The <i>Galleria mellonella</i>-Enteropathogenic <i>Escherichia coli</i> Model System: Characterization of Pathogen Virulence and Insect Immune Responses

Chen, Robin Y; Keddie, B. A.

doi:10.1093/jisesa/ieab046

Cited by 16 publications

(10 citation statements)

References 84 publications

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“…In addition to this, three more groups of 10 larvae were injected with non-pathogenic E. coli ATCC25922 to evaluate whether larvae were killed by non-infection related reactions, with 10 µL of PBS to measure any lethal effects due to physical injury and positive control inoculation with pathogenic strain KP-2 belonging to ST-131. Results were analyzed by Kaplan-Meier survival curves (GraphPad Prism statistics software) ( 36 ).…”

Section: Methodsmentioning

confidence: 99%

Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant

Abdullah,

Almusallam,

et al. 2023

Microbiol Spectr

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Carbapenem resistance among Enterobacterales has become a global health concern. Clinical Escherichia coli isolates producing the metallo β-lactamase NDM have been isolated from two hospitals in Faisalabad, Pakistan. These E. coli strains were characterized by MALDI-TOF, PCR, antimicrobial susceptibility testing, XbaI and S1 nuclease pulsed-field gel electrophoresis (PFGE), conjugation assay, DNA hybridization, whole genome sequencing, bioinformatic analysis, and Galleria mellonella experiments. Thirty-four bla NDM producing E. coli strains were identified among 52 nonduplicate carbapenem-resistant strains. More than 90% of the isolates were found to be multidrug resistant by antimicrobial susceptibility testing. S1 PFGE confirmed the presence of bla NDM gene on plasmids ranging from 40 kbps to 250 kbps, and conjugation assays demonstrated transfer frequencies of bla NDM harboring plasmids ranging from 1.59 × 10 −1 to 6.46 × 10 −8 per donor. Whole genome sequencing analysis revealed bla NDM-5 as the prominent NDM subtype with the highest prevalence of bla OXA-1 , bla CTX-M-15 , aadA2 , aac(6')-Ib-cr , and tet(A ) associated resistant determinants. E. coli sequence types: ST405, ST361, and ST167 were prominent, and plasmid Inc types: FII, FIA, FIB, FIC, X3, R, and Y, were observed among all isolates. The genetic environment of bla NDM region on IncF plasmids included partial IS Aba125 , the bleomycin ble gene, and a class I integron. The virulence genes terC, traT, gad, fyuA, irp2, capU, and sitA were frequently observed, and G. mellonella experiments showed that virulence correlated with the number of virulence determinants. A strong infection control management in the hospital is necessary to check the emergence of carbapenem resistance in Gram-negative bacteria. IMPORTANCE We describe a detailed analysis of highly resistant clinical E. coli isolates from two tertiary care centers in Pakistan including carbapenem resistance as well as common co-resistance mechanisms. South Asia has a huge problem with highly resistant E. coli . However, we find that though these isolates are very difficult to treat they are of low virulence. Thus the Western world has an increasing problem with virulent E. coli that are mostly of low antibiotic resistance, whereas, South Asia has an increasing problem with highly resistant E. coli that are of low virulence potential. These observations allow us to start to devise methodologies to limit both virulence and resistance and combat problems in developing nations as well as the Western world.

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Section: Methodsmentioning

confidence: 99%

Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant

Abdullah,

Almusallam,

et al. 2023

Microbiol Spectr

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“…It is important to mention that G. mellonella in vivo model has been used to study E. coli pathogenicity. Based on this model, it is possible to evaluate the immune innate response of G. mellonella against bacteria, namely, melanization and antimicrobial peptides (AP) production (such as cecropin and gloverin) [ 45 ]. To achieve a more close experimental design to the porcine model, pig serum has been also studied, since it contains a variety of AP, inhibiting bacterial cell growth and constituting a host's innate immune system against bacterial infection [ 46 ].…”

Section: Discussionmentioning

confidence: 99%

Effect of phage vB_EcoM_FJ1 on the reduction of ETEC O9:H9 infection in a neonatal pig cell line

