scholarly journals Novel Roles for the AIDA Adhesin from Diarrheagenic Escherichia coli: Cell Aggregation and Biofilm Formation

2004 ◽  
Vol 186 (23) ◽  
pp. 8058-8065 ◽  
Author(s):  
Orla Sherlock ◽  
Mark A. Schembri ◽  
Andreas Reisner ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Mammalian Cells ◽  
Gastrointestinal Disease ◽  
Cell Aggregation ◽  
Bacterial Attachment ◽  
E Coli ◽  
Abiotic Surfaces ◽  
Broad Variety ◽  
Self Association

ABSTRACT Diarrhea-causing Escherichia coli strains are responsible for numerous cases of gastrointestinal disease and constitute a serious health problem throughout the world. The ability to recognize and attach to host intestinal surfaces is an essential step in the pathogenesis of such strains. AIDA is a potent bacterial adhesin associated with some diarrheagenic E. coli strains. AIDA mediates bacterial attachment to a broad variety of human and other mammalian cells. It is a surface-displayed autotransporter protein and belongs to the selected group of bacterial glycoproteins; only the glycosylated form binds to mammalian cells. Here, we show that AIDA possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrate that intercellular AIDA-AIDA interaction is responsible for bacterial autoaggregation. Interestingly, AIDA-expressing cells can interact with antigen 43 (Ag43)-expressing cells, which is indicative of an intercellular AIDA-Ag43 interaction. Additionally, AIDA expression dramatically enhances biofilm formation by E. coli on abiotic surfaces in flow chambers.

scholarly journals The TibA Adhesin/Invasin from Enterotoxigenic Escherichia coli Is Self Recognizing and Induces Bacterial Aggregation and Biofilm Formation

2005 ◽  
Vol 73 (4) ◽  
pp. 1954-1963 ◽  
Author(s):  
Orla Sherlock ◽  
Rebecca Munk Vejborg ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Mammalian Cells ◽  
Gastrointestinal Disease ◽  
Bacterial Attachment ◽  
Enterotoxigenic Escherichia Coli ◽  
E Coli ◽  
Abiotic Surfaces ◽  
Self Association ◽  
Exclusive Group

ABSTRACT Escherichia coli strains are responsible for many cases of gastrointestinal disease and represent a serious health problem worldwide. An essential step in the pathogenesis of such strains involves recognition and attachment to host intestinal surfaces. TibA is a potent bacterial adhesin associated with a number of enterotoxigenic E. coli strains and mediates bacterial attachment to a variety of human cells; additionally, it promotes invasion of such cells. This adhesin is a surface-displayed autotransporter protein and belongs to the exclusive group of bacterial glycoproteins; only the glycosylated form confers binding to and invasion of mammalian cells. Here we characterized TibA and showed that it possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrated that intercellular TibA-TibA interaction is responsible for bacterial autoaggregation. Also, TibA expression significantly enhances biofilm formation by E. coli on abiotic surfaces.

scholarly journals UpaG, a New Member of the Trimeric Autotransporter Family of Adhesins in Uropathogenic Escherichia coli

2008 ◽  
Vol 190 (12) ◽  
pp. 4147-4161 ◽  
Author(s):  
Jaione Valle ◽  
Amanda N. Mabbett ◽  
Glen C. Ulett ◽  
Alejandro Toledo-Arana ◽  
Karine Wecker ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Cell Aggregation ◽  
Phylogenetic Groups ◽  
E Coli ◽  
Ecm Proteins ◽  
Abiotic Surfaces ◽  
Specific Affinity ◽  
Specific Virulence

ABSTRACT The ability of Escherichia coli to colonize both intestinal and extraintestinal sites is driven by the presence of specific virulence factors, among which are the autotransporter (AT) proteins. Members of the trimeric AT adhesin family are important virulence factors for several gram-negative pathogens and mediate adherence to eukaryotic cells and extracellular matrix (ECM) proteins. In this study, we characterized a new trimeric AT adhesin (UpaG) from uropathogenic E. coli (UPEC). Molecular analysis of UpaG revealed that it is translocated to the cell surface and adopts a multimeric conformation. We demonstrated that UpaG is able to promote cell aggregation and biofilm formation on abiotic surfaces in CFT073 and various UPEC strains. In addition, UpaG expression resulted in the adhesion of CFT073 to human bladder epithelial cells, with specific affinity to fibronectin and laminin. Prevalence analysis revealed that upaG is strongly associated with E. coli strains from the B2 and D phylogenetic groups, while deletion of upaG had no significant effect on the ability of CFT073 to colonize the mouse urinary tract. Thus, UpaG is a novel trimeric AT adhesin from E. coli that mediates aggregation, biofilm formation, and adhesion to various ECM proteins.

