The demand for rapid, consistent and easy-to-use techniques for detecting and identifying pathogens in various areas, such as clinical diagnosis, the pharmaceutical industry, environmental science and food inspection, is very important. In this study, the reference strains of six food-borne pathogens, namely, Escherichia coli 0157: H7 ATCC 43890, Cronobacter sakazakii ATCC 29004, Salmonella Typhimurium ATCC 43971, Staphylococcus aureus KCCM 40050, Bacillus subtilis ATCC 14579, and Listeria monocytogenes ATCC 19115, were chosen for scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) analysis. In our study, the time-consuming sample preparation step for the microbial analysis under SEM was avoided, which makes this detection process notably rapid. Samples were loaded onto a 0.01-µm-thick silver (Ag) foil surface to avoid any charging effect. Two different excitation voltages, 10 kV and 5 kV, were used to determine the elemental information. Information obtained from SEM-EDX can distinguish individual single cells and detect viable and nonviable microorganisms. This work demonstrates that the combination of morphological and elemental information obtained from SEM-EDX analysis with the help of principal component analysis (PCA) enables the rapid identification of single microbial cells without following time-consuming microbiological cultivation methods. Rapidly detecting and identifying biological threat microorganisms without traditional culture or chemical-based methods are highly important. The widely used identification techniques are nucleic acid-based, biosensor-based and immunologically based techniques. Real-time PCR multiplex PCR, loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification (NASBA), and oligonucleotide DNA microarray are examples of some common nucleic acid-based identification techniques 1-4. These techniques have higher sensitivity, specificity, and reliability and can detect multiple pathogens in an automated manner with several constraints, such as sensitivity to PCR inhibitors and complicated primer design, and the methods cannot differentiate viable and nonviable cells 5,6. All of these nucleic acid-based techniques are slow processes that require 4-72 h to detect microbes 1-6. Electrochemical, optical and mass-based biosensors are commonly used to detect microbes. These automated, label-free, real-time detection processes can handle a large number of samples. Biosensor-based processes have several drawbacks, such as long incubation time, numerous washing steps, low specificity, interference with the food matrix and unsuitability for lesser cells 7-10. The lateral flow immunoassay and enzyme-linked immunosorbent assay (ELISA) are two immunological-based detection techniques with several advantages and disadvantages; most importantly, these techniques are also slow processes that require 3-10 h 11,12. These limitations increase the overall cost of the detection process due to costly logistics trails and restrict autonomous operation 13,14. ...

A submerged dielectric barrier discharge plasma reactor (underwater DBD) has been used on Escherichia coli O157:H7 (ATCC 35150). Plasma treatment was carried out using clean dry air gas to investigate the individual effects of the radicals produced by underwater DBD on an E. coli O157:H7 suspension (8.0 log CFU/ml). E. coli O157:H7 was reduced by 6.0 log CFU/ml for 2 min of underwater DBD plasma treatment. Optical Emission Spectra (OES) shows that OH and NO (α, β) radicals, generated by underwater DBD along with ozone gas. E. coli O157:H7 were reduced by 2.3 log CFU/ml for 10 min of underwater DBD plasma treatment with the terephthalic acid (TA) OH radical scavenger solution, which is significantly lower (3.7 log CFU/ml) than the result obtained without using the OH radical scavenger. A maximum of 1.5 ppm of ozone gas was produced during the discharge of underwater DBD, and the obtained reduction difference in E.coli O157:H7 in presence and in absence of ozone gas was 1.68 log CFU/ml. The remainder of the 0.62 log CFU/ml reduction might be due to the effect of the NO (α, β) radicals or due to the combined effect of all the radicals produced by underwater DBD. A small amount of hydrogen peroxide was also generated but does not play any role in E. coli O157:H7 inactivation.

