scholarly journals Mat fimbriae promote biofilm formation by meningitis-associated Escherichia coli

Microbiology ◽  
2010 ◽  
Vol 156 (8) ◽  
pp. 2408-2417 ◽  
Author(s):  
Timo A. Lehti ◽  
Philippe Bauchart ◽  
Johanna Heikkinen ◽  
Jörg Hacker ◽  
Timo K. Korhonen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Early Stage ◽  
Surface Expression ◽  
Growth Media ◽  
Biofilm Growth ◽  
Abiotic Surfaces ◽  
K 12 ◽  
Biofilm Development

The mat (or ecp) fimbrial operon is ubiquitous and conserved in Escherichia coli, but its functions remain poorly described. In routine growth media newborn meningitis isolates of E. coli express the meningitis-associated and temperature-regulated (Mat) fimbria, also termed E. coli common pilus (ECP), at 20 °C, and here we show that the six-gene (matABCDEF)-encoded Mat fimbria is needed for temperature-dependent biofilm formation on abiotic surfaces. The matBCDEF deletion mutant of meningitis E. coli IHE 3034 was defective in an early stage of biofilm development and consequently unable to establish a detectable biofilm, contrasting with IHE 3034 derivatives deleted for flagella, type 1 fimbriae or S-fimbriae, which retained the wild-type biofilm phenotype. Furthermore, induced production of Mat fimbriae from expression plasmids enabled biofilm-deficient E. coli K-12 cells to form biofilm at 20 °C. No biofilm was detected with IHE 3034 or MG1655 strains grown at 37 °C. The surface expression of Mat fimbriae and the frequency of Mat-positive cells in the IHE 3034 population from 20 °C were high and remained unaltered during the transition from planktonic to biofilm growth and within the matured biofilm community. Considering the prevalence of the highly conserved mat locus in E. coli genomes, we hypothesize that Mat fimbria-mediated biofilm formation is an ancestral characteristic of E. coli.

scholarly journals Role of DLP12 lysis genes in Escherichia coli biofilm formation

Microbiology ◽  
2011 ◽  
Vol 157 (6) ◽  
pp. 1640-1650 ◽  
Author(s):  
Faustino A. Toba ◽  
Mitchell G. Thompson ◽  
Bryan R. Campbell ◽  
Lauren M. Junker ◽  
Karl-Gustav Rueggeberg ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Biofilm Formation ◽  
Wall Stress ◽  
Wild Type ◽  
Lambda Phage ◽  
Kinase Gene ◽  
Abiotic Surfaces ◽  
K 12 ◽  
Biofilm Development

Phages have recently been implicated as important in biofilm development, although the mechanisms whereby phages impact biofilms remain unclear. One defective lambdoid phage carried by Escherichia coli K-12 is DLP12. Among the genes found in DLP12 are essD, ybcS and rzpD/rzoD, which are homologues of the Lambda phage genes encoding cell-lysis proteins (S, R and Rz/Rz1). The role that these DLP12 lysis genes play in biofilm formation was examined in deletion mutants of E. coli PHL628, a curli-overproducing, biofilm-forming K-12 derivative. Strains lacking essD, ybcS and rzpD/rzoD were unable to form wild-type biofilms. While all mutants were compromised in attachment to abiotic surfaces and aggregated less well than the wild-type, the effect of the essD knockout on biofilm formation was less dramatic than that of deleting ybcS or rzpD/rzoD. These results were consistent with electron micrographs of the mutants, which showed a decreased number of curli fibres on cell surfaces. Also consistent with this finding, we observed that expression from the promoter of csgB, which encodes the curli subunits, was downregulated in the mutants. As curli production is transcriptionally downregulated in response to cell wall stress, we challenged the mutants with SDS and found them to be more sensitive to the detergent than the wild-type. We also examined the release of 14C-labelled peptidoglycan from the mutants and found that they did not lose labelled peptidoglycan to the same extent as the wild-type. Given that curli production is known to be suppressed by N-acetylglucosamine 6-phosphate (NAG-6P), a metabolite produced during peptidoglycan recycling, we deleted nagK, the N-acetylglucosamine kinase gene, from the lysis mutants and found that this restored curli production. This suggested that deletion of the lysis genes affected cell wall status, which was transduced to the curli operon by NAG-6P via an as yet unknown mechanism. These observations provide evidence that the S, R and Rz/Rz1 gene homologues encoded by DLP12 are not merely genetic junk, but rather play an important, though undefined, role in cell wall maintenance.

