scholarly journals Identification and Characterization of a Peculiarvtx2-Converting Phage Frequently Present in Verocytotoxin-Producing Escherichia coli O157 Isolated from Human Infections

2014 ◽  
Vol 82 (7) ◽  
pp. 3023-3032 ◽  
Author(s):  
Rosangela Tozzoli ◽  
Laura Grande ◽  
Valeria Michelacci ◽  
Rosa Fioravanti ◽  
David Gally ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Determinants ◽  
Differential Distribution ◽  
Content Type ◽  
Phage Types ◽  
Genes Encoding ◽  
Enterocyte Effacement ◽  
Human Infections ◽  
Identification And Characterization

ABSTRACTCertain verocytotoxin-producingEscherichia coli(VTEC) O157 phage types (PTs), such as PT8 and PT2, are associated with severe human infections, while others, such as PT21, seem to be restricted to cattle. In an attempt to delve into the mechanisms underlying such a differential distribution of PTs, we performed microarray comparison of human PT8 and animal PT21 VTEC O157 isolates. The main differences observed were in thevtx2-converting phages, with the PT21 strains bearing a phage identical to that present in the reference strain EDL933, BP933W, and all the PT8 isolates displaying lack of hybridization in some regions of the phage genome. We focused on the region spanning thegamandcIIgenes and developed a PCR tool to investigate the presence of PT8-like phages in a panel of VTEC O157 strains belonging to different PTs and determined that avtx2phage reacting with the primers deployed, which we named Φ8, was more frequent in VTEC O157 strains from human disease than in bovine strains. No differences were observed in the production of the VT2 mRNA when Φ8-positive strains were compared with VTEC O157 possessing BP933W. Nevertheless, we show that thegam-cIIregion of phage Φ8 might carry genetic determinants downregulating the transcription of the genes encoding the components of the type III secretion system borne on the locus of enterocyte effacement pathogenicity island.

scholarly journals Identification and Characterization of Spontaneous Deletions within the Sp11-Sp12 Prophage Region of Escherichia coli O157:H7 Sakai

2013 ◽  
Vol 79 (6) ◽  
pp. 1934-1941 ◽  
Author(s):  
Chun Chen ◽  
Carrie R. Lewis ◽  
Kakolie Goswami ◽  
Elisabeth L. Roberts ◽  
Chitrita DebRoy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Type Iii Secretion ◽  
Genomic Region ◽  
Escherichia Coli O157 ◽  
Content Type ◽  
E Coli ◽  
Food Borne ◽  
Genes Encoding ◽  
Identification And Characterization

ABSTRACTProphages make up 12% of the enterohemorrhagicEscherichia coligenome and play prominent roles in the evolution and virulence of this food-borne pathogen. Acquisition and loss of and rearrangements within prophage regions are the primary causes of differences in pulsed-field gel electrophoresis (PFGE) patterns among strains ofE. coliO157:H7. Sp11 and Sp12 are two tandemly integrated and putatively defective prophages carried byE. coliO157:H7 strain Sakai. In this study, we identified 3 classes of deletions that occur within the Sp11-Sp12 region, at a frequency of ca. 7.74 × 10−4. One deletion resulted in a precise excision of Sp11, and the other two spanned the junction of Sp11 and Sp12. All deletions resulted in shifts in the XbaI fragment pattern observed by PFGE. We sequenced the inducible prophage pool of Sakai but did not identify any mature phage particles corresponding to either Sp11 or Sp12. Deletions containingpchBandpsrC, which are Sp11-carried genes encoding proteins known or suspected to regulate type III secretion, did not affect the secretion levels of the EspA or EspB effector. Alignment of the Sp11-Sp12 DNA sequence with its corresponding regions in otherE. coliO157:H7 and O55:H7 strains suggested that homologous recombination rather than integrase-mediated excision is the mechanism behind these deletions. Therefore, this study provides a mechanism behind the previously observed genetic instability of this genomic region ofE. coliO157:H7.

scholarly journals Characterization of Epidemic IncI1-Iγ Plasmids Harboring Ambler Class A and C Genes in Escherichia coli and Salmonella enterica from Animals and Humans

2015 ◽  
Vol 59 (9) ◽  
pp. 5357-5365 ◽  
Author(s):  
Hilde Smith ◽  
Alex Bossers ◽  
Frank Harders ◽  
Guanghui Wu ◽  
Neil Woodford ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Salmonella Enterica ◽  
Phylogenetic Trees ◽  
Functional Study ◽  
Content Type ◽  
Gene Associations ◽  
Genes Encoding ◽  
Marked Resistance

