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Applied and Environmental Microbiology, March 2008, p. 1350-1356, Vol. 74, No. 5
0099-2240/08/$08.00+0     doi:10.1128/AEM.02254-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Molecular Structure and Transferability of Tn1546-Like Elements in Enterococcus faecium Isolates from Clinical, Sewage, and Surface Water Samples in Iran{triangledown}

M. Talebi,1 M. R. Pourshafie,1* M. Katouli,2 and R. Möllby3

Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran,1 Faculty of Science, Health and Education, University of the Sunshine Coast, Maroochydore, Queensland 4558, Australia,2 Microbiology, Tumor and Cell Biology Centre, Karolinska Institutet, S-171 77 Stockholm, Sweden3

Received 3 October 2007/ Accepted 30 December 2007

The molecular structure and transferability of Tn1546 in 143 vancomycin-resistant Enterococcus faecium (VREF) isolates obtained from patients (n = 49), surface water (n = 28), and urban and hospital sewage (n = 66) in Tehran, Iran, were investigated. Molecular characterization of Tn1546 elements in vanA VREF was performed using a combination of restriction fragment length polymorphism analysis and DNA sequencing of the internal PCR fragments of vanA transposons. Long-PCR amplification showed that the molecular size of Tn1546 elements varied from 10.8 to 12.8 kb. The molecular analysis of Tn1546 showed that 45 isolates (31.5%) harbored a deletion/mutation upstream from nucleotide 170. No horizontal transfer of Tn1546 was observed following filter-mating conjugation with these isolates. Nevertheless, the rates of transferability for other isolates were 10–5 to 10–6 per donor. Insertion sequences IS1216V and IS1542 were present in 103 (72%) and 138 (96.5%) of the isolates, respectively. The molecular analysis of Tn1546 elements resulted in three genomic organizations. The genomic organization lineage 1 was dominated by the isolates from clinical samples (3.4%), lineage 2 was dominated mostly by sewage isolates (24.5%), and lineage 3 contained isolates obtained from all sources (72.1%). The genetic diversity determined using pulsed-field gel electrophoresis (PFGE) revealed a single E. faecium clone, designated 44, which was common to the samples obtained from clinical specimens and hospital and municipal sewage. Furthermore, the results suggest that lineage 3 Tn1546 was highly disseminated among our enterococcal isolates in different PFGE patterns.

* Corresponding author. Mailing address: Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran. Phone: (98) 21 66405535. Fax: (98) 21 66405535. E-mail: pour{at}pasteur.ac.ir

{triangledown} Published ahead of print on 11 January 2008.

Applied and Environmental Microbiology, March 2008, p. 1350-1356, Vol. 74, No. 5
0099-2240/08/$08.00+0     doi:10.1128/AEM.02254-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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