scholarly journals Bundle-Forming Pili and EspA Are Involved in Biofilm Formation by Enteropathogenic Escherichia coli

2006 ◽  
Vol 188 (11) ◽  
pp. 3952-3961 ◽  
Author(s):  
Cristiano G. Moreira ◽  
Kelli Palmer ◽  
Marvin Whiteley ◽  
Marcelo P. Sircili ◽  
Luiz R. Trabulsi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Signaling Mechanism ◽  
Bacterial Aggregation ◽  
Abiotic Surfaces ◽  
Genes Encoding ◽  
Continuous Culture System ◽  
Static Conditions ◽  
Biofilm Development ◽  
Isogenic Mutants

ABSTRACT Microcolony formation is one of the initial steps in biofilm development, and in enteropathogenic Escherichia coli (EPEC) it is mediated by several adhesins, including the bundle-forming pilus (BFP) and the EspA filament. Here we report that EPEC forms biofilms on plastic under static conditions and a flowthrough continuous culture system. The abilities of several EPEC isogenic mutants to form biofilms were assessed. Adhesins such as BFP and EspA, important in microcolony formation on epithelial cells, are also involved in bacterial aggregation during biofilm formation on abiotic surfaces. Mutants that do not express BFP or EspA form more-diffuse biofilms than does the wild type. We also determined, using gfp transcriptional fusions, that, consistent with the role of these adhesins in biofilms, the genes encoding BFP and EspA are expressed during biofilm formation. Finally, expression of espA is controlled by a quorum-sensing (QS) regulatory mechanism, and the EPEC qseA QS mutant also forms altered biofilms, suggesting that this signaling mechanism plays an important role in EPEC biofilm development. Taken together, these studies allowed us to propose a model of EPEC biofilm formation.

scholarly journals Regulation of type 1 fimbriae synthesis and biofilm formation by the transcriptional regulator LrhA of Escherichia coli

Microbiology ◽  
2005 ◽  
Vol 151 (10) ◽  
pp. 3287-3298 ◽  
Author(s):  
Caroline Blumer ◽  
Alexandra Kleefeld ◽  
Daniela Lehnen ◽  
Margit Heintz ◽  
Ulrich Dobrindt ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Invertible Element ◽  
Transcriptional Regulator ◽  
Host Cells ◽  
Yeast Cells ◽  
Type 1 Fimbriae ◽  
Biofilm Development

Type 1 fimbriae of Escherichia coli facilitate attachment to the host mucosa and promote biofilm formation on abiotic surfaces. The transcriptional regulator LrhA, which is known as a repressor of flagellar, motility and chemotaxis genes, regulates biofilm formation and expression of type 1 fimbriae. Whole-genome expression profiling revealed that inactivation of lrhA results in an increased expression of structural components of type 1 fimbriae. In vitro, LrhA bound to the promoter regions of the two fim recombinases (FimB and FimE) that catalyse the inversion of the fimA promoter, and to the invertible element itself. Translational lacZ fusions with these genes and quantification of fimE transcript levels by real-time PCR showed that LrhA influences type 1 fimbrial phase variation, primarily via activation of FimE, which is required for the ON-to-OFF transition of the fim switch. Enhanced type 1 fimbrial expression as a result of lrhA disruption was confirmed by mannose-sensitive agglutination of yeast cells. Biofilm formation was stimulated by lrhA inactivation and completely suppressed upon LrhA overproduction. The effects of LrhA on biofilm formation were exerted via the changed levels of surface molecules, most probably both flagella and type 1 fimbriae. Together, the data show a role for LrhA as a repressor of type 1 fimbrial expression, and thus as a regulator of the initial stages of biofilm development and, presumably, bacterial adherence to epithelial host cells also.

scholarly journals Immunoglobulin-Mediated Agglutination of and Biofilm Formation by Escherichia coli K-12 Require the Type 1 Pilus Fiber