scholarly journals Sab, a Novel Autotransporter of Locus of Enterocyte Effacement-Negative Shiga-Toxigenic Escherichia coli O113:H21, Contributes to Adherence and Biofilm Formation

2009 ◽  
Vol 77 (8) ◽  
pp. 3234-3243 ◽  
Author(s):  
Sylvia Herold ◽  
James C. Paton ◽  
Adrienne W. Paton
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Gastrointestinal Disease ◽  
Pcr Analysis ◽  
Systemic Complications ◽  
E Coli ◽  
Acid Protein ◽  
Life Threatening ◽  
Uremic Syndrome ◽  
Enterocyte Effacement

ABSTRACT Shiga-toxigenic Escherichia coli (STEC) strains cause serious gastrointestinal disease, which can lead to potentially life-threatening systemic complications such as hemolytic-uremic syndrome. Although the production of Shiga toxin has been considered to be the main virulence trait of STEC for many years, the capacity to colonize the host intestinal epithelium is a crucial step in pathogenesis. In this study, we have characterized a novel megaplasmid-encoded outer membrane protein in locus of enterocyte effacement (LEE)-negative O113:H21 STEC strain 98NK2, termed Sab (for STEC autotransporter [AT] contributing to biofilm formation). The 4,296-bp sab gene encodes a 1,431-amino-acid protein with the features of members of the AT protein family. When expressed in E. coli JM109, Sab contributed to the diffuse adherence to human epithelial (HEp-2) cells and promoted biofilm formation on polystyrene surfaces. A 98NK2 sab deletion mutant was also defective in biofilm formation relative to its otherwise isogenic wild-type parent, and this was complemented by transformation with a sab-carrying plasmid. Interestingly, an unrelated O113:H21 STEC isolate that had a naturally occurring deletion in sab was similarly defective in biofilm formation. PCR analysis indicated that sab is present in LEE-negative STEC strains belonging to serotypes/groups O113:H21, O23, and O82:H8. These findings raise the possibility that Sab may contribute to colonization in a subset of LEE-negative STEC strains.

scholarly journals 2603. Biofilm Formation as a Predictive Marker of Prognosis for Escherichia coli Sepsis

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S904-S905 ◽  
Author(s):  
Kermit Zhang ◽  
Daniella Schneider ◽  
Rakesh Biswas ◽  
Mariana Gomez de la Espriella ◽  
Jayasimha Rao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Bacterial Attachment ◽  
Fisher Exact Test ◽  
Positive Finding ◽  
Continuous Variables ◽  
Clinical Patient ◽  
E Coli ◽  
Risk Of Death ◽  
Increased Risk

Abstract Background Escherichia coli is the Gram-negative organism most commonly associated with bloodstream infections and death due to sepsis. Timely administration of appropriate antibiotic(s) plays a significant role in improving patient outcomes. E. coli expresses virulence factors (VFs) such as biofilm formation and motility phenotypes which play a role in bacterial attachment and dissemination by enabling immune system evasion and host migration. The role of these VFs in bacteremia prognosis is not well characterized. Our study aims to evaluate the clinical characteristics and outcomes of E. coli bacteremia patients specifically in relation to biofilm forming isolates. Methods 91 E. coli bacteremia clinical isolates were consecutively collected from patients between 2013 to 2015. Virulence factor phenotypes were determined by in vitro biofilm formation, motility, and milk hydrolysis. Clinical patient data associated with the isolates were abstracted from the electronic medical records database and blinded from research team throughout characterization. Descriptive statistics were used for clinical variables and analyzed in a dichotomized fashion based on biofilm formation. The chi-square or Fisher exact test were used for categorical data and the Mann–Whitney U or Student T-test for continuous variables as appropriate. Results Of the 91 isolates, 41 had a biofilm-forming phenotype. Of the 87 isolates tested for milk hydrolysis and motility a positive finding was seen in 61 (70%) and 67(77%) isolates, respectively. In the multivariate model, patients with E.coli bacteremia from biofilm producing isolates were at increased risk of death or going into hospice during that hospitalization. ([OR],9.8; 95% CI, 1.1,88.7, P = 0.041) Conclusion Patients with biofilm-forming E. coli bacteremia had worse clinical outcomes than their non-biofilm forming counterparts suggesting that this phenotype leads to a more pathogenic organism. A prospective study to confirm this finding is needed as is the design of rapid diagnostics to promptly identify this phenotype in septic patients. Disclosures All authors: No reported disclosures.

