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Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat
Published on: October 29, 2014
Characterizing the pathotype of neonatal meningitis causing Escherichia coli (NMEC)
D S S Wijetunge1, S Gongati2, C DebRoy3
1Department of Veterinary and Biomedical Sciences, Pennsylvania State University, 115 Henning Bldg, University Park, Pennsylvania, USA. dsw179@psu.edu.
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.
Insights
Neonatal meningitis-causing Escherichia coli (NMEC) strains can be distinguished from commensal E. coli using virulence genotyping and phylogrouping. These methods identify key genetic markers and phylogenetic groups specific to NMEC, aiding in pathotype definition.
Area of Science:
- Microbiology
- Infectious Diseases
- Genetics
Background:
- Neonatal meningitis-causing Escherichia coli (NMEC) is a leading Gram-negative pathogen in newborns.
- Strain diversity in NMEC virulence factors complicates pathotype definition.
Purpose of the Study:
- To identify genotypic and phenotypic markers distinguishing NMEC from commensal E. coli.
- To aid in defining the NMEC pathotype.
Main Methods:
- Comparative analysis of 53 NMEC and 48 fecal E. coli (HFEC) isolates.
- Utilized phenotypic methods: serotyping, serum bactericidal assay, biofilm assay, antimicrobial susceptibility, and cell invasion assays.
- Employed genotypic methods: phylogrouping, virulence genotyping, and pulsed-field gel electrophoresis.
Main Results:
- NMEC predominantly belongs to phylogenetic group B2, while HFEC are mainly in groups A and D.
- Specific O and H serotypes (O1, O8, O18, H7) were prevalent in NMEC but absent or rare in HFEC.
- NMEC strains showed higher prevalence of virulence genes (kpsII, K1, neuC, iucC, sitA, vat) and biofilm formation compared to HFEC.
Conclusions:
- Virulence genotyping and phylogrouping are valuable tools for defining the NMEC pathotype.
- These genetic approaches can differentiate pathogenic NMEC from commensal E. coli strains.
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