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Applied and Environmental Microbiology, October 2006, p. 6544-6553, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.00749-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Pheromone-Responsive Conjugative Vancomycin Resistance Plasmids in Enterococcus faecalis Isolates from Humans and Chicken Feces

Suk-Kyung Lim,1,3 Koichi Tanimoto,2 Haruyoshi Tomita,1 and Yasuyoshi Ike1,2*

Department of Bacteriology and Bacterial Infection Control,1 Laboratory of Bacterial Drug Resistance, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan,2 National Veterinary Research and Quarantine Service (NVRQS) 480, Anyang 6 Dong, Manangu, Anyang City 430-824, Korea3

Received 31 March 2006/ Accepted 14 July 2006

The drug resistances and plasmid contents of a total of 85 vancomycin-resistant enterococcus (VRE) strains that had been isolated in Korea were examined. Fifty-four of the strains originated from samples of chicken feces, and 31 were isolated from hospital patients in Korea. Enterococcus faecalis KV1 and KV2, which had been isolated from a patient and a sample of chicken feces, respectively, were found to carry the plasmids pSL1 and pSL2, respectively. The plasmids transferred resistances to vancomycin, gentamicin, kanamycin, streptomycin, and erythromycin to E. faecalis strains at a high frequency of about 10–3 per donor cell during 4 hours of broth mating. E. faecalis strains containing each of the pSL plasmids formed clumps after 2 hours of incubation in broth containing E. faecalis FA2-2 culture filtrate (i.e., the E. faecalis sex pheromone), and the plasmid subsequently transferred to the recipient strain in a 10-min short mating in broth, indicating that the plasmids are responsive to E. faecalis pheromones. The pSL plasmids did not respond to any of synthetic pheromones for the previously characterized plasmids. The pheromone specific for pSL plasmids has been designated cSL1. Southern hybridization analysis showed that specific FspI fragments from each of the pSL plasmids hybridized with the aggregation substance gene (asa1) of the pheromone-responsive plasmid pAD1, indicating that the plasmids had a gene homologous to asa1. The restriction maps of the plasmids were identical, and the size of the plasmids was estimated to be 128.1 kb. The plasmids carried five drug resistance determinants for vanA, ermB, aph(3'), aph(6'), and aac(6')/aph(2'), which encode resistance to vancomycin, erythromycin, kanamycin, streptomycin, and gentamicin/kanamycin, respectively. Nucleotide sequence analyses of the drug resistance determinants and their flanking regions are described in this report. The results described provide evidence for the exchange of genetic information between human and animal (chicken) VRE reservoirs and suggest the potential for horizontal transmission of multiple drug resistance, including vancomycin resistance, between farm animals and humans via a pheromone-responsive conjugative plasmid.

* Corresponding author. Mailing address: Department of Bacteriology and Bacterial Infection Control and Laboratory of Bacterial Drug Resistance, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan. Phone: 81-27-220-7990. Fax: 81-27-220-7990. E-mail: yasuike{at}med.gunma-u.ac.jp.

Applied and Environmental Microbiology, October 2006, p. 6544-6553, Vol. 72, No. 10
0099-2240/06/$08.00+0     doi:10.1128/AEM.00749-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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