Recent advances in the degradation efficacy and mechanisms of mycotoxins in food by atmospheric cold plasma.

Food contaminated by mycotoxins has become a worldwide public problem with political and economic implications. Although a variety of traditional methods have been used to eliminate mycotoxins from agri-foods, the results have been somewhat less than satisfactory. As an emerging non-thermal processing technology, atmospheric cold plasma (ACP) has great potential for food decontamination. Herein, this review mainly presents the degradation efficiency of ACP on mycotoxins in vitro and agri-foods as well as its possible degradation mechanisms. Meanwhile, ACP effects on food quality, factors affecting the degradation efficiency and the toxicity of degradation products are also discussed. According to the literatures, ACP could efficiently degrade many mycotoxins (e.g., aflatoxin, deoxynivalenol, zearalenone, ochratoxin A, fumonisin, and T-2 toxin) both in vitro and various foods (e.g., hazelnut, peanut, maize, rice, wheat, barley, oat flour, and date palm fruit) with little effects on the nutritional and sensory properties of food. The degradation efficacy was dependent on many factors including ACP treatment parameter, working gas, mycotoxin property, and food substrate. The mycotoxin degradation by ACP was mainly attributed to the reactive oxygen and nitrogen species in ACP, which can damage the chemical bonds of mycotoxins, consequently reducing the toxicity of mycotoxins.

Effects of low-energy N(+)-beam implantation on root growth in Arabidopsis seedlings.

The effects of ion implantation on the morphology changes and biological responses of plants are dependent on implantation doses. Previous studies mainly focus on the application of ion-beam technology in genetic mutation. Our knowledge regarding the mechanism underlying the plant growth inhibition induced by ion implantation remains limited. In this study, we explore the responses of root growth to low-energy N(+)-beam implantation using implanted Arabidopsis seeds. Our results showed that the root and root tip length were obviously reduced by implantation with large doses of low-energy N(+) beam. The analysis of confocal images showed that ion implantation reduced the cell viability and cell division activity in root meristem. The production rate of superoxide radical (O2(•-)) and contents of hydrogen peroxide (H2O2) in roots under ion implantation were markedly higher than those of controls. Transcriptional expression analysis of selected genes revealed that Arabidopsis RBOH genes associated with reactive oxygen species (ROS) production were significantly up-regulated in roots in response to ion implantation. The activities of antioxidant enzymes were also induced by ion implantation. Moreover, ROS scavenging obviously enhanced cell viability and cell division in response to ion implantation and alleviated the root growth inhibition of the implanted seedlings. Our results suggest that the overproduction of ROS induced by ion implantation is involved in the inhibitory effect of low-energy ion beam on root growth by affecting the cell viability and cell division of root meristem in Arabidopsis seedlings.

Pretreatment with low-dose gamma irradiation enhances tolerance to the stress of cadmium and lead in Arabidopsis thaliana seedlings.

Heavy metals are important environmental pollutants with negative impact on plant growth and development. To investigate the physiological and molecular mechanisms of heavy metal stress mitigated by low-dose gamma irradiation, the dry seeds of Arabidopsis thaliana were exposed to a Cobalt-60 gamma source at doses ranging from 25 to 150Gy before being subjected to 75µM CdCl2 or 500µM Pb(NO3)2. Then, the growth parameters, and physiological and molecular changes were determined in response to gamma irradiation. Our results showed that 50-Gy gamma irradiation gave maximal beneficial effects on the germination index and root length in response to cadmium/lead stress in Arabidopsis seedlings. The hydrogen peroxide and malondialdehyde contents in seedlings irradiated with 50-Gy gamma rays under stress were significantly lower than those of controls. The antioxidant enzyme activities and proline levels in the irradiated seedlings were significantly increased compared with the controls. Furthermore, a transcriptional expression analysis of selected genes revealed that some components of heavy metal detoxification were stimulated by low-dose gamma irradiation under cadmium/lead stress. Our results suggest that low-dose gamma irradiation alleviates heavy metal stress, probably by modulating the physiological responses and gene expression levels related to heavy metal resistance in Arabidopsis seedlings.

Physiological and molecular characterization of the enhanced salt tolerance induced by low-dose gamma irradiation in Arabidopsis seedlings.

