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Influence of the design of fresh-cut food washing tanks on the growth kinetics of Pseudomonas fluorescens biofilms.

ABSTRACT: Mitigation of cross-contamination of fresh-cut food products at the washing step was studied by investigating how the vat design would affect the biofilm contamination surfaces. Hygienic design features such as no horizontal surfaces and only open angles exceeding 100° were proposed. The flow organization (velocity streamlines, wall shear stresses, and dynamics of the flow) was identified by means of computational fluid dynamics (CFD) calculation. Pseudomonas fluorescens PF1 biofilm growth kinetics were then mapped. The change in some geometrical features induced a better flow organization reducing "dead zones". This significantly changed the biofilm growth kinetics, delaying the detection of biofilms from 20 hr to 24 hr. Critical areas such as welds, corners, and interfaces appeared far less prone to strong bacterial development. This would mean milder or less chemicals required at the washing step and faster and easier cleaning.

SUBMITTER: Bouvier L

PROVIDER: S-EPMC8164039 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Glutaraldehyde resistance mechanisms in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms

Project description:The objective of the current study was to understand the glutaraldehyde resistance mechanisms in P. fluorescens and P. aeruginosa biofilms. Glutaraldehyde is a common biocide used in various industries to control the microbial growth. Recent reports of emergence of glutaraldehyde resistance in several bacterial species motivated this study to understand the genetic factors responsible got glutaraldehyde resistance. Using a combination of phenotypic assays, chemical genetic assays and RNA-seq, we demonstrate that novel efflux pump, polyamine biosynthesis, lipid biosynthesis and phosphonate degradation play significant role in glutaraldehyde resistance and post-glutaraldehyde recovery of Psudomonad biofilms.

2015-04-06 | GSE64448 | GEO

Efflux as a glutaraldehyde resistance mechanism in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms.

Project description:A major challenge in microbial biofilm control is biocide resistance. Phenotypic adaptations and physical protective effects have been historically thought to be the primary mechanisms for glutaraldehyde resistance in bacterial biofilms. Recent studies indicate the presence of genetic mechanisms for glutaraldehyde resistance, but very little is known about the contributory genetic factors. Here, we demonstrate that efflux pumps contribute to glutaraldehyde resistance in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms. The RNA-seq data show that efflux pumps and phosphonate degradation, lipid biosynthesis, and polyamine biosynthesis metabolic pathways were induced upon glutaraldehyde exposure. Furthermore, chemical inhibition of efflux pumps potentiates glutaraldehyde activity, suggesting that efflux activity contributes to glutaraldehyde resistance. Additionally, induction of known modulators of biofilm formation, including phosphonate degradation, lipid biosynthesis, and polyamine biosynthesis, may contribute to biofilm resistance and resilience. Fundamental understanding of the genetic mechanism of biocide resistance is critical for the optimization of biocide use and development of novel disinfection strategies. Our results reveal genetic components involved in glutaraldehyde resistance and a potential strategy for improved control of biofilms.

| S-EPMC4432172 | biostudies-literature

Self-Adaptation of <i>Pseudomonas fluorescens</i> Biofilms to Hydrodynamic Stress.

Project description:In some conditions, bacteria self-organize into biofilms, supracellular structures made of a self-produced embedding matrix, mainly composed of polysaccharides, DNA, proteins, and lipids. It is known that bacteria change their colony/matrix ratio in the presence of external stimuli such as hydrodynamic stress. However, little is still known about the molecular mechanisms driving this self-adaptation. In this work, we monitor structural features of Pseudomonas fluorescens biofilms grown with and without hydrodynamic stress. Our measurements show that the hydrodynamic stress concomitantly increases the cell density population and the matrix production. At short growth timescales, the matrix mediates a weak cell-cell attractive interaction due to the depletion forces originated by the polymer constituents. Using a population dynamics model, we conclude that hydrodynamic stress causes a faster diffusion of nutrients and a higher incorporation of planktonic bacteria to the already formed microcolonies. This results in the formation of more mechanically stable biofilms due to an increase of the number of crosslinks, as shown by computer simulations. The mechanical stability also relies on a change in the chemical compositions of the matrix, which becomes enriched in carbohydrates, known to display adhering properties. Overall, we demonstrate that bacteria are capable of self-adapting to hostile hydrodynamic stress by tailoring the biofilm chemical composition, thus affecting both the mesoscale structure of the matrix and its viscoelastic properties that ultimately regulate the bacteria-polymer interactions.

