Bacteraemia caused by Escherichia coli is a growing problem with a significant mortality. The factors that influence the acquisition and outcome of these infections are not clear. Here, we have linked detailed genetic data from the whole-genome sequencing of 162 bacteraemic isolates collected in Scotland, UK, in 2013–2015, with clinical data in order to delineate bacterial and host factors that influence the acquisition in hospital or the community, outcome and antibiotic resistance. We identified four major sequence types (STs) in these isolates: ST131, ST69, ST73 and ST95. Nearly 50 % of the bacteraemic isolates had a urinary origin. ST69 was genetically distinct from the other STs, with significantly less sharing of accessory genes and with a distinct plasmid population. Virulence genes were widespread and diversely distributed between the dominant STs. ST131 was significantly associated with hospital-associated infections (HAIs), and ST69 with those from the community. However, there was no association of ST with outcome, although patients with HAI had a higher immediate mortality compared to those with community-associated infections (CAIs). Genome-wide association studies revealed genes involved in antibiotic persistence as significantly associated with HAIs and those encoding elements of a type VI secretion system with CAIs. Antibiotic resistance was common, and there were networks of correlated resistance genes and phenotypic antibiotic resistance. This study has revealed the complex interactions between the genotype of E. coli and its ability to cause bacteraemia, and some of the determinants influencing hospital or community acquisition. In part, these are shaped by antibiotic usage, but strain-specific factors are also important.
Blood stream invasion by Escherichia coli is the commonest cause of bacteremia in the UK and elsewhere with an attributable mortality of about 15–20 %; antibiotic resistance to multiple agents is common in this microbe and is associated with worse outcomes. Genes conferring antimicrobial resistance, and their frequent location on horizontally transferred genetic elements is well-recognised, but the origin of these determinants, and their ability to be maintained and spread within clinically-relevant bacterial populations is unclear. Here, we set out to examine the distribution of antimicrobial resistance genes in chromosomes and plasmids of 16 bloodstream isolates of E. coli from patients within Scotland, and how these genes are maintained and spread. Using a combination of short and long-read whole genome sequencing methods, we were able to assemble complete sequences of 44 plasmids, with 16 Inc group F and 20 col plasmids; antibiotic resistance genes located almost exclusively within the F group. bla CTX-M15 genes had re-arranged in some strains into the chromosome alone (five strains), while others contained plasmid copies alone (two strains). Integrons containing multiple antibiotic genes were widespread in plasmids, notably many with a dfrA7 gene encoding resistance to trimethoprim, thus linking trimethoprim resistance to the other antibiotic resistance genes within the plasmids. This will allow even narrow spectrum antibiotics such as trimethoprim to act as a selective agent for plasmids containing antibiotic resistance genes mediating much broader resistance, including blaCTX-M15. To our knowledge, this is the first analysis to provide complete sequence data of chromosomes and plasmids in a collection of pathogenic human bloodstream isolates of E. coli . Our findings reveal the interplay between plasmids and integrative and conjugative elements in the maintenance and spread of antibiotic resistance genes within pathogenic E. coli .
Bacterial type III secretion systems (T3SSs) play an important role in pathogenesis of Gram-negative infections. enteropathogenic and enterohemorrhagic Escherichia coli contain a well-defined T3SS but in addition a second T3SS termed E. coli T3SS 2 (ETT2) has been described in a number of strains of E. coli. the majority of pathogenic E. coli contain elements of a genetic locus encoding ETT2, but which has undergone significant mutational attrition rendering it without predicted function. Only a very few strains have been reported to contain an intact ETT2 locus. To investigate the occurrence of the ETT2 locus in strains of human pathogenic E. coli, we carried out genomic sequencing of 162 isolates obtained from patient blood cultures in Scotland. We found that 22 of 26 sequence type (ST) 69 isolates from this collection contained an intact ETT2 together with an associated eip locus which encodes putative secreted ETT2 effectors as well as eilA, a gene encoding a putative transcriptional regulator of ETT2 associated genes. Using a reporter gene for eilA activation, we defined conditions under which this gene was differentially activated. Analysis of published E. coli genomes with worldwide representation showed that ST69 contained an intact ETT2 in these strains as well. The conservation of the genes encoding ETT2 in human pathogenic ST69 strains strongly suggests it has importance in infection, although its exact functional role remains obscure. Pathogenic bacteria possess a number of different secretion systems that facilitate host infection as well as interbacterial competition 1. One of these is the type III secretion system (T3SS), which is found in a number of different Gram-negative pathogens and is key to the ability of these microbes to cause disease 2-4. Broadly, T3SS comprise two elements: a highly conserved multiprotein structural complex that forms the conduit between the bacterial and the host cell; and various effector proteins that are translocated through this channel. Genes encoding the T3SS channel, or needle complex, are contained within pathogenicity islands comprised of a single cluster of genes 5,6. Genes encoding effectors are more widely spread within the genome and vary greatly between different bacterial species. Certain strains of Escherichia coli possess a well-defined T3SS, notably enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC). This T3SS is encoded on the locus of enterocyte effacement (LEE) and in concert with its secreted effectors, produces the characteristic attaching and effacing lesions that mediate close attachment of the pathogen with the intestinal epithelial wall 7. Whole genome sequencing of strains of EHEC revealed the presence of a putative additional T3SS 8,9 , which has been termed E. coli T3SS 2 (ETT2). The gene cluster encoding this additional T3SS shows significant homology to the SPI-1 T3SS of Salmonella serotype Typihimurium 10,11. However, unlike the LEE, the T3SS first described in EHEC and EPEC, there do not appear to be any putative effe...
We report a rare case of severe, non-contact lens-related Corynebacterium bovis corneal infection on a background of viral keratitis, resulting in corneal abscess formation with subsequent corneal perforation. An 89-year-old Caucasian lady presented with a significant epithelial defect and a dense stromal infiltrates on a background of herpes zoster keratitis, ultimately resulting in corneal perforation. Enrichment culture obtained from corneal scraping isolated the unusual organism Corynebacterium bovis. This was treated with a combination of culture-directed, targeted course of antibiotics and surgical interventions. To the best of our knowledge, this is the first reported case of profuse bacterial keratitis secondary to Corynebacterium bovis infiltration, on a background of viral keratitis, resulting in corneal abscess formation and subsequent perforation. This report highlights this rare bacterium's characteristics including its pathogenicity in causing severe corneal disease, particularly in immunosuppressed environments such as in this case, apparent antibiotic sensitivities & resistance, and potential transmission route.
A case of Listeria monocytogenes induced spontaneous bacterial peritonitis (SBP) is reported in a patient with primary biliary cirrhosis. It is an indolent illness and may not show a neutrophil reaction in peritoneal fluid. Enrichment broth was required to isolate L monocytogenes in the patient. This is not routinely used in the UK and therefore isolates may be missed. L monocytogenes remains sensitive to ampicillin, penicillin and gentamicin, but is resistant to cephalosporin antibiotics. The rising incidence of listeriosis in the population suggests that the incidence of SBP from L monocytogenes is likely to increase.
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