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Applied and Environmental Microbiology, May 2007, p. 3307-3319, Vol. 73, No. 10
0099-2240/07/$08.00+0     doi:10.1128/AEM.02239-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

VanA-Type Enterococci from Humans, Animals, and Food: Species Distribution, Population Structure, Tn1546 Typing and Location, and Virulence Determinants{triangledown}

F. Biavasco,1* G. Foglia,1 C. Paoletti,1 G. Zandri,1 G. Magi,1 E. Guaglianone,2 A. Sundsfjord,3 C. Pruzzo,4 G. Donelli,2 and B. Facinelli1

Institute of Microbiology and Biomedical Sciences, Polytechnic University of Marche, Ancona, Italy,1 Department of Technologies and Health, Istituto Superiore di Sanità, Rome, Italy,2 Department of Microbiology and Virology, University of Tromsø, Tromsø, Norway,3 Department of Biology, University of Genoa, Genoa, Italy4

Received 22 September 2006/ Accepted 1 March 2007

VanA-type human (n = 69), animal (n = 49), and food (n = 36) glycopeptide-resistant enterococci (GRE) from different geographic areas were investigated to study their possible reservoirs and transmission routes. Pulsed-field gel electrophoresis (PFGE) revealed two small genetically related clusters, M39 (n = 4) and M49 (n = 13), representing Enterococcus faecium isolates from animal and human feces and from clinical and fecal human samples. Multilocus sequence typing showed that both belonged to the epidemic lineage of CC17. purK allele analysis of 28 selected isolates revealed that type 1 was prevalent in human strains (8/11) and types 6 and 3 (14/15) were prevalent in poultry (animals and meat). One hundred and five of the 154 VanA GRE isolates, encompassing different species, origins, and PFGE types, were examined for Tn1546 type and location (plasmid or chromosome) and the incidence of virulence determinants. Hybridization of S1- and I-CeuI-digested total DNA revealed a plasmid location in 98% of the isolates. Human intestinal and animal E. faecium isolates bore large (>150 kb) vanA plasmids. Results of PCR-restriction fragment length polymorphism and sequencing showed the presence of prototype Tn1546 in 80% of strains and the G-to-T mutation at position 8234 in three human intestinal and two pork E. faecium isolates. There were no significant associations (P > 0.5) between Tn1546 type and GRE source or enterococcal species. Virulence determinants were detected in all reservoirs but were significantly more frequent (P < 0.02) among clinical strains. Multiple determinants were found in clinical and meat Enterococcus faecalis isolates. The presence of indistinguishable vanA elements (mostly plasmid borne) and virulence determinants in different species and PFGE-diverse populations in the presence of host-specific purK housekeeping genes suggested that all GRE might be potential reservoirs of resistance determinants and virulence traits transferable to human-adapted clusters.

* Corresponding author. Mailing address: Institute of Microbiology and Biomedical Sciences, Polytechnic University of Marche, Via Ranieri, Monte d'Ago, 60131 Ancona, Italy. Phone: 39 071 2204 637. Fax: 39 071 2204 622. E-mail: f.biavasco{at}univpm.it

{triangledown} Published ahead of print on 9 March 2007.

Applied and Environmental Microbiology, May 2007, p. 3307-3319, Vol. 73, No. 10
0099-2240/07/$08.00+0     doi:10.1128/AEM.02239-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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