Barrier Discharge Atmospheric Cold Plasma Research Articles

Resistance of Alicyclobacillus acidoterrestris spores to atmospheric cold plasma: Insights from sporulation temperature and mechanism analysis

Environmental conditions during sporulation, such as temperature, pH value, and mineral content, can significantly influence the formation production and resistance of spores. In this study, we investigated the sporulation capacity and resistance of Alicyclobacillus acidoterrestris (AAT) spores obtained at different temperatures (25 °C, 35 °C, 45 °C) to Dielectric Barrier Discharge Atmospheric cold plasma (DBD-ACP, 20 and 30 kV for 2–8 min). The results indicated that the efficiency of sporulation was highest at the optimum temperature (45 °C), deviation from optimal temperature would have a negative effect on sporulation. Otherwise, sporicidal efficiency of DBD-ACP increased with longer treatment time and lower sporulation temperatures, suggesting that AAT spores exhibit weaker resistance to DBD-ACP at lower sporulation temperature. Results from the release of DPA, and electron microscopic observations confirmed that DBD-ACP caused disruption of the cortex and membrane structure of AAT spores, leading to the release of intracellular contents and subsequent spore's death. Further analysis through proteomics revealed intriguing insights into the differences between spores formed at 25 °C and those at 45 °C. Specifically, spores formed at 25 °C exhibited lower expression levels of proteins related to sporulation, intracellular energy metabolism, membrane transport, and significant down-regulation of proteins involved in peptidoglycan and spore coat formation compared to those at 45 °C. These alterations likely contribute to the development of a thinner and looser spore structure, rendering it less effective in defending, i.e. less resistant to DBD-ACP treatment. Industrial relevanceAAT spores' resistance to pasteurization and persistence in NFC juices impact juice quality and safety. This study investigates the resistance of AAT spore to DBD-ACP treatment under different sporulation temperatures. Insights into sporicidal behavior of DBD-ACP and the effect of sporulation temperature on resistance provide essential knowledge for optimizing juice processing and enhancing microbial safety. The findings offer practical guidance for the juice industry to develop improved preservation strategies, ensuring longer shelf life and consumer satisfaction.

Combined of plasma-activated water and dielectric barrier discharge atmospheric cold plasma treatment improves the characteristic flavor of Asian sea bass (Lates calcarifer) through facilitating lipid oxidation

To explore the combination effects of plasma-activated water and dielectric barrier discharge (PAW-DBD) cold plasma treatment on the formation of volatile flavor and lipid oxidation in Asian sea bass (ASB), the volatile flavor compounds and lipid profiles were characterized by gas chromatography-ion mobility spectrometry and LC-MS-based lipidomics analyses. In total, 38 volatile flavor compound types were identified, and the PAW-DBD group showed the most kinds of volatile components with a significant (p < 0.05) higher content in aldehydes, ketones, and alcohols. A total of 1500 lipids was detected in lipidomics analysis, phosphatidylcholine was the most followed by triglyceride. The total saturated fatty acids content in PAW-DBD group increased by 105.02 μg/g, while the total content of unsaturated fatty acids decreased by 275.36 μg/g. It can be concluded that the PAW-DBD processing increased both the types and amounts of the volatile flavor in ASB and promoted lipid oxidation by altering lipid profiles.

Proteomics analysis reveals the inactivation mechanisms of Alicyclobacillus acidoterrestris spores by atmospheric cold plasma

SummaryAlicyclobacillus acidoterrestris is a typical endospore‐forming microorganism, whose spores are able to germinate in Not From Concentrate juice causing its spoilage. In this work, dielectric barrier discharge atmospheric cold plasma (DBD‐ACP) at the voltage of 30 kV was used to inactivate Alicyclobacillus acidoterrestris spores in NFC juice (2–8 min). The results showed spores reduced nearly 3‐log CFU mL−1 after 8 min treatment, and the morphology of spores were severely disrupted, causing the release of their contents. Furthermore, proteomic analysis showed totals of 122 and 170 different expression proteins were detected in group C (control)/Ct1 (30 kV, 4 min) and C/Ct2 (30 kV, 8 min), respectively, indicating that metabolic activities of spores including amino acid and energy metabolic, protein biosynthesis, DNA replication and spore germination are inhibited. This study confirms the inactivation of Alicyclobacillus acidoterrestris spores by DBD‐ACP and provides some new perspectives on the inactivation mechanisms of spores by DBD‐ACP.

