Relationship between heat-labile enterotoxin secretion capacity and virulence in wild type porcine-origin enterotoxigenic Escherichia coli strains

Prageeth Wijemanne, Jun Xing, Emil M. Berberov, David B. Marx, David H. Francis, Rodney A. Moxley

Research output: Contribution to journalArticle

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Abstract

Heat-labile enterotoxin (LT) is an important virulence factor secreted by some strains of enterotoxigenic Escherichia coli (ETEC). The prototypic human-origin strain H10407 secretes LT via a type II secretion system(T2SS). We sought to determine the relationship between the capacity to secrete LT and virulence in porcine-origin wild type (WT) ETEC strains. Sixteen WT ETEC strains isolated from cases of severe diarrheal disease were analyzed by GM1ganglioside enzyme-linked immunosorbent assay tomeasure LT concentrations in culture supernatants. All strains had detectable LT in supernatants by 2 h of culture and 1 strain, which was particularly virulent in gnotobiotic piglets (3030-2), had the highest LT secretion level all porcine-origin WT strains tested (P<0.05). The level of LT secretion (concentration in supernatants at 6-h culture) explained 92% of the variation in time-to-a-moribund-condition (R2 = 0.92, P<0.0001) in gnotobiotic piglets inoculated with either strain 3030-2, or an ETEC strain of lesser virulence (2534-86), or a non-enterotoxigenicWT strain (G58-1). All 16 porcine ETEC strains were positive by PCR analysis for the T2SS genes, gspD and gspK, and bioinformatic analysis of 4 porcine-origin strains for which complete genomic sequences were available revealed a T2SS with a high degree of homology to that of H10407.MaximumLikelihood phylogenetic trees constructed using T2SS genes gspC, gspD, gspE and homologs showed that strains 2534-86 and 3030-2 clustered together in the same clade with other porcine-origin ETEC strains in the database, UMNK88 and UMN18. Protein modeling of the ATPase gene (gspE) further revealed a direct relationship between the predicted ATP-binding capacities and LT secretion levels as follows: H10407, -8.8 kcal/mol and 199 ng/ml; 3030-2, -8.6 kcal/mol and 133 ng/ml; and 2534-86, -8.5 kcal/mol and 80 ng/ml. This study demonstrated a direct relationship between predicted ATP-binding capacity of GspE and LT secretion, and between the latter and virulence.

Original languageEnglish (US)
Article numbere0117663
JournalPloS one
Volume10
Issue number3
DOIs
StatePublished - Mar 13 2015

Fingerprint

Enterotoxigenic Escherichia coli
enterotoxigenic Escherichia coli
Enterotoxins
enterotoxins
Escherichia coli
Virulence
virulence
Swine
Hot Temperature
secretion
heat
swine
Germ-Free Life
Adenosine Triphosphate
Genes
Virulence Factors
Computational Biology
binding capacity
Adenosine Triphosphatases
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Relationship between heat-labile enterotoxin secretion capacity and virulence in wild type porcine-origin enterotoxigenic Escherichia coli strains. / Wijemanne, Prageeth; Xing, Jun; Berberov, Emil M.; Marx, David B.; Francis, David H.; Moxley, Rodney A.

In: PloS one, Vol. 10, No. 3, e0117663, 13.03.2015.

Research output: Contribution to journalArticle

Wijemanne, Prageeth ; Xing, Jun ; Berberov, Emil M. ; Marx, David B. ; Francis, David H. ; Moxley, Rodney A. / Relationship between heat-labile enterotoxin secretion capacity and virulence in wild type porcine-origin enterotoxigenic Escherichia coli strains. In: PloS one. 2015 ; Vol. 10, No. 3.
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