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Applied and Environmental Microbiology, January 2004, p. 61-68, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.61-68.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Localization of the Insertion Site and Pathotype Determination of the Locus of Enterocyte Effacement of Shiga Toxin-Producing Escherichia coli Strains

Yolande Bertin,1 Karima Boukhors,1 Valerie Livrelli,2 and Christine Martin1*

Laboratoire de Microbiologie, Centre de Recherche INRA de Clermont-Ferrand-Theix, 63122 St-Genès Chapanelle,1 Groupe de Recherche Pathogénie Bactérienne Intestinale, Faculté de Pharmacie, Université d'Auvergne Clermont 1, 63001 Clermont-Ferrand Cedex, France2

Received 15 July 2003/ Accepted 10 October 2003

Of 220 Shiga toxin-producing Escherichia coli (STEC) strains collected in central France from healthy cattle, food samples, and asymptomatic children, 12 possessed the eae gene included in the locus of enterocyte effacement (LEE) pathogenicity island. Based on gene typing, we observed 7 different eae espA espB tir pathotypes among the 12 STEC strains and described the new espAßv variant. As previously observed, the O157 serogroup is associated with eae{gamma}, O26 is associated with eaeß, and O103 is associated with eae{varepsilon}. However, the unexpected eae{zeta} allele was detected in 5 of the 12 isolates. PCR amplification and pulsed-field gel electrophoresis using the I-CeuI endonuclease followed by Southern hybridization indicated that the LEE was inserted in the vicinity of the selC (three isolates), pheU (two isolates), or pheV (six isolates) tRNA gene. Six isolates harbored two or three of these tRNA loci altered by the insertion of integrase genes (CP4-int and/or int-phe), suggesting the insertion of additional foreign DNA fragments at these sites. In spite of great genetic diversity of LEE pathotypes and LEE insertion sites, bovine strains harbor alleles of LEE genes that are frequently found in clinical STEC strains isolated from outbreaks and sporadic cases around the world, underscoring the potential risk of the bovine strains on human health.

* Corresponding author. Mailing address: Laboratoire de Microbiologie, INRA de Clermont-Ferrand-Theix, 63122 St-Genès Champanelle, France. Phone: (33) 4 73 62 42 47. Fax: (33) 4 73 63 45 81. E-mail: cmartin{at}clermont.inra.fr.

Applied and Environmental Microbiology, January 2004, p. 61-68, Vol. 70, No. 1
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.1.61-68.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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