Isolation of an Escherichia coli strain mutant unable to form biofilm on polystyrene and to adhere to human pneumocyte cells: involvement of tryptophanase

2002 ◽  
Vol 48 (2) ◽  
pp. 132-137 ◽  
Author(s):  
P Di Martino ◽  
A Merieau ◽  
R Phillips ◽  
N Orange ◽  
C Hulen
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
A549 Cells ◽  
Coli Strain ◽  
Gene Encoding ◽  
Cell Monolayers ◽  
E Coli ◽  
Transposon Insertion ◽  
Abiotic Surfaces ◽  
Tryptophanase Activity

Escherichia coli adherence to biotic and abiotic surfaces constitutes the first step of infection by promoting colonization and biofilm formation. The aim of this study was to gain a better understanding of the relationship between E. coli adherence to different biotic surfaces and biofilm formation on abiotic surfaces. We isolated mutants defective in A549 pneumocyte cells adherence, fibronectin adherence, and biofilm formation by random transposition mutagenesis and sequential passages over A549 cell monolayers. Among the 97 mutants tested, 80 were decreased in biofilm formation, 8 were decreased in A549 cells adherence, 7 were decreased in their adherence to fibronectin, and 17 had no perturbations in either of the three phenotypes. We observed a correlation between adherence to fibronectin or A549 cells and biofilm formation, indicating that biotic adhesive factors are involved in biofilm formation by E. coli. Molecular analysis of the mutants revealed that a transposon insertion in the tnaA gene encoding for tryptophanase was associated with a decrease in both A549 cells adherence and biofilm formation by E. coli. The complementation of the tnaA mutant with plasmid-located wild-type tnaA restored the tryptophanase activity, epithelial cells adherence, and biofilm formation on polystyrene. The possible mechanism of tryptophanase involvement in E. coli adherence and biofilm formation is discussed.Key words: Escherichia coli, biofilm, adherence, A549 cells, fibronectin, tryptophanase.

scholarly journals Genotypic and phenotypic characteristics associated with biofilm formation in Escherichia coli and Salmonella spp. isolated from ulam in Terengganu

Food Research ◽  
2019 ◽  
Vol 4 (1) ◽  
pp. 91-101
Author(s):  
Anis, A. Bahri ◽  
Wan Zawiah Wan Abdullah ◽  
Mohd Nizam Lani ◽  
W. Salleh ◽  
Rozila Alias
Keyword(s):  
Escherichia Coli ◽  
Disease Transmission ◽  
Congo Red ◽  
Biofilm Formation ◽  
Initial Step ◽  
Bacterial Attachment ◽  
Salmonella Spp ◽  
Pellicle Formation ◽  
E Coli ◽  
Fresh Vegetables

Nowadays, the foodborne outbreaks associated with fresh produces, including ulam, are increasing worldwide. The biofilm formation or bacterial attachment to plant surface is the initial step towards the contamination in fresh produce. The biofilm phenotype of bacteria grown on congo red agar is termed as red, dry and rough (rdar) morphotype. The binding of congo red dye with both biological proteins and inert surfaces is due to the presence of curli fimbriae and cellulose as the main extracellular components. The objective of this study was to determine the rdar morphotypes, biofilm ability and the role of csgA gene of Escherichia coli and Salmonella spp. isolated from ulam or Malaysian herbs. A total of 29 isolates, including 23 E. coli and 6 Salmonella spp. were analyzed for their ability to produce biofilm by colony morphology test, microtiter plate biofilm assay and qualitative biofilm test (pellicle formation). The presence of the csgA gene of E. coli was identified by PCR, which demonstrated the potential gene that able to produce curli fimbriae. Results revealed that 16 (69.6%) E. coli isolates were categorized as strong biofilm producers, 2 (8.7%) as moderate biofilm producers, 3 (13%) as weak biofilm producers, whereas 2 (8.7%) as negative biofilm producers (did not produce biofilm). While 4 (66.7%) Salmonella spp. isolates were identified as strong biofilm producers, 1 (16.7%) as moderate biofilm producers and 1 (16.7%) as negative biofilm producers. Majority of the E. coli strains (69.6%) were identified as strong biofilm producers and able to express rdar morphotypes. The ability of the of E. coli and Salmonella spp. isolates to form biofilm reveals the ability of these isolates to persist on the fresh vegetables and become hosts for the disease transmission to humans or/and animals.