ABSTRACTThe aim of the study was to identify the plasmid-encoded factors contributing to the emergence and spread of epidemic IncI1-Iγ plasmids obtained fromEscherichia coliandSalmonella entericaisolates from animal and human reservoirs. For this, 251 IncI1-Iγ plasmids carrying various extended-spectrum β-lactamase (ESBL) or AmpC β-lactamase genes were compared using plasmid multilocus sequence typing (pMLST). Thirty-two of these plasmids belonging to different pMLST types were sequenced using Roche 454 and Illumina platforms. Epidemic IncI1-Iγ plasmids could be assigned to various dominant clades, whereas rarely detected plasmids clustered together as a distinct clade. Similar phylogenetic trees were obtained using only the plasmid backbone sequences, showing that the differences observed between the plasmids belonging to distinct clades resulted mainly from differences between their backbone sequences. Plasmids belonging to the various clades differed particularly in the presence/absence of genes encoding partitioning and addiction systems, which contribute to stable inheritance during cell division and plasmid maintenance. Despite this, plasmids belonging to the various phylogenetic clades also showed marked resistance gene associations, indicating the circulation of successful plasmid-gene combinations. The variation intraYandexcAgenes found in IncI1-Iγ plasmids is conserved within pMLST sequence types and plays a role in incompatibility, although functional study is needed to elucidate the role of these genes in plasmid epidemiology.

scholarly journals Characterization of Multidrug-Resistant Escherichia coli Isolates from Animals Presenting at a University Veterinary Hospital

2011 ◽  
Vol 77 (20) ◽  
pp. 7104-7112 ◽  
Author(s):  
Maria Karczmarczyk ◽  
Yvonne Abbott ◽  
Ciara Walsh ◽  
Nola Leonard ◽  
Séamus Fanning
Keyword(s):  
Escherichia Coli ◽  
Molecular Mechanisms ◽  
Gene Array ◽  
Multidrug Resistant ◽  
Genetic Determinants ◽  
Gene Encoding ◽  
Content Type ◽  
E Coli ◽  
Class 1

ABSTRACTIn this study, we examined molecular mechanisms associated with multidrug resistance (MDR) in a collection ofEscherichia coliisolates recovered from hospitalized animals in Ireland. PCR and DNA sequencing were used to identify genes associated with resistance. Class 1 integrons were prevalent (94.6%) and contained gene cassettes recognized previously and implicated mainly in resistance to aminoglycosides, β-lactams, and trimethoprim (aadA1,dfrA1-aadA1,dfrA17-aadA5,dfrA12-orfF-aadA2,blaOXA-30-aadA1,aacC1-orf1-orf2-aadA1,dfr7). Class 2 integrons (13.5%) contained thedfrA1-sat1-aadA1gene array. The most frequently occurring phenotypes included resistance to ampicillin (97.3%), chloramphenicol (75.4%), florfenicol (40.5%), gentamicin (54%), neomycin (43.2%), streptomycin (97.3%), sulfonamide (98.6%), and tetracycline (100%). The associated resistance determinants detected includedblaTEM,cat,floR,aadB,aphA1,strA-strB,sul2, andtet(B), respectively. TheblaCTX-M-2gene, encoding an extended-spectrum β-lactamase (ESβL), andblaCMY-2, encoding an AmpC-like enzyme, were identified in 8 and 18 isolates, respectively. The mobility of the resistance genes was demonstrated using conjugation assays with a representative selection of isolates. High-molecular-weight plasmids were found to be responsible for resistance to multiple antimicrobial compounds. The study demonstrated that animal-associated commensalE. coliisolates possess a diverse repertoire of transferable genetic determinants. Emergence of ESβLs and AmpC-like enzymes is particularly significant. To our knowledge, theblaCTX-M-2gene has not previously been reported in Ireland.

scholarly journals Functional Characterization of EscK (Orf4), a Sorting Platform Component of the Enteropathogenic Escherichia coli Injectisome

2016 ◽  
Vol 199 (1) ◽  
Author(s):  
Eduardo Soto ◽  
Norma Espinosa ◽  
Miguel Díaz-Guerrero ◽  
Meztlli O. Gaytán ◽  
José L. Puente ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Type Iii Secretion ◽  
Diarrheal Disease ◽  
Functional Characterization ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Effector Proteins ◽  
Content Type ◽  
Enterocyte Effacement

