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. 2001 Jul;49(1):47–55. doi: 10.1136/gut.49.1.47

Escherichia coli strains colonising the gastrointestinal tract protect germfree mice against Salmonella typhimurium infection

S Hudault 1, J Guignot 1, A Servin 1
PMCID: PMC1728375  PMID: 11413110

Abstract

BACKGROUND—Escherichia coli is part of the normal gastrointestinal microflora which exerts a barrier effect against enteropathogens. Several E coli strains develop a protective effect against other Enterobacteriaceae.
AIMS—Two E coli strains, EM0, a human faecal strain, and JM105 K-12 were tested for their ability to prevent in vivo and in vitro infection by Salmonella typhimurium C5.
METHODS—Inhibition of C5 cell invasion by E coli was investigated in vitro using Caco-2/TC7 cells. The protective effect of E coli was examined in vivo in germfree or conventional C3H/He/Oujco mice orally infected by the lethal strain C5.
RESULTS—EMO expresses haemolysin and cytotoxic necrotising factor in vitro. In vitro, the two strains did not prevent the growth of C5 by secreted microcins or modified cell invasion of C5. In vivo, establishment of EM0 or JM105 in the gut of germfree mice resulted in a significant increase in the number of surviving mice: 11/12 and 9/12, respectively, at 58 days after infection (2×106/mouse) versus 0/12 in control germfree group at 13 days after infection. Colonisation level and translocation rate of C5 were significantly reduced during the three days after infection. In contrast, no reduction in faecal C5 excretion was observed in C5 infected conventional mice (1×108/mouse) receiving the EM0 or JM105 cultures daily.
CONCLUSIONS—Establishment of E coli strains, which do not display antimicrobial activity, protects germfree mice against infection and delays the establishment of C5 in the gut. Possible mechanisms of defence are discussed.


Keywords: Escherichia coli; gastrointestinal infection; Salmonella; germfree mice; bacterial antagonism

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Figure 1  .

Figure 1  

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Polymerase chain reaction (PCR) amplification of hly, cnf, and cdt from E coli EM0 strain. Lane 1: 100 bp molecular weight marker; lanes 2-4: hly primers; lanes 5-7: cnf primers; lanes 8-10: cdt primers; lane 11: 1 kb molecular weight markers; lanes 3, 6, and 9: E coli EM0; lanes 4, 7, and 10: E coli JM105; lane 2: E coli SE124, hly+; lane 5: E coli J96, cnf +; lane 8: E coli DH5αpOMEO1, cdt+. The E coli strain EM0 showed specific amplification with the hly and cnf primers whereas the E coli JM105 strain showed no amplification.

Figure 2  .

Figure 2  

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Survival of control germfree mice (n=12) and ex germfree mice monoassociated with E coli JM105 (n=12) or EM0 (n=12), after S typhimurium C5 oral infection. C5 oral infection (2×106 cfu/mice) occurred eight days after inoculation of E coli. Significant difference between control germfree group and EM0 and JM105 diassociated groups (p<0.0001).

Figure 3  .

Figure 3  

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Faecal excretion of S typhimurium C5 from conventional mice treated daily with E coli strains EM0 and JM105, after C5 oral infection. Control and treated mice were infected with a single dose of S typhimurium C5 (1×108 cfu/mouse) on day 0. The treatment consisted of 0.2 ml of LB broth (control) or E coli culture (2×108 cfu/mouse) and was given on day 0 and then daily for seven days after infection. Infection and treatment were achieved using a gastric probe.

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