Details

Autor(en) / Beteiligte

• Quilès, Fabienne
• Saadi, Souhir
• Francius, Grégory
• Bacharouche, Jalal
• Humbert, François

Titel

In situ and real time investigation of the evolution of a Pseudomonas fluorescens nascent biofilm in the presence of an antimicrobial peptide

Ist Teil von

• Biochimica et biophysica acta, 2016-01, Vol.1858 (1), p.75-84

Ort / Verlag

Netherlands: Elsevier B.V

Erscheinungsjahr

2016

Quelle

EZB-FREE-00999 freely available EZB journals

Beschreibungen/Notizen

• Against the increase of bacterial resistance to traditional antibiotics, antimicrobial peptides (AMP) are considered as promising alternatives. Bacterial biofilms are more resistant to antibiotics that their planktonic counterpart. The purpose of this study was to investigate the action of an AMP against a nascent bacterial biofilm. The activity of dermaseptin S4 derivative S4(1–16)M4Ka against 6h-old Pseudomonas fluorescens biofilms was assessed by using a combination of Attenuated Total Reflectance–Fourier Transform InfraRed (ATR–FTIR) spectroscopy in situ and in real time, fluorescence microscopy using the Baclight™ kit, and Atomic Force Microscopy (AFM, imaging and force spectroscopy). After exposure to the peptide at three concentrations, different dramatic and fast changes over time were observed in the ATR–FTIR fingerprints reflecting a concentration-dependent action of the AMP. The ATR–FTIR spectra revealed major biochemical and physiological changes, adsorption/accumulation of the AMP on the bacteria, loss of membrane lipids, bacterial detachment, bacterial regrowth, or inhibition of biofilm growth. AFM allowed estimating at the nanoscale the effect of the AMP on the nanomechanical properties of the sessile bacteria. The bacterial membrane elasticity data measured by force spectroscopy were consistent with ATR–FTIR spectra, and they allowed suggesting a mechanism of action of this AMP on sessile P. fluorescens. The combination of these three techniques is a powerful tool for in situ and in real time monitoring the activity of AMPs against bacteria in a biofilm. [Display omitted] •In situ investigation of action of a dermaseptin derivative on a bacterial biofilm•Dramatic and fast changes over time were observed in the ATR–FTIR fingerprints.•AFM allowed estimating bacterial membrane elasticities without and with the peptide.•Concentration-dependent mechanisms are suggested from complementary methods.