The Effect of Atmospheric Dielectric Barrier Discharge Cold Plasma Treatment on the Nutritional and Physicochemical Characteristics of Various Legumes.

High activity of lipoxygenase (LOX) has been identified as a primary cause of oxidative rancidity in legumes. In this study, the application of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) (5 W, 10 min) resulted in an obvious decrease in LOX activity in mung bean (MB), kidney bean (KB), and adzuki bean (AB) flours by 36.96%, 32.49%, and 28.57%, respectively. Moreover, DBD-ACP induced significant increases (p < 0.05) in content of soluble dietary fiber, saturated fatty acids, and methionine. The starch digestibility of legumes was changed, evidenced by increased (p < 0.05) slowly digestible starch and rapidly digestible starch, while resistant starch decreased. Furthermore, DBD-ACP treatment significantly affected (p < 0.05) the hydration and thermal characteristics of legume flours, evidenced by the increased water absorption index (WAI) and gelatinization temperature, and the decreased swelling power (SP) and gelatinization enthalpy (ΔH). Microscopic observations confirmed that DBD-ACP treatment caused particle aggregation.

Effects of a cold plasma-assisted shrimp processing chain on biochemical and sensory quality alterations in Pacific white shrimps (Penaeus vannamei).

In this paper, remodeling the shrimp processing chain and the effects of the transformation on the biochemical and sensory qualities of fresh Pacific white shrimp (Penaeus vannamei) under refrigeration storage were investigated. In the proposed model, a dielectric barrier discharge atmospheric cold plasma pretreatment step using a 60 kV source for 60, 90, 120, and 150 s was introduced after the first and second wash followed by refrigeration storage at 4 ± 1 °C for 12 days. Chemical, biochemical, and sensory attributes of the shrimp were monitored and compared with those of shrimp processed through the traditional method without atmospheric cold plasma pretreatment (control). Incorporating minimal dielectric barrier discharge atmospheric cold plasma pretreatment step had more desirable quality outcomes characterized by low malondialdehyde concentration, low volatile nitrogen products content, and comparable proximate composition. Texture, pH, and color were remarkably retained at 120 and 150 s of atmospheric cold plasma pretreatment and protein degradation was negligible up to 90 s than at 120 and 150 s of pretreatment. We conclude that remodeling the shrimp processing chain through incorporating minimal dielectric barrier discharge atmospheric cold plasma pretreatment with key considerations on operation parameters can maximize the beneficial biochemical and sensory quality outcomes while minimizing the negative impacts associated with traditional shrimp processing.

Contactless flash sintering based on cold plasma

We report on flash sintering using a contactless approach. The process is based on dielectric barrier discharge atmospheric cold plasma which allows flash sintering without external preheating. The evenly spread cold plasma plume contributed to homogenous densification of zirconia samples, instead arc discharge resulted in un-sintered and cracked material. Final density and grain size could be controlled by adjusting current and discharge time. A 7 mm diameter disc was sintered up to 99% in 30 s by passing a current of 15 mA (0.39 mA/mm2). The proposed technique is industrially scalable and offers a reliable replacement to platinum electrodes.

