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JAC Advance Access published online on November 14, 2008
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dkn464
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© The Author 2008. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Original research

High in vitro antimicrobial activity of synthetic antimicrobial peptidomimetics against staphylococcal biofilms

Kristina Flemming1, Claus Klingenberg1,2,*, Jorun Pauline Cavanagh2, Merethe Sletteng1,2, Wenche Stensen3,4, John Sigurd Svendsen3,4 and Trond Flægstad1,2

1 Department of Paediatrics, University Hospital of North-Norway, Tromsø, Norway 2 Department of Paediatrics, Institute of Clinical Medicine, University of Tromsø, Tromsø, Norway 3 Department of Chemistry, Faculty of Science, University of Tromsø, Tromsø, Norway 4 Lytix Biopharma AS, Tromsø, Norway

Received 20 August 2008; returned 15 September 2008; revised 1 October 2008; accepted 7 October 2008

* Correspondence address. Department of Paediatrics, University Hospital of North-Norway, N-9038 Tromsø, Norway. Tel: +47-77669845; Fax: +47-77626369; E-mail: claus.klingenberg{at}unn.no

Objectives: The aim of the study was to investigate the antimicrobial effect of different antibiotics and synthetic antimicrobial peptidomimetics (SAMPs) on staphylococcal biofilms.

Methods: Biofilms of six staphylococcal strains (two Staphylococcus haemolyticus, two Staphylococcus epidermidis and two Staphylococcus aureus isolates) were grown for 24 h in microtitre plates. They were washed and treated for 24 h with different concentrations of linezolid, tetracycline, rifampicin and vancomycin and four different SAMPs. After treatment, the redox indicator Alamar Blue was used to quantify metabolic activity of bacteria in biofilms, and confocal laser scanning microscopy with LIVE/DEAD staining was used to further elucidate any effects.

Results: At MIC levels, rifampicin and tetracycline showed a marked reduction of metabolic activity in the S. epidermidis and S. haemolyticus biofilm. Linezolid had a moderate effect and vancomycin had a poor effect. MIC x10 and MIC x100 improved the antimicrobial activity of all antibiotics, especially vancomycin. However, metabolic activity was not completely suppressed in strong biofilm-producing strains. At MIC x10, the three most effective SAMPs (Ltx5, Ltx9 and Ltx10) were able to completely eliminate metabolic activity in the S. epidermidis and S. haemolyticus biofilms, which was also confirmed by complete cell death using confocal laser scanning microscopy investigations. Although none of the Ltx SAMPs could fully suppress metabolic activity in the S. aureus biofilm, their effect was superior to all tested antibiotics.

Conclusions: SAMPs had superior antimicrobial activity in staphylococcal biofilms compared with conventional antibiotics and are potential new therapeutic agents for biofilm-associated infections.

Key Words: Alamar Blue , biofilm inhibitory concentrations , staphylococci


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