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      Leaf Surface Topography Contributes to the Ability of Escherichia coli on Leafy Greens to Resist Removal by Washing, Escape Disinfection With Chlorine, and Disperse Through Splash


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          The attachment of foodborne pathogens to leaf surfaces is a complex process that involves multiple physical, chemical, and biological factors. Here, we report the results from a study designed to specifically determine the contribution of spinach leaf surface topography as it relates to leaf axis (abaxial and adaxial) and leaf age (15, 45, and 75 days old) to the ability of Escherichia coli to resist removal by surface wash, to avoid inactivation by chlorine, and to disperse through splash impact. We used fresh spinach leaves, as well as so-called “replicasts” of spinach leaf surfaces in the elastomer polydimethylsiloxane to show that leaf vein density correlated positively with the failure to recover E. coli from surfaces, not only using a simple water wash and rinse, but also a more stringent wash protocol involving a detergent. Such failure was more pronounced when E. coli was surface-incubated at 24°C compared to 4°C, and in the presence, rather than absence, of nutrients. Leaf venation also contributed to the ability of E. coli to survive a 50 ppm available chlorine wash and to laterally disperse by splash impact. Our findings suggest that the topographical properties of the leafy green surface, which vary by leaf age and axis, may need to be taken into consideration when developing prevention or intervention strategies to enhance the microbial safety of leafy greens.

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          Most cited references78

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          Use of the green fluorescent protein (Gfp) from the jellyfish Aequorea victoria is a powerful method for nondestructive in situ monitoring, since expression of green fluorescence does not require any substrate addition. To expand the use of Gfp as a reporter protein, new variants have been constructed by the addition of short peptide sequences to the C-terminal end of intact Gfp. This rendered the Gfp susceptible to the action of indigenous housekeeping proteases, resulting in protein variants with half-lives ranging from 40 min to a few hours when synthesized in Escherichia coli and Pseudomonas putida. The new Gfp variants should be useful for in situ studies of temporal gene expression.
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              Fitness of human enteric pathogens on plants and implications for food safety.

              The continuous rise in the number of outbreaks of foodborne illness linked to fresh fruit and vegetables challenges the notion that enteric pathogens are defined mostly by their ability to colonize the intestinal habitat. This review describes the epidemiology of produce-associated outbreaks of foodborne disease and presents recently acquired knowledge about the behavior of enteric pathogens on plants, with an emphasis on Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. The growth and survival of enteric pathogens on plants are discussed in the light of knowledge and concepts in plant microbial ecology, including epiphytic fitness, the physicochemical nature of plant surfaces, biofilm formation, and microbe-microbe and plant-microbe interactions. Information regarding the various stresses that affect the survival of enteric pathogens and the molecular events that underlie their interactions in the plant environment provides a good foundation for assessing their role in the infectious dose of the pathogens when contaminated fresh produce is the vehicle of illness.

                Author and article information

                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                17 July 2020
                : 11
                : 1485
                [1] 1Department of Plant Pathology, University of California, Davis , Davis, CA, United States
                [2] 2Departamento de Microbiología, Instituto de Hortofruticultura Subtropical y Mediterránea “La Mayora”, Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Universidad de Málaga , Málaga, Spain
                [3] 3Department of Biomedical Engineering, University of California, Davis , Davis, CA, United States
                [4] 4Department of Materials Science and Engineering, University of California, Davis , Davis, CA, United States
                Author notes

                Edited by: Wei Zhang, Illinois Institute of Technology, United States

                Reviewed by: Hao Feng, University of Illinois at Urbana-Champaign, United States; Maria T. Brandl, Agricultural Research Service (USDA), United States; Maria Isabel Gil, Consejo Superior de Investigaciones Científicas (CSIC), Spain; Jitendra Patel, Beltsville Agricultural Research Center (USDA-ARS), United States

                *Correspondence: Johan H. J. Leveau, jleveau@ 123456ucdavis.edu

                This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology

                Copyright © 2020 Doan, Antequera-Gómez, Parikh and Leveau.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                : 11 January 2020
                : 08 June 2020
                Page count
                Figures: 8, Tables: 0, Equations: 0, References: 87, Pages: 14, Words: 0
                Funded by: National Institute of Food and Agriculture 10.13039/100005825
                Original Research

                Microbiology & Virology
                phyllosphere,phyllotelma,pdms,topomimetic,leaf replicasting,food safety
                Microbiology & Virology
                phyllosphere, phyllotelma, pdms, topomimetic, leaf replicasting, food safety


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