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Applied and Environmental Microbiology, September 2009, p. 5779-5786, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00507-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Contributions of O Island 48 to Adherence of Enterohemorrhagic Escherichia coli O157:H7 to Epithelial Cells In Vitro and in Ligated Pig Ileal Loops{triangledown}

Xianhua Yin,1,4 Roger Wheatcroft,1 James R. Chambers,1 Bianfang Liu,2 Jing Zhu,3 and Carlton L. Gyles4*

Guelph Food Research Centre, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Ontario, Canada N1G 5C9,1 College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi Province 712100, People's Republic of China,2 College of Life Science and Biotechnology, Shanghai Jiao Tong University, Minhang Campus, 800 Dongchuan Road, Minhang 200240, Shanghai, People's Republic of China,3 Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2W14

Received 1 March 2009/ Accepted 17 July 2009

O island 48 (OI-48) of Escherichia coli consists of three functional gene clusters that encode urease, tellurite resistance (Ter), and putative adhesins Iha and AIDA-1. The functions of these clusters in enterohemorrhagic E. coli (EHEC) O157:H7 infection are unknown. Deletion mutants for these three regions were constructed and evaluated for their ability to adhere to epithelial cells in vitro and in ligated pig ileal loops. Deletion of the Ter gene cluster reduced the ability of the organism to adhere to and form large clusters on IPEC-J2 and HEp-2 cells. Complementation of the mutation by introducing the wild-type ter genes restored adherence and large-cluster formation. Tests in ligated pig ileal loops showed a decrease in colonization by the Ter-negative mutant, but the difference was not significant compared to colonization by the wild type (26.4% ± 21.2% versus 40.1% ± 19.1%; P = 0.168). The OI-48 aidA gene deletion had no effect on adherence in vitro or in vivo. Deletion of the iha and ureC genes had no effect on adherence in vitro but significantly reduced the colonization of EHEC O157:H7 in the ligated pig intestine. These data suggest that Ter, Iha, and urease may contribute to EHEC O157:H7 pathogenesis by promoting adherence of the pathogen to the host intestinal epithelium.

* Corresponding author. Mailing address: Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2W1. Phone: (519) 824-4120, ext. 54657. Fax: (519) 824-5930. E-mail: cgyles{at}uoguelph.ca

{triangledown} Published ahead of print on 24 July 2009.

Applied and Environmental Microbiology, September 2009, p. 5779-5786, Vol. 75, No. 18
0099-2240/09/$08.00+0     doi:10.1128/AEM.00507-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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