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      Impact of chlorine dioxide disinfection of irrigation water on the epiphytic bacterial community of baby spinach and underlying soil

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          Abstract

          The contamination of pathogenic bacteria through irrigation water is a recognized risk factor for fresh produce. Irrigation water disinfection is an intervention strategy that could be applied to reduce the probability of microbiological contamination of crops. Disinfection treatments should be applied ensuring minimum effective doses, which are efficient in inhibiting the microbial contamination while avoiding formation and accumulation of chemical residues. Among disinfection technologies available for growers, chlorine dioxide (ClO 2) represents, after sodium hypochlorite, an alternative disinfection treatment, which is commercially applied by growers in the USA and Spain. However, in most of the cases, the suitability of this treatment has been tested against pathogenic bacteria and low attention have been given to the impact of chemical residues on the bacterial community of the vegetable tissue. The aim of this study was to (i) to evaluate the continual application of chlorine dioxide (ClO 2) as a water disinfection treatment of irrigation water during baby spinach growth in commercial production open fields, and (ii) to determine the subsequent impact of these treatments on the bacterial communities in water, soil, and baby spinach. To gain insight into the changes in the bacterial community elicited by ClO 2, samples of treated and untreated irrigation water as well as the irrigated soil and baby spinach were analyzed using Miseq® Illumina sequencing platform. Next generation sequencing and multivariate statistical analysis revealed that ClO 2 treatment of irrigation water did not affect the diversity of the bacterial community of water, soil and crop, but significant differences were observed in the relative abundance of specific bacterial genera. This demonstrates the different susceptibility of the bacteria genera to the ClO 2 treatment. Based on the obtained results it can be concluded that the phyllosphere bacterial community of baby spinach was more influenced by the soil bacteria community rather than that of irrigation water. In the case of baby spinach, the use of low residual ClO 2 concentrations (approx. 0.25 mg/L) to treat irrigation water decreased the relative abundance of Pseudomonaceae (2.28-fold) and Enterobacteriaceae (2.5-fold) when comparing treated versus untreated baby spinach. Members of these two bacterial families are responsible for food spoilage and foodborne illnesses. Therefore, a reduction of these bacterial families might be beneficial for the crop and for food safety. In general it can be concluded that the constant application of ClO 2 as a disinfection treatment for irrigation water only caused changes in two bacterial families of the baby spinach and soil microbiota, without affecting the major phyla and classes. The significance of these changes in the bacterial community should be further evaluated.

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          Microbial contributions to climate change through carbon cycle feedbacks.

          There is considerable interest in understanding the biological mechanisms that regulate carbon exchanges between the land and atmosphere, and how these exchanges respond to climate change. An understanding of soil microbial ecology is central to our ability to assess terrestrial carbon cycle-climate feedbacks, but the complexity of the soil microbial community and the many ways that it can be affected by climate and other global changes hampers our ability to draw firm conclusions on this topic. In this paper, we argue that to understand the potential negative and positive contributions of soil microbes to land-atmosphere carbon exchange and global warming requires explicit consideration of both direct and indirect impacts of climate change on microorganisms. Moreover, we argue that this requires consideration of complex interactions and feedbacks that occur between microbes, plants and their physical environment in the context of climate change, and the influence of other global changes which have the capacity to amplify climate-driven effects on soil microbes. Overall, we emphasize the urgent need for greater understanding of how soil microbial ecology contributes to land-atmosphere carbon exchange in the context of climate change, and identify some challenges for the future. In particular, we highlight the need for a multifactor experimental approach to understand how soil microbes and their activities respond to climate change and consequences for carbon cycle feedbacks.
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            Bacterial Communities Associated with the Surfaces of Fresh Fruits and Vegetables

            Fresh fruits and vegetables can harbor large and diverse populations of bacteria. However, most of the work on produce-associated bacteria has focused on a relatively small number of pathogenic bacteria and, as a result, we know far less about the overall diversity and composition of those bacterial communities found on produce and how the structure of these communities varies across produce types. Moreover, we lack a comprehensive view of the potential effects of differing farming practices on the bacterial communities to which consumers are exposed. We addressed these knowledge gaps by assessing bacterial community structure on conventional and organic analogs of eleven store-bought produce types using a culture-independent approach, 16 S rRNA gene pyrosequencing. Our results demonstrated that the fruits and vegetables harbored diverse bacterial communities, and the communities on each produce type were significantly distinct from one another. However, certain produce types (i.e., sprouts, spinach, lettuce, tomatoes, peppers, and strawberries) tended to share more similar communities as they all had high relative abundances of taxa belonging to the family Enterobacteriaceae when compared to the other produce types (i.e., apples, peaches, grapes, and mushrooms) which were dominated by taxa belonging to the Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria phyla. Although potentially driven by factors other than farming practice, we also observed significant differences in community composition between conventional and organic analogs within produce types. These differences were often attributable to distinctions in the relative abundances of Enterobacteriaceae taxa, which were generally less abundant in organically-grown produce. Taken together, our results suggest that humans are exposed to substantially different bacteria depending on the types of fresh produce they consume with differences between conventionally and organically farmed varieties contributing to this variation.
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              Baseline survey of the anatomical microbial ecology of an important food plant: Solanum lycopersicum (tomato)