Ferreira

Silva

Almeida

et al. 2023

Vet Res

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Enterotoxigenic Escherichia coli (ETEC) colonizes the intestine of young pigs causing severe diarrhoea and consequently bringing high production costs. The rise of antibiotic selective pressure together with ongoing limitations on their use, demands new strategies to tackle this pathology. The pertinence of using bacteriophages as an alternative is being explored, and in this work, the efficacy of phage vB_EcoM_FJ1 (FJ1) in reducing the load of ETEC EC43-Ph (serotype O9:H9 expressing the enterotoxin STa and two adhesins F5 and F41) was assessed. Foreseeing the oral application on piglets, FJ1 was encapsulated on calcium carbonate and alginate microparticles, thus preventing phage release under adverse conditions of the simulated gastric fluid (pH 3.0) and allowing phage availability in simulated intestinal fluid (pH 6.5). A single dose of encapsulated FJ1, provided to IPEC-1 cultured cells (from intestinal epithelium of piglets) previously infected by EC43, provided bacterial reductions of about 99.9% after 6 h. Although bacteriophage-insensitive mutants (BIMs) have emerged from treatment, the consequent fitness costs associated with this new phenotype were demonstrated, comparatively to the originating strain. The higher competence of the pig complement system to decrease BIMs’ viability, the lower level of colonization of IPEC-1 cells observed with these mutants, and the increased survival rates and health index recorded in infected Galleria mellonella larvae supported this observation. Most of all, FJ1 established a proof-of-concept of the efficiency of phages to fight against ETEC in piglet intestinal cells.

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“…The pS88 plasmid conjugation strategy was based on the presence of the ColV-encoding cva operon on the pS88 plasmid of bovine AE-STEC and EPEC O80:H2 [ 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 ]. The recipient strain was the E. coli K12 DH10B strain harboring the recombinant plasmid pAuto-ColV-Switch1.0 carrying a synthetic ColV-encoding locus (received from Syngulon Company, Seraing, Belgium; ).…”

Section: Methodsmentioning

confidence: 99%

“…Besides their low cost and the possibility of testing multiple groups of larvae in a short time, G. mellonella larvae possess an innate immune system similar to mammals and can be maintained at 37 °C, like the bacterial pathogens of mammals [ 25 , 29 ]. So, it was recently observed that the STX2d phage and the type 3 secretion system (T3SS) contribute to the virulence of AE-STEC O80:H2 and EPEC O127:H6, respectively, in G. mellonella larvae [ 25 , 30 , 31 ], while the contribution of the T3SS to the virulence of AE-STEC O157:H7 is less clear [ 32 , 33 ]. However, no study has been performed yet to assess the role of the pS88 plasmids of AE-STEC and EPEC O80:H2.…”

Section: Introductionmentioning

confidence: 99%

Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage

Ikeda,

Laforêt,

Antoine

et al. 2023

Veterinary Sciences

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The invasiveness properties of Shigatoxigenic and enteropathogenic Escherichia coli (STEC and EPEC) O80:H2 in humans and calves are encoded by genes located on a pS88-like ColV conjugative plasmid. The main objectives of this study in larvae of the Galleria mellonella moth were therefore to compare the virulence of eight bovine STEC and EPEC O80:H2, of two E. coli pS88 plasmid transconjugant and STX2d phage transductant K12 DH10B, of four E. coli O80:non-H2, and of the laboratory E. coli K12 DH10B strains. Thirty larvae per strain were inoculated in the last proleg with 10 μL of tenfold dilutions of each bacterial culture corresponding to 10 to 106 colony-forming units (CFUs). The larvae were kept at 37 °C and their mortality rate was followed daily for four days. The main results were that: (i) not only the STEC and EPEC O80:H2, but also different E. coli O80:non-H2 were lethal for the larvae at high concentrations (from 104 to 106 CFU) with some variation according to the strain; (ii) the Stx2d toxin and partially the pS88 plasmid were responsible for the lethality caused by the E. coli O80:H2; (iii) the virulence factors of E. coli O80:non-H2 were not identified. The general conclusions are that, although the Galleria mellonella larvae represent a useful first-line model to study the virulence of bacterial pathogens, they are more limited in identifying their actual virulence properties.

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The Galleria mellonella-Enteropathogenic Escherichia coli Model System: Characterization of Pathogen Virulence and Insect Immune Responses

Cited by 16 publications

References 84 publications

Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant

Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant

Effect of phage vB_EcoM_FJ1 on the reduction of ETEC O9:H9 infection in a neonatal pig cell line

Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage

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The Galleria mellonella-Enteropathogenic Escherichia coli Model System: Characterization of Pathogen Virulence and Insect Immune Responses

Cited by 16 publications

References 84 publications

Whole genome-based genetic insights of bla NDM producing clinical E. coli isolates in hospital settings of Pakistant

Whole genome-based genetic insights of bla NDM producing clinical E. coli isolates in hospital settings of Pakistant

Effect of phage vB_EcoM_FJ1 on the reduction of ETEC O9:H9 infection in a neonatal pig cell line

Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage

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Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant

Whole genome-based genetic insights of bla _NDM producing clinical E. coli isolates in hospital settings of Pakistant