Isolation of a Lysogenic Bacteriophage Carrying thestx1OX3 Gene, Which Is Closely Associated with Shiga Toxin-Producing Escherichia coli Strains from Sheep and Humans

ABSTRACT A specific PCR for the detection of a variant of the gene encoding Shiga toxin 1 (stx1) calledstx1OX3 (GenBank accession no. Z36901 ) was developed. The PCR was used to investigate 148 Stx1-producing Escherichia colistrains from human patients (n = 72), cattle (n = 27), sheep (n = 48), and a goat (n = 1) for the presence of thestx1OX3 gene. Thestx1OX3 gene was present in 38 Shiga toxin-producing E. coli (STEC) strains from sheep belonging to serogroups O5, O125, O128, O146, and OX3 but was absent from Stx1-positive ovine STEC O91 strains. Thestx1OX3 gene was also detected in 22 STEC strains from humans with nonbloody diarrhea and from asymptomatic excreters. Serotypes O146:H21 and O128:H2 were most frequently associated withstx1OX3-carrying STEC from sheep and humans. In contrast, Stx1-producing STEC strains from cattle and goats and 50 STEC strains from humans were all negative for the stx1OX3 gene. Thestx1OX3-negative strains belonged to 13 serotypes which were different from those of thestx1OX3-positive STEC strains. Moreover, the stx1OX3gene was not associated with STEC belonging to enterohemorrhagicE. coli (EHEC) serogroups O26, O103, O111, O118, O145, and O157. A bacteriophage carrying thestx1OX3 gene (phage 6220) was isolated from a human STEC O146:H21 strain. The phage was able to lysogenize laboratory E. coli K-12 strain C600. Phage 6220 shared a similar morphology and a high degree of DNA homology with Stx2-encoding phage 933W, which originates from EHEC O157. In contrast, few similarities were found between phage 6220 and Stx1-encoding bacteriophage H-19B from EHEC O26.

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