Laboratory adapted <i><scp>E</scp>scherichia coli</i><scp>K</scp>‐12 becomes a pathogen of <i><scp>C</scp>aenorhabditis elegans</i> upon restoration of <scp>O</scp> antigen biosynthesis

Browning, Douglas F.; Wells, Timothy J.; França, Fernanda L. S.; Morris, Faye C.; Sevastsyanovich, Yanina R.; Bryant, Jack A.; Johnson, Matthew D.; Lund, Peter A.; Cunningham, Adam F.; Hobman, Jon L.; May, Robin C.; Webber, Mark A.; Henderson, Ian R.

doi:10.1111/mmi.12144

Cited by 73 publications

(63 citation statements)

References 55 publications

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“…Similarly, expression of OAg by the apathogenic E. coli strain, K-12, rendered this organism fully virulent in the same model, underlining the importance of this surface structure to the colonization of an invertebrate host [14]. …”

Section: Introductionmentioning

confidence: 99%

LPS Structure and PhoQ Activity Are Important for Salmonella Typhimurium Virulence in the Gallleria mellonella Infection Model

et al. 2013

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The larvae of the wax moth, Galleria mellonella , have been used experimentally to host a range of bacterial and fungal pathogens. In this study we evaluated the suitability of G . mellonella as an alternative animal model of Salmonella infection. Using a range of inoculum doses we established that the LD50 of Salmonella Typhimurium strain NCTC 12023 was 3.6 × 103 bacteria per larva. Further, a set of isogenic mutant strains depleted of known virulence factors was tested to identify determinants essential for S . Typhimurium pathogenesis. Mutants depleted of one or both of the type III secretion systems encoded by Salmonella Pathogenicity Islands 1 and 2 showed no virulence defect. In contrast, we observed reduced pathogenic potential of a phoQ mutant indicating an important role for the PhoPQ two-component signal transduction system. Lipopolysaccharide (LPS) structure was also shown to influence Salmonella virulence in G . mellonella . A waaL (rfaL) mutant, which lacks the entire O-antigen (OAg), was virtually avirulent, while a wzz ST/wzz fepE double mutant expressing only a very short OAg was highly attenuated for virulence. Furthermore, shortly after infection both LPS mutant strains showed decreased replication when compared to the wild type in a flow cytometry-based competitive index assay. In this study we successfully established a G . mellonella model of S . Typhimurium infection. By identifying PhoQ and LPS OAg length as key determinants of virulence in the wax moth larvae we proved that there is an overlap between this and other animal model systems, thus confirming that the G . mellonella infection model is suitable for assessing aspects of Salmonella virulence function.

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Section: Introductionmentioning

confidence: 99%

LPS Structure and PhoQ Activity Are Important for Salmonella Typhimurium Virulence in the Gallleria mellonella Infection Model

et al. 2013

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“…This deletion appears to have occurred through recombination of the ISE3C element with the wzy gene, which encodes the O-antigen polymerase, resulting in truncation of wzy and loss of the wzx and wbuS genes encoding the O-antigen flippase and a glycosyltransferase, respectively [25–27]. The loss of O-antigen expression in E. coli is a common feature of strains cultured in the laboratory for extended periods of time [28, 29]. …”

Section: Resultsmentioning

confidence: 99%

Sequencing a piece of history: complete genome sequence of the original Escherichia coli strain

et al. 2017

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In 1885, Theodor Escherich first described the Bacillus coli commune, which was subsequently renamed Escherichia coli. We report the complete genome sequence of this original strain (NCTC 86). The 5 144 392 bp circular chromosome encodes the genes for 4805 proteins, which include antigens, virulence factors, antimicrobial-resistance factors and secretion systems, of a commensal organism from the pre-antibiotic era. It is located in the E. coli A subgroup and is closely related to E. coli K-12 MG1655. E. coli strain NCTC 86 and the non-pathogenic K-12, C, B and HS strains share a common backbone that is largely co-linear. The exception is a large 2 803 932 bp inversion that spans the replication terminus from gmhB to clpB. Comparison with E. coli K-12 reveals 41 regions of difference (577 351 bp) distributed across the chromosome. For example, and contrary to current dogma, E. coli NCTC 86 includes a nine gene sil locus that encodes a silver-resistance efflux pump acquired before the current widespread use of silver nanoparticles as an antibacterial agent, possibly resulting from the widespread use of silver utensils and currency in Germany in the 1800s. In summary, phylogenetic comparisons with other E. coli strains confirmed that the original strain isolated by Escherich is most closely related to the non-pathogenic commensal strains. It is more distant from the root than the pathogenic organisms E. coli 042 and O157 : H7; therefore, it is not an ancestral state for the species.

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“…In one study, it was demonstrated that Salmonella Typhimurium required an intact LPS to resist the immune response, persist and multiply within G. mellonella [52]. A recent study by Browning and others (2013) showed the essential nature of O-antigen production as a key virulence determinant mediating killing of Caenorhabditis elegans worms by E. coli [53]. Browning demonstrated that regeneration of the O-antigen biosynthesis cluster renders E. coli K-12 strain MG1655 pathogenic in C. elegans .…”

Section: Discussionmentioning

confidence: 99%

Galleria mellonella Infection Model Demonstrates High Lethality of ST69 and ST127 Uropathogenic E. coli

Alghoribi

Gibreel

Dodgson³

et al. 2014

PLoS ONE

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Galleria mellonella larvae are an alternative in vivo model for investigating bacterial pathogenicity. Here, we examined the pathogenicity of 71 isolates from five leading uropathogenic E. coli (UPEC) lineages using G. mellonella larvae. Larvae were challenged with a range of inoculum doses to determine the 50% lethal dose (LD50) and for analysis of survival outcome using Kaplan-Meier plots. Virulence was correlated with carriage of a panel of 29 virulence factors (VF). Larvae inoculated with ST69 and ST127 isolates (104 colony-forming units/larvae) showed significantly higher mortality rates than those infected with ST73, ST95 and ST131 isolates, killing 50% of the larvae within 24 hours. Interestingly, ST131 isolates were the least virulent. We observed that ST127 isolates are significantly associated with a higher VF-score than isolates of all other STs tested (P≤0.0001), including ST69 (P<0.02), but one ST127 isolate (strain EC18) was avirulent. Comparative genomic analyses with virulent ST127 strains revealed an IS1 mediated deletion in the O-antigen cluster in strain EC18, which is likely to explain the lack of virulence in the larvae infection model. Virulence in the larvae was not correlated with serotype or phylogenetic group. This study illustrates that G. mellonella are an excellent tool for investigation of the virulence of UPEC strains. The findings also support our suggestion that the incidence of ST127 strains should be monitored, as these isolates have not yet been widely reported, but they clearly have a pathogenic potential greater than that of more widely recognised clones, including ST73, ST95 or ST131.

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Laboratory adapted Escherichia coliK‐12 becomes a pathogen of Caenorhabditis elegans upon restoration of O antigen biosynthesis

Cited by 73 publications

References 55 publications

LPS Structure and PhoQ Activity Are Important for Salmonella Typhimurium Virulence in the Gallleria mellonella Infection Model

LPS Structure and PhoQ Activity Are Important for Salmonella Typhimurium Virulence in the Gallleria mellonella Infection Model

Sequencing a piece of history: complete genome sequence of the original Escherichia coli strain

Galleria mellonella Infection Model Demonstrates High Lethality of ST69 and ST127 Uropathogenic E. coli

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