In-package atmospheric cold plasma treatment of bulk grape tomatoes for microbiological safety and preservation

Effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Salmonella and the storability of grape tomato were investigated. Grape tomatoes, with or without inoculation with Salmonella, were packaged in a polyethylene terephthalate (PET) commercial clamshell container and cold plasma-treated at 35 kV at 1.1 A for 3 min using a DACP system equipped with a pin-type high-voltage electrode. DACP treatment inactivated Salmonella (p < 0.05) without altering the color or firmness of the grape tomatoes (p > 0.05). DACP treatment inactivated Salmonella uniformly in both layers of the double-layer configuration of the grape tomatoes regardless of the position of the tomatoes in each layer. Salmonella was most efficiently inactivated when the headspace to tomato volume ratio of the container was highest. Integration of rolling of tomatoes during treatment significantly increased the Salmonella reduction rates from 0.9 ± 0.2 log CFU/tomato to 3.3 ± 0.5 log CFU/tomato in the double-layer configuration of the tomato samples. Rolling-integrated DACP also initially reduced the number of total mesophilic aerobic bacteria and yeast and molds in the double-layer configuration of tomato samples by 1.3 ± 0.3 and 1.5 ± 0.2 log CFU/tomato, respectively. DACP treatment effectively reduced the growth of Salmonella and indigenous microorganisms at 10 and 25 °C, and did not influence the surface color, firmness, weight loss, lycopene concentration and residual ascorbic acid of grape tomatoes during storage at 10 and 25 °C. DACP treatment holds promise as a post-packaging process for improving microbial safety against Salmonella and storability of fresh grape tomatoes.

Assessment of the disinfection capacity and eco-toxicological impact of atmospheric cold plasma for treatment of food industry effluents

Generation of wastewater is one of the main environmental sustainability issues across food sector industries. The constituents of food process effluents are often complex and require high energy and processing for regulatory compliance. Wastewater streams are the subject of microbiological and chemical criteria, and can have a significant eco-toxicological impact on the aquatic life. Thus, innovative treatment approaches are required to mitigate environmental impact in an energy efficient manner. Here, dielectric barrier discharge atmospheric cold plasma (ACP) was evaluated for control of key microbial indicators encountered in food industry effluent. This study also investigated the eco-toxicological impact of cold plasma treatment of the effluents using a range of aquatic bioassays. Continuous ACP treatment was applied to synthetic dairy and meat effluents. Microbial inactivation showed treatment time dependence with significant reduction in microbial populations within 120 s, and to undetectable levels after 300 s. Post treatment retention time emerged as critical control parameter which promoted ACP bacterial inactivation efficiency. Moreover, ACP treatment for 20 min achieved significant reduction (≥2 Log10) in Bacillus megaterium endospores in wastewater effluent. Acute aquatic toxicity was assessed using two fish cell lines (PLHC-1 and RTG-2) and a crustacean model (Daphnia magna). Untreated effluents were toxic to the aquatic models, however, plasma treatment limited the toxic effects. Differing sensitivities were observed to ACP treated effluents across the different test bio-assays in the following order: PLHC-1 > RTG-2 ≥ D. magna; with greater sensitivity retained to plasma treated meat effluent than dairy effluent. The toxic effects were dependent on concentration and treatment time of the ACP treated effluent; with 30% cytotoxicity in D. magna and fish cells observed after 24 h of exposure to ACP treated effluent for concentrations up to 5%. The findings suggest the need to employ wider variety of aquatic organisms for better understanding and complete toxicity evaluation of long-term effects.The study demonstrates the potential to tailor ACP system parameters to control pertinent microbial targets (mono/poly-microbial, vegetative or spore form) found in complex and nutritious wastewater effluents whilst maintaining a safe eco-toxicity profile for aquatic species.

The potential of atmospheric air cold plasma for control of bacterial contaminants relevant to cereal grain production

