Extraintestinal pathogenic Escherichia coli O1:K1:H7/NM from human and avian origin: detection of clonal groups B2 ST95 and D ST59 with different host distribution

<p>Abstract</p> <p>Background</p> <p>Extraintestinal pathogenic <it>Escherichia coli </it>(ExPEC) strains of serotype O1:K1:H7/NM are frequently implicated in neonatal meningitis, urinary tract infections and septicemia in humans. They are also commonly isolated from colibacillosis in poultry. Studies to determine the similarities of ExPEC from different origins have indicated that avian strains potentially have zoonotic properties.</p> <p>Results</p> <p>A total of 59 ExPEC O1:K1:H7/NM isolates (21 from avian colibacillosis, 15 from human meningitis, and 23 from human urinary tract infection and septicemia) originated from four countries were characterized by phylogenetic PCR grouping, Multilocus Sequence Typing (MLST), Pulsed Field Gel Electrophoresis (PFGE) and genotyping based on several genes known for their association with ExPEC or avian pathogenic <it>Escherichia coli </it>(APEC) virulence.</p> <p>APEC and human ExPEC isolates differed significantly in their assignments to phylogenetic groups, being phylogroup B2 more prevalent among APEC than among human ExPEC (95% vs. 53%, <it>P </it>= 0.001), whereas phylogroup D was almost exclusively associated with human ExPEC (47% vs. 5%, <it>P </it>= 0.0000). Seven virulence genes showed significant differences, being <it>fimAv</it>_MT78and <it>sat </it>genes linked to human isolates, while <it>papGII</it>, <it>tsh</it>, <it>iron</it>, <it>cvaC </it>and <it>iss </it>were significantly associated to APEC. By MLST, 39 of 40 ExPEC belonging to phylogroup B2, and 17 of 19 belonging to phylogroup D exhibited the Sequence Types (STs) ST95 and ST59, respectively. Additionally, two novel STs (ST1013 and ST1006) were established. Considering strains sharing the same ST, phylogenetic group, virulence genotype and PFGE cluster to belong to the same subclone, five subclones were detected; one of those grouped six strains of human and animal origin from two countries.</p> <p>Conclusion</p> <p>Present results reveal that the clonal group B2 O1:K1:H7/NM ST95, detected in strains of animal and human origin, recovered from different dates and geographic sources, provides evidence that some APEC isolates may act as potential pathogens for humans and, consequently, poultry as a foodborne source, suggesting no host specificity for this type of isolates. A novel and important finding has been the detection of the clonal group D O1:K1:H7/NM ST59 almost exclusively in humans, carrying pathogenic genes linked to the phylogenetic group D. This finding would suggest D O1:K1:H7/NM ST59 as a host specific pathotype for humans.</p

Full Sequence and Comparative Analysis of the Plasmid pAPEC-1 of Avian Pathogenic E. coli χ7122 (O78∶K80∶H9)

(APEC), are very diverse. They cause a complex of diseases in Human, animals, and birds. Even though large plasmids are often associated with the virulence of ExPEC, their characterization is still in its infancy., are also present in the sequence of pAPEC-1. The comparison of the pAPEC-1 sequence with the two available plasmid sequences reveals more gene loss and reorganization than previously appreciated. The presence of pAPEC-1-associated genes is assessed in human ExPEC by PCR. Many patterns of association between genes are found.The pathotype typical of pAPEC-1 was present in some human strains, which indicates a horizontal transfer between strains and the zoonotic risk of APEC strains. ColV plasmids could have common virulence genes that could be acquired by transposition, without sharing genes of plasmid function

Spleen transcriptome response to infection with avian pathogenic Escherichia coli in broiler chickens

<p>Abstract</p> <p>Background</p> <p>Avian pathogenic <it>Escherichia coli </it>(APEC) is detrimental to poultry health and its zoonotic potential is a food safety concern. Regulation of antimicrobials in food-production animals has put greater focus on enhancing host resistance to bacterial infections through genetics. To better define effective mechanism of host resistance, global gene expression in the spleen of chickens, harvested at two times post-infection (PI) with APEC, was measured using microarray technology, in a design that will enable investigation of effects of vaccination, challenge, and pathology level.</p> <p>Results</p> <p>There were 1,101 genes significantly differentially expressed between severely infected and non-infected groups on day 1 PI and 1,723 on day 5 PI. Very little difference was seen between mildly infected and non-infected groups on either time point. Between birds exhibiting mild and severe pathology, there were 2 significantly differentially expressed genes on day 1 PI and 799 on day 5 PI. Groups with greater pathology had more genes with increased expression than decreased expression levels. Several predominate immune pathways, Toll-like receptor, Jak-STAT, and cytokine signaling, were represented between challenged and non-challenged groups. Vaccination had, surprisingly, no detectible effect on gene expression, although it significantly protected the birds from observable gross lesions. Functional characterization of significantly expressed genes revealed unique gene ontology classifications during each time point, with many unique to a particular treatment or class contrast.</p> <p>Conclusions</p> <p>More severe pathology caused by APEC infection was associated with a high level of gene expression differences and increase in gene expression levels. Many of the significantly differentially expressed genes were unique to a particular treatment, pathology level or time point. The present study not only investigates the transcriptomic regulations of APEC infection, but also the degree of pathology associated with that infection. This study will allow for greater discovery into host mechanisms for disease resistance, providing targets for marker assisted selection and advanced drug development.</p