              Background Research to understand and control microbiological risks associated with the consumption of fresh fruits and vegetables has examined many environments in the farm to fork continuum. An important data gap however, that remains poorly studied is the baseline description of microflora that may be associated with plant anatomy either endemically or in response to environmental pressures. Specific anatomical niches of plants may contribute to persistence of human pathogens in agricultural environments in ways we have yet to describe. Tomatoes have been implicated in outbreaks of Salmonella at least 17 times during the years spanning 1990 to 2010. Our research seeks to provide a baseline description of the tomato microbiome and possibly identify whether or not there is something distinctive about tomatoes or their growing ecology that contributes to persistence of Salmonella in this important food crop. Results DNA was recovered from washes of epiphytic surfaces of tomato anatomical organs; leaves, stems, roots, flowers and fruits of Solanum lycopersicum (BHN602), grown at a site in close proximity to commercial farms previously implicated in tomato-Salmonella outbreaks. DNA was amplified for targeted 16S and 18S rRNA genes and sheared for shotgun metagenomic sequencing. Amplicons and metagenomes were used to describe “native” bacterial microflora for diverse anatomical parts of Virginia-grown tomatoes. Conclusions Distinct groupings of microbial communities were associated with different tomato plant organs and a gradient of compositional similarity could be correlated to the distance of a given plant part from the soil. Unique bacterial phylotypes (at 95% identity) were associated with fruits and flowers of tomato plants. These include Microvirga, Pseudomonas, Sphingomonas, Brachybacterium, Rhizobiales, Paracocccus, Chryseomonas and Microbacterium. The most frequently observed bacterial taxa across aerial plant regions were Pseudomonas and Xanthomonas. Dominant fungal taxa that could be identified to genus with 18S amplicons included Hypocrea, Aureobasidium and Cryptococcus. No definitive presence of Salmonella could be confirmed in any of the plant samples, although 16S sequences suggested that closely related genera were present on leaves, fruits and roots.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: MethodologyRole: Writing – original draft
                Role: ConceptualizationRole: Writing – review & editing
                Role: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                18 July 2018
                2018
                : 13
                : 7
                : e0199291
                Affiliations
                [1 ] Research Group on Quality, Safety and Bioactivity of Plant Foods, CEBAS-CSIC, Campus Universitario de Espinardo, Murcia, Spain
                [2 ] Department of Plant Science, University of California, One Shields Avenue, Mann Laboratory, Davis, CA, United States of America
                Wageningen Universiteit en Researchcentrum, NETHERLANDS
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0002-5622-4332
                Article
                PONE-D-17-42033
                10.1371/journal.pone.0199291
                6051574
                30020939
                548b7038-ab58-4e41-8a9f-02b52a7aeaba
                © 2018 Truchado et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 29 November 2017
                : 5 June 2018
                Page count
                Figures: 3, Tables: 3, Pages: 17
                Funding
                Funded by: MINECO
                Award ID: AGL2016-75878-R
                Award Recipient :
                Funded by: MINECO
                Award ID: AGL2013-48529-R
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100008678, Center for Produce Safety;
                Award ID: 2015-374
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100008678, Center for Produce Safety;
                Award ID: 2017-01
                Award Recipient :
                Funded by: MINECO
                Award ID: IJCI-2014-20932
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100007801, Fundación Séneca;
                Award ID: 19900/GERM/15
                Funded by: funder-id http://dx.doi.org/10.13039/501100006003, Fundación General CSIC;
                Award ID: Proyecto Intramural 201670E056
                Award Recipient :
                Authors are thankful for the financial support from the Center for Produce Safety Grant Agreement (Projects 2015-374 and 2017-01 to A.A.) and the MINECO (Projects AGL2013-48529-R and AGL2016-75878-R to M.I.G.). P. Truchado is holder of a Juan de la Cierva incorporation contract from the MINECO (IJCI-2014-20932). Support provided by the Fundación Séneca (19900/GERM/15) and Proyecto Intramural (201670E056 to M.I.G.) is also appreciated. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Flowering Plants
                Spinach
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Vegetables
                Spinach
                Biology and Life Sciences
                Agriculture
                Agricultural Methods
                Agricultural Irrigation
                Biology and Life Sciences
                Agriculture
                Agricultural Soil Science
                Ecology and Environmental Sciences
                Soil Science
                Agricultural Soil Science
                Biology and Life Sciences
                Agriculture
                Crop Science
                Crops
                Vegetable Crops
                Biology and Life Sciences
                Agriculture
                Crop Science
                Crops
                Medicine and Health Sciences
                Health Care
                Sanitization
                Disinfection
                Medicine and Health Sciences
                Infectious Diseases
                Infectious Disease Control
                Sanitization
                Disinfection
                Medicine and Health Sciences
                Public and Occupational Health
                Preventive Medicine
                Sanitization
                Disinfection
                Biology and Life Sciences
                Organisms
                Bacteria
                Actinobacteria
                Biology and Life Sciences
                Organisms
                Bacteria
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                All relevant data are within the paper and its Supporting Information files.

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