This work is concerned with investigating the effect of substrate hydrophobicity and zeta potential on the dynamics and kinetics of the initial stages of bacterial adhesion. For this purpose, bacterial pathogens Staphylococcus aureus and Escherichia coli O157:H7 were inoculated on the substrates coated with thin thiol layers (i.e., 1-octanethiol, 1-decanethiol, 1-octadecanethiol, 16-mercaptohexadecanoic acid, and 2-aminoethanethiol hydrochloride) with varying hydrophobicity and surface potential. The time-resolved adhesion data revealed a transformation from an exponential dependence to a square root dependence on time upon changing the substrate from hydrophobic or hydrophilic with a negative zeta potential value to hydrophilic with a negative zeta potential for both pathogens. The dewetting of extracellular polymeric substances (EPS) produced by E. coli O157:H7 was more noticeable on hydrophobic substrates, compared to that of S. aureus, which is attributed to the more amphiphilic nature of staphylococcal EPS. The interplay between the timescale of EPS dewetting and the inverse of the adhesion rate constant modulated the distribution of E. coli O157:H7 within microcolonies and the resultant microcolonial morphology on hydrophobic substrates. Observed trends in the formation of bacterial monolayers rather than multilayers and microcolonies rather than isolated and evenly spaced bacterial cells could be explained by a colloidal model considering van der Waals and electrostatic double-layer interactions only after introducing the contribution of elastic energy due to adhesion-induced deformations at intercellular and substrate-cell interfaces. The gained knowledge is significant in the context of identifying surfaces with greater risk of bacterial contamination and guiding the development of novel surfaces and coatings with superior bacterial antifouling characteristics.

24 This study demonstrates the rose essential oil component (EOC) geraniol can be loaded into 25 polymeric nanoparticles (NPs) with sustainable release profile. Geraniol-loaded NPs were 26 prepared by flash nanoprecipitation and characterized for size, encapsulation efficiency, payload 27 release during storage, inhibition of Escherichia coli O157:H7 and Salmonella enterica 28 Typhimurium in vitro and on spinach surfaces, and NP-assisted transport of EOC into cellular 29 membranes. Adjusting concentrations of stabilizing polymer, Pluronic® F-127, and geraniol 30 produced NPs ranging in size from 26 to 412 nm. Antimicrobial NPs inhibited S. Typhimurium 31 and E. coli O157:H7 growth at 0.25 and 0.2 wt.%, respectively. Geraniol-loaded NPs displayed 32 sustained release with a time constant of 24 hr, maintaining their anti-pathogenic properties over 33 a prolonged time period. Pathogen reductions on treated spinach surfaces ranged from 0.3 to 4.2 34 log 10 CFU/cm 2 . Antimicrobial NPs may be useful for post-harvest decontamination of foods such 35 as fresh produce from cross-contaminating microbial pathogens. 36 37

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This study aimed to quantify survival in Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium isolates on melon rind surface samples achieved by sanitizer treatment under three differing melon contamination and sanitization scenarios. Sanitizing treatments consisted of the plant-derived antimicrobial (PDA) essential oil component (EOC) geraniol (0.5 wt.%) entrapped in the polymeric surfactant Pluronic F-127 (GNP), 0.5 wt.% unencapsulated geraniol (UG), 200 mg/L hypochlorous acid at pH 7.0 (HOCl), and a sterile distilled water wash (CON). The experimental contamination and sanitization scenarios tested were: (1) pathogen inoculation preceded by treatment; (2) the pathogen was inoculated onto samples twice with sanitizing treatment applied in between inoculation events; or (3) pathogen inoculation followed by sanitizing treatment. Reductions in the numbers of surviving pathogens were dependent on the sanitizing treatment, the storage period, or the interaction of these effects. GNP treatment provided the greatest reductions in surviving pathogen counts on melon rinds, but these did not regularly statistically differ from those achieved by HOCl or UG treatment. GNP treatment provided the best pathogen control under differing conditions of pre- and/or post-harvest cross-contamination and can be applied to reduce the risk of pathogen transmission on melon rinds.

Protecting fresh-packed produce microbiological safety against pre- and post-harvest microbial pathogen contamination requires innovative antimicrobial strategies. Although largely ignored in the scientific literature, there exists the potential for gross failure in food safety protection of fresh fruits and vegetables leading to opportunity for multiple produce contamination events to occur during production and post-harvest handling of food crops. The primary objective of this research was to determine the efficacy of plant-derived antimicrobial-loaded nanoparticles to reduce Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium on spinach leaf surfaces whilst simulating multiple pathogen contamination events (pre-harvest and post-harvest). Spinach samples were inoculated with a blend of E. coli O157:H7 and S. Typhimurium, each diluted to ~8.0 log10 CFU/mL. The inoculated samples were then submerged in solutions containing nanoparticles loaded with geraniol (GPN; 0.5 wt.% geraniol), unencapsulated geraniol (UG; 0.5 wt.%), or 200 ppm chlorine (HOCl; pH 7.0), with untreated samples serving for controls. Following antimicrobial treatment application, samples were collected for surviving pathogen enumeration or were placed under refrigeration (5°C) for up to 10 days, with periodic enumeration of pathogen loads. After 3 days of refrigerated storage, all samples were removed, aseptically opened and subjected to a second inoculation with both pathogens. Treatment of spinach surfaces with encapsulated geraniol reduced both pathogens to non-detectable numbers within 7 days of refrigerated storage, even with a second contamination event occurring 3 days after experiment initiation. Similar results were observed with the UG treatment, except that upon recontamination at day 3, a higher pathogen load was detected on UG-treated spinach vs. GPN-treated spinach. These data fill a research gap by providing a novel tool to reduce enteric bacterial pathogens on spinach surfaces despite multiple contamination events, a potential food safety risk for minimally processed edible produce.