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Applied and Environmental Microbiology, August 2009, p. 4975-4983, Vol. 75, No. 15
0099-2240/09/$08.00+0     doi:10.1128/AEM.00297-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Adherence of Escherichia coli O157:H7 Mutants In Vitro and in Ligated Pig Intestines{triangledown}

Xianhua Yin,1,5 James R. Chambers,1 Roger Wheatcroft,1 Roger P. Johnson,2 Jing Zhu,3 Bianfang Liu,4 and Carlton L. Gyles5*

Guelph Food Research Centre, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Ontario, Canada N1G 5C9,1 Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Guelph, Ontario, Canada N1G 3W4,2 College of Life Science and Biotechnology, Shanghai Jiao Tong University; 800 Dongchuan Road, Minhang Campus, Minhang 200240, Shanghai, People's Republic of China,3 College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi Province 712100, People's Republic of China,4 Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2W15

Received 5 February 2009/ Accepted 6 June 2009

There are contradictory literature reports on the role of verotoxin (VT) in adherence of enterohemorrhagic Escherichia coli O157:H7 (O157 EHEC) to intestinal epithelium. There are reports that putative virulence genes of O island 7 (OI-7), OI-15, and OI-48 of this pathogen may also affect adherence in vitro. Therefore, mutants of vt2 and segments of OI-7 and genes aidA15 (gene from OI-15) and aidA48 (gene from OI-48) were generated and evaluated for adherence in vitro to cultured human HEp-2 and porcine jejunal epithelial (IPEC-J2) cells and in vivo to enterocytes in pig ileal loops. VT2-negative mutants showed significant decreases in adherence to both HEp-2 and IPEC-J2 cells and to enterocytes in pig ileal loops; complementation only partially restored VT2 production but fully restored the adherence to the wild-type level on cultured cells. Deletion of OI-7 and aidA48 had no effect on adherence, whereas deletion of aidA15 resulted in a significant decrease in adherence in pig ileal loops but not to the cultured cells. This investigation supports the findings that VT2 plays a role in adherence, shows that results obtained in adherence of E. coli O157:H7 in vivo may differ from those obtained in vitro, and identified AIDA-15 as having a role in adherence of E. coli O157:H7.

* 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 12 June 2009.

Applied and Environmental Microbiology, August 2009, p. 4975-4983, Vol. 75, No. 15
0099-2240/09/$08.00+0     doi:10.1128/AEM.00297-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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