Now showing 1 - 9 of 9
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Effects of cold plasma on wheat grain microbiome and antimicrobial efficacy against challenge pathogens and their resistance

2020-12-16, Los, Agata, Ziuzina, Dana, Boehm, Daniela, Bourke, Paula

The safety and quality of cereal grain supplies are adversely impacted by microbiological contamination, with novel interventions required to maximise whole grains safety and stability. The microbiological contaminants of wheat grains and the efficacy of Atmospheric Cold Plasma (ACP) for potential to control these risks were investigated. The evaluations were performed using a contained reactor dielectric barrier discharge (DBD) system; samples were treated for 0–20 min using direct and indirect plasma exposure. Amplicon-based metagenomic analysis using bacterial 16S rRNA gene and fungal 18S rRNA gene with internal transcribed spacer (ITS) region was performed to characterize the change in microbial community composition in response to ACP treatment. The antimicrobial efficacy of ACP against a range of bacterial and fungal contaminants of wheat, was assessed to include individual isolates from grains as challenge pathogens. ACP influenced wheat microbiome composition, with a higher microbial diversity as well as abundance found on the untreated control grain samples. Culture and genomic approaches revealed different trends for mycoflora detection and control. A challenge study demonstrated that using direct mode of plasma exposure with 20 min of treatment significantly reduced the concentration of all pathogens. Overall, reduction levels for B. atrophaeus vegetative cells were higher than for all fungal species tested, whereas B. atrophaeus spores were the most resistant to ACP among all microorganisms tested. Of note, repeating sub-lethal plasma treatment did not induce resistance to ACP in either B. atrophaeus or A. flavus spores. ACP process control could be tailored to address diverse microbiological risks for grain stability and safety.

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Cold plasma inactivation of bacterial biofilms and reduction of quorum sensing regulated virulence factors

2015-09-21, Ziuzina, Dana, Boehm, Daniela, Patil, Sonal, Cullen, P. J., Bourke, Paula

The main objectives of this work were to investigate the effect of atmospheric cold plasma (ACP) against a range of microbial biofilms commonly implicated in foodborne and healthcare associated human infections and against P. aeruginosa quorum sensing (QS)-regulated virulence factors, such as pyocyanin, elastase (Las B) and biofilm formation capacity post-ACP treatment. The effect of processing factors, namely treatment time and mode of plasma exposure on antimicrobial activity of ACP were also examined. Antibiofilm activity was assessed for E. coli, L. monocytogenes and S. aureus in terms of reduction of culturability and retention of metabolic activity using colony count and XTT assays, respectively. All samples were treated 'inpack' using sealed polypropylene containers with a high voltage dielectric barrier discharge ACP generated at 80 kV for 0, 60, 120 and 300 s and a post treatment storage time of 24 h. According to colony counts, ACP treatment for 60 s reduced populations of E. coli to undetectable levels, whereas 300 s was necessary to significantly reduce populations of L. monocytogenes and S. aureus biofilms. The results obtained from XTT assay indicated possible induction of viable but non culturable state of bacteria. With respect to P. aeruginosa QS-related virulence factors, the production of pyocyanin was significantly inhibited after short treatment times, but reduction of elastase was notable only after 300 s and no reduction in actual biofilm formation was achieved post-ACP treatment. Importantly, reduction of virulence factors was associated with reduction of the cytotoxic effects of the bacterial supernatant on CHO-K1 cells, regardless of mode and duration of treatment. The results of this study point to ACP technology as an effective strategy for inactivation of established biofilms and may play an important role in attenuation of virulence of pathogenic bacteria. Further investigation is warranted to propose direct evidence for the inhibition of QS and mechanisms by which this may occur.

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Efficacy of Cold Plasma for Direct Deposition of Antibiotics as a Novel Approach for Localized Delivery and Retention of Effect

2019-12-20, Los, Agata, Ziuzina, Dana, Boehm, Daniela, Bourke, Paula, et al.

Antimicrobial coating of medical devices has emerged as a potentially effective tool to prevent or ameliorate device-related infections. In this study the plasma deposition process for direct deposition of pharmaceutical drugs on to a range of surfaces and the retention of structure function relationship and antimicrobial efficacy against mono-species biofilms were investigated. Two selected sample antibiotics—ampicillin and gentamicin, were deposited onto two types of surfaces—polystyrene microtiter plates and stainless steel coupons. The antimicrobial efficacy of the antibiotic-coated surfaces was tested against challenge populations of both planktonic and sessile Escherichia coli and Pseudomonas aeruginosa, with responses monitored for up to 14 days. The plasma deposition process bonded the antibiotic to the surfaces, with localized retention of antibiotic activity. The antibiotics deposited on the test surfaces retained a good efficacy against planktonic cells, and importantly prevented biofilm formation of attached cells for up to 96 h. The antibiotic rapidly eluted from the surface of antibiotic-coated surfaces to the surrounding medium, with retention of effect in this surrounding milieu for up to 2 weeks. Control experiments established that there was no independent antimicrobial or growth promoting effect of the plasma deposition process, where there was no antibiotic in the helium plasma assisted delivery stream. Apart from the flexibility offered through deposition on material surfaces, there was no additive or destructive effect associated with the helium assisted plasma deposition process on the antibiotic. The plasma assisted process was a viable mean of coating clinically relevant materials and developing innovative functional materials with retention of antibiotic activity, without employing a linker or plasma modified polymer, thus minimizing bio-compatibility issues for medical device materials. This offers potential to prevent or control instrumented or non-permanent device associated infection localized to the surgical or implant site.