ABSTRACT The type III secretion system (T3SS) is a supramolecular machine used by many bacterial pathogens to translocate effector proteins directly into the eukaryotic host cell cytoplasm. Enteropathogenic Escherichia coli (EPEC) is an important cause of infantile diarrheal disease in underdeveloped countries. EPEC virulence relies on a T3SS encoded within a chromosomal pathogenicity island known as the locus of enterocyte effacement (LEE). In this work, we pursued the functional characterization of the LEE-encoded protein EscK (previously known as Orf4). We provide evidence indicating that EscK is crucial for efficient T3S and belongs to the SctK (OrgA/YscK/MxiK) protein family, whose members have been implicated in the formation of a sorting platform for secretion of T3S substrates. Bacterial fractionation studies showed that EscK localizes to the inner membrane independently of the presence of any other T3SS component. Combining yeast two-hybrid screening and pulldown assays, we identified an interaction between EscK and the C-ring/sorting platform component EscQ. Site-directed mutagenesis of conserved residues revealed amino acids that are critical for EscK function and for its interaction with EscQ. In addition, we found that T3S substrate overproduction is capable of compensating for the absence of EscK. Overall, our data suggest that EscK is a structural component of the EPEC T3SS sorting platform, playing a central role in the recruitment of T3S substrates for boosting the efficiency of the protein translocation process. IMPORTANCE The type III secretion system (T3SS) is an essential virulence determinant for enteropathogenic Escherichia coli (EPEC) colonization of intestinal epithelial cells. Multiple EPEC effector proteins are injected via the T3SS into enterocyte cells, leading to diarrheal disease. The T3SS is encoded within a genomic pathogenicity island termed the locus of enterocyte effacement (LEE). Here we unravel the function of EscK, a previously uncharacterized LEE-encoded protein. We show that EscK is central for T3SS biogenesis and function. EscK forms a protein complex with EscQ, the main component of the cytoplasmic sorting platform, serving as a docking site for T3S substrates. Our results provide a comprehensive functional analysis of an understudied component of T3SSs.

scholarly journals Genetic Diversity and Virulence Potential of Shiga Toxin-Producing Escherichia coli O113:H21 Strains Isolated from Clinical, Environmental, and Food Sources

2014 ◽  
Vol 80 (15) ◽  
pp. 4757-4763 ◽  
Author(s):  
Peter C. H. Feng ◽  
Sabine Delannoy ◽  
David W. Lacher ◽  
Luis Fernando dos Santos ◽  
Lothar Beutin ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Diversity ◽  
Shiga Toxin ◽  
Food Sources ◽  
Content Type ◽  
Virulence Potential ◽  
Uremic Syndrome ◽  
Enterocyte Effacement ◽  
Human Infections ◽  
Pcr Microarray

ABSTRACTShiga toxin-producingEscherichia colistrains of serotype O113:H21 have caused severe human diseases, but they are unusual in that they do not produce adherence factors coded by the locus of enterocyte effacement. Here, a PCR microarray was used to characterize 65 O113:H21 strains isolated from the environment, food, and clinical infections from various countries. In comparison to the pathogenic strains that were implicated in hemolytic-uremic syndrome in Australia, there were no clear differences between the pathogens and the environmental strains with respect to the 41 genetic markers tested. Furthermore, all of the strains carried only Shiga toxin subtypes associated with human infections, suggesting that the environmental strains have the potential to cause disease. Most of the O113:H21 strains were closely related and belonged in the same clonal group (ST-223), but CRISPR analysis showed a great degree of genetic diversity among the O113:H21 strains.

scholarly journals Identification and Characterization of a Novel FosA7 Member from Fosfomycin-Resistant Escherichia coli Clinical Isolates from Canadian Hospitals

2020 ◽  
Vol 65 (1) ◽  
pp. e00865-20
Author(s):  
Kieran A. Milner ◽  
Denice C. Bay ◽  
David Alexander ◽  
Andrew Walkty ◽  
James A. Karlowsky ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Clinical Isolates ◽  
Phenotypic Characterization ◽  
Content Type ◽  
E Coli ◽  
Fosfomycin Resistance ◽  
Identification And Characterization

ABSTRACTHere, we characterize the fosA genes from three Escherichia coli clinical isolates recovered from Canadian patients. Each fosA sequence was individually overexpressed in E. coli BW25113, and antimicrobial susceptibility testing was performed to assess their role in fosfomycin resistance. The findings from this study identify and functionally characterize FosA3, FosA8, and novel FosA7 members and highlight the importance of phenotypic characterization of fosA genes.

scholarly journals Pas, a Novel Protein Required for Protein Secretion and Attaching and Effacing Activities of Enterohemorrhagic Escherichia coli

1998 ◽  
Vol 180 (17) ◽  
pp. 4370-4379 ◽  
Author(s):  
Andreas U. Kresse ◽  
Kai Schulze ◽  
Christina Deibel ◽  
Frank Ebel ◽  
Manfred Rohde ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Protein Secretion ◽  
Enterohemorrhagic Escherichia Coli ◽  
Host Cells ◽  
Reading Frame ◽  
E Coli ◽  
Genes Encoding ◽  
Enterocyte Effacement ◽  
Putative Member