It has been established that gamma rays at low doses stimulate the tolerance to salt stress in plants. However, our knowledge regarding the molecular mechanism underlying the enhanced salt tolerance remains limited. In this study, we found that 50-Gy gamma irradiation presented maximal beneficial effects on germination index and root length in response to salt stress in Arabidopsis seedlings. The contents of H2O2 and MDA in irradiated seedlings under salt stress were significantly lower than those of controls. The activities of antioxidant enzymes and proline levels in the irradiated seedlings were markedly increased compared with the controls. Furthermore, transcriptional expression analysis of selected genes revealed that some components of salt stress signaling pathways were stimulated by low-dose gamma irradiation under salt stress. Our results suggest that gamma irradiation at low doses alleviates the salt stress probably by modulating the physiological responses as well as stimulating the stress signal transduction in Arabidopsis seedlings.

Application of atmospheric cold plasma for zearalenone detoxification in cereals: Kinetics, mechanisms, and cytotoxicity analysis.

INTRODUCTION: Zearalenone (ZEN) is one of the most widely contaminated mycotoxins in world, posing a severe threat to human and animal health. Atmospheric cold plasma (ACP) holds great penitential in mycotoxin degradation. OBJECTIVES: This study aimed to investigate the degradation efficiency and mechanisms of ACP on ZEN as well as the cytotoxicity of ZEN degradation products by ACP. Additionally, this study also investigated the degradation efficiency of ACP on ZEN in cereals and its effect on cereal quality. METHODS: The degradation efficiency and products of ZEN by ACP was analyzed by HPLC and LC-MS/MS. The human normal liver cells and mice were employed to assess the cytotoxicity of ZEN degradation products. The ZEN artificially contaminated cereals were used to evaluate the feasibility of ACP detoxification in cereals. RESULTS: The results showed that the degradation rate of ZEN was 96.18 % after 30-W ACP treatment for 180 s. The degradation rate was dependent on the discharge power, and treatment time and distance. Four major ZEN degradation products were produced after ACP treatment due to the oxidative destruction of CC double bond, namely C18H22O7 (m/z = 351.19), C18H22O8 (m/z = 367.14), C18H22O6 (m/z = 335.14), and C17H20O6 (m/z = 321.19). L02 cell viability was increased from 52.4 % to 99.76 % with ACP treatment time ranging from 0 to 180 s. Mice results showed significant recovery of body weight and depth of colonic crypts as well as mitigation of glomerular and liver damage. Additionally, ACP removed up to 50.55 % and 58.07 % of ZEN from wheat and corn. CONCLUSIONS: This study demonstrates that ACP could efficiently degrade ZEN in cereals and its cytotoxicity was significantly reduced. Therefore, ACP is a promising effective method for ZEN detoxification in cereals to ensure human and animal health. Future study needs to develop large-scale ACP device with high degradation efficiency.

Inhibition of Fungal Growth and Aflatoxin B1 Synthesis in Aspergillus flavus by Plasma-Activated Water.

The gaseous reactive oxygen/nitrogen species (RONS) generated by cold atmospheric plasma (CAP) can effectively inactivate Aspergillus flavus (A. flavus) and prolong the shelf-life of food. Plasma-activated water (PAW) is the extension of cold plasma sterilization technology. Without the limitation of a plasma device, PAW can be applied to more scenarios of food decontamination. However, the efficacy of PAW as a carrier of RONS for eradicating A. flavus or inhibiting its growth remains unclear. In this study, the immediate fungicidal effect and long-term inhibitory effect of PAW on A. flavus were investigated. The results demonstrated that 60-min instant-prepared PAW could achieve a 3.22 log reduction CFU/mL of A. flavus and the fungicidal efficacy of PAW gradually declined with the extension of storage time. Peroxynitrite (ONOO-/ONOOH) played a crucial role in this inactivation process, which could damage the cell wall and membrane structure, disrupt intracellular redox homeostasis, and impair mitochondrial function, ultimately leading to fungal inactivation. In addition to the fungicidal effect, PAW also exhibited fungistatic properties and inhibited the synthesis of aflatoxin B1 (AFB1) in A. flavus. By analyzing the cellular antioxidant capacity, energy metabolism, and key gene expression in the AFB1 synthesis pathway, it was discovered that PAW can significantly reduce ATP levels, while increasing SOD and CAT activity during 5-d cultivation. Meanwhile, PAW effectively suppressed the expression of genes related to AFB1 synthesis.

Comparative analysis of helium and air surface micro-discharge plasma treatment on the microbial reduction and quality attributes of beef slices.

