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      Biofilm formation by Listeria monocytogenes on stainless steel surface and biotransfer potential

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          Abstract

          An experimental model was proposed to study biofilm formation by Listeria monocytogenes ATCC 19117 on AISI 304 (#4) stainless steel surface and biotransfer potential during this process. In this model, biofilm formation was conducted on the surface of stainless steel coupons, set on a stainless steel base with 4 divisions, each one supporting 21 coupons. Trypic Soy Broth was used as bacterial growth substrate, with incubation at 37 ºC and stirring of 50 rpm. The number of adhered cells was determined after 3, 48, 96, 144, 192 and 240 hours of biofilm formation and biotransfer potential from 96 hours. Stainless steel coupons were submitted to Scanning Electron Microscopy (SEM) after 3, 144 and 240 hours. Based on the number of adhered cells and SEM, it was observed that L. monocytogenes adhered rapidly to the stainless steel surface, with mature biofilm being formed after 240 hours. The biotransfer potential of bacterium to substrate occurred at all the stages analyzed. The rapid capacity of adhesion to surface, combined with biotransfer potential throughout the biofilm formation stages, make L. monocytogenes a potential risk to the food industry. Both the experimental model developed and the methodology used were efficient in the study of biofilm formation by L. monocytogenes on stainless steel surface and biotransfer potential.

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          Experimental reproducibility in flow-chamber biofilms.

          The structural organization of microbial communities is influenced by many factors, e.g. nutrient composition, shear stress and temperature. This paper presents a general method for quantitative comparison of biofilm structures and assessment of experimental reproducibility between independent biofilm experiments. By using a novel computer program, COMSTAT, biofilm structures of Pseudomonas aeruginosa and an isogenic rpoS mutant were quantified. The strains were tagged with the green fluorescent protein (GFP) and grown in flow chambers with a defined minimal medium as substrate. Three independent rounds of biofilm experiments were performed and in each round, each of the two variants was grown in two separate channels. Nine image stacks were acquired in each channel 146 h after inoculation. An analysis of variance model incorporating the factors experiment round, bacterial strain, channel number and image stack number was used to analyse the data calculated by COMSTAT. Experimental reproducibility was verified by estimating the magnitude of the variance of the effects round (sigma(2)R) and the interaction between bacterial strain and round (sigma(2)BR). Mean thickness of the wild-type and rpoS mutant biofilms was estimated at 6.31 microm (SE 0.81 microm) and 16.85 microm (SE 0.87 microm), respectively.
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            Structure and functional characteristics of bacterial biofilms in fluid processing operations.

            Bacterial biofilms create a number of serious problems for industrial fluid processing operations. Mechanical blockages, impedance of heat transfer processes, and biodeterioration of the components of metallic and polymeric systems result in billions of dollars in losses each year. Product spoilage and possible risks to public health are also consequences of biofilm-mediated contamination. Fundamentally, these biofouling activities can be described in terms of the physicochemical properties that are associated with bacterial metabolism and biofilm development. Treatment of biofouling is also complicated by the unique structural attributes of biofilms: extracellular polymeric substances create diffusional barriers to antimicrobial agents, protecting labile cellular targets from both oxidizing and nonoxidizing compounds. The mechanisms associated with the initial events of bacterial adhesion to engineered surfaces and subsequent fouling of biofilm formation are poorly understood. However, studies of bacterial biofilm architecture have been greatly facilitated by the application of confocal laser microscopy, scanning or transmission electron microscopy, and Fourier transform infrared spectroscopy. This paper reviews the genesis of biofilm formation and describes the influence of structure on biofouling activities in industrial fluid handling systems.
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              Measurement of polysaccharides and proteins in biofilm extracellular polymers

              The structure of biofilm extracellular polymers (ECPs) was studied by measuring their polysaccharide and protein spatial distributions along biofilm depth. Biofilm was collected from two aerobic heterotrophic biofilm reactors, which were seeded with Sphingomonas sp. and Sphingomonas sp. plus mixed liquor, respectively, and operated under toxic organic (in this case, azo dye) degrading conditions. Complete mixing conditions in the two reactors were verified by measuring water content, and polysaccharide and protein quantities from three vertical sampling positions over time. Experimental results showed that: (1) the biofilm water content of either reactor did not change with sample position or biofilm age, with an average biofilm water content in both reactors of 97%; (2) polysaccharides and proteins in the ECPs did not change with sample position; (3) the profiles of polysaccharides and proteins along the biofilm depth showed a stratified biofilm structure, with their ratio (proteins/polysaccharides) being relatively stable over the depth. Oxygen and substrate transport and interactions among species were considered to be the main reasons for producing such a non-uniform biofilm structure; and (4) Sphingomonas sp. could not compete well with microorganisms derived from the mixed liquor of a wastewater treatment plant aeration basin.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                bjm
                Brazilian Journal of Microbiology
                Braz. J. Microbiol.
                Sociedade Brasileira de Microbiologia (São Paulo )
                1678-4405
                March 2010
                : 41
                : 1
                : 97-106
                Affiliations
                [1 ] Universidade Federal de Lavras Brazil
                [2 ] Universidade Federal de Lavras Brazil
                Article
                S1517-83822010000100016
                10.1590/S1517-83822010000100016
                1b0d9dda-7b6c-4b59-bcfd-84892caca7ed

                http://creativecommons.org/licenses/by/4.0/

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                Product

                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1517-8382&lng=en
                Categories
                MICROBIOLOGY

                Microbiology & Virology
                Listeria monocytogenes,biofilm,biotransfer potential
                Microbiology & Virology
                Listeria monocytogenes, biofilm, biotransfer potential

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