scholarly journals Inhibition of L. monocytogenes Biofilm Formation by the Amidase Domain of the Phage vB_LmoS_293 Endolysin

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 722 ◽  
Author(s):  
Pennone ◽  
Sanz-Gaitero ◽  
O’Connor ◽  
Coffey ◽  
Jordan ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Food Production ◽  
Production Environment ◽  
E Coli ◽  
Abiotic Surfaces ◽  
Progeny Phage ◽  
Production Environments ◽  
Peptidoglycan Cell Wall ◽  
Different Temperatures ◽  
Serotype 4B

Listeria monocytogenes is a ubiquitous Gram-positive bacterium that is a major concern for food business operators because of its pathogenicity and ability to form biofilms in food production environments. Bacteriophages (phages) have been evaluated as biocontrol agents for L. monocytogenes in a number of studies and, indeed, certain phages have been approved for use as anti-listerial agents in food processing environments (ListShield and PhageGuard Listex). Endolysins are proteins produced by phages in the host cell. They cleave the peptidoglycan cell wall, thus allowing release of progeny phage into the environment. In this study, the amidase domain of the phage vB_LmoS_293 endolysin (293-amidase) was cloned and expressed in Escherichia. coli (E. coli). Muralytic activity at different concentrations, pH and temperature values, lytic spectrum and activity against biofilms was determined for the purified 293-amidase protein. The results showed activity on autoclaved cells at three different temperatures (20 °C, 37 °C and 50 °C), with a wider specificity (L. monocytogenes 473 and 3099, a serotype 4b and serogroup 1/2b-3b-7, respectively) compared to the phage itself, which targets only L. monocytogenes serotypes 4b and 4e. The protein also inhibits biofilm formation on abiotic surfaces. These results show the potential of using recombinant antimicrobial proteins against pathogens in the food production environment.

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.

Phenotypic Characterization of Salmonella Isolated from Food Production Environments Associated with Low–Water Activity Foods

2013 ◽  
Vol 76 (9) ◽  
pp. 1488-1499 ◽  
Author(s):  
SARAH FINN ◽  
JAY C. D. HINTON ◽  
PETER McCLURE ◽  
ALÉJANDRO AMÉZQUITA ◽  
MARTA MARTINS ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Food Production ◽  
Phenotypic Characterization ◽  
Minimal Risk ◽  
Limited Information ◽  
Planktonic Cells ◽  
Cellulose Production ◽  
Production Chain ◽  
Production Environments

Salmonella can survive for extended periods of time in low-moisture environments posing a challenge for modern food production. This dangerous pathogen must be controlled throughout the production chain with a minimal risk of dissemination. Limited information is currently available describing the behavior and characteristics of this important zoonotic foodborne bacterium in low-moisture food production environments and in food. In our study, the phenotypes related to low-moisture survival of 46 Salmonella isolates were examined. Most of the isolates in the collection could form biofilms under defined laboratory conditions, with 57% being positive for curli fimbriae production and 75% of the collection positive for cellulose production, which are both linked with stronger biofilm formation. Biocides in the factory environment to manage hygiene were found to be most effective against planktonic cells but less so when the same bacteria were surface dried or present as a biofilm. Cellulose-producing isolates were better survivors when exposed to a biocide compared with cellulose-negative isolates. Examination of Salmonella growth of these 18 serotypes in NaCl, KCl, and glycerol found that glycerol was the least inhibitory of these three humectants. We identified a significant correlation between the ability to survive in glycerol and the ability to survive in KCl and biofilm formation, which may be important for food safety and the protection of public health.

scholarly journals Biofilm Formation in a Hydrodynamic Environment by Novel FimH Variants and Ramifications for Virulence

2001 ◽  
Vol 69 (3) ◽  
pp. 1322-1328 ◽  
Author(s):  
Mark A. Schembri ◽  
Per Klemm
Keyword(s):  
Urinary Tract ◽  
Biofilm Formation ◽  
Random Mutagenesis ◽  
Cumulative Effect ◽  
Growth Conditions ◽  
E Coli ◽  
Enrichment Technique ◽  
Naturally Occurring ◽  
Abiotic Surfaces ◽  
Hydrodynamic Environment