This work aimed to compare the effects of helium and air surface micro-discharge (SMD) plasma on the microbial safety and quality of beef tissues. For the beef tissue model, the concentration and diffusion depth of hydroxyl radical and ozone have different change patterns over plasma treatment time and distance in helium and air SMD plasma. The inactivation efficiency of helium plasma depended on the plasma treatment time and distance, while the inactivation efficiency of air plasma only depended on the treatment time. For the fresh beef slices, air SMD plasma treatment exhibited a higher antimicrobial activity against S. aureus and E. coli than helium SMD plasma treatment (1.5 versus 0.9; 0.9 versus 0.28 log CFU/g at 10 min). However, air SMD plasma treatment caused more adverse effects on beef quality, leading to a smooth surface, extensive lipid oxidation, protein structure damage, low pH and discoloration compared to helium SMD plasma treatment. This work provides valuable guidelines for the working gas choice in the practical application of plasma to meat decontamination.

Simultaneous removal of norfloxacin and chloramphenicol using cold atmospheric plasma jet (CAPJ): Enhanced performance, synergistic effect, plasma-activated water (PAW) contribution, mechanism and toxicity evaluation.

The extensive abuse and inadvertent discharge of various antibiotics into the environment has become a serious problem for posing a big threat to human health. In order to deal with this problem, we utilized cold atmospheric plasma jet (CAPJ) to treat two different antibiotics, namely, norfloxacin and chloramphenicol, and investigated the efficiencies and corresponding mechanisms for removing the mixed antibiotics. In the application of the CAPJ technique, we made use of not only the direct plasma processing, but also the indirect plasma-activated water (PAW) treatment. The efficiency for mixed antibiotics treatment was considerably enhanced as compared to the efficiency for treatment of single antibiotics. The contributions from the CAPJ-induced reactive oxygen/nitrogen species (RONS) were examined, showing that ·OH and 1O2 played a major role in the degradation of norfloxacin and chloramphenicol in the direct plasma treatment, while 1O2 played a major role in the PAW treatment. The bio-toxicity evaluation was also provided to verify the ecological safety of the CAPJ treatment. As such, this work has not only showed the effectiveness of CAPJ treatment of mixed antibiotics, but also elucidated the mechanisms for the enhanced treatment efficiency, which may provide a new solution for treatment of antibiotics in the environment.

Reactive oxygen species may be involved in the distinctive biological effects of different doses of 12C6+ ion beams on Arabidopsis.

Introduction: Heavy ion beam is a novel approach for crop mutagenesis with the advantage of high energy transfer line density and low repair effect after injury, however, little investigation on the biological effect on plant was performed. 50 Gy irradiation significantly stimulated the growth of Arabidopsis seedlings, as indicated by an increase in root and biomass, while 200 Gy irradiation significantly inhibited the growth of seedlings, causing a visible decrease in plant growth. Methods: The Arabidopsis seeds were irradiated by 12C6+. Monte Carlo simulations were used to calculate the damage to seeds and particle trajectories by ion implantation. The seed epidermis received SEM detection and changes in its organic composition were detected using FTIR. Evidence of ROS and antioxidant systems were analyzed. RNA-seq and qPCR were used to detect changes in seedling transcript levels. Results and discussion: Monte Carlo simulations revealed that high-dose irradiation causes various damage. Evidence of ROS and antioxidant systems implies that the emergence of phenotypes in plant cells may be associated with oxidative stress. Transcriptomic analysis of the seedlings demonstrated that 170 DEGs were present in the 50 Gy and 200 Gy groups and GO enrichment indicated that they were mainly associated with stress resistance and cell wall homeostasis. Further GO enrichment of DEGs unique to 50 Gy and 200 Gy revealed 58 50Gy-exclusive DEGs were enriched in response to oxidative stress and jasmonic acid entries, while 435 200 Gy-exclusive DEGs were enriched in relation to oxidative stress, organic cyclic compounds, and salicylic acid. This investigation advances our insight into the biological effects of heavy ion irradiation and the underlying mechanisms.

Degradation of tetracycline by atmospheric-pressure non-thermal plasma: Enhanced performance, degradation mechanism, and toxicity evaluation.

Tetracycline is a common antibiotic and is often carelessly released into the natural environment, thus constantly posing potential threats to the environment. Currently, due to lack of effective methods to remove it from the environmental water system, researchers are still exploring new ways to deal with tetracycline. In this work, we employed atmospheric-pressure non-thermal plasma (NTP) to treat tetracycline in water and investigated the involved degradation mechanism. The enhanced degradation efficiency was acquired and investigated, and the degradation mechanism by the plasma-generated active species were explored. The tetracycline degradation pathways via especially the interactions with plasma-generated hydroxyl radical and ozone were examined by virtue of UV spectroscopy, three-dimensional fluorescence spectroscopy, high performance liquid chromatography-mass spectrometry (HPLC-MS), together with the assistance of theoretical simulations. Moreover, the toxicological evaluation of NTP treatment of tetracycline was also provided, which confirmed that the biological toxicity of tetracycline degradation products was negligible. Therefore, this work provides not only the effective way of treating antibiotics by engineered plasma technology, but also the insights into the mechanisms of degradation of antibiotics by NTP.

