Engineered water nanostructures (EWNS) synthesized utilizing electrospray and ionization of water, have been, recently, shown to be an effective, green, antimicrobial platform for surface and air disinfection, where reactive oxygen species (ROS), generated and encapsulated within the particles during synthesis, were found to be the main inactivation mechanism. Herein, the antimicrobial potency of the EWNS was further enhanced by integrating electrolysis, electrospray and ionization of de-ionized water in the EWNS synthesis process. Detailed physicochemical characterization of these enhanced EWNS (eEWNS) was performed using state-of-the-art analytical methods and has shown that, while both size and charge remain similar to the EWNS (mean diameter of 13 nm and charge of 13 electrons), they possess a three times higher ROS content. The increase of the ROS content as a result of the addition of the electrolysis step before electrospray and ionization led to an increased antimicrobial ability as verified by E. coli inactivation studies using stainless steel coupons. It was shown that a 45-min exposure to eEWNS resulted in a 4-log reduction as opposed to a 1.9-log reduction when exposed to EWNS. In addition, the eEWNS were assessed for their potency to inactivate natural microbiota (total viable and yeast and mold counts), as well as, inoculated E. coli on the surface of fresh organic blackberries. The results showed a 97% (1.5-log) inactivation of the total viable count, a 99% (2-log) reduction in the yeast and mold count and a 2.5-log reduction of the inoculated E. coli after 45 min of exposure, without any visual changes to the fruit. This enhanced antimicrobial activity further underpins the EWNS platform as an effective, dry and chemical free approach suitable for a variety of food safety applications and could be ideal for delicate fresh produce that cannot withstand the classical, wet disinfection treatments.

The disinfection efficacy of slightly acidic electrolyzed water (SAEW) has been recognized in food industry. However, the application of single SAEW limited its disinfection potential. The efficacy of the two-step disinfection with SAEW for the reduction of L. monocytogenes contamination on different food raw materials was evaluated compared to the one-step disinfection with SAEW in this study. Results demonstrated that SAEW could reduce both the natural aerobic bacterial count and inoculated L. monocytogenes population on endive and chicken immediately after processing and SAEW with approximately 60 mg/L of available chlorine concentration (ACC) had equal or higher antibacterial efficacy compared to NaClO solutions with approximately 150 mg/L of ACC. Moreover, SAEW treatments could control the growth of microbial populations of L. monocytogenes during storage and the efficacy on the microbial reduction was associated to the initial populations. In addition, the SAEW-treated food raw materials were stored and disinfected again after storage and the results showed that the two-step disinfection method could decrease the survival populations of L. monocytogenes by 24.8%–99.6% compared to the one-step disinfection of SAEW. Therefore, considering storage habits, the two-step disinfection of SAEW may be a better choice in the disinfection of non-consumed-directly food raw materials.

Treatment of fresh fruits and vegetables with electrolyzed water (EW) has been shown to kill or reduce foodborne pathogens. We evaluated the efficacy of EW in killing Escherichia coli O157:H7 on iceberg lettuce, cabbage, lemons, and tomatoes by using washing and/or chilling treatments simulating those followed in some food service kitchens. Greatest reduction levels on lettuce were achieved by sequentially washing with 14-A (amperage) acidic EW (AcEW) for 15 or 30 s followed by chilling in 16-A AcEW for 15 min. This procedure reduced the pathogen by 2.8 and 3.0 log CFU per leaf, respectively, whereas washing and chilling with tap water reduced the pathogen by 1.9 and 2.4 log CFU per leaf. Washing cabbage leaves for 15 or 30 s with tap water or 14-A AcEW reduced the pathogen by 2.0 and 3.0 log CFU per leaf and 2.5 to 3.0 log CFU per leaf, respectively. The pathogen was reduced by 4.7 log CFU per lemon by washing with 14-A AcEW and 4.1 and 4.5 log CFU per lemon by washing with tap water for 15 or 30 s. A reduction of 5.3 log CFU per lemon was achieved by washing with 14-A alkaline EW for 15 s prior to washing with 14-A AcEW for 15 s. Washing tomatoes with tap water or 14-A AcEW for 15 s reduced the pathogen by 6.4 and 7.9 log CFU per tomato, respectively. Application of AcEW using procedures mimicking food service operations should help minimize cross-contamination and reduce the risk of E. coli O157:H7 being present on produce at the time of consumption.

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0.95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.

The antimicrobial effect of water extracts of sumac (Rhus coriaria L.) at concentrations of 0.1%, 0.5%, 1.0%, 2.5% and 5.0% (w/v), non-neutralized and after neutralization to pH 7.2 +/- 0.1, was studied on the growth of 12 bacterial strains (six Gram positive strains and six Gram negative strains), mostly food borne including pathogens. It was found to be effective against all the test organisms with Gram positive strains being more sensitive than Gram negative strains. Significant differences (P<0.01) were found among the bacteria and between the non-neutralized and neutralized extracts with non-neutralized being more effective against all the bacteria. The minimal inhibitory concentration (MIC) of the extract for each bacterial strain was studied by a gradient plate method. Among the Gram positive organisms, Bacillus species (Bacillus cereus, Bacillus megaterium, Bacillus subtilis, and Bacillus thuringiensis) were found to be the most sensitive showing MICs of 0.25-0.32% (after 24 h incubation) followed by Staphylococcus aureus (0.49%), while Listeria monocytogenes was found to be the least sensitive demonstrating a MIC of 0.67%. Of the Gram negative organisms, Salmonella enteritidis was found to be the most resistant with a MIC of 0.67% followed by Escherichia coli Type I, E. coli O157:H7, Proteus vulgaris and Hafnia alvei having MICs of 0.63%, 0.60%, 0.55% and 0.45%, respectively; whereas Citrobacter freundii was found to be the least resistant surviving up to 0.42%. Some loss of antimicrobial activity was, however, observed after incubation for 3 days. Bacteriostatic/bactericidal effects of sumac, as studied by enumerating survival by the viable count technique after 1 h direct contact of each microorganism with various concentrations of sumac extract, revealed a 4-5 log cycle reduction in Bacillus spp. and 2-3 log cycle reduction in other bacteria tested with 1.0% sumac extract.