ABSTRACT Enterohemorrhagic Escherichia coli (EHEC) exhibits a pattern of localized adherence to host cells, with the formation of microcolonies, and induces a specific histopathological phenotype collectively known as the attaching and effacing lesion. The genes encoding the products responsible for this phenotype are located on a 35-kb pathogenicity island designated the locus of enterocyte effacement, which is also shared by enteropathogenic E. coli. We have identified an open reading frame (ORF) which is located upstream of the espA, espB, andespD genes on the complementary strand and which exhibits high homology to the genes spiB fromSalmonella, yscD from Yersinia, andpscD from Pseudomonas. Localization studies showed that the encoded product is present in the cytoplasmic and inner membrane fractions of EHEC. The construction and characterization of a recombinant clone containing an in-frame deletion of this ORF demonstrated that the encoded product is a putative member of a type III system required for protein secretion. Disruption of this ORF, designated pas (protein associated with secretion), abolished the secretion of Esp proteins. The mutant adhered only poorly and lost its capacities to trigger attaching and effacing activity and to invade HeLa cells. These results demonstrate that Pas is a virulence-associated factor that plays an essential role in EHEC pathogenesis.

scholarly journals Biofilm-Forming Abilities of Shiga Toxin-Producing Escherichia coli Isolates Associated with Human Infections

2015 ◽  
Vol 82 (5) ◽  
pp. 1448-1458 ◽  
Author(s):  
Philippe Vogeleer ◽  
Yannick D. N. Tremblay ◽  
Grégory Jubelin ◽  
Mario Jacques ◽  
Josée Harel
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Food Industry ◽  
Enzymatic Digestion ◽  
Human Infection ◽  
Genetic Determinants ◽  
Independent Manner ◽  
Content Type ◽  
Human Infections

ABSTRACTForming biofilms may be a survival strategy of Shiga toxin-producingEscherichia colito enable it to persist in the environment and the food industry. Here, we evaluate and characterize the biofilm-forming ability of 39 isolates of Shiga toxin-producingEscherichia coliisolates recovered from human infection and belonging to seropathotypes A, B, or C. The presence and/or production of biofilm factors such as curli, cellulose, autotransporter, and fimbriae were investigated. The polymeric matrix of these biofilms was analyzed by confocal microscopy and by enzymatic digestion. Cell viability and matrix integrity were examined after sanitizer treatments. Isolates of the seropathotype A (O157:H7 and O157:NM), which have the highest relative incidence of human infection, had a greater ability to form biofilms than isolates of seropathotype B or C. Seropathotype A isolates were unique in their ability to produce cellulose and poly-N-acetylglucosamine. The integrity of the biofilms was dependent on proteins. Two autotransporter genes,ehaBandespP, and two fimbrial genes,z1538andlpf2, were identified as potential genetic determinants for biofilm formation. Interestingly, the ability of several isolates from seropathotype A to form biofilms was associated with their ability to agglutinate yeast in a mannose-independent manner. We consider this an unidentified biofilm-associated factor produced by those isolates. Treatment with sanitizers reduced the viability of Shiga toxin-producingEscherichia colibut did not completely remove the biofilm matrix. Overall, our data indicate that biofilm formation could contribute to the persistence of Shiga toxin-producingEscherichia coliand specifically seropathotype A isolates in the environment.

scholarly journals Identification and Characterization of Conjugative Plasmids That Encode Ciprofloxacin Resistance inSalmonella

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
Kaichao Chen ◽  
Ning Dong ◽  
Shaohua Zhao ◽  
Lizhang Liu ◽  
Ruichao Li ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Infection Control ◽  
Conjugative Plasmid ◽  
Content Type ◽  
E Coli ◽  
Conjugative Plasmids ◽  
Ciprofloxacin Resistance ◽  
Identification And Characterization

ABSTRACTThis study aimed to characterize novel conjugative plasmids that encode transferable ciprofloxacin resistance inSalmonella. In this study, 157 nonduplicatedSalmonellaisolates were recovered from food products, of which 55 were found to be resistant to ciprofloxacin. Interestingly, 37 of the 55 CiprSalmonellaisolates (67%) did not harbor any mutations in the quinolone resistance-determining regions (QRDR). SixSalmonellaisolates were shown to carry two novel types of conjugative plasmids that could transfer the ciprofloxacin resistance phenotype toEscherichia coliJ53 (azithromycin resistant [Azir]). The first type of conjugative plasmid belonged to the ∼110-kb IncFIB-type conjugative plasmids carryingqnrB-bearing andaac(6′)-Ib-cr-bearing mobile elements. Transfer of the plasmid betweenE. coliandSalmonellacould confer a ciprofloxacin MIC of 1 to 2 μg/ml. The second type of conjugative plasmid belonged to ∼240-kb IncH1/IncF plasmids carrying a single PMQR gene,qnrS. Importantly, this type of conjugative ciprofloxacin resistance plasmid could be detected in clinicalSalmonellaisolates. The dissemination of these conjugative plasmids that confer ciprofloxacin resistance poses serious challenges to public health andSalmonellainfection control.

Sign in / Sign up

Export Citation Format

Share Document