2004 ◽  
Vol 72 (4) ◽  
pp. 1929-1938 ◽  
Author(s):  
Paul E. Orndorff ◽  
Aditya Devapali ◽  
Sarah Palestrant ◽  
Aaron Wyse ◽  
Mary Lou Everett ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Immunoglobulin A ◽  
Minor Component ◽  
Secretory Iga ◽  
Bacterial Surface ◽  
E Coli ◽  
Type 1 Pili ◽  
K 12

ABSTRACT The binding of human secretory immunoglobulin A (SIgA), the primary immunoglobulin in the gut, to Escherichia coli is thought to be dependent on type 1 pili. Type 1 pili are filamentous bacterial surface attachment organelles comprised principally of a single protein, the product of the fimA gene. A minor component of the pilus fiber (the product of the fimH gene, termed the adhesin) mediates attachment to a variety of host cell molecules in a mannose inhibitable interaction that has been extensively described. We found that the aggregation of E. coli K-12 by human secretory IgA (SIgA) was dependent on the presence of the pilus fiber, even in the absence of the mannose specific adhesin or in the presence of 25 mM α-CH3Man. The presence of pilus without adhesin also facilitated SIgA-mediated biofilm formation on polystyrene, although biofilm formation was stronger in the presence of the adhesin. IgM also mediated aggregation and biofilm formation in a manner dependent on pili with or without adhesin. These findings indicate that the pilus fiber, even in the absence of the adhesin, may play a role in biologically important processes. Under conditions in which E. coli was agglutinated by SIgA, the binding of SIgA to E. coli was not increased by the presence of the pili, with or without adhesin. This observation suggests that the pili, with or without adhesin, affect factors such as cell surface rigidity or electrostatic repulsion, which can affect agglutination but which do not necessarily determine the level of bound immunoglobulin.

Cellular chain formation in Escherichia coli biofilms

Microbiology ◽  
2009 ◽  
Vol 155 (5) ◽  
pp. 1407-1417 ◽  
Author(s):  
Rebecca Munk Vejborg ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Immunofluorescence Microscopy ◽  
Asymptomatic Bacteriuria ◽  
Type I ◽  
Chain Formation ◽  
Biofilm Growth ◽  
E Coli ◽  
Type I Fimbriae ◽  
K 12

In this study we report on a novel structural phenotype in Escherichia coli biofilms: cellular chain formation. Biofilm chaining in E. coli K-12 was found to occur primarily by clonal expansion, but was not due to filamentous growth. Rather, chain formation was the result of intercellular interactions facilitated by antigen 43 (Ag43), a self-associating autotransporter (SAAT) protein, which has previously been implicated in auto-aggregation and biofilm formation. Immunofluorescence microscopy suggested that Ag43 was concentrated at or near the cell poles, although when the antigen was highly overexpressed, a much more uniform distribution was seen. Immunofluorescence microscopy also indicated that other parameters, including dimensional constraints (flow, growth alongside a surface), may also affect the final biofilm architecture. Moreover, chain formation was affected by other surface structures; type I fimbriae expression significantly reduced cellular chain formation, presumably by steric hindrance. Cellular chain formation did not appear to be specific to E. coli K-12. Although many urinary tract infection (UTI) isolates were found to form rather homogeneous, flat biofilms, three isolates, including the prototypic asymptomatic bacteriuria strain, 83972, formed highly elaborate cellular chains during biofilm growth in human urine. Combined, these results illustrate the diversity of biofilm architectures that can be observed even within a single microbial species.

scholarly journals Biofilm Formation and Dispersal under the Influence of the Global Regulator CsrA of Escherichia coli

2002 ◽  
Vol 184 (1) ◽  
pp. 290-301 ◽  
Author(s):  
Debra W. Jackson ◽  
Kazushi Suzuki ◽  
Lawrence Oakford ◽  
Jerry W. Simecka ◽  
Mark E. Hart ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Rna Binding ◽  
Glycogen Synthase ◽  
Regulatory Protein ◽  
Ectopic Expression ◽  
Knockout Mutant ◽  
E Coli ◽  
K 12 ◽  
Biofilm Development

