Use of MALDI-TOF mass spectrometry to analyze the molecular profile of Pseudomonas aeruginosa biofilms grown on glass and plastic surfaces

• Published in: Microbial pathogenesis 2015-09, Vol.86, p.32-37
• Main Authors: Pereira, Flávio D.E.S., Bonatto, Cínthia C., Lopes, Cláudio A.P., Pereira, Alex L., Silva, Luciano P.
• Format: Article
• Language: English
• Subjects: Atomic force microscopy; Bacterial Proteins - analysis; Biofilm maturation; Biofilms - growth & development; Environmental Microbiology; Glass; MALDI profiling; Microscopy, Atomic Force; Microscopy, Electron, Scanning; Phenotype; Polypropylenes; Proteome - analysis; Pseudomonas aeruginosa; Pseudomonas aeruginosa - chemistry; Pseudomonas aeruginosa - growth & development; Pseudomonas aeruginosa - physiology; Scanning electron microscopy; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1016/j.micpath.2015.07.005

Biofilms are microbial sessile communities attached to surfaces that are known for causing many medical problems. A bacterial biofilm of clinical relevance is formed by the gram-negative bacteria Pseudomonas aeruginosa. During the formation of a biofilm, the initial adhesion of the cells is of crucial importance, and the characteristics of the contact surface have great influence on this step. In the present study, we aimed to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling as a new methodology to monitor P. aeruginosa biofilm development. Biofilms were grown within polypropylene tubes containing a glass slide, and were harvested after 3, 5, 7, 9, or 12 days of inoculation. Planktonic cells were obtained separately by centrifugation as control. Two independent MALDI-TOF experiments were performed, one by collecting biofilms from both the glass slide and the polypropylene tube internal surface, and the other by acquiring biofilms from these surfaces separately. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were used to evaluate the morphological progression of the biofilm. The molecular results showed that MALDI profiling is able not only to distinguish between different biofilm stages, but it is also appropriate to indicate when the biofilm cells are released at the dispersion stage, which occurred first on polypropylene surface. Finally, the present study pointed out that MALDI profiling may emerge as a promising tool for the clinical diagnostic and prognostic workup of biofilms formation and control. •MALDI-TOF mass spectrometry is able to distinguish the different stages of the biofilms.•The molecular profile of Pseudomonas aeruginosa is changed according to the type of support.•MALDI-TOF mass spectrometry is effective in distinction between planktonic cells and biofilms.