Xinyu Liao

Synergetic effects of ultrasound and slightly acidic electrolyzed water against Staphylococcus aureus evaluated by flow cytometry and electron microscopy

This study evaluated the synergetic effects of ultrasound and slightly acidic electrolyzed water (SAEW) on the inactivation of Staphylococcus aureus using flow cytometry and electron microscopy. The individual ultrasound treatment for 10min only resulted in 0.36logCFU/mL reductions of S. aureus, while the SAEW treatment alone for 10min resulted in 3.06logCFU/mL reductions. The log reductions caused by combined treatment were enhanced to 3.68logCFU/mL, which were greater than the sum of individual treatments. This phenomenon was referred to as synergistic effects. FCM analysis distinguished live and dead cells as well as revealed dynamic changes in the physiological states of S. aureus after different treatments. The combined treatment greatly reduced the number of viable but nonculturable (VBNC) bacteria to 0.07%; in contrast, a single ultrasound treatment for 10min induced the formation of VBNC cells to 45.75%. Scanning and transmission electron microscopy analysis revealed that greater damage to the appearance and ultrastructure of S. aureus were achieved after combined ultrasound-SAEW treatment compared to either treatment alone. These results indicated that combining ultrasound with SAEW is a promising sterilization technology with potential uses for environmental remediation and food preservation.

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.

Analysis of Staphylococcus aureus cell viability, sublethal injury and death induced by synergistic combination of ultrasound and mild heat

This study was designed to investigate the combined effects of ultrasound and mild heat on the viability of S. aureus in association with the cell membrane integrity and intracellular enzyme activity. Cells were treated by ultrasound under 55°C for 3, 5, 7, 10, and 15min. The dynamic changes of S. aureus cell viability, sublethal injury and death were evaluated using flow cytometric assay. Microscopies were applied to identify the morphological appearance, ultrastructure and topography of S. aureus. The results showed the membrane damage was synchronous with esterase inhibition during the exposure to sonication, leading to the immediate lethal effect. On the other hand, bacteria under the mild heat at 55°C were inactivated via a sublethal injury process. The different lethal modes were observed between sonication and mild heat treatments, which could synergistically inactivate S. aureus. The antibacterial value of thermo-sonication was greater than the sum of the individual treatments. The thermo-sonication combination synergistically reduced the number of sublethal cells and also resulted in severe cell damage.

Application of a Dielectric Barrier Discharge Atmospheric Cold Plasma (Dbd-Acp) for Eshcerichia Coli Inactivation in Apple Juice: Inactivation of E. coli by cold plasma…

Atmospheric cold plasma (ACP) is a promising non-thermal technology in food industry. In this study, a dielectric barrier discharge (DBD)-ACP exhibited strong bactericidal effect on Escherichia coli in apple juice. Under a 30 to 50 W input power, less than 40 s treatment time was required for DBD-ACP to result in 3.98 to 4.34 log CFU/mL reduction of E. coli in apple juice. The inactivation behavior of ACP on E. coli was well described by the Weibull model. During the treatment, the cell membrane of E. coli was damaged severely by active species produced by plasma, such as hydrogen peroxide, ozone and nitrate. In addition, the ACP exposure had slight effect on the °Brix, pH, titratable acidity (TA), color values, total phenolic content, and antioxidant capacity of apple juice. However, higher level of DBD-ACP treatment, 50 W for more than 10 s in this case, resulted in significant change of the pH, TA, color and total phenolic content of apple juice. The results in this study have provided insight in potential use of DBD-ACP as an alternative to thermal processing for fruit juices in food industry. PRACTICAL APPLICATION Escherichia coli O157:H7 in apple juice is a potential risk for public health. This study demonstrated that 30 s cold plasma treatment resulted in more than 4 log CFU/mL reduction under 50 W, while the quality attributes of apple juice were not significantly affected. Therefore, cold plasma technology is a promising alternative substitute of traditional thermal processing for juice pasteurization.

Bacterial spore inactivation induced by cold plasma

ABSTRACT Cold plasma has emerged as a non-thermal technology for microbial inactivation in the food industry over the last decade. Spore-forming microorganisms pose challenges for microbiological safety and for the prevention of food spoilage. Inactivation of spores induced by cold plasma has been reported by several studies. However, the exact mechanism of spore deactivation by cold plasma is poorly understood; therefore, it is difficult to control this process and to optimize cold plasma processing for efficient spore inactivation. In this review, we summarize the factors that affect the resistance of spores to cold plasma, including processing parameters, environmental elements, and spore properties. We then describe possible inactivation targets in spore cells (e.g., outer structure, DNA, and metabolic proteins) that associated with inactivation by cold plasma according to previous studies. Kinetic models of the sporicidal activity of cold plasma have also been described here. A better understanding of the interaction between spores and cold plasma is essential for the development and optimization of cold plasma technology in food the industry.

Understanding the Impact of Nonthermal Plasma on Food Constituents and Microstructure—A Review

Nonthermal plasma (NTP) is superior to thermal technologies as a technique that provides a satisfactory microbial safety and maintains reasonable standards in food quality attributes. Currently, the effects of NTP on some food components is regarded as beneficial, such as effects on starch and protein modification. For other food components, such as lipid oxidation, NTP is regarded as an undesirable treatment because it leads to quality deterioration and formation of off-flavor. An overview of the basic principles of NTP and food microstructure in relation to NTP-treated food and the underlying mechanisms are discussed. The review further highlights the latest research on plasma application in food and the related impact on food matrices. Efforts were made to outline the research findings in terms of NTP application on foods with an emphasis on the impacts on the food microstructure and their related qualities. In this review, the industrial capacity of NTP to improve the functional properties of starch, proteins, and lipids as well as provide little or no alteration in food quality compared to other technologies are emphasized. Some oxidative breakdown in relation to starch, proteins, and lipids are discussed and documented in this paper as a review of representative available publications.