Sequencing and functional annotation of avian pathogenic Escherichia coli serogroup O78 strains reveals the evolution of E. coli lineages pathogenic for poultry via distinct mechanisms

Avian pathogenic Escherichia coli (APEC) causes respiratory and systemic disease in poultry. Sequencing of a multilocus sequence type 95 (ST95) serogroup O1 strain previously indicated that APEC resembles E. coli causing extraintestinal human diseases. We sequenced the genomes of two strains of another dominant APEC lineage (ST23 serogroup O78 strains χ7122 and IMT2125) and compared them to each other and to the reannotated APEC O1 sequence. For comparison, we also sequenced a human enterotoxigenic E. coli (ETEC) strain of the same ST23 serogroup O78 lineage. Phylogenetic analysis indicated that the APEC O78 strains were more closely related to human ST23 ETEC than to APEC O1, indicating that separation of pathotypes on the basis of their extraintestinal or diarrheagenic nature is not supported by their phylogeny. The accessory genome of APEC ST23 strains exhibited limited conservation of APEC O1 genomic islands and a distinct repertoire of virulence-associated loci. In light of this diversity, we surveyed the phenotype of 2,185 signature-tagged transposon mutants of χ7122 following intra-air sac inoculation of turkeys. This procedure identified novel APEC ST23 genes that play strain- and tissue-specific roles during infection. For example, genes mediating group 4 capsule synthesis were required for the virulence of χ7122 and were conserved in IMT2125 but absent from APEC O1. Our data reveal the genetic diversity of E. coli strains adapted to cause the same avian disease and indicate that the core genome of the ST23 lineage serves as a chassis for the evolution of E. coli strains adapted to cause avian or human disease via acquisition of distinct virulence genes

Characterizing the pathotype of neonatal meningitis causing <i>Escherichia coli</i> (NMEC)

Background Neonatal meningitis-causing Escherichia coli (NMEC) is the predominant Gram-negative bacterial pathogen associated with meningitis in newborn infants. High levels of heterogeneity and diversity have been observed in the repertoire of virulence traits and other characteristics among strains of NMEC making it difficult to define the NMEC pathotype. The objective of the present study was to identify genotypic and phenotypic characteristics of NMEC that can be used to distinguish them from commensal E. coli. Methods A total of 53 isolates of NMEC obtained from neonates with meningitis and 48 isolates of fecal E. coli obtained from healthy individuals (HFEC) were comparatively evaluated using five phenotypic (serotyping, serum bactericidal assay, biofilm assay, antimicorbial susceptibility testing, and in vitro cell invasion assay) and three genotypic (phylogrouping, virulence genotyping, and pulsed-field gel electrophoresis) methods. Results A majority (67.92 %) of NMEC belonged to B2 phylogenetic group whereas 59 % of HFEC belonged to groups A and D. Serotyping revealed that the most common O and H types present in NMEC tested were O1 (15 %), O8 (11.3 %), O18 (13.2 %), and H7 (25.3 %). In contrast, none of the HFEC tested belonged to O1 or O18 serogroups. The most common serogroup identified in HFEC was O8 (6.25 %). The virulence genotyping reflected that more than 70 % of NMEC carried kpsII, K1, neuC, iucC, sitA, and vat genes with only less than 27 % of HFEC possessing these genes. All NMEC and 79 % of HFEC tested were able to invade human cerebral microvascular endothelial cells. No statistically significant difference was observed in the serum resistance phenotype between NMEC and HFEC. The NMEC strains demonstrated a greater ability to form biofilms in Luria Bertani broth medium than did HFEC (79.2 % vs 39.9 %). Conclusion The results of our study demonstrated that virulence genotyping and phylogrouping may assist in defining the potential NMEC pathotype

The GimA Locus of Extraintestinal Pathogenic E. coli: Does Reductive Evolution Correlate with Habitat and Pathotype?