ABSTRACT Type 1 fimbriae are surface-located adhesion organelles ofEscherichia coli that are directly associated with virulence of the urinary tract. They mediated-mannose-sensitive binding to different host surfaces by way of the minor fimbrial component FimH. Naturally occurring variants of FimH that bind strongly to terminally exposed monomannose residues have been associated with a pathogenicity-adaptive phenotype that enhances E. coli colonization of extraintestinal locations such as the urinary tract. The FimH adhesin also promotes biofilm formation in a mannose-inhibitable manner on abiotic surfaces under static growth conditions. In this study, we used random mutagenesis combined with a novel selection-enrichment technique to specifically identify mutations in the FimH adhesin that confer onE. coli the ability to form biofilms under hydrodynamic flow (HDF) conditions. We identified three FimH variants from our mutant library that could mediate an HDF biofilm formation phenotype to various degrees. This phenotype was induced by the cumulative effect of multiple changes throughout the receptor-binding region of the protein. Two of the HDF biofilm-forming FimH variants were insensitive to mannose inhibition and represent novel phenotypes not previously identified in naturally occurring isolates. Characterization of our enriched clones revealed some similarities to amino acid alterations that occur in urinary tract infection (UTI) strains. Subsequent screening of a selection of UTI FimH variants demonstrated that they too could promote biofilm formation on abiotic surfaces under HDF conditions. Interestingly, the same correlation was not observed for commensal FimH variants. FimH is a multifaceted protein prone to rapid microevolution. In addition to its previously documented roles in adherence and invasion, we have now demonstrated its function in biofilm formation on abiotic surfaces subjected to HDF conditions. The study indicates that UTI FimH variants possess adaptations that enhance biofilm formation and suggests a novel role for FimH in UTIs associated with medical implants such as catheters.

scholarly journals Potentially Pathogenic Escherichia coli Can Form a Biofilm under Conditions Relevant to the Food Production Chain

2013 ◽  
Vol 80 (7) ◽  
pp. 2042-2049 ◽  
Author(s):  
Live L. Nesse ◽  
Camilla Sekse ◽  
Kristin Berg ◽  
Karianne C. S. Johannesen ◽  
Heidi Solheim ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Food Production ◽  
Bacterial Cells ◽  
Production Chain ◽  
Content Type ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Different Temperatures ◽  
Pathogenic Serotypes ◽  
Sheep Feces

ABSTRACTThe biofilm-producing abilities of potentially human-pathogenic serotypes ofEscherichia colifrom the ovine reservoir were studied at different temperatures and on different surfaces. A possible influence of the hydrophobicity of the bacterial cells, as well as the presence of two virulence factors, the Shiga toxin-encoding (Stx) bacteriophage and theeaegene, was also studied. A total of 99E. coliisolates of serotypes O26:H11, O103:H2, and O103:H25 isolated from sheep feces were included. The results show that isolates of all threeE. coliserotypes investigated can produce biofilm on stainless steel, glass, and polystyrene at 12, 20, and 37°C. There was a good general correlation between the results obtained on the different surfaces.E. coliO103:H2 isolates produced much more biofilm than those of the other two serotypes at all three temperatures. In addition, isolates of serotype O26:H11 produced more biofilm than those of O103:H25 at 37°C. The hydrophobicity of the isolates varied between serotypes and was also influenced by temperature. The results strongly indicated that hydrophobicity influenced the attachment of the bacteria rather than their ability to form biofilm once attached. Isolates with theeaegene produced less biofilm at 37°C than isolates without this gene. The presence of a Stx bacteriophage did not influence biofilm production. In conclusion, our results show that potentially human-pathogenicE. colifrom the ovine reservoir can form biofilm on various surfaces and at several temperatures relevant for food production and handling.

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 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 Application of Whole Genome Sequencing to Aid in Deciphering the Persistence Potential of Listeria monocytogenes in Food Production Environments

2021 ◽  
Vol 9 (9) ◽  
pp. 1856
Author(s):  
Natalia Unrath ◽  
Evonne McCabe ◽  
Guerrino Macori ◽  
Séamus Fanning
Keyword(s):  
Listeria Monocytogenes ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Biofilm Formation ◽  
Food Production ◽  
Mortality Rates ◽  
Foodborne Illness ◽  
Whole Genome ◽  
Bioinformatic Tools ◽  
Production Environments

Listeria monocytogenes is the etiological agent of listeriosis, a foodborne illness associated with high hospitalizations and mortality rates. This bacterium can persist in food associated environments for years with isolates being increasingly linked to outbreaks. This review presents a discussion of genomes of Listeria monocytogenes which are commonly regarded as persisters within food production environments, as well as genes which are involved in mechanisms aiding this phenotype. Although criteria for the detection of persistence remain undefined, the advent of whole genome sequencing (WGS) and the development of bioinformatic tools have revolutionized the ability to find closely related strains. These advancements will facilitate the identification of mechanisms responsible for persistence among indistinguishable genomes. In turn, this will lead to improved assessments of the importance of biofilm formation, adaptation to stressful conditions and tolerance to sterilizers in relation to the persistence of this bacterium, all of which have been previously associated with this phenotype. Despite much research being published around the topic of persistence, more insights are required to further elucidate the nature of true persistence and its implications for public health.

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.

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