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Appl. Environ. Microbiol. doi:10.1128/AEM.02937-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Shiga Toxin Gene Loss and Transfer In Vitro and In Vivo During Enterohemorrhagic Escherichia coli O26 Infection in Humans

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

^* To whom correspondence should be addressed. Email: mbiela{at}uni-muenster.de.

	Abstract

Escherichia coli serogroup O26 consists of enterohemorrhagic E. coli (EHEC) and atypical enteropathogenic E. coli (aEPEC). The former produce Shiga toxins (Stx), major determinants of EHEC pathogenicity, encoded by bacteriophages; the latter are Stx-negative. We have isolated EHEC O26 from initial (early in illness) stools and aEPEC O26 from subsequent stools of patients, 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 if EHEC O26 can lose stx genes and whether or not 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 occured in 10% to 14% of colonies tested. Conversely, Stx2- and, to a lesser extent, Stx1-encoding bacteriophages from EHEC O26 lysogenized aEPEC O26, converting them to EHEC. 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 inter-conversions 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.

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