A collection of 266 faecal isolates of Escherichia coli from humans was assayed for the production of mitomycin C-inducible bacteriocins and screened using a PCR-based method for the presence of eleven colicins and seven microcins. Eight different colicins were detected and all seven microcins. Of the strains examined, 38 % produced a bacteriocin, 24 % produced a colicin and 20 % produced a microcin. Of the 102 bacteriocin-producing strains, 42 % produced one type of bacteriocin, 41 % produced two, 16 % produced three and one strain was found to produce four different bacteriocins. Strains producing more than one bacteriocin were more likely to be members of E. coli genetic group B2 and less likely to belong to genetic groups A or D. Several of the bacteriocins were found to co-occur in a strain more often than would be expected by chance: microcins H47 and M; colicin Ia and microcin V; colicins B and M; colicins E1 and M; colicins E1 and Ia. No bacteriocins released as a consequence of cell lysis were found to co-associate more often than expected by chance. Three non-mutually exclusive hypotheses are presented that might explain the high frequency of multiple bacteriocin production in E. coli strains: (1) expanded killing range, (2) expanded receptor repertoire and (3) fitness benefits in different environments.

Escherichia coli is mostly a commensal of birds and mammals, including humans, where it can act as an opportunistic pathogen. It is also found in water and sediments. We investigated the phylogeny, genetic diversification, and habitat-association of 1,294 isolates representative of the phylogenetic diversity of more than 5,000 isolates from the Australian continent. Since many previous studies focused on clinical isolates, we investigated mostly other isolates originating from humans, poultry, wild animals and water. These strains represent the species genetic diversity and reveal widespread associations between phylogroups and isolation sources. The analysis of strains from the same sequence types revealed very rapid change of gene repertoires in the very early stages of divergence, driven by the acquisition of many different types of mobile genetic elements. These elements also lead to rapid variations in genome size, even if few of their genes rise to high frequency in the species. Variations in genome size are associated with phylogroup and isolation sources, but the latter determine the number of MGEs, a marker of recent transfer, suggesting that gene flow reinforces the association of certain genetic backgrounds with specific habitats. After a while, the divergence of gene repertoires becomes linear with phylogenetic distance, presumably reflecting the continuous turnover of mobile element and the occasional acquisition of adaptive genes. Surprisingly, the phylogroups with smallest genomes have the highest rates of gene repertoire diversification and fewer but more diverse mobile genetic elements. This suggests that smaller genomes are associated with higher, not lower, turnover of genetic information. Many of these genomes are from freshwater isolates and have peculiar traits, including a specific capsule, suggesting adaptation to this environment. Altogether, these data contribute to explain why epidemiological clones tend to emerge from specific phylogenetic groups in the presence of pervasive horizontal gene transfer across the species.

No abstract

The gut microbiota is important in maintaining human health, but numerous factors have the potential to alter its composition. Our aim was to examine the impact of a standard bowel preparation on the intestinal microbiota using two different techniques. Fifteen subjects undergoing colonoscopy consumed a bowel preparation comprised of 10 mg bisacodyl and 2 L polyethylene glycol. The microbiota of stool samples, collected one month before, one week before (pre-colonoscopy), and one week, one month, and three to six months after colonoscopy (post-colonoscopy) was evaluated. Two samples were taken three to six months apart from five healthy subjects who did not undergo colonoscopy. Universal primers targeting the V2–V3 region of the 16S rRNA gene were used to PCR amplify all samples for denaturing gradient gel electrophoresis (PCR-DGGE). Pre- and post-colonoscopy samples were compared using Dice’s similarity coefficients. Three samples from ten subjects who underwent colonoscopy, and both samples from the five subjects who didn’t, were used for high-throughput sequencing of the V1–V3 region of the 16S rRNA gene. Samples were curated and analysed in Mothur. Results of the DGGE analyses show that the fecal microbiota of a small number of subjects had short-term changes. High-throughput sequencing results indicated that the variation between the samples of subjects who underwent colonoscopy was no greater than the variation observed between samples from subjects who did not. We conclude that bowel preparation does not have a lasting effect on the composition of the intestinal microbiota for the majority of subjects.

