Vibartional Micro- and Nanospectroscopy of Burkholderia contaminans Biofilms

CECILIA BEATRIZ FÍGOLI; MAREN STÄMMLER; LAURA BELTINA LEON; CHRISTOPH SCHAUDINN; ALEJANDRA BOSCH; MICHAEL LAUE; PETER LASCH

Berlín

Workshop; FTIR Spectroscopy in Microbiological and Medical Diagnostics; 2017

Robert Koch Institute

Among the Burkholderia cepacia complex species, B. contaminans represents the most frequently recovered from patients with cystic fibrosis (CF) in Argentina, and its incidence is currently increasing in Portugal and Spain [1]. Although B. contaminans respiratory tract infection may be transient, its acquisition most frequently results in a chronic infection with unfavorable clinical manifestation and a gradual decline in lung function [2]. The adaptation strategies leading to the establishment of these bacteria in CF lungs are still unknown; nevertheless biofilm formation may be playing a role in protecting bacteria from antimicrobial treatments and host immune system. A biofilm is characterized by aggregated cells surrounded by a self-produced matrix which contain polysaccharides, and other components such as eDNA, proteins, and lipopolysaccharides [3]. In this work we present data obtained from Burkholderia contaminans LMG 23361 biofilms produced under batch conditions on CaF2 windows. A tandem imaging analysis by optical light microscopy, atomic force microscopy (AFM), FTIR hyperspectral imaging, confocal Raman microspectroscopy (CRM), confocal laser scan microscopy (CLSM) and scanning electron microscopy (SEM) was developed. The combined application of these techniques allowed studying the physical and chemical heterogeneity of bacterial biofilms at the micro- and nanoscale levels. This novel correlative approach enabled us to get insights into the spatial phenotypic heterogeneity of biofilms, to analyze the individual cell phenotypes and to characterize the biofilm´s matrix composition with very high spatial resolution.