Abstract The aim of this work was to investigate the efficacy of dielectric barrier discharge atmospheric cold plasma (DBD ACP) against bacteria associated with grains quality and safety. ACP inactivation efficacy was tested against biofilms formed by different strains of E. coli , Bacillus and Lactobacillus in grain model media and against B. atrophaeus endospores either in grain media or attached on abiotic surfaces. Effects were dependent on bacterial strain, media composition and mode of ACP exposure. ACP treatment for 5 min reduced E. coli spp., B. subtilis and Lactobacillus spp. biofilms by > 3 log 10 , whereas insignificant reductions were achieved for B. atrophaeus . ACP treatment of 5–20 min reduced B. atrophaeus spores in liquids by > 5 log 10 . Treatment for 30 min reduced spores on hydrophobic surface by > 6 log 10 , whereas maximum of 4.4 log reductions were achieved with spores attached to hydrophilic surface. Microscopy demonstrated that ACP caused significant damage to spores. In package ACP treatment has potential to inactivate grain contaminants in the form of biofilms, as well as spores and vegetative cells. Industrial relevance This study demonstrates that ACP technology is a promising tool for effective bio-decontamination which offers a wide range of possible applications including inactivation of microorganisms on cereal grains. However, due to the nature of the microbial contamination of grains and complex grain structures it may be necessary to optimise the potential for surface inactivation at several stages of grain processing and storage to enhance ACP efficacy against bacterial endospores.

Lethal and Sublethal Effect of a Dielectric Barrier Discharge Atmospheric Cold Plasma on Staphylococcus aureus

The lethal and sublethal effect of a dielectric barrier discharge atmospheric cold plasma on Staphylococcus aureus were investigated by nonselective and selective media. The inactivation level and sublethally injured proportions of S. aureus cells by dielectric barrier discharge atmospheric cold plasma depended on the treatment times (0, 5, 15, and 45 s), applied input powers (40, 50, and 60 W), and gap distances (2, 4, and 6 mm). Under weak conditions (e.g., 5 s, 40 W, and 6 mm), the occurrence of inactivation and sublethal injury of S. aureus showed a relatively low level. Strong treatment (e.g., 45 s, 60 W, and 2 mm) resulted in highly an inactivated rate, but with small fractions of sublethally injured S. aureus cells. Mild action of cold plasma induced both a large inactivation and a sublethal injury of S. aureus . Therefore, it is significant to optimize the processing of cold plasma sterilization in practice to ensure food safety.

Controlling Brochothrix thermosphacta as a spoilage risk using in-package atmospheric cold plasma

Brochothrix thermosphacta is the predominant spoilage microorganism in meat and its control in processing environments is important to maintain meat product quality. Atmospheric cold plasma is of interest for control of pathogenic and spoilage microorganisms in foods. This study ascertained the potential of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) for control of B. thermosphacta, taking microbial and food environment factors into consideration, and investigated the shelf-life of lamb chop after in-package plasma treatment in modified atmosphere. Community profiling was used to assess the treatment effects on the lamb microflora.ACP treatment (80 kV) for 30s inactivated B. thermosphacta populations below detection levels in PBS, while 5 min treatment achieved a 2 Log cycle reduction using a complex meat model medium and attached cells. The antimicrobial efficacy of plasma was reduced but still apparent on lamb chop surface-inoculated with high concentrations of B. thermosphacta. Lamb chop treated under modified atmosphere exhibited reduced microbial growth over the product shelf-life and community profiling showed no evident changes to the microbial populations after the treatment. The overall results indicated potential of ACP to enhance microbial control leading to meat storage life extension through adjusting the modality of treatment.

In-package inhibition of E. coli O157:H7 on bulk Romaine lettuce using cold plasma

Dielectric barrier discharge atmospheric cold plasma (DACP) treatment was evaluated for the inactivation of Escherichia coli O157:H7, surface morphology, color, carbon dioxide generation, and weight loss of bulk Romaine lettuce in a commercial plastic clamshell container. The lettuce samples were packed in a model bulk packaging configuration (three rows with either 1, 3, 5, or 7 layers) in the container and treated by DACP (42.6 kV, 10 min). DACP treatment reduced the number of E. coli O157:H7 in the leaf samples in the 1-, 3-, and 5-layer configurations by 0.4–0.8 log CFU/g lettuce, with no significant correlation to the sample location (P > 0.05). In the largest bulk stacking with 7 layers, a greater degree of reduction (1.1 log CFU/g lettuce) was observed at the top layer, but shaking the container increased the uniformity of the inhibition. DACP did not significantly change the surface morphology, color, respiration rate, or weight loss of the samples, nor did these properties differ significantly according to their location in the bulk stack. DACP treatment inhibited E. coli O157:H7 on bulk lettuce in clamshell containers in a uniform manner, without affecting the physical and biological properties and thus holds promise as a post-packaging process for fresh and fresh-cut fruits and vegetables.

