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Applied and Environmental Microbiology, July 2008, p. 4247-4255, Vol. 74, No. 14
0099-2240/08/$08.00+0     doi:10.1128/AEM.02474-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genotypic Diversity, Antimicrobial Resistance, and Virulence Factors of Human Isolates and Probiotic Cultures Constituting Two Intraspecific Groups of Enterococcus faecium Isolates

Vaccine and Infectious Disease Institute (VIDI), Laboratory of Medical Microbiology, University of Antwerp, Wilrijk, Belgium,¹ Laboratory of Microbiology, Faculty of Sciences,² BCCM/LMG Bacteria Collection, Ghent University, Ghent, Belgium,³ Robert Koch Institute, Wernigerode, Germany⁴

Received 2 November 2007/ Accepted 10 May 2008

The intraspecific relationships among a collection of Enterococcus faecium isolates comprising probiotic cultures and human clinical isolates were investigated through the combined use of two high-resolution DNA-fingerprinting techniques. In addition, the incidences of antimicrobial resistance and virulence traits were investigated. A total of 128 E. faecium isolates from human clinical or nonclinical sources or used as probiotic cultures were subjected to fluorescent amplified fragment length polymorphism (FAFLP) fingerprinting and pulsed-field gel electrophoresis (PFGE) analysis of SmaI macrorestriction patterns. Susceptibilities to 16 antimicrobial agents were tested using broth microdilution, and the presence of the corresponding resistance genes was investigated using PCR. Multiplex PCR was used to detect the presence of the enterococcal virulence genes asa1, gelE, cylA, esp, and hyl. The results of the study showed that two intraspecific genomic groups (I and II) were obtained in FAFLP analysis. PFGE analysis demonstrated high variability within these two groups but also indicated that some probiotic cultures were indistinguishable and that a number of clinical isolates may be reisolations of commercial probiotic cultures. Compared to group II, which contained the majority of the probiotic isolates and fewer human clinical isolates, higher phenotypic and genotypic resistance frequencies were observed in group I. Two probiotic isolates were phenotypically resistant to erythromycin, one of which contained an erm(B) gene that was not transferable to enterococcal recipients. None of the probiotic E. faecium isolates demonstrated the presence of the tested virulence genes. The previously reported observation that E. faecium consists of two intraspecific genomic groups was further substantiated by FAFLP fingerprinting of 128 isolates. In combination with antimicrobial resistance and virulence testing, this grouping might represent an additional criterion in assessing the safety of new potential probiotic E. faecium isolates.

Published ahead of print on 16 May 2008.