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Infection and Immunity, March 2005, p. 1836-1846, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1836-1846.2005

Role for Flagella but Not Intimin in the Persistent Infection of the Gastrointestinal Tissues of Specific-Pathogen-Free Chicks by Shiga Toxin-Negative Escherichia coli O157:H7

Angus Best,1 Roberto M. La Ragione,1 A. Robin Sayers,2 and Martin J. Woodward1*

Department of Food and Environmental Safety,1 Centre for Epidemiology and Risk Analysis, Veterinary Laboratories Agency (Weybridge), New Haw, Addlestone, Surrey, United Kingdom2

Received 16 March 2004/ Returned for modification 24 April 2004/ Accepted 30 September 2004

Shiga toxin (Stx)-positive Escherichia coli O157:H7 readily colonize and persist in specific-pathogen-free (SPF) chicks, and we have shown that an Stx-negative E. coli O157:H7 isolate (NCTC12900) readily colonizes SPF chicks for up to 169 days after oral inoculation at 1 day of age. However, the role of intimin in the persistent colonization of poultry remains unclear. Thus, to investigate the role of intimin and flagella, which is a known factor in the persistence of non-O157 E. coli in poultry, isogenic single- and double-intimin and aflagellar mutants were constructed in E. coli O157:H7 isolate NCTC12900. These mutants were used to inoculate (105 CFU) 1-day-old SPF chicks. In general, significant attenuation of the aflagellate and intimin-aflagellate mutants, but not the intimin mutant, was noted at similar time points between 22 and 92 days after inoculation. The intimin-deficient mutant was still being shed at the end of the experiment, which was 211 days after inoculation, 84 days more than the wild type. Shedding of the aflagellar and intimin-aflagellar mutants ceased 99 and 113 days after inoculation, respectively. Histological analysis of gastrointestinal tissues from inoculated birds gave no evidence for true microcolony formation by NCTC12900 or intimin and aflagellar mutants to epithelial cells. However, NCTC12900 mutant derivatives associated with the mucosa were observed as individual cells and/or as large aggregates. Association with luminal contents was also noted. These data suggest that O157 organisms do not require intimin for the persistent colonization of chickens, whereas flagella do play a role in this process.

* Corresponding author. Mailing address: Department of Food and Environmental Safety, VLA (Weybridge), New Haw, Addlestone, Surrey KT15 3NB, United Kingdom. Phone: 01932 357582. Fax: 01932 347046. E-mail: m.j.woodward{at}vla.defra.gsi.gov.uk.

Editor: J. B. Bliska

Infection and Immunity, March 2005, p. 1836-1846, Vol. 73, No. 3
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.3.1836-1846.2005




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