Study of the synergistic effect of singlet oxygen with other plasma-generated ROS in fungi inactivation during water disinfection.

Cold atmospheric plasma (CAP) possesses the ability of high-efficiency disinfection. It is reported that mixtures of reactive oxygen species (ROS) including ·OH, 1O2, O2- and H2O2 generated from CAP have better antimicrobial ability than mimicked solution of mixture of single ROS type, but the reason is not clear. In this study, CAP was applied to treat yeasts in water in order to investigate the fungal inactivation efficiency and mechanism. The results showed that plasma treatment for 5 min could result in >2-log reduction of yeast cells, and application of varied ROS scavengers could significantly increase the yeast survival rate, indicating that ·OH and 1O2 played the pivotal role in yeast inactivation. Moreover, the synergistic effect of 1O2 with other plasma-generated ROS was revealed. 1O2 could diffuse into cells and induce the depolarization of mitochondrial membrane potential (MMP), and different levels of MMP depolarization determined different cell death modes. Mild damage of mitochondria during short-term plasma treatment could lead to apoptosis. For long-term plasma treatment, the cell membrane could be severely damaged by the plasma-generated ·OH, so a large amount of 1O2 could induce more depolarization of MMP, leading to increase of intracellular O2- and Fe2+ which subsequently caused cell inactivation. 1O2 could also induce protein aggregation and increase of RIP1/RIP3 necrosome, leading to necroptosis. With participation of 1O2, endogenous ·OH could also be generated via Fenton and Haber-Weiss reactions during plasma treatment, which potentiated necroptosis. Adding l-His could mitigate membrane damage, inhibit the drop of MMP and the formation of necrosome, and thus prevent the happening of necroptosis. These findings may deepen the understanding of plasma sterilization mechanisms and provide guidance for microbial killing in the environment.

Subcellular mechanism of microbial inactivation during water disinfection by cold atmospheric-pressure plasma.

Although the identification of effective reactive oxygen species (ROS) generated by plasma has been extensively studied, yet the subcellular mechanism of microbial inactivation has never been clearly elucidated in plasma disinfection processes. In this study, subcellular mechanism of yeast cell inactivation during plasma-liquid interaction was revealed in terms of comprehensive factors including cell morphology, membrane permeability, lipid peroxidation, membrane potential, intracellular redox homeostasis (intracellular ROS and H2O2, and antioxidant system (SOD, CAT and GSH)), intracellular ionic equilibrium (intracellular H+ and K+) and energy metabolism (mitochondrial membrane potential, intracellular Ca2+ and ATP level). The ROS analysis show that ·OH, 1O2, ·O2-and H2O2 were generated in this plasma-liquid interaction system and ·O2-served as the precursor of 1O2. Additionally, the solution pH was reduced. Plasma can effectively inactivate yeast cells mainly via apoptosis by damaging cell membrane, intracellular redox and ion homeostasis and energy metabolism as well as causing DNA fragmentation. ROS scavengers (l-His, d-Man and SOD) and pH buffer (phosphate buffer solution, PBS) were employed to investigate the role of five antimicrobial factors (·OH, 1O2, ·O2-, H2O2 and low pH) in plasma sterilization. Results show that they have different influences on the aforementioned cell physiological activities. The ·OH and 1O2 contributed most to the yeast inactivation. The ·OH mainly attacked cell membrane and increased cell membrane permeability. The disturb of cell energy metabolism was mainly attributed to 1O2. The damage of cell membrane as well as extracellular low pH could break the intracellular ionic equilibrium and further reduce cell membrane potential. The remarkable increase of intracellular H2O2 was mainly due to the influx of extracellular H2O2 via destroyed cell membrane, which played a little role in yeast inactivation during 10-min plasma treatment. These findings provide comprehensive insights into the antimicrobial mechanism of plasma, which can promote the development of plasma as an alternative water disinfection strategy.

Visible-light-driven photocatalytic inactivation of S. aureus in aqueous environment by hydrophilic zinc oxide (ZnO) nanoparticles based on the interfacial electron transfer in S. aureus/ZnO composites.