| S-EPMC7835673 | biostudies-literature

Glutaraldehyde resistance mechanisms in Pseudomonas fluorescens and Pseudomonas aeruginosa biofilms

2015-04-06 | E-GEOD-64448 | biostudies-arrayexpress

Characterization of structures in biofilms formed by a Pseudomonas fluorescens isolated from soil.

Project description:BackgroundMicrobial biofilms represent an incompletely understood, but fundamental mode of bacterial growth. These sessile communities typically consist of stratified, morphologically-distinct layers of extracellular material, where numerous metabolic processes occur simultaneously in close proximity. Limited reports on environmental isolates have revealed highly ordered, three-dimensional organization of the extracellular matrix, which may hold important implications for biofilm physiology in vivo.ResultsA Pseudomonas spp. isolated from a natural soil environment produced flocculent, nonmucoidal biofilms in vitro with unique structural features. These mature biofilms were made up of numerous viable bacteria, even after extended culture, and contained up to 50% of proteins and accumulated 3% (by dry weight) calcium, suggesting an important role for the divalent metal in biofilm formation. Ultrastructurally, the mature biofilms contained structural motifs consisting of dense, fibrillary clusters, nanofibers, and ordered, honeycomb-like chambers enveloped in thin sheets.ConclusionMature biofilms contained living bacteria and were structurally, chemically, and physiologically heterogeneous. The principal architectural elements observed by electron microscopy may represent useful morphological clues for identifying bacterial biofilms in vivo. The complexity and reproducibility of the structural motifs observed in bacterial biofilms appear to be the result of organized assembly, suggesting that this environmental isolate may possess ecological advantages in its natural habitat.

| S-EPMC2697165 | biostudies-literature

Wetting of Dehydrated Hydrophilic <i>Pseudomonas fluorescens</i> Biofilms under the Action of External Body Forces.

Project description:Wetting of dehydrated Pseudomonas fluorescens biofilms grown on glass substrates by an external liquid is employed as a means to investigate the complex morphology of these biofilms along with their capability to interact with external fluids. The porous structure left behind after dehydration induces interesting droplet spreading on the external surface and imbibition into pores upon wetting. Static contact angles and volume loss by imbibition measured right upon droplet deposition indicate that biofilms of higher incubation times show a higher porosity and effective hydrophilicity. Furthermore, during subsequent rotation tests, using Kerberos device, these properties dictate a peculiar forced wetting/spreading behavior. As rotation speed increases a long liquid tail forms progressively at the rear part of the droplet, which stays pinned at all times, while only the front part of the droplet depins and spreads. Interestingly, the experimentally determined retention force for the onset of droplet sliding on biofilm external surface is lower than that on pure glass. An effort is made to describe such complex forced wetting phenomena by presenting apparent contact angles, droplet length, droplet shape contours, and edges position as obtained from detailed image analysis.

| S-EPMC8459453 | biostudies-literature

Genes Involved in Biofilm Matrix Formation of the Food Spoiler Pseudomonas fluorescens PF07

