation (EBI), atmospheric-pressure plasma (APP), ultrasound (US), high-pressure processing (HPP), supercritical fluids (SCF), pulsed electric fields (PEF), protective cultures (PC), and membrane filtration (MF). It evaluates technical requirements and the impacts on food quality of liquid, semi-solid, and solid food matrices. No single method is universally applicable, but complement the toolbox of non-thermal decontamination methods. Key contrasts include deep penetration by EBI versus surface/clarity limitations of UV. HPP’s uniform bulk inactivation but limited sporicidal activity and potential off-flavor formation. APP and US generating reactive species or cavitation that can alter texture and color. SCF’s microbial inactivation coupled with extraction of desirable components. PEF’s suitability for flowing liquids and potential electrophoretic effects on proteins. MF’s efficacy for microbial removal in liquids but inapplicability to solids and PCs’ targeted biological control that may modify sensory profiles. Broader commercial adoption will require matrix-specific validation (microbial log-reductions and quality metrics), quantitative mechanistic studies (e.g. reactive oxygen/nitrogen species, extraction yields, molecular changes), and pilot-scale, interdisciplinary scale-up with economic assessment to produce standardized, product-tailored protocols.

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Foodborne illness caused by bacterial pathogens is a global health concern and results in millions of infections annually. Therefore, food products typically undergo several processing stages, including sanitation steps, before being distributed in an attempt to remove pathogens. However, many sanitation methods have compounding effects on the color, texture, flavor, and nutritional quality of the product or do not effectively reduce the pathogens that food can be exposed to. Some bacterial pathogens particularly possess traits and tactics that make them even more difficult to mitigate such as biofilm formation. Non-thermal plasma sanitation techniques, including plasma-treated water (PTW), have proven to be promising methods that significantly reduce pathogenic bacteria that food is exposed to. Published work reveals that PTW can effectively mitigate both gram-positive and gram-negative bacterial biofilms. This study presents a novel analysis of the differences in antimicrobial effects of PTW treatment between biofilm-forming gram-positive bacteria, commonly associated with foodborne illness, that are sporulating (Bacillus cereus) and non-sporulating (Listeria monocytogenes). After treatment with PTW, the results suggest the following hypotheses: (1) that the non-sporulating species experiences less membrane damage but a greater reduction in metabolic activity, leading to a possible viable but non-culturable (VBNC) state, and (2) that the sporulating species undergoes spore formation, which may subsequently convert into vegetative cells over time. PTW treatment on gram-positive bacterial biofilms that persist in food processing environments proves to be effective in reducing the proliferating abilities of the bacteria. However, the variance in PTW’s effects on metabolic activity and cell vitality between sporulating and non-sporulating species suggest that other survival tactics might be induced. This analysis further informs the application of PTW in food processing as an effective sanitation method.

Plasma-treated water (PTW) recently entered science as a sanitizing agent, which possess the capability for on-demand production. It offers interesting possibilities for sustainable and resource-saving applications in healthcare and food production. The present study monitors the impact of PTW on suspended cells before the biofilm formation of the putrefactive bacterium B. subtilis. Light and electron microscope imaging captures the maturing of growing biofilms within the first 24 h. Microbiological assays (proliferation, LIVE/DEAD, and XTT), which mirror the proliferation of the bacterium, the metabolic activity, and the integrity of the cell membrane, underpinning the metabolic response of still-suspended cells. B. subtilis cells without any treatment build up a resistive biofilm within the 24 h. Cells that remain in the supernatant predominantly appear as monomers or dimers. Treated B. subtilis cells have hampered biofilm formation and were not able to build up a confluent growing biofilm within the first 24 h. Moreover, the microscopic observation of PTW-treated suspension showed cellular aggregates with an unusually high connectivity of the individual cells. The findings suggest this cellular reaction as a counter measure against the adverse impact of PTW treatment. The experiments show the adverse impact of PTW on B. subtilis–biofilm formation and the phenomenological reaction of B. subtilis.

The consumer demand for organic food including apples is increasing worldwide. Despite favorable environmental and health benefits, organic farming bears also disadvantages like high amounts of fruit losses due to storage rot. A novel treatment with plasma-processed air (PPA) to sanitize organic apples is investigated. The plasma source for the generation of PPA was operated at a frequency of 2.45 GHz, a power output of 1.1 kW and a gas flow of 18 standard liters per minute. The antimicrobial efficiency of the PPA was tested on the natural load of organic apples (cultivar Natyra) with a load ranging from 10⁴ to 10⁶ CFU/mL in an experimental laboratory setup. A larger application was applied on artificially inoculated (Pseudomonas fluorescens~10⁸ CFU/mL and Pezicula malicorticis~10⁶ CFU/mL) organic apples to test the up-scalability of the PPA treatment. The apples were photographically documented and their texture was analyzed during the 26-day storage phase to investigate the influence of the PPA treatment on the appearance of the apples. The laboratory experiments resulted in a log₁₀-reduction of one to two log₁₀ levels compared to untreated and compressed-air-treated apples. For apples inoculated with P. fluorescens, the up-scaled procedure resulted in up to four levels of log₁₀ reduction. In apples inoculated with P. malicorticis, the up-scaled procedure resulted in no reduction. This indicates that the application of PPA to organic apples can be effective for bacteria but needs to be optimized for fungi. Therefore, further testing is needed to validate the results.

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.

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.

Methods, which use an indirect plasma treatment for the inactivation of microorganisms in foods, claim a vastly growing field of research. This paper presents a method that uses plasma-processed air (PPA) as a sanitizer. In addition to a sanitation concept for the decontamination of produce in the value chain, the presented method offers a possible application as an “in-process” surface sanitation. PPA provides antimicrobial-potent species, which are predominantly reactive nitrogen species (RNS); this has an outstanding groove penetration property. In an experimental approach, surfaces, made from materials, which are frequently used for the construction of food-processing plants, were inoculated with different microorganisms. Listeria monocytogenes (ATCC 15313), Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 10538), Salmonella enterica subsp. enterica serovar Typhimurium (ATCC 43971), and Salmonella enterica subsp. enterica serovar Enteritidis (ATCC 13076) are all microorganisms that frequently appear in foods and possess the risk for cross-contamination from the plant to the produce or vice versa. The contaminated samples were treated for various treatment times (1–5 min) with PPA of different antimicrobial potencies. Subsequently, the microbial load on the specimens was determined and compared with the load of untreated samples. As a result, reduction factors (RF) up to several log₁₀-steps were obtained. Although surface and the bacterial strain showed an influence on the RF, the major influence was seen by a prolongation of the treatment time and an increase in the potency of the PPA.

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.

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.