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JAC Advance Access published online on June 25, 2008
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkn265
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© The Author 2008. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Original research

Relevance of hot spots in the evolution and transmission of Tn1546 in glycopeptide-resistant Enterococcus faecium (GREF) from broiler origin

Lourdes Garcia-Migura*, Henrik Hasman, Christina Svendsen and Lars Bogø Jensen

National Food Institute, DTU, Antimicrobial Resistance Unit, Bülowsvej 27, DK-1790 Copenhagen V, Denmark

Received 7 March 2008; returned 29 April 2008; revised 28 May 2008; accepted 1 June 2008

* Corresponding author. Tel: +45-7234-6401; Fax: +45-7234-6001; E-mail: lgmi{at}food.dtu.dk

Objectives: Glycopeptide-resistant enterococci are still present within the broiler sector, despite the EU ban of avoparcin more than a decade ago. In the present study, we have developed a rapid method for screening the flanking regions at the integration point of Tn1546 in glycopeptide-resistant Enterococcus faecium isolated from broiler farms.

Methods: Total DNA was digested, ligated and amplified using primers from inside Tn1546. The resulting amplicons were purified and sequenced. Two new primers were designed based on obtained sequences.

Results: Two main insertion points have been repeatedly found in isolates from the UK (n = 150). The first insertion point revealed that 25 isolates harboured Tn1546 positioned in a sequence with 96% homology to a streptomycin adenyltransferase gene (AY604739 [GenBank] ) from a Staphylococcus intermedius plasmid. At this insertion point, a direct repeat (GTCCT) was duplicated as previously described, indicating transposition at the target site. Furthermore, this ‘hot spot’ was also detected in isolates from Norway (2/8) and Denmark (17/20). The second insertion point detected in 45 isolates from the UK revealed integration into an Inc18-like plasmid, most likely by a process of target site recombination.

Conclusions: The presence of a common insertion point for isolates from different geographical areas could suggest the insertion of Tn1546 by transposition in a plasmid-specific site, followed by genetic rearrangement both inside the transposon and in the flanking regions.

Key Words: streptomycin adenyltransferase genes , Inc18 plasmids , transposons , E. faecium


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