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Applied and Environmental Microbiology, May 2007, p. 3144-3150, Vol. 73, No. 10
0099-2240/07/$08.00+0     doi:10.1128/AEM.02937-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Shiga Toxin Gene Loss and Transfer In Vitro and In Vivo during Enterohemorrhagic Escherichia coli O26 Infection in Humans{triangledown}

Martina Bielaszewska,1* Rita Prager,2 Robin Köck,1 Alexander Mellmann,1 Wenlan Zhang,1 Helmut Tschäpe,2 Phillip I. Tarr,3 and Helge Karch1

Institute for Hygiene and the National Consulting Laboratory on Hemolytic Uremic Syndrome, University of Münster, Robert-Koch-Str. 41, 48149 Münster, Germany,1 National Reference Center for Salmonella and Other Enteric Pathogens, Robert Koch Institute, Branch Wernigerode, Burgstr. 37, 38855 Wernigerode, Germany,2 Departments of Pediatrics and Molecular Microbiology, Washington University School of Medicine, 660 S. Euclid Ave., St. Louis, Missouri3

Received 19 December 2006/ Accepted 6 March 2007

Escherichia coli serogroup O26 consists of enterohemorrhagic E. coli (EHEC) and atypical enteropathogenic E. coli (aEPEC). The former produces Shiga toxins (Stx), major determinants of EHEC pathogenicity, encoded by bacteriophages; the latter is Stx negative. We have isolated EHEC O26 from patient stools early in illness and aEPEC O26 from stools later in illness, and vice versa. Intrapatient EHEC and aEPEC isolates had quite similar pulsed-field gel electrophoresis (PFGE) patterns, suggesting that they might have arisen by conversion between the EHEC and aEPEC pathotypes during infection. To test this hypothesis, we asked whether EHEC O26 can lose stx genes and whether aEPEC O26 can be lysogenized with Stx-encoding phages from EHEC O26 in vitro. The stx2 loss associated with the loss of Stx2-encoding phages occurred in 10% to 14% of colonies tested. Conversely, Stx2- and, to a lesser extent, Stx1-encoding bacteriophages from EHEC O26 lysogenized aEPEC O26 isolates, converting them to EHEC strains. In the lysogens and EHEC O26 donors, Stx2-converting bacteriophages integrated in yecE or wrbA. The loss and gain of Stx-converting bacteriophages diversifies PFGE patterns; this parallels findings of similar but not identical PFGE patterns in the intrapatient EHEC and aEPEC O26 isolates. EHEC O26 and aEPEC O26 thus exist as a dynamic system whose members undergo ephemeral interconversions via loss and gain of Stx-encoding phages to yield different pathotypes. The suggested occurrence of this process in the human intestine has diagnostic, clinical, epidemiological, and evolutionary implications.

* Corresponding author: Mailing address: Institut für Hygiene, Universität Münster, Robert-Koch-Str. 41, 48149 Münster, Germany. Phone: 49-251/980-2849. Fax: 49-251/980-2868. E-mail: mbiela{at}uni-muenster.de

{triangledown} Published ahead of print on 30 March 2007.

Applied and Environmental Microbiology, May 2007, p. 3144-3150, Vol. 73, No. 10
0099-2240/07/$08.00+0     doi:10.1128/AEM.02937-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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