Sung-Wook Choi

Herein, we investigated the potential of plasma-activated water (PAW) as a wash solution for the microbial decontamination of cherry tomatoes. We analyzed the efficacy of PAW as a bactericidal agent based on reactive species and pH. Immersion for 5 min in PAW15 (generated via plasma activation for 15 min) was determined as optimal for microbial decontamination of fresh produce. The decontamination efficacy of PAW15 exceeded those of mimic solutions with equivalent reactive species concentrations and pH (3.0 vs. 1.7 log reduction), suggesting that the entire range of plasma-derived reactive species participates in decontamination rather than a few reactive species. PAW15-washing treatment achieved reductions of 6.89 ± 0.36, 7.49 ± 0.40, and 5.60 ± 0.05 log₁₀ CFU/g in the counts of Bacillus cereus, Salmonella sp., and Escherichia coli O157:H7, respectively, inoculated on the surface of cherry tomatoes, with none of these strains detected in the wash solution. During 6 days of 25 °C storage post-washing, the counts of aerobic bacteria, yeasts, and molds were below the detection limit. However, PAW15 did not significantly affect the viability of RAW264.7 cells. These results demonstrate that PAW effectively inactivates microbes and foodborne pathogens on the surface of cherry tomatoes and in the wash solution. Thus, PAW could be used as an alternative wash solution in the fresh produce industry without cross-contamination during washing and environmental contamination by foodborne pathogens or potential risks to human health.

Chitosan (CTS)/gelatin (GEL)/poly(vinyl alcohol) (PVA)-based composite films with different concentrations of Duchesnea indica extract (DIE) (6.25 and 25 mg/mL), an antimicrobial agent, were manufactured using a casting technique. Results indicated that elongation at break decreased as DIE was added at higher concentrations. Composite films showed no significant differences in thickness, tensile strength, and water vapor permeability. Scanning electron microscopy images revealed that DIE was successfully incorporated into film matrices to interact with polymers. The addition of DIE to the film inhibited the growth of S. aureus by up to 4.9 log CFU/mL. The inhibitory effect on S. aureus using DIE-incorporated coating applied to strawberries was greatest at room temperature storage for 24 h only when it was coated twice or more. The maximum inhibition in strawberries was 2.5 log CFU/g when they were coated twice and 3.2 log CFU/g when they were coated three times. The results of this study suggest that DIE could be used as a natural antimicrobial agent, and DIE-integrated CTS/GEL/PVA films or coatings have potential as a food packaging alternative for preventing foodborne pathogen contamination.

Aptamer-based methods for detecting pesticides are more efficient than antibody-based methods by high thermal stability, low molecular weight, easy modification, and low cost. In this study, the systematic evolution of ligands by exponential enrichment (SELEX) process, combined with next-generation sequencing (NGS), was performed to select aptamers specific to the pesticide, diazinon, which was fixed on a sol–gel-coated nanoporous-anodized aluminum oxide membrane to overcome the immobilization effect of general method and simplify the elution step. The frequency of specific nucleotide sequences obtained after SELEX rounds was directly analyzed using NGS to eliminate the time-consuming cloning process used in the general SELEX methods. Nine sequences with the highest frequency after SELEX round 10 followed by NGS were selected and tested to derive their binding affinity with the target, diazinon, through circular dichroism (CD) spectrophotometry. The CD signal difference of the aptamer candidates ranged from 0.13 to 2.242 mdeg between diazinon-only treated and diazinon-aptamer-treated samples at a wavelength near 270 nm. Aptamer D-4, which had the highest binding affinity from CD spectrophotometry analysis, showed no cross-reactivity with non-target pesticides, such as baycarb, bifenthrin, and pyridaben, but interacted with the other pesticides, fipronil and 2-phenylphenol. Therefore, an aptamer was effectively screened by selection of high-frequency candidates after SELEX-NGS followed by CD analysis with the highest difference signal. A follow-up study is needed to confirm whether the proposed SELEX process combined with NGS for the discovery of aptamers for new targets can further shorten the SELEX cycle by reducing the number of SELEX rounds to 10 or less.

