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Applied and Environmental Microbiology, September 1999, p. 3855-3861, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Transduction of Enteric Escherichia coli Isolates with a Derivative of Shiga Toxin 2-Encoding Bacteriophage phi 3538 Isolated from Escherichia coli O157:H7

Herbert Schmidt,1,* Martina Bielaszewska,2 and Helge Karch1

Institut für Hygiene und Mikrobiologie der Universität Würzburg, 97080 Würzburg, Germany,1 and Institute of Medical Microbiology, The 2nd Medical Faculty, Charles University, 15006 Prague, Czech Republic2

Received 19 May 1999/Accepted 7 July 1999

We investigated the ability of a detoxified derivative of a Shiga toxin 2 (Stx2)-encoding bacteriophage to infect and lysogenize enteric Escherichia coli strains and to develop infectious progeny from such lysogenized strains. The stx2 gene of the patient E. coli O157:H7 isolate 3538/95 was replaced by the chloramphenicol acetyltransferase (cat) gene from plasmid pACYC184. Phage phi 3538(Delta stx2::cat) was isolated after induction of E. coli O157:H7 strain 3538/95 with mitomycin. A variety of strains of enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Stx-producing E. coli (STEC), enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC), and E. coli from the physiological stool microflora were infected with phi 3538(Delta stx2::cat), and plaque formation and lysogenic conversion of wild-type E. coli strains were investigated. With the exception of one EIEC strain, none of the E. coli strains supported the formation of plaques when used as indicators for phi 3538(Delta stx2::cat). However, 2 of 11 EPEC, 11 of 25 STEC, 2 of 7 EAEC, 1 of 3 EIEC, and 1 of 6 E. coli isolates from the stool microflora of healthy individuals integrated the phage in their chromosomes and expressed resistance to chloramphenicol. Following induction with mitomycin, these lysogenic strains released infectious particles of phi 3538(Delta stx2::cat) that formed plaques on a lawn of E. coli laboratory strain C600. The results of our study demonstrate that phi 3538(Delta stx2::cat) was able to infect and lysogenize particular enteric strains of pathogenic and nonpathogenic E. coli and that the lysogens produced infectious phage progeny. Stx-encoding bacteriophages are able to spread stx genes among enteric E. coli strains.

* Corresponding author. Mailing address: Institut für Hygiene und Mikrobiologie der Universität Würzburg, Josef-Schneider-Str. 2, D-97080 Würzburg, Germany. Phone: 49/931/201-5160. Fax: 49/931/201-3445. E-mail: hschmidt{at}hygiene.uni-wuerzburg.de.

Applied and Environmental Microbiology, September 1999, p. 3855-3861, Vol. 65, No. 9
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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