Project description:The extracellular matrix is essential for the biofilm formation of food spoilers. Pseudomonas fluorescens PF07 is a previous isolate from spoiled marine fish; however, the genes involved in the extracellular matrix formation of PF07 biofilms remain poorly defined. In this study, PF07 formed a wrinkled macrocolony biofilm through the high production of extracellular matrix. The genes involved in biofilm matrix formation and regulation were screened and identified by RNA-seq-dependent transcriptomic analysis and gene knock-out analysis. The macrocolony biofilms of PF07 grown for 5 days (PF07_5d) were compared with those grown for 1 day (PF07_1d). A total of 1,403 genes were significantly differentially expressed during biofilm formation. These mainly include the genes related to biofilm matrix proteins, polysaccharides, rhamnolipids, secretion system, biofilm regulation, and metabolism. Among them, functional amyloid genes fapABCDE were highly upregulated in the mature biofilm, and the operon fapA-E had a –24/–12 promoter dependent on the sigma factor RpoN. Moreover, the RNA-seq analyses of the rpoN mutant, compared with PF07, revealed 159 genes were differentially expressed in the macrocolony biofilms, and fapA-E genes were positively regulated by RpoN. In addition, the deletion mutants of fapC, rpoN, and brfA (a novel gene coding for an RpoN-dependent transcriptional regulator) were defective in forming mature macrocolony biofilms, solid surface-associated (SSA) biofilms, and pellicles, and they showed significantly reduced biofilm matrices. The fap genes were significantly downregulated in ΔbrfA, as in ΔrpoN. These findings suggest that the functional amyloid Fap is the main component of PF07 biofilm matrices, and RpoN may directly regulate the transcription of fap genes, in conjunction with BrfA. These genes may serve as potential molecular targets for screening new anti-biofilm agents or for biofilm detection in food environments.

| S-EPMC9207406 | biostudies-literature

The influence of postharvest UV-C treatment on anthocyanin biosynthesis in fresh-cut red cabbage.

Project description:Red cabbage (Brassica oleracea L. var. capitata f. rubra DC.) is a fresh edible vegetable consumed globally that contains high levels of antioxidant compounds including anthocyanins. In this study, fresh-cut red cabbage was treated with different Ultraviolet-C (UV-C) dosages. Fifteen cyanidin derivatives were observed in UV-C treated fresh-cut red cabbage; four of these were anthocyanins absent in control samples. The optimum dose of UV-C for enhancing total anthocyanin content in fresh-cut red cabbage was 3.0 kJ/m2. Different UV-C irradiation doses resulted in miscellaneous responses for each of the anthocyanin compounds, and these alterations appeared to be dose-dependent. The expression of genes relating to anthocyanin metabolism was altered by UV-C irradiation. For example, genes for biosynthetic enzymes including glycosyltransferase and acyltransferase, as well as R2R3 MYB transcription factors (production of anthocyanin pigment 1 and MYB114), had strongly increased expression following UV-C treatment. These results are in accord with the roles of these gene products in anthocyanin metabolism. This is, to the authors' knowledge, the first report demonstrating that UV-C treatment can increase the antioxidant activity in fresh-cut red cabbage in storage. Moreover, our detailed phytochemical and gene expression analysis establish specific roles for both anthocyanins and metabolism genes in this process.

| S-EPMC5507880 | biostudies-literature

Immobilization of the Aspartate Ammonia-Lyase from Pseudomonas fluorescens R124 on Magnetic Nanoparticles: Characterization and Kinetics.

Project description:Aspartate ammonia-lyases (AALs) catalyze the non-oxidative elimination of ammonia from l-aspartate to give fumarate and ammonia. In this work the AAL coding gene from Pseudomonas fluorescens R124 was identified, isolated, and cloned into the pET-15b expression vector and expressed in E. coli. The purified enzyme (PfAAL) showed optimal activity at pH 8.8, Michaelis-Menten kinetics in the ammonia elimination from l-aspartate, and no strong dependence on divalent metal ions for its activity. The purified PfAAL was covalently immobilized on epoxy-functionalized magnetic nanoparticles (MNP), and effective kinetics of the immobilized PfAAL-MNP was compared to the native solution form. Glycerol addition significantly enhanced the storability of PfAAL-MNP. Inhibiting effect of the growing viscosity (modulated by addition of glycerol or glucose) on the enzymatic activity was observed for the native and immobilized form of PfAAL, as previously described for other free enzymes. The storage stability and recyclability of PfAAL-MNP is promising for further biocatalytic applications.

| S-EPMC9307013 | biostudies-literature

Influence of the hydrodynamic environment on quorum sensing in Pseudomonas aeruginosa biofilms.

Project description:We provide experimental and modeling evidence that the hydrodynamic environment can impact quorum sensing (QS) in a Pseudomonas aeruginosa biofilm. The amount of biofilm biomass required for full QS induction of the population increased as the flow rate increased.

| S-EPMC2168657 | biostudies-literature

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