Inactivation of Escherichia coli O157:H7 and Aerobic Microorganisms in Romaine Lettuce Packaged in a Commercial Polyethylene Terephthalate Container Using Atmospheric Cold Plasma

The effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7 and aerobic microorganisms in romaine lettuce packaged in a conventional commercial plastic container were evaluated during storage at 4°C for 7 days. Effects investigated included the color, carbon dioxide (CO2) generation, weight loss, and surface morphology of the lettuce during storage. Romaine lettuce pieces, with or without inoculation with a cocktail of three strains of E. coli O157:H7 (~6 log CFU/g of lettuce), were packaged in a polyethylene terephthalate commercial clamshell container and treated at 34.8 kV at 1.1 kHz for 5 min by using a DACP treatment system equipped with a pin-type high-voltage electrode. Romaine lettuce samples were analyzed for inactivation of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds, color, CO2 generation, weight loss, and surface morphology during storage at 4°C for 7 days. The DACP treatment reduced the initial counts of E. coli O157:H7 and total aerobic microorganisms by ~1 log CFU/g, with negligible temperature change from 24.5 ± 1.4°C to 26.6 ± 1.7°C. The reductions in the numbers of E. coli O157:H7, total mesophilic aerobes, and yeasts and molds during storage were 0.8 to 1.5, 0.7 to 1.9, and 0.9 to 1.7 log CFU/g, respectively. DACP treatment, however, did not significantly affect the color, CO2 generation, weight, and surface morphology of lettuce during storage (P > 0.05). Some mesophilic aerobic bacteria were sublethally injured by DACP treatment. The results from this study demonstrate the potential of applying DACP as a postpackaging treatment to decontaminate lettuce contained in conventional plastic packages without altering color and leaf respiration during posttreatment cold storage.

Dielectric barrier discharge atmospheric cold plasma inhibits Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus in Romaine lettuce

The present study investigated the effects of dielectric barrier discharge atmospheric cold plasma (DACP) treatment on the inactivation of Escherichia coli O157:H7, Salmonella, Listeria monocytogenes, and Tulane virus (TV) on Romaine lettuce, assessing the influences of moisture vaporization, modified atmospheric packaging (MAP), and post-treatment storage on the inactivation of these pathogens. Romaine lettuce was inoculated with E. coli O157:H7, Salmonella, L. monocytogenes (~6logCFU/g lettuce), or TV (~2logPFU/g lettuce) and packaged in either a Petri dish (diameter: 150mm, height: 15mm) or a Nylon/polyethylene pouch (152×254mm) with and without moisture vaporization. Additionally, a subset of pouch-packaged leaves was flushed with O2 at 5% or 10% (balance N2). All of the packaged lettuce samples were treated with DACP at 34.8kV for 5min and then analyzed either immediately or following post-treatment storage for 24h at 4°C to assess the inhibition of microorganisms. DACP treatment inhibited E. coli O157:H7, Salmonella, L. monocytogenes, and TV by 1.1±0.4, 0.4±0.3, 1.0±0.5logCFU/g, and 1.3±0.1logPFU/g, respectively, without environmental modifications of moisture or gas in the packages. The inhibition of the bacteria was not significantly affected by packaging type or moisture vaporization (p>0.05) but a reduced-oxygen MAP gas composition attenuated the inhibition rates of E. coli O157:H7 and TV. L. monocytogenes continued to decline by an additional 0.6logCFU/g in post-treatment cold storage for 24h. Additionally, both rigid and flexible conventional plastic packages appear to be suitable for the in-package decontamination of lettuce with DACP.