ABSTRACT The predominant mode of growth of bacteria in the environment is within sessile, matrix-enclosed communities known as biofilms. Biofilms often complicate chronic and difficult-to-treat infections by protecting bacteria from the immune system, decreasing antibiotic efficacy, and dispersing planktonic cells to distant body sites. While the biology of bacterial biofilms has become a major focus of microbial research, the regulatory mechanisms of biofilm development remain poorly defined and those of dispersal are unknown. Here we establish that the RNA binding global regulatory protein CsrA (carbon storage regulator) of Escherichia coli K-12 serves as both a repressor of biofilm formation and an activator of biofilm dispersal under a variety of culture conditions. Ectopic expression of the E. coli K-12 csrA gene repressed biofilm formation by related bacterial pathogens. A csrA knockout mutation enhanced biofilm formation in E. coli strains that were defective for extracellular, surface, or regulatory factors previously implicated in biofilm formation. In contrast, this csrA mutation did not affect biofilm formation by a glgA (glycogen synthase) knockout mutant. Complementation studies with glg genes provided further genetic evidence that the effects of CsrA on biofilm formation are mediated largely through the regulation of intracellular glycogen biosynthesis and catabolism. Finally, the expression of a chromosomally encoded csrA′-′lacZ translational fusion was dynamically regulated during biofilm formation in a pattern consistent with its role as a repressor. We propose that global regulation of central carbon flux by CsrA is an extremely important feature of E. coli biofilm development.

scholarly journals Abstracts of an international conference “Biofilms II: Attachment and detachment in pure and mixed cultures”, held at the Leipziger Kubus UFZ Centre for Environmental Research, Leipzig, Germany, from 23 March to 24 March 2006

Biofilms ◽  
2005 ◽  
Vol 2 (4) ◽  
pp. 245-273
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Three Dimensional ◽  
Conjugative Plasmid ◽  
Environmental Research ◽  
Biofilm Growth ◽  
Biofilm Detachment ◽  
Biofilm Development

The effect of growth and detachment on formation of large-scale biofilm structureBiofilm cohesive energy density determination using a novel atomic force microscopy methodologyFluorescence correlation spectroscopy under two-photon excitation for the study of diffusion and reactivity of bacteriophage inside bacterial biofilmsBiothermodynamic characterization and dynamic analysis of biofilms using calorimetryBiomimetic antifouling coatings for sensor surfaces for water monitoring: performance control in defined biofilm cultures and under real environmental conditionsThe contribution of rpos to formation of Escherichia coli biofilmsSynergistic effects in mixed Escherichia coli biofilms: conjugative plasmid transfer drives biofilm expansionThe universal stress protein PA3309 in Pseudomonas aeruginosa is induced in biofilmsExtracellular polymeric substances from biofilms on membranes in waste-water treatment plantsBiofilm-to-planktonic cell yield: a strategy for proliferationPhysiological and phylogenetic characterization of the dispersed and loosely attached fraction of activated sludge flocsTowards a deterministic model of biofilm detachment: an experimental studyEffect of backwash on the characteristics of biofilm in a biological activated filter reactor using elemental sulfur particlesProcess performance and biomass properties in membrane-aerated bioreactorsBioaugmentation via conjugation in biofilms treating 3-chloroaniline: effects of selective pressureEffect of phosphorus on biofilm growth in a completely mixed biofilm reactorImpacts of biofilm development on reactive transport in porous media under variable flow regimensInfluence of biofilms on colloid mobility in the subsurfaceBiofilms in amendable in situ microcosms indicate relevant electron acceptor processes at a BTEX-contaminated aquiferFunctional biodiversity of complex biofilms grown on polychlorinated biphenyl oilIdentification and characterization of biofilm formation phenotypes of several clinically relevant Streptococcus pyogenes serotype strainsSelected probiotic bacteria disrupt biofilm development of vancomycin-resistant Enterococcus faeciumComparison of the extracellular polymeric substances of Candida albicans and Candida dubliniensis biofilmsInfluence of quorum-sensing regulated production of an antimicrobial component by Serratia plymuthica on establishment of dual species biofilms with Escherichia coliBiofilm formation by the thermophilic and cellulolytic actinomycete Thermobifida fuscaBiomonitoring of bacterial contamination on different surfaces of food-processing machinesRole of the flagella during the adhesion of Listeria monocytogenes EGD-e to inert surfaces after cultivation at different pHs and temperaturesAdhesion of Saccharomyces cerevisiae to stainless steel: influence of surface propertiesInvestigating the mechanical strength of biofilms with fluid dynamic gaugingThree-dimensional biofilm model with individual cells and continuum extracellular polymeric substances matrixA three-dimensional computer model analysis of four hypothetical biofilm detachment mechanismsModelling biofilm growth, detachment and fluid flow in a cross-section of tube reactorsBiofilm games