Effect of preliminary stresses on the resistance of Escherichia coli and Staphylococcus aureus toward non-thermal plasma (NTP) challenge

As the development of hurdle technology, cross-protection of various stresses for pathogens posed the potential risk to food safety and public health. This study tried to explore various preliminary stresses including acidity, osmosis, oxidation, heat and cold on the resistance of microbial cells toward the non-thermal plasma (NTP) exposure. The results indicated that short-term (4h) exposure of Staphylococcus aureus and Escherichia coli to acidity, osmosis, oxidation, heat and cold stresses did not lead to the resistance to the subsequent NTP treatment. On the contrary, acidity, osmosis and heat preadaptation increased the vulnerability of E. coli cells to NTP treatment. After exposing S. aureus to osmosis, oxidation, heat and cold stress for longer period (24h), the reduction level showed significantly (P<0.05) higher. Interestingly, long-term (24h) preliminary exposure of acidic stress exhibited protective effect for S. aureus against the following NTP exposure with less damage in cell membrane integrity, membrane potential and intracellular enzyme activity. It might be due to the protein production for oxidative stress response during preliminary acidic adaptation. In general, the obtained result helped to grasp better understanding of the microbial stress response to NTP treatment and provided insight for the future research in order to accelerate the development of NTP technology in food industry.

Effects of Nonthermal Plasma Technology on Functional Food Components: NTP effects on functional components…

Understanding the impact of nonthermal plasma (NTP) technology on key nutritional and functional food components is of paramount importance for the successful adoption of the technology by industry. NTP technology (NTPT) has demonstrated marked antimicrobial efficacies with good retention of important physical, chemical, sensory, and nutritional parameters for an array of food products. This paper presents the influence of NTPT on selected functional food components with a focus on low-molecular-weight bioactive compounds and vitamins. We discuss the mechanisms of bioactive compound alteration by plasma-reactive species and classify their influence on vitamins and their antioxidant capacities. The impact of NTP on specific bioactive compounds depends both on plasma properties and the food matrix. Induced changes are mainly associated with oxidative degradation and cleavage of double bonds in organic compounds. The effects reported to date are mainly time-dependent increases in the concentrations of polyphenols, vitamin C, or increases in antioxidant activity. Also, improvement in the extraction efficiency of polyphenols is observed. The review highlights future research needs regarding the complex mechanisms of interaction with plasma species. NTP is a novel technology that can both negatively and positively affect the functional components in food.

Combating Staphylococcus aureus and its methicillin resistance gene (mecA) with cold plasma

The increase in antibiotic resistance has become a global challenge to public health. In this study, an atmospheric cold plasma (ACP) system was applied for combating methicillin-resistant Staphylococcus aureus (MRSA) and its methicillin resistance gene (mecA) during food wastewater treatment. The plate count and flow cytometry methods were employed to estimate the damage in MRSA induced by plasma treatment. A quantitative real-time PCR (qPCR) method was used to assess the plasma-induced degradation of the mecA genes. The inactivation of MRSA and degradation of extracellular (e-) and intracellular (i-)mecA genes were investigated in phosphate buffered solution as a function of plasma exposure. A relatively low plasma influence of 0.12 kJ/cm2 accounted for 5-log MRSA and 1.4-log e-mecA genes reduction, while only around 0.19-log degradation for i-mecA genes. As the plasma intensity was accumulated to 0.35 kJ/cm2, the reduction of e- and i-mecA genes was increased to 2.6 and 0.8 logs, respectively. The degradation of i-mecA genes was much slower than that of e-mecA genes due to the protective effects of the outer envelopes or intracellular components against plasma. The matrix effect of wastewater effluents shielded both antibiotic resistance bacteria (ARB) and antibiotic resistance genes (ARGs) from plasma disinfection, which led to a lower degradation efficacy. Our results could support the development and optimization of plasma-based wastewater treatment.

Ultrasound-induced Escherichia coli O157:H7 cell death exhibits physical disruption and biochemical apoptosis

Ultrasound has attracted great interest of both industry and scientific communities for its potential use as a physical processing and preservation tool. In this study, Escherichia coli O157:H7 was selected as the model microbe to investigate the ultrasound-induced cell death. Slight variations in membrane potential and ion exchanges across membrane induced by low-intensity ultrasound increased the membrane permeability of E. coli O157:H7, and this reversible sublethal effect can preserve the viability of E. coli O157:H7 and meanwhile be beneficial for bioprocessing application. In comparison, high-intensity ultrasound resulted in irreversible lethal effect on E. coli O157:H7, which can be applied in the field of microbial inactivation. In addition, both low- and high-intensity ultrasound induced either physical destruction or trigger genetically encoded apoptosis of E. coli O157:H7. Accumulation of reactive oxygen species and decrease of adenosine tri-phosphate might be related to the physiological and biochemical hallmarks of apoptosis, including exposed phosphatidylserine and activated caspases in E. coli O157:H7. The result provides novel insight into the mechanisms of non-thermal physical treatment on the inactivation of bacteria and lays foundation for the further research on the cell signaling and metabolic pathway in apoptotic bacteria.