Identification of a genomic cluster related to hypersecretion of intestinal mucus and mucinolytic activity of atypical enteropathogenic Escherichia coli (aEPEC)

Front Cell Infect Microbiol. 2024 Dec 4:14:1393369. doi: 10.3389/fcimb.2024.1393369. eCollection 2024.

Abstract

Enteropathogenic Escherichia coli (EPEC) strains are subdivided into typical (tEPEC) and atypical (aEPEC) according to the presence or absence of a virulence-associated plasmid called pEAF. Our research group has previously demonstrated that two aEPEC strains, 0421-1 and 3991-1, induce an increase in mucus production in a rabbit ileal loop model in vivo. This phenomenon was not observed with a tEPEC prototype strain. Few studies on aEPEC strains evaluating their capacity to induce intestinal mucus hypersecretion were done. This study aimed to investigate aEPEC strains regarding their genotypic and phenotypic characteristics, their ability to alter mucus production in an in vivo intestinal infection model, and their potential mucinolytic activity. To investigate the relationship between strains 0421-1 and 3991-1 and 11 other aEPEC strains, their serotypes, sequence types (ST), and virulence factors (VF), several sequencing and genomic analyses were carried out. The study also involved researching the reproduction of mucus hypersecretion in rabbits in vivo. We found that the two mucus-inducing strains and two other strains (1582-4 and 2531-13) shared the same phylogroup (A), ST (378), serotype (O101/O162:H33), and intimin subtype (ι2), were phylogenetically related, and induced mucus hypersecretion in vivo. A wide diversity of VFs was found among the strains, confirming their genomic heterogeneity. However, among the genes studied, no unique virulence factor or gene set was identified exclusively in the mucus-inducing strains, suggesting the multifactorial nature of this phenomenon. The two strains (1582-4 and 2531-13) closely related to the two aEPEC strains that induced mucus production in vivo also induced the phenomenon. The investigation of the mucinolytic activity revealed that all aEPEC strains used mucins as their carbon sources. Ten of the 13 aEPEC strains could cross a mucin layer, and only four adhered better to agar containing mucin than to agar without mucin. The present study paves the way for subsequent investigations into the molecular mechanisms regarding cellular interactions and responses, as well as the correlation between virulence factors and the induction of mucus production/expression during aEPEC infections.

Keywords: atypical EPEC; genomic analysis; mucinases; mucus hypersecretion; rabbit ileal loop; virulence factors.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Enteropathogenic Escherichia coli* / genetics
  • Enteropathogenic Escherichia coli* / metabolism
  • Enteropathogenic Escherichia coli* / pathogenicity
  • Escherichia coli Infections* / microbiology
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Genome, Bacterial
  • Genotype
  • Intestinal Mucosa / metabolism
  • Intestinal Mucosa / microbiology
  • Mucins / metabolism
  • Mucus* / metabolism
  • Mucus* / microbiology
  • Multigene Family
  • Phylogeny
  • Plasmids / genetics
  • Rabbits
  • Serogroup
  • Virulence / genetics
  • Virulence Factors* / genetics
  • Virulence Factors* / metabolism

Substances

  • Virulence Factors
  • Escherichia coli Proteins
  • Mucins

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) thematic research grant number 2017/14821-7 for TG; and fellowship to LT grant number 2023/03097-7, JP-S 2022/12006-2, and AS 2023/05910-7; LT received a scholarship and TK a research grant from the National Council for Science and Technological Development (CNPq) [grants 168193/2018-3 and 306396/2020-3, respectively], and AS and FS received postdoctoral fellowships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Capes) [grants: 88882.306532/2018-01 and 88887.464416/2019-00, respectively] under financial code 001. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.