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Journal of Bacteriology, December 2004, p. 8287-8294, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8287-8294.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Isolation of Escherichia coli Bacteriophages from the Stool of Pediatric Diarrhea Patients in Bangladesh

Sandra Chibani-Chennoufi,1 Josette Sidoti,1 Anne Bruttin,1 Marie-Lise Dillmann,1 Elizabeth Kutter,2 Firdausi Qadri,3 Shafiqul Alam Sarker,3 and Harald Brüssow1*

Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, Lausanne, Switzerland,1 Evergreen State College, Olympia, Washington,2 ICDDRB Centre for Health and Population Research, Dhaka, Bangladesh3

Received 17 June 2004/ Accepted 13 September 2004

A 3-week coliphage survey was conducted in stool samples from 140 Bangladeshi children hospitalized with severe diarrhea. On the Escherichia coli indicator strain K803, all but one phage isolate had 170-kb genomes and the morphology of T4 phage. In spot tests, the individual T4-like phages infected up to 27 out of 40 diarrhea-associated E. coli, representing 22 O serotypes and various virulence factors; only five of them were not infected by any of these new phages. A combination of diagnostic PCR based on g32 (DNA binding) and g23 (major capsid protein) and Southern hybridization revealed that half were T-even phages sensu strictu, while the other half were pseudo-T-even or even more distantly related T4-like phages that failed to cross-hybridize with T4 or between each other. Nineteen percent of the acute stool samples yielded T4-like phages, and the prevalence was lower in convalescent stool samples. T4-like phages were also isolated from environmental and sewage water, but with low frequency and low titers. On the enteropathogenic E. coli strain O127:K63, 14% of the patients yielded phage, all of which were members of the phage family Siphoviridae with 50-kb genomes, showing the morphology of Jersey- and beta-4 like phages and narrow lytic patterns on E. coli O serotypes. Three siphovirus types could be differentiated by lack of cross-hybridization. Only a few stool samples were positive on both indicator strains. Phages with closely related restriction patterns and, in the case of T4-like phages, identical g23 gene sequences were isolated from different patients, suggesting epidemiological links between the patients.

* Corresponding author. Mailing address: Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland. Phone: 41 21 785 86 76. Fax: 41 21 785 85 49. E-mail: harald.bruessow{at}rdls.nestle.com.

Journal of Bacteriology, December 2004, p. 8287-8294, Vol. 186, No. 24
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.24.8287-8294.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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