Sol-gel-based mesopores allow the entry of target small molecules retained in their cavity and aptamers to bind to target molecules. Herein, sol-gel-based materials are applied to screen-selective aptamers for small molecules, such as pesticides. To enhance the efficiency of aptamer screening using a sol-gel, it is necessary to increase the binding surface. In this study, we applied the sol-gel to an anodized aluminum oxide (AAO) membrane, and the morphological features were observed via electron microscopy after spin coating. The binding and elution processes were conducted and confirmed by fluorescence microscopy and polymerase chain reaction. The sol-gel coating on the AAO membrane formed a hollow nanocolumn structure. A diazinon-binding aptamer was bound to the diazinon-containing sol-gel-coated AAO membrane, and the bound aptamer was effectively retrieved from the sol-gel matrix by thermal elution. As a proof of concept, a sol-gel-coated AAO disc was mounted on the edge of a pipette tip, and the feasibility of the prepared platform for the systematic evolution of ligands by exponential enrichment (SELEX) of the aptamer binding was also confirmed. The proposed approach will be applied to an automated SELEX cycle using an automated dispenser, such as a pipetting robot, in the near future.

In this study, we describe the development of an automated immunomagnetic separation device combined with real-time polymerase chain reaction (PCR) for detecting foodborne bacteria. Immunomagnetic separation (IMS) is a well-known method for the separation and concentration of target bacteria from a large volume of food samples. Magnetic beads functionalized with an antibody provide selectivity for target bacteria such as Escherichia coli O157:H7. Moreover, compared to conventional methods, real-time PCR enables high-sensitivity detection of target bacteria. The method proposed in this study involves three steps: (1) pre-enrichment, (2) automated IMS and concentration of target bacteria, and (3) detection of target bacteria by real-time PCR. Using food samples with a working sample volume as large as 250 mL, the whole process only requires 3 h. As a result, target bacteria in the range of 10¹–10² colony-forming units per mg or g of sample can be detected in food samples, such as milk, ground beef, and cabbage, by using the proposed approach. We anticipate that the automated IMS system combined with real-time PCR will contribute to the development of a fully automated system for detecting foodborne bacteria and serve as a multi-tester for a variety of bacterial strains in the capacity of a sample-to-answer device in the near future.

The food industry requires rapid and simple detection methods for preventing harm from pathogenic bacteria. Until now, various technologies used to detect foodborne bacteria were time-consuming and laborious. Therefore, we have developed an automated immunomagnetic separation combined with a colorimetric assay for the rapid detection of E. coli O157:H7 in food samples. The colorimetric detection method using enzymatic reaction is fascinating because of its simplicity and rapidity and does not need sophisticated devices. Moreover, the proposed procedures for the detection of bacteria in food take less than 3 h including pre-enrichment, separation and detection steps. First, target-specific immunomagnetic beads were introduced to contaminated milk in a pre-enrichment step. Second, the pre-enriched sample solution containing target bacteria bound on immunomagnetic beads was injected into an automated pretreatment system. Subsequently, the immunomagnetic beads along with target bacteria were separated and concentrated into a recovery tube. Finally, released β-galactosidase from E. coli O157:H7 after lysis was reacted with chlorophenol red β-galactopyranoside (CPRG) used as a substrate and the colorimetric change of CPRG was determined by absorbance measuring or the naked eye. By the proposed approach in this study, we could detect 3 × 10² CFU/mL of E. coli O157:H7 from a milk sample within 3 h.

We experimentally modulate the refractive index and the absorption coefficient of an SU-8 dry film in the terahertz region by UV light (362 nm) exposure with time dependency. Consequently, the refractive index of SU-8 film is increased by approximately 6% after UV light exposure. Moreover, the absorption coefficient also changes significantly. Using the reflective terahertz imaging technique, in addition, we can read security information printed by UV treatment on an SU-8 film that is transparent in the visible spectrum. From these results, we successfully demonstrate security printing and reading by using photoresist materials and the terahertz technique. This investigation would provide a new insight into anti-counterfeiting applications in fields that need security.

Fabrication of 1D gold nanoparticle (NP) arrays by crack templated reduction lithography (CTRL) is demonstrated. The gold NPs are synthesized in situ on the cracks in a polydimethoxysiloxane (PDMS) substrate without additional reducing/stabilizing agents. Upon directionally bending the substrate, various array patterns formed on the PDMS rubber.