Inactivation of Spoilage Bacteria in Package by Dielectric Barrier Discharge Atmospheric Cold Plasma—Treatment Time Effects

The objective was to investigate the effect of treatment time of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) on inactivation of spoilage bacteria, Pseudomonas fluorescens and Macrococcus caseolyticus. P. fluorescens and M. caseolyticus were isolated from spoiled chicken carcasses and suspended in liquid media in packages before the samples were treated with DBD-ACP at 55 kV for different times. Ozone concentrations in package headspace and pH changes in bacterial suspensions were measured to assess plasma formation and antimicrobial activity inside packages. Colony formation was used to indicate the inactivation efficacy. Ozone concentrations reached 1000 ppm after a 3-min treatment. pH dropped by more than 0.25 units in bacterial suspensions immediately after a 6-min treatment and 0.70 units after the 6-min treatment and 24-h storage at 4 °C. Populations of both bacteria significantly reduced as the DBD-ACP treatment time increased. The treatment for less than 1.5 min resulted in more than 1 log reductions in both bacteria, which were similar to each other. However, treatments for additional 1.5 min reduced P. fluorescens and M. caseolyticus populations by more than 4 and 2.5 log cycles, respectively. The difference in bacterial kills between the two types of bacteria extended to the 6-min treatments. Our results demonstrate that treatment times significantly affect the inactivation of spoilage bacteria by DBD-ACP system. After the treatment times are extended for more than 1.5 min, the in-package DBD-ACP treatment has more effects on Gram-negative spoilage bacteria P. fluorescens than Gram-positive spoilage bacteria M. caseolyticus.

Cytotoxic and mutagenic potential of solutions exposed to cold atmospheric plasma.

The exposure of aqueous solutions to atmospheric plasmas results in the generation of relatively long-lived secondary products such as hydrogen peroxide which are biologically active and have demonstrated anti-microbial and cytotoxic activity. The use of plasma-activated solutions in applications such as microbial decontamination or anti-cancer treatments requires not only adequate performance on target cells but also a safe operating window regarding the impact on surrounding tissues. Furthermore the generation of plasma-activated fluids needs to be considered as a by-stander effect of subjecting tissue to plasma discharges. Cytotoxicity and mutagenicity assays using mammalian cell lines were used to elucidate the effects of solutions treated with di-electric barrier discharge atmospheric cold plasma. Plasma-treated PBS inhibited cell growth in a treatment time-dependent manner showing a linear correlation to the solutions’ peroxide concentration which remained stable over several weeks. Plasma-treated foetal bovine serum (FBS) acting as a model for complex bio-fluids showed not only cytotoxic effects but also exhibited increased mutagenic potential as determined using the mammalian HPRT assay. Further studies are warranted to determine the nature, causes and effects of the cyto- and genotoxic potential of solutions exposed to plasma discharges to ensure long-term safety of novel plasma applications in medicine and healthcare.

Atmospheric cold plasma inactivation of Escherichia coli in liquid media inside a sealed package

The main objective of this study was to determine the inactivation efficacy of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) generated inside a sealed package for Escherichia coli ATCC 25922. A plasma discharge was generated between two circular aluminium electrodes at 40 kV. E. coli suspensions (10(7) CFU ml(-1)) in either maximum recovery diluent (MRD) or phosphate buffered saline (PBS) were treated in a 96-well microtitre plate inside a sealed package. The effects of treatment time, post-treatment storage time, either direct or indirect samples exposure to the plasma discharge and suspension media were studied. Regardless of the media tested, 20 s of direct and 45 s of indirect plasma treatment resulted in complete bacterial inactivation (7 log CFU ml(-1)). At the lower plasma treatment times (10-30 s) investigated, the effects of suspension media and mode of exposure on the inactivation efficacy were evident. The inactivation efficacy was also influenced by the post-treatment storage time. It was demonstrated that the novel DBD-ACP can inactivate high concentrations of E. coli suspended in liquids within sealed packages in seconds. A key advantage of this in-package nonthermal novel disinfection approach is the elimination of post-processing contamination.