IbeA (invasion of brain endothelium), which is located on a genomic island termed GimA, is involved in the pathogenesis of several extraintestinal pathogenic E. coli (ExPEC) pathotypes, including newborn meningitic E. coli (NMEC) and avian pathogenic E. coli (APEC). To unravel the phylogeny of GimA and to investigate its island character, the putative insertion locus of GimA was determined via Long Range PCR and DNA-DNA hybridization in 410 E. coli isolates, including APEC, NMEC, uropathogenic (UPEC), septicemia-associated E. coli (SEPEC), and human and animal fecal isolates as well as in 72 strains of the E. coli reference (ECOR) collection. In addition to a complete GimA (∼20.3 kb) and a locus lacking GimA we found a third pattern containing a 342 bp remnant of GimA in this strain collection. The presence of GimA was almost exclusively detected in strains belonging to phylogenetic group B2. In addition, the complete GimA was significantly more frequent in APEC and NMEC strains while the GimA remnant showed a higher association with UPEC strains. A detailed analysis of the ibeA sequences revealed the phylogeny of this gene to be consistent with that obtained by Multi Locus Sequence Typing of the strains. Although common criteria for genomic islands are partially fulfilled, GimA rather seems to be an ancestral part of phylogenetic group B2, and it would therefore be more appropriate to term this genomic region GimA locus instead of genomic island. The existence of two other patterns reflects a genomic rearrangement in a reductive evolution-like manner

Identification et caractérisation d'un îlot génomique d'une souche d'Escherichia coli pathogène aviaire

Certaines souches d'E. coli sont pathogènes et provoquent des infections intestinales ou extra-intestinales chez l'homme et les animaux. Chez les volailles, les APEC (Avian Pathogenic E. coli), provoquent une infection systémique à point de départ respiratoire pouvant conduire à la mort de l'animal. Plusieurs facteurs de virulence ont été identifiés pour ces souches, mais ne suffisent pas à expliquer tout le processus infectieux. L'identification de nouveaux facteurs de virulence devrait permettre de mieux comprendre le schéma infectieux de ces souches. Les facteurs de virulence bactériens sont fréquemment situés au sein d'éléments mobiles (plasmides, transposons ou îlots de pathogénicité (PAIs)). Les PAIs sont souvent associés à leurs extrémités au loci d'ARNt. Nous avons mis en évidence chez la souche APEC BEN2908, un nouvel îlot génomique nommé AGI-3 pour "APEC Genomic Island 3" inséré au locus selC et possédant la plupart des caractéristiques des PAIs. AGI-3 est encadré par des séquences directes répétées, possède des gènes codant pour des éléments de mobilité potentiels et est de taille importante (49 600 pb). Cet îlot est composé de 49 ORFs, dont 3, aec35 à aec37, semblent impliqués dans le métabolisme des sucres. Par comparaison du phénotype de la souche sauvage et de son mutant isogénique délété des ORFs aec35-37, nous avons mis en évidence l'implication de ce groupe de gènes dans l'assimilation des hydrates de carbone. Des reproductions expérimentales de la colibacillose chez le poulet ont permis de déterminer leur implication dans les étapes précoces de l'infection. La présence d'une intégrase de phage ainsi que des sites att indiquent qu'AGI-3 a probablement été acquis par transfert horizontal. En faveur de cette hypothèse, nous avons montré qu'AGI-3 pouvait exister sous forme épisomale dans le cytoplasme bactérien et être transféré à une souche d'E. coli K-12 par conjugaison. Ces données montrent qu'AGI-3 est un vecteur potentiel de dissémination de gènes de virulence entre souches bactériennes.Escherichia coli is also of causing a variety of intestinal or extraintestinal infections in humans and animals. In poultry flocks APEC (Avian pathogenic E. coli) are involved in a systemic infection initiated in the respiratory tract that can be lethal for the animals. Several bacterial factors have been associated with their virulence, however the mechanisms underlying pathogenicity are not fully understood yet. Identification of new factors involved in the pathogenesis of APEC should lead to a better understanding of the infectious process. In bacteria, virulence factors are often located in mobile genetic elements (transposons or pathogenicity islands (PAIs)). PAIs are often associated with tRNA loci. We identified a new PAI in the APEC strain BEN2908, that we named AGI-3 for "APEC genomic island 3".AGI-3 is located at the 3'-end of the selC tRNA and possesses several characteristics of PAIs (flanked by direct repeats, presence of genes coding for putative mobile elements and large size 49 600 pb). AGI-3 shows a mosaic structure. It is composed of 49 ORFs three of them, aec35 to aec37, seem to be implicated in sugar metabolism. Comparing the abilities of the wild type strain BEN2908 and of the isogenic mutant, deleted for the three genes, to assimilate carbohydrates and to induce colibacillosis in a chicken experimental model, we demonstrated the implication of the cluster in the uptake of seven carbohydrates and in pathogenesis at the early steps of the infection. The presence of a putative active integrase gene as well as att sites suggested that AGI-3 could have been acquired by horizontal gene transfer. We demonstrated that AGI-3 is able to excise from the chromosome and to persist in the bacterial cytoplasm as a circular form and be transferred to an E. coli K-12 strain by conjugation. These data show that AGI-3 could be a potential vector for dissemination of virulence genes between bacterial strains.TOURS-BU Sciences Pharmacie (372612104) / SudocSudocFranceF