scholarly journals Flagellar Motility Is Critical for Listeria monocytogenes Biofilm Formation

2007 ◽  
Vol 189 (12) ◽  
pp. 4418-4424 ◽  
Author(s):  
Katherine P. Lemon ◽  
Darren E. Higgins ◽  
Roberto Kolter
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
Molecular Mechanisms ◽  
Early Stage ◽  
Food Contamination ◽  
Initial Surface ◽  
Flagellar Motility ◽  
Surface Attachment ◽  
Abiotic Surfaces ◽  
Biofilm Development

ABSTRACT The food-borne pathogen Listeria monocytogenes attaches to environmental surfaces and forms biofilms that can be a source of food contamination, yet little is known about the molecular mechanisms of its biofilm development. We observed that nonmotile mutants were defective in biofilm formation. To investigate how flagella might function during biofilm formation, we compared the wild type with flagellum-minus and paralyzed-flagellum mutants. Both nonmotile mutants were defective in biofilm development, presumably at an early stage, as they were also defective in attachment to glass during the first few hours of surface exposure. This attachment defect could be significantly overcome by providing exogenous movement toward the surface via centrifugation. However, this centrifugation did not restore mature biofilm formation. Our results indicate that it is flagellum-mediated motility that is critical for both initial surface attachment and subsequent biofilm formation. Also, any role for L. monocytogenes flagella as adhesins on abiotic surfaces appears to be either minimal or motility dependent under the conditions we examined.

scholarly journals Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure

mBio ◽  
2013 ◽  
Vol 4 (5) ◽  
Author(s):  
Chia Hung ◽  
Yizhou Zhou ◽  
Jerome S. Pinkner ◽  
Karen W. Dodson ◽  
Jan R. Crowley ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Extracellular Matrix ◽  
Biofilm Formation ◽  
Developmental Stages ◽  
Three Dimensional ◽  
Bacterial Biofilms ◽  
Content Type ◽  
Abiotic Surfaces ◽  
Biochemical Analyses ◽  
Biofilm Development

ABSTRACTBacterial biofilms are ubiquitous in nature, and their resilience is derived in part from a complex extracellular matrix that can be tailored to meet environmental demands. Although common developmental stages leading to biofilm formation have been described, how the extracellular components are organized to allow three-dimensional biofilm development is not well understood. Here we show that uropathogenicEscherichia coli(UPEC) strains produce a biofilm with a highly ordered and complex extracellular matrix (ECM). We used electron microscopy (EM) techniques to image floating biofilms (pellicles) formed by UPEC. EM revealed intricately constructed substructures within the ECM that encase individual, spatially segregated bacteria with a distinctive morphology. Mutational and biochemical analyses of these biofilms confirmed curli as a major matrix component and revealed important roles for cellulose, flagella, and type 1 pili in pellicle integrity and ECM infrastructure. Collectively, the findings of this study elucidated that UPEC pellicles have a highly organized ultrastructure that varies spatially across the multicellular community.IMPORTANCEBacteria can form biofilms in diverse niches, including abiotic surfaces, living cells, and at the air-liquid interface of liquid media. Encasing these cellular communities is a self-produced extracellular matrix (ECM) that can be composed of proteins, polysaccharides, and nucleic acids. The ECM protects biofilm bacteria from environmental insults and also makes the dissolution of biofilms very challenging. As a result, formation of biofilms within humans (during infection) or on industrial material (such as water pipes) has detrimental and costly effects. In order to combat bacterial biofilms, a better understanding of components required for biofilm formation and the ECM is required. This study defined the ECM composition and architecture of floating pellicle biofilms formed byEscherichia coli.

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