Competition for proline between indigenous Escherichia coli and E. coli O157:H7 in gnotobiotic mice associated with infant intestinal microbiota and its contribution to the colonization resistance against E. coli O157:H7

Previously, we produced two groups of gnotobiotic mice, GB-3 and GB-4, which showed different responses to Escherichia coli O157:H7 challenge. E. coli O157:H7 was eliminated from GB-3, whereas GB-4 mice became carriers. It has been reported that the lag time of E. coli O157:H7 growth in 50% GB-3 caecal suspension was extended when compared to GB-4 caecal suspension. In this study, competition for nutrients between intestinal microbiota of GB-3 and GB-4 mice and E. coli O157:H7 was examined. Amino acid concentrations in the caecal contents of GB-3 and GB-4 differed, especially the concentration of proline. The supplementation of proline into GB-3 caecal suspension decreased the lag time of E. coli O157:H7 growth in vitro. When E. coli O157:H7 was cultured with each of the strains used to produce GB-3 mice in vitro, 2 strains of E. coli (proline consumers) out of 5 enterobacteriaceae strains strongly suppressed E. coli O157:H7 growth and the suppression was attenuated by the addition of proline into the medium. These results indicate that competition for proline with indigenous E. coli affected the growth of E. coli O157:H7 in vivo and may contribute to E. coli O157:H7 elimination from the intestine.

[1]  Y. Benno,et al.  Comparison of fecal microflora of elderly persons in rural and urban areas of Japan , 1989, Applied and environmental microbiology.

[2]  K. G. Hardy Plasmids : a practical approach , 1987 .

[3]  S. Biesterveld,et al.  Survival of Escherichia coli O157:H7 ATCC 43895 in a Model Apple Juice Medium with Different Concentrations of Proline and Caffeic Acid , 2001, Applied and Environmental Microbiology.

[4]  T. Mitsuoka Significance of dietary modulation of intestinal flora and intestinal environment. , 2000 .

[5]  I. Makino,et al.  Antibacterial action of bile acids against Helicobacter pylori and changes in its ultrastructural morphology: effect of unconjugated dihydroxy bile acid , 1999, Journal of Gastroenterology.

[6]  J. Que,et al.  Factors responsible for increased susceptibility of mice to intestinal colonization after treatment with streptomycin , 1986, Infection and immunity.

[7]  H. Masaki,et al.  Prevalence of enteric bacteria that inhibit growth of enterohaemorrhagic Escherichia coli O157 in humans , 2006, Epidemiology and Infection.

[8]  B. Oudega,et al.  Methods for studying colicins and their plasmids , 1987 .

[9]  S. Kamiya,et al.  Growth inhibition of Clostridium difficile by intestinal flora of infant faeces in continuous flow culture. , 1994, Journal of medical microbiology.

[10]  H. Guiot Role of competition for substrate in bacterial antagonism in the gut , 1982, Infection and immunity.

[11]  K. Wilson,et al.  Role of competition for nutrients in suppression of Clostridium difficile by the colonic microflora , 1988, Infection and immunity.

[12]  K. Itoh,et al.  Effect of organic acids on inhibition of Escherichia coli O157:H7 colonization in gnotobiotic mice associated with infant intestinal microbiota , 2007, Antonie van Leeuwenhoek.

[13]  F. Gavini,et al.  Differences in the Distribution of Bifidobacterial and Enterobacterial Species in Human Faecal Microflora of Three Different (Children, Adults, Elderly) Age Groups , 2001 .

[14]  R. Carman,et al.  Ciprofloxacin at low levels disrupts colonization resistance of human fecal microflora growing in chemostats. , 2004, Regulatory toxicology and pharmacology : RTP.

[15]  R. Roberts,et al.  Evaluation of colicins for inhibitory activity against diarrheagenic Escherichia coli strains, including serotype O157:H7 , 1996, Applied and environmental microbiology.

[16]  Y. Benno,et al.  THE INTESTINAL MICROFLORA OF INFANTS; COMPOSITION OF FECAL FLORA IN BREAST AND BOTTLE FED INFANTS , 1985 .

[17]  R. Tanaka,et al.  Distribution of Bifidobacterial Species in Human Intestinal Microflora Examined with 16S rRNA-Gene-Targeted Species-Specific Primers , 1999, Applied and Environmental Microbiology.

[18]  L. Rice,et al.  Inhibition of vancomycin-resistant enterococci by an in vitro continuous-flow competitive exclusion culture containing human stool flora. , 2001, The Journal of infectious diseases.

[19]  A. Seto,et al.  Selected faecal bacteria and nutrients essential for antagonism of Salmonella typhimurium in anaerobic continuous flow cultures. , 1991, Journal of medical microbiology.

[20]  R. Freter,et al.  Mechanisms That Control Bacterial Populations in Continuous-Flow Culture Models of Mouse Large Intestinal Flora , 1983, Infection and immunity.

[21]  Y. Benno,et al.  The Intestinal Microflora of Infants: Composition of Fecal Flora in Breast‐Fed and Bottle‐Fed Infants , 1984, Microbiology and immunology.

[22]  J. Nicoli,et al.  Antagonism against Vibrio cholerae by diffusible substances produced by bacterial components of the human faecal microbiota. , 2001, Journal of medical microbiology.

[23]  D. Grainger,et al.  A microtitre format assay for proline in human serum or plasma. , 2004, Clinica chimica acta; international journal of clinical chemistry.