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Cold plasma for insect pest control: Tribolium castaneum mortality and defense mechanisms in response to treatment

2021-10, Ziuzina, Dana, Van Cleynenbreugel, Robin, Tersaruolo, Claudio, Bourke, Paula

The insecticidal properties and mechanisms of high-voltage air-based atmospheric cold plasma using a contained dielectric barrier discharge reactor were investigated against Tribolium castaneum as an important bio-contaminant in stored grains spoilage. The mortality of 95.0%–100% for preadult stages can be achieved within seconds of treatment, but longer plasma exposure (5 min) is required to kill adult insects. Cold plasma treatment reduces both the respiration rate and the weight of insects and affects the levels of oxidative stress markers in adult populations. Sufficient toxicity is achievable through plasma process control in air to address the range of insect lifecycle stages that are disease vectors and pose risks for grain stability in storage. Balancing insecticidal activity with grains' quality retention can provide a route to sustainable integrated pest management.

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Degradation kinetics of cold plasma-treated antibiotics and their antimicrobial activity

2019-03-08, Sarangapani, Chaitanya, Ziuzina, Dana, Behan, Patrice, Bourke, Paula, et al.

Antibiotics, such as ofloxacin (OFX) and ciprofloxacin (CFX), are often detected in considerable concentrations in both wastewater effluents and surface water. This poses a risk to non-target organisms and to human health. The aim of this work was to study atmospheric cold plasma (ACP) degradation of antibiotics in water and meat effluent and to explore any residual antimicrobial activity of samples submitted to the plasma process. The results revealed that ACP successfully degraded the studied antibiotics and that the reaction mechanism is principally related to attack by hydroxyl radicals and ozone. According to the disk diffusion assay, the activity of both antibiotics was considerably reduced by the plasma treatment. However, a microdilution method demonstrated that CFX exhibited higher antimicrobial activity after ACP treatment than the corresponding control revealing a potentially new platform for future research to improve the efficiency of conventional antibiotic treatments. Importantly, short-term exposures to sub-lethal concentrations of the antibiotic equally reduced bacterial susceptibility to both ACP treated and untreated CFX. As a remediation process, ACP removal of antibiotics in complex wastewater effluents is possible. However, it is recommended that plasma encompass degradant structure activity relationships to ensure that biological activity is eliminated against non-target organisms and that life cycle safety of antibiotic compounds is achieved.

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Controlling microbial safety challenges of meat using high voltage atmospheric cold plasma

2016-06-22, Han, L., Ziuzina, Dana, Heslin, Caitlin, Bourke, Paula, et al.

Atmospheric cold plasma (ACP) is a non-thermal technology, effective against a wide range of pathogenic microorganisms. Inactivation efficacy results from plasma generated reactive species. These may interact with any organic components in a test matrix including the target microorganism, thus food components may exert a protective effect against the antimicrobial mode of action. The effect of an in-package high voltage ACP process applied in conjunction with common meat processing MAP gas compositions as well as bacteria type and meat model media composition have been investigated to determine the applicability of this technology for decontamination of safety challenges associated with meat products. E. coli, L. monocytogenes, and S. aureus in PBS were undetectable after 60 s of treatment at 80 kVRMS in air, while ACP treatment of the contaminated meat model required post-treatment refrigeration to retain antimicrobial effect. The nutritive components in the meat model exerted a protective effect during treatment, where 300 s ACP exposure yielded a maximum reduction of 1.5 log using a high oxygen atmosphere, whilst using air and high nitrogen atmospheres yielded lower antimicrobial efficacy. Furthermore, an ROS assay was performed to understand the protective effects observed using the meat model. This revealed that nutritive components inhibited penetration of ROS into bacterial cells. This knowledge can assist the optimization of meat decontamination using ACP technology where interactions with all components of the food matrix require evaluation.

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Investigation of a large gap cold plasma reactor for continuous in-package decontamination of fresh strawberries and spinach

2020-01, Ziuzina, Dana, Misra, N. N., Han, L., Bourke, Paula, et al.