The degradation of two organophosphates, chlorpyrifos and diazinon, in water using microplasma equipment to produce ozone and the identification of their products were studied by using liquid chromatography–mass spectrometry. The organophosphates gradually decreased with time and were completely removed after 10 min, and diazinon was degraded at a relatively fast rate compared to chlorpyrifos. The products formed during the process were identified and determined with accurate mass measurements and tandem mass spectrometry spectra, providing reliable structural determination. Chlorpyrifos oxon was formed through the oxidation of chlorpyrifos, followed by the formation of 3,5,6‐trichloro‐2‐pyridinol and diethyl phosphate by hydrolysis. Diazinon formed various products through more complicated degradation processes than those of chlorpyrifos. The major products of diazinon degradation were 2‐isopropyl‐6‐methyl‐4‐pyrimidinol and diethyl phosphate by hydrolysis after oxidation, exhibiting diazoxon as an intermediate at trace levels. Direct hydrolysis of diazinon also occurred, producing diethyl thiophosphate, which was observed at a low concentration for a transient time and exhibited a less favorable process than sequential oxidation and hydrolysis. The other products, hydroxy diazinons and hydroxy‐2‐isopropyl‐6‐methyl‐4‐pyrimidinols, formed by hydroxylation, were also identified, but they were present in low amounts. Degradation mechanisms of chlorpyrifos and diazinon were proposed with the quantitatively evaluated products.

A submerged dielectric barrier discharge plasma reactor (underwater DBD) has been used to inactivate biofilm produced by three different food-borne pathogens, namely Escherichia coli O157:H7 (ATCC 438), Cronobacter sakazakii (ATCC 29004), and Staphylococcus aureus (KCCM 40050). The inactivation that were obtained after 90 minutes of plasma operation were found to measure 5.50 log CFU/coupon, 6.88 log CFU/coupon and 4.20 log CFU/coupon for Escherichia coli O157:H7 (ATCC 438), Cronobacter sakazakii (ATCC 29004), and Staphylococcus aureus (KCCM 40050), respectively. Secondary Electron Images (SEI) obtained from Field Emission Scanning Electron Microscopy (FE-SEM) show the biofilm morphology and its removal trend by plasma operation at different time intervals. An attenuated total reflectance Fourier transform infrared (ATR-FTIR) measurement was performed to elucidate the biochemical changes that occur on the bacterial cell and extracellular polymeric substance (EPS) of biofilm during the plasma inactivation process. The ATR-FTIR measurement shows the gradual reduction of carbohydrates, proteins, and lipid and DNA peak regions with increased plasma exposure time. The presence of an EPS layer on the upper surface of the biofilm plays a negative and significant role in its removal from stainless steel (SS) coupons.

This study was performed to assess the effect of plasma-discharged water recycling technology as irrigation water on soybean sprout production. Two different types of irrigation water were used individually for cultivation, including plasma discharged water as a source of oxides of nitrogen and tap water, irrigation water was recycled for every 30 minutes. Plasma discharged irrigation water reduced overall 4.3 log CFU/ml aerobic microbe and 7.0 log CFU/ml of artificially inoculated S. Typhimurium within 5 minutes and 2 minutes, respectively, therefore sprout production occurs in a hygienic environment. Using of plasma-discharged water for cultivation, increases the amount of ascorbate, asparagine, and γ-aminobutyric acid (GABA) significantly (p < 0.05), in the part of cotyledon and hypocotyl of soybean sprout during 1 to 4 days of farming. A NO scavenger, 2-(4-carboxy-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxy-3-oxide (cPTIO), was added in irrigation water to elucidate the roles of the oxides of nitrogen such as NO3−, NO2− generated in plasma discharged water. It was observed that all three nutrients decreased in the cotyledon part, whereas ascorbate and GABA contents increased in the hypocotyl and radicle part of bean sprout for the same duration of farming. The addition of NO scavenger in the irrigation water also reduced growth and overall yield of the soybean sprouts. A recycling water system with plasma-discharged water helped to reduce the amount of water consumption and allowed soybean sprouts growth in a hygienic environment during the hydroponic production.