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Applied and Environmental Microbiology, January 2005, p. 159-168, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.159-168.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Persistence of Animal and Human Glycopeptide-Resistant Enterococci on Two Norwegian Poultry Farms Formerly Exposed to Avoparcin Is Associated with a Widespread Plasmid-Mediated vanA Element within a Polyclonal Enterococcus faecium Population

P. J. Johnsen,1* J. I. Østerhus,1 H. Sletvold,2 M. Sørum,3 H. Kruse,4 K. Nielsen,2 G. S. Simonsen,1,5 and A. Sundsfjord1,5*

Department of Microbiology and Virology, Faculty of Medicine, University and University Hospital of North Norway,1 Department of Pharmacy, Faculty of Medicine, University of Tromsø, Tromsø,2 National Veterinary Institute,3 Norwegian Zoonosis Center,4 Norwegian Institute of Public Health, Oslo, Norway5

Received 20 April 2004/ Accepted 23 August 2004

The evolutionary processes responsible for the long-term persistence of glycopeptide-resistant Enterococcus faecium (GREF) in nonselective environments were addressed by genetic analyses of E. faecium populations in animals and humans on two Norwegian poultry farms that were previously exposed to avoparcin. A total of 222 fecal GREF (n = 136) and glycopeptide-susceptible (n = 86) E. faecium (GSEF) isolates were obtained from farmers and poultry on three separate occasions in 1998 and 1999. Pulsed-field gel electrophoresis (PFGE) and plasmid DNA analyses discerned 22 GREF and 32 GSEF PFGE types within shifting polyclonal animal and human E. faecium populations and indicated the presence of transferable plasmid-mediated vanA resistance, respectively. Examples of dominant, persistent GREF PFGE types supported the notion that environmentally well-adapted GREF types may counteract the reversal of resistance. PFGE analyses, sequencing of the purK housekeeping gene, and partial typing of vanA-containing Tn1546 suggested a common animal and human reservoir of glycopeptide resistance. Inverse PCR amplification and sequence analyses targeting the right end of the Tn1546-plasmid junction fragment strongly indicated the presence of a common single Tn1546-plasmid-mediated element in 20 of 22 GREF PFGE types. This observation was further strengthened by vanY-vanZ hybridization analyses of plasmid DNAs as well as the finding of a physical linkage between Tn1546 and a putative postsegregation killing system for seven GREF PFGE types. In conclusion, our observations suggest that the molecular unit of persistence of glycopeptide resistance is a common mobile plasmid-mediated vanA-containing element within a polyclonal GREF population that changes over time. In addition, we propose that "plasmid addiction systems" may contribute to the persistence of GREF in nonselective environments.

* Corresponding author. Mailing address: Department of Microbiology and Virology, Faculty of Medicine, University of Tromsø, N-9037 Tromsø, Norway. Phone: 47 77644663. Fax: 47 77645350. E-mail for P. J. Johnsen: paalj@fagmed.uit.no. E-mail for A. Sundsfjord: arnfinn.sundsfjord{at}fagmed.uit.no.

Applied and Environmental Microbiology, January 2005, p. 159-168, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.159-168.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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