The aim of this work was to investigate the efficacy of a large gap atmospheric cold plasma (ACP) generated with an open-air high-voltage dielectric barrier discharge (DBD) pilot-scale reactor, operated in either static (batch) or continuous mode for produce decontamination and quality retention. Significant reductions in the bacterial populations inoculated on the strawberries and spinach were obtained after the static mode of ACP treatment with 2.0 and 2.2 log10 CFU/ml reductions for E. coli and 1.3 and 1.7 log10 CFU/ml reductions for L. innocua, respectively. Continuous treatment was effective against L. innocua inoculated on strawberries, with 3.8 log10 CFU/ml reductions achieved. No significant differences in colour, firmness, pH or total soluble solids (TSS) was observed between control and ACP-treated samples with the effects of treatment retained during the shelf-life period. The pilot-scale atmospheric air plasma reactor retained the strawberry quality characteristics in tandem with useful antimicrobial efficacy. Industrial relevance: This in-package plasma technology approach is a low-power, water-free, non-thermal, post-package treatment. Generating cold plasma discharges inside food packages achieved useful antimicrobial effects on fresh produce. Depending on the bacterial type, produce and mode of ACP treatment significant reductions in the populations of pathogenic microorganisms attached to the fresh produce was achieved within 2.5 min of treatment. The principal technical advantages include contaminant control, quality retention, mitigation of re-contamination and crucially the retention of bactericidal reactive gas molecules in the food package volume, which then revert back to the original gas.

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High voltage atmospheric cold air plasma control of bacterial biofilms on fresh produce

2019-03-16, Patange, Apurva, Boehm, Daniela, Ziuzina, Dana, Cullen, P. J., Gilmore, Brendan F., Bourke, Paula

Atmospheric cold plasma (ACP) offers great potential for decontamination of food borne pathogens. This study examined the antimicrobial efficacy of ACP against a range of pathogens of concern to fresh produce comparing planktonic cultures, monoculture biofilms (Escherichia coli, Salmonella enterica, Listeria monocytogenes, Pseudomonas fluorescens) and mixed culture biofilms (Listeria monocytogenes and Pseudomonas fluorescens). Biotic and abiotic surfaces commonly occurring in the fresh food industry were investigated. Microorganisms showed varying susceptibility to ACP treatment depending on target and process factors. Bacterial biofilm populations treated with high voltage (80 kV) ACP were reduced significantly (p < 0.05) in both mono- and mixed species biofilms after 60 s of treatment and yielded non-detectable levels after extending treatment time to 120 s. However, an extended time was required to reduce the challenge mixed culture biofilm of L. monocytogenes and P. fluorescens inoculated on lettuce, which was dependent on biofilm formation conditions and substrate. Contained treatment for 120 s reduced L. monocytogenes and P. fluorescens inoculated as mixed cultures on lettuce (p < 0.05) by 2.2 and 4.2 Log 10 CFU/ml respectively. When biofilms were grown at 4 °C on lettuce, there was an increased resistance to ACP treatment by comparison with biofilm grown at temperature abuse conditions of 15 °C. Similarly, L. monocytogenes and P. fluorescens exposed to cold stress (4 °C) for 1 h demonstrated increased tolerance to ACP treatment compared to non-stressed cells. These finding demonstrates that bacterial form, mono versus mixed challenges as well as environmental stress conditions play an important role in ACP inactivation efficacy.

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Assessing the Biological Safety of Atmospheric Cold Plasma Treated Wheat Using Cell and Insect Models

2020-07-08, Los, Agata, Ziuzina, Dana, Van Cleynenbreugel, Robin, Boehm, Daniela, Bourke, Paula

Atmospheric cold plasma (ACP) is under investigation for an extensive range of biocontrol applications in food biosystems. However, the development of a novel intervention technology requires a thorough evaluation of the potential for negative effects and the implications for the human and animal food chains' safety. The evaluations were performed using a contained, high-voltage, dielectric barrier discharge plasma system. The cytotoxicity of two types of food models-a liquid model (wheat model medium (WMM)) vs. a solid model (wheat grain extract (WGE)) was compared in vitro using the mammalian cell line CHO-K1. The residual toxicity of ACP treatment of grains for food purposes was assessed using the invertebrate model Tribolium castaneum, by feeding the beetles with flour produced from ACP-treated wheat grains. The cytotoxic effects and changes in the chemistry of the ACP-treated samples were more pronounced in samples treated in a liquid form as opposed to actual wheat grains. The feeding trial using T. castaneum demonstrated no negative impacts on the survivability or weight profiles of insects. Investigations into the interactions of plasma-generated species with secondary metabolites in the food matrices are necessary to ensure the safety of plasma for food applications.