23Escherichia coli is a commensal of birds and mammals, including humans. It can act as an 24 opportunistic pathogen and is also found in water and sediments. Since most population 25 studies have focused on clinical isolates, we studied the phylogeny, genetic diversification, 26 and habitat-association of 1,294 isolates representative of the phylogenetic diversity of more 27 than 5,000, mostly non-clinical, isolates originating from humans, poultry, wild animals and 28 water sampled from the Australian continent. These strains represent the species diversity 29 and show large variations in gene repertoires within sequence types. Recent gene transfer is 30 driven by mobile elements and determined by habitat sharing and by phylogroup 31 membership, suggesting that gene flow reinforces the association of certain genetic 32 backgrounds with specific habitats. The phylogroups with smallest genomes had the highest 33 rates of gene repertoire diversification and fewer but more diverse mobile genetic elements, 34 suggesting that smaller genomes are associated with higher, not lower, turnover of genetic 35 information. Many of these small genomes were in freshwater isolates suggesting that some 36 lineages are specifically adapted to this environment. Altogether, these data contribute to 37 explain why epidemiological clones tend to emerge from specific phylogenetic groups in the 38 presence of pervasive horizontal gene transfer across the species. 39 40 circulation of strains and the high plasticity of their genomes have not erased the 68 associations of certain clades with certain isolation sources. In consequence, such 69 associations might reflect local adaptation 16,45 , which would suggest frequent genetic 70 interactions between the novel adaptive changes and the strains' genomic background. 71Understanding how the evolution of gene repertoires is shaped by population structure and 72 habitats requires large-scale comparative genomics of samples with diverse sources of 73 isolation representative of natural populations of E. coli. Most of the efforts of genome 74 sequencing have been devoted to study pathogenic lineages and very few genomic data are 75 available for commensal strains, especially in wild animals, and environmental strains. Here, 76we analysed the genomes of a large collection of E. coli strains collected across many 77 human, domestic and wild animal and environmental sources in different geographic 78 locations from the Australian continent. This collection is dominated by non-clinical isolates, 79 corresponding to the main habitats of the species. We sought to understand the dynamics of 80 the evolution of gene repertoires and how it was driven by mobile genetic elements. The 81 analysis of the isolation sources in the light of phylogenetic structure and genome variation 82 suggests that adaptation varies with the habitat and the phylogenomic background. This 83 contributes to explain why known epidemiological clones of the species emerge from specific 84 phylogenetic groups, even though virulen...

SummaryEscherichia coli is a widespread commensal of the vertebrate intestinal tract. Until recently, no strong association between a particular clone and a given host species has been found. However, members of the B2 subgroup VIII clone with an O81 serotype appear to be human host specific. To determine the degree of host specificity exhibited by this clone, a PCR-based assay was used to screen 723 faecal and clinical isolates from humans, and 904 faecal isolates from animals. This clone was not detected among the animal isolates, but was discovered in people living in Africa, Europe and South America. The clone is rarely isolated from people suffering from intestinal or extraintestinal disease and is avirulent in a mouse model of extraintestinal infection. Fine-scale epidemiological analysis suggests that this clone is competitively dominant relative to other members of the B2 phylogenetic group and that it has increased in frequency over the past 20 years. This clone appears to be a good candidate for use as a probiotic, and may be suitable as an indicator of human faecal contamination in microbial source tracking studies.

Escherichia coli clonal complex 95 represents a cosmopolitan, genetically diverse lineage, and the extensive substructure observed in this lineage is epidemiologically and clinically relevant. The frequency with which CC95 strains are responsible for extraintestinal infection appears to have been stable over the past 15 years. However, the different subgroups identified within this lineage have an epidemic structure depending on the host, sample, continent, and time. Thus, the evolution and spread of strains belonging to CC95 are very different from those of another cosmopolitan human-associated clonal complex, CC131, which has increased significantly in frequency as a cause of extraintestinal infection over the past 15 years due to the evolution and spread of two very closely related, nearly monomorphic lineages.

Previous studies examining the clonal diversity of Escherichia coli populations within humans have been based on faecal isolates. In this study E. coli were isolated from biopsies taken from the terminal ileum, ascending, transverse and descending colon, and rectum of 69 individuals. Multiple isolates from each biopsy were characterized using Rep-PCR. An average of 3.5 genotypes were recovered per host, and in hosts with two or more strains, the phylogroup membership of the second most abundant strain was significantly more likely to be the same as the dominant strain. There was no indication of a non-random distribution of E. coli phylogroups among the regions of the lower intestine. In hosts with multiple genotypes, as defined by Repetitive extragenic palindromic-PCR, genotypes were non-randomly distributed among gut regions in over half the individuals. The phylogroup membership of an individual's numerically dominant strain explained some of the variation in the extent to which strains within an individual were heterogeneously distributed, with most heterogeneity observed when the numerically dominant strain belonged to phylogroups E or F, and the least when the dominant strain belonged to phylogroup B2. The results of this study support previous studies on pigs that demonstrated faecal sampling underestimates the genotype diversity present within a host.