scholarly journals Conditional Function of Autoaggregative Protein Cah and Common cah Mutations in Shiga Toxin-Producing Escherichia coli

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Michelle Qiu Carter ◽  
Maria T. Brandl ◽  
Indira T. Kudva ◽  
Robab Katani ◽  
Matthew R. Moreau ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Calcium Binding ◽  
Biofilm Formation ◽  
Bacterial Attachment ◽  
Ecological Niches ◽  
Loss Of Function ◽  
Host Colonization ◽  
Content Type ◽  
E Coli

ABSTRACTCah is a calcium-binding autotransporter protein involved in autoaggregation and biofilm formation. Althoughcahis widespread in Shiga toxin-producingEscherichia coli(STEC), we detected mutations incahat a frequency of 31.3% in this pathogen. In STEC O157:H7 supershedder strain SS17, a large deletion results in a smaller coding sequence, encoding a protein lacking the C-terminal 71 amino acids compared with Cah in STEC O157:H7 strain EDL933. We examined the function of Cah in biofilm formation and host colonization to better understand the selective pressures forcahmutations. EDL933-Cah played a conditional role in biofilm formationin vitro: it enhancedE. coliDH5α biofilm formation on glass surfaces under agitated culture conditions that prevented autoaggregation but inhibited biofilm formation under hydrostatic conditions that facilitated autoaggregation. This function appeared to be strain dependent since Cah-mediated biofilm formation was diminished when an EDL933cahgene was expressed in SS17. Deletion ofcahin EDL933 enhanced bacterial attachment to spinach leaves and altered the adherence pattern of EDL933 to bovine recto-anal junction squamous epithelial (RSE) cells. In contrast, intransexpression of EDL933cahin SS17 increased its attachment to leaf surfaces, and in DH5α, it enhanced its adherence to RSE cells. Hence, the ecological function of Cah appears to be modulated by environmental conditions and other bacterial strain-specific properties. Considering the prevalence ofcahin STEC and its role in attachment and biofilm formation,cahmutations might be selected in ecological niches in which inactivation of Cah would result in an increased fitness in STEC during colonization of plants or animal hosts.IMPORTANCEShiga toxin-producingEscherichia coli(STEC) harbors genes encoding diverse adhesins, and many of these are known to play an important role in bacterial attachment and host colonization. We demonstrated here that the autotransporter protein Cah confers onE. coliDH5α cells a strong autoaggregative phenotype that is inversely correlated with its ability to form biofilms and plays a strain-specific role in plant and animal colonization by STEC. Althoughcahis widespread in the STEC population, we detected a mutation rate of 31.3% incah, which is similar to that reported forrpoSandfimH. The formation of cell aggregates due to increased bacterium-to-bacterium interactions may be disadvantageous to bacterial populations under conditions that favor a planktonic state in STEC. Therefore, a loss-of-function mutation incahis likely a selective trait in STEC when autoaggregative properties become detrimental to bacterial cells and may contribute to the adaptability of STEC to fluctuating environments.

scholarly journals Characterization of IcmF of the type VI secretion system in an avian pathogenic Escherichia coli (APEC) strain

Microbiology ◽  
2011 ◽  
Vol 157 (10) ◽  
pp. 2954-2962 ◽  
Author(s):  
Fernanda de Pace ◽  
Jacqueline Boldrin de Paiva ◽  
Gerson Nakazato ◽  
Marcelo Lancellotti ◽  
Marcelo Palma Sircili ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Legionella Pneumophila ◽  
Biofilm Formation ◽  
Secretion System ◽  
Type Vi Secretion System ◽  
E Coli ◽  
Type Vi Secretion ◽  
Abiotic Surfaces ◽  
Pathogenic Escherichia Coli ◽  
Zoonotic Risk

The intracellular multiplication factor (IcmF) protein is a component of the recently described type VI secretion system (T6SS). IcmF has been shown to be required for intra-macrophage replication and inhibition of phagosome–lysosome fusion in Legionella pneumophila. In Vibrio cholerae it is involved in motility, adherence and conjugation. Given that we previously reported that two T6SS genes (hcp and clpV) contribute to the pathogenesis of a septicaemic strain (SEPT362) of avian pathogenic Escherichia coli (APEC), we investigated the function of IcmF in this strain. Further elucidation of the virulence mechanisms of APEC is important because this pathogen is responsible for financial losses in the poultry industry, and is closely related to human extraintestinal pathogenic E. coli (ExPEC) strains, representing a potential zoonotic risk, as well as serving as a reservoir of virulence genes. Here we show that an APEC icmF mutant has decreased adherence to and invasion of epithelial cells, as well as decreased intra-macrophage survival. The icmF mutant is also defective for biofilm formation on abiotic surfaces. Additionally, expression of the flagella operon is decreased in the icmF mutant, leading to decreased motility. The combination of these phenotypes culminates in this mutant being altered for infection in chicks. These results suggest that IcmF in APEC may play a role in disease, and potentially also in the epidemiological spread of this pathogen through enhancement of biofilm formation.

