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JAC Advance Access published online on August 13, 2003
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkg372
© 2003 by The British Society for Antimicrobial Chemotherapy
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© 2003 The British Society for Antimicrobial Chemotherapy

Original article

Antimicrobial activity of SMAP-29 against the Bacteroides fragilis group and clostridia

Alessandra Arzese 1 *, Barbara Skerlavaj 2 , Linda Tomasinsig 3 , Renato Gennaro 4 , and Margherita Zanetti 3

1 Institute of Microbiology, Udine Medical School, University of Udine, p.zza S.M. Misericordia 1, 33100 Udine
2 Department of Biomedical Sciences and Technology, Udine Medical School, University of Udine, p.zza S.M. Misericordia 1, 33100 Udine
3 Department of Biomedical Sciences and Technology, Udine Medical School, University of Udine, p.zza S.M. Misericordia 1, 33100 Udine; National Laboratory CIB, AREA Science Park, Padriciano-Trieste
4 Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Italy

* Corresponding author. E-mail: alessandra.arzese{at}drmm.uniud.it.

Received 20 January 2003 ; revised 11 June 2003 ; accepted 18 June 2003

Abstract

Objectives: The cathelicidin-derived peptide SMAP-29 exerts rapid and broad-spectrum antimicrobial activity against aerobic bacteria and fungi. In this study, the effects of the peptide against the Bacteroides fragilis group, including antibiotic-resistant isolates, Clostridium perfringens and Clostridium difficile reference and clinical isolates, were investigated.

Methods: The microbicidal activity of SMAP-29 against eight reference and 100 clinical anaerobic strains from a national collection was assessed using a microdilution susceptibility assay, and by determining the killing kinetics on selected strains. The killing mechanism was investigated further by means of a two-colour fluorescent permeabilization assay, and by scanning electron microscopy (SEM).

Results: The Bacteroides fragilis group, Clostridium reference strains and most clinical isolates were inhibited in vitro by 1-2 µM (3.2-6.4 mg/L) SMAP-29, and killed by 1.5- to 2-fold higher peptide concentrations. The anaerobic bacterial cells were 90%-100% permeabilized within 2 h of exposure to bactericidal concentrations of the peptide. The SEM images of bacteria exposed to SMAP-29 provide morphological evidence that the envelope is an important target of the bactericidal activity of this peptide. These results are consistent with earlier studies indicating that SMAP-29 kills aerobic bacteria with a membranolytic mechanism, and suggest that both aerobic and anaerobic bacteria share surface features that are targeted by this peptide.

Conclusions: These studies demonstrate that the spectrum of antibacterial activity of SMAP-29 includes the B. fragilis group and Clostridium species, and encourage further investigations of the therapeutic potential of this peptide.

Keywords: antimicrobial peptide, cathelicidin, anaerobic bacteria, membrane permeabilization
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