Waterborne diseases caused by pathogenic microorganisms pose severe threats to human health. ZnO nanoparticles (NPs) hold great potentials as an effective, economical and eco-friendly method for water disinfection, but the exact antimicrobial mechanism of ZnO NPs under visible-light illumination is still not clear. Herein, we investigate the visible-light-driven photocatalytic inactivation mechanism of amino-functionalized hydrophilic ZnO (AH-ZnO) NPs against Staphylococcus aureus (S. aureus) in aqueous environment from the perspective of electron transfer theory. The results show that the antibacterial effects of AH-ZnO NPs are dependent on the AH-ZnO NPs concentration and treatment time. The bulk ORP value and released Zn2+ concentration in AH-ZnO NPs solutions increase with AH-ZnO NPs concentration. The SEM and intracellular protein leakage results indicate that AH-ZnO NPs can adhere to S. aureus surface without causing obvious cell membrane disruption. The photoluminescence (PL) intensity and fluorescence lifetime of AH-ZnO NPs are remarkedly decreased after adding S. aureus, which confirms the electron transfer from S. aureus to AH-ZnO NPs. Moreover, the ΔPL intensity is closely correlated with the inactivation efficiency, demonstrating that the interfacial electron transfer in S. aureus/AH-ZnO NPs composites contributes to the antibacterial activity, which is speculated to disrupt the normal respiratory electron transfer chain of S. aureus, thereby causing intracellular ROS generation, cell membrane depolarization and eventually apoptosis-like death.

A systematic study of the antimicrobial mechanisms of cold atmospheric-pressure plasma for water disinfection.

Waterborne diseases caused by pathogenic microorganisms pose a severe threat to human health. Cold atmospheric-pressure plasma (CAP) has recently gained much interest as a promising fast, effective, economical and eco-friendly method for water disinfection. However, the antimicrobial mechanism of CAP in aqueous environments is still not clearly understood. Herein, we investigate the role of several short-lived reactive oxygen species (ROS) and cellular responses in the CAP inactivation of yeast cells in water. The results show that singlet oxygen (1O2), hydroxyl radical (OH) and superoxide anion (O2-) are generated in this plasma-water system, and O2- served as the precursor of OH. The 5-min plasma treatment resulted in the effective inactivation (more than 2-log reduction) of yeast cells in water. The ROS scavengers significantly increased the survival ratio in the following order: water < D-Man (scavenging OH) < SOD (scavenging O2-) < L-His (scavenging 1O2), indicating that 1O2 contributes the most to the yeast inactivation. In addition, the acidic pH had a synergetic antimicrobial effect with ROS against the yeast cells. During the CAP inactivation process, yeast cells underwent apoptosis in the first 3 min due to the accumulation of intracellular ROS, mitochondrial dysfunction and intracellular acidification, later followed by necrosis under longer exposure times, attributed to the destruction of the cell membrane. Additionally, L-His could switch the cell fate from necrosis to apoptosis through mitigating plasma-induced oxidative stress, indicating that the level of oxidative stress is a critical factor for cell death fate determination. These findings provide comprehensive insights into the antimicrobial mechanism of CAP, which can promote the development of CAP as an alternative water disinfection strategy.

ROS and ABA signaling are involved in the growth stimulation induced by low-dose gamma irradiation in Arabidopsis seedling.

It has been well established that gamma rays at low doses have stimulatory effects on plant growth and development. However, our knowledge regarding the molecular mechanism underlying the growth stimulation remains limited. In this study, we report the role of reactive oxygen species (ROS) and abscisic acid (ABA) in the growth stimulation using irradiated Arabidopsis seeds. The results indicated that 50 Gy gamma irradiation presented maximal beneficial effects on germination index, root length, and fresh weight. The contents of hydrogen peroxide (H2O2) and activities of antioxidant enzymes under gamma irradiation were markedly higher than those of controls. ROS scavenging significantly suppressed the growth of the irradiated plants. Furthermore, endogenous ABA was induced under low-dose gamma irradiation. The growth stimulation and elevated H2O2 level were affected in the irradiated ABA-deficient mutant aba2-1 compared with the mutant control. Transcriptional expression analysis of selected genes revealed that several genes for ABA biosynthesis were upregulated, and the genes for ABA catabolic pathway and transport were differentially regulated in response to low-dose gamma irradiation. Our results suggest that ROS and ABA signaling play an essential role in the stimulatory effects of low-dose gamma irradiation and that ROS, as secondary molecules, mediate ABA signal transduction under irradiation in response to stress factors during plant growth.