scholarly journals Biofilm Formation on Biotic and Abiotic Surfaces in the Presence of Antimicrobials by Escherichia coli Isolates from Cases of Bovine Mastitis

2014 ◽  
Vol 80 (19) ◽  
pp. 6136-6145 ◽  
Author(s):  
Vitor O. Silva ◽  
Larissa O. Soares ◽  
Abelardo Silva Júnior ◽  
Hilário C. Mantovani ◽  
Yung-Fu Chang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Expression Profiles ◽  
Bovine Mastitis ◽  
Pcr Analysis ◽  
Content Type ◽  
E Coli ◽  
Biofilm Production ◽  
Abiotic Surfaces

ABSTRACTEscherichia coliis a highly adaptive microorganism, and its ability to form biofilms under certain conditions can be critical for antimicrobial resistance. The adhesion of fourE. coliisolates from bovine mastitis to bovine mammary alveolar (MAC-T) cells, biofilm production on a polystyrene surface, and the expression profiles of the genesfliC,csgA,fimA, andluxSin the presence of enrofloxacin, gentamicin, co-trimoxazole, and ampicillin at half of the MIC were investigated. Increased adhesion ofE. coliisolates in the presence of antimicrobials was not observed; however, increased internalization of some isolates was observed by confocal microscopy. All of the antimicrobials induced the formation of biofilms by at least one isolate, whereas enrofloxacin and co-trimoxazole decreased biofilm formation by at least one isolate. Quantitative PCR analysis revealed that all four genes were differentially expressed when bacteria were exposed to subinhibitory concentrations of antimicrobials, with expression altered on the order of 1.5- to 22-fold. However, it was not possible to associate gene expression with induction or reduction of biofilm formation in the presence of the antimicrobials. Taken together, the results demonstrate that antimicrobials could induce biofilm formation by some isolates, in addition to inducing MAC-T cell invasion, a situation that might occurin vivo, potentially resulting in a bacterial reservoir in the udder, which might explain some cases of persistent mastitis in herds.

scholarly journals Label-free quantitative proteomic analysis of the inhibition effect of Lactobacillus rhamnosus GG on Escherichia coli biofilm formation in co-culture

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Huiyi Song ◽  
Ni Lou ◽  
Jianjun Liu ◽  
Hong Xiang ◽  
Dong Shang
Keyword(s):  
Escherichia Coli ◽  
Proteomic Analysis ◽  
Biofilm Formation ◽  
Lactobacillus Rhamnosus ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Label Free ◽  
Quantitative Proteomic Analysis ◽  
E Coli ◽  
Protein Ubiquitination

Abstract Background Escherichia coli (E. coli) is the principal pathogen that causes biofilm formation. Biofilms are associated with infectious diseases and antibiotic resistance. This study employed proteomic analysis to identify differentially expressed proteins after coculture of E. coli with Lactobacillus rhamnosus GG (LGG) microcapsules. Methods To explore the relevant protein abundance changes after E. coli and LGG coculture, label-free quantitative proteomic analysis and qRT-PCR were applied to E. coli and LGG microcapsule groups before and after coculture, respectively. Results The proteomic analysis characterised a total of 1655 proteins in E. coli K12MG1655 and 1431 proteins in the LGG. After coculture treatment, there were 262 differentially expressed proteins in E. coli and 291 in LGG. Gene ontology analysis showed that the differentially expressed proteins were mainly related to cellular metabolism, the stress response, transcription and the cell membrane. A protein interaction network and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis indicated that the differentiated proteins were mainly involved in the protein ubiquitination pathway and mitochondrial dysfunction. Conclusions These findings indicated that LGG microcapsules may inhibit E. coli biofilm formation by disrupting metabolic processes, particularly in relation to energy metabolism and stimulus responses, both of which are critical for the growth of LGG. Together, these findings increase our understanding of the interactions between bacteria under coculture conditions.

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