Vibrational Micro- and Nanospectroscopy of Burkholderia contaminans Biofilms

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

Dublin

Conferencia; 10th Conferencia SPEC 2018; 2018

International Society of Clinical Spectroscopy

Among the Burkholderia cepacia complex species, B. contaminans represents one of the most frequently recovered species from patients with cystic fibrosis (CF) in Argentina, Spain and Portugal. [1] Although B. contaminans respiratory tract infections may be transient, its acquisition results in many cases in a chronic infection with unfavorable clinical manifestation and a gradual decline of lung function.[2] The adaptation strategies leading to the establishment of these bacteria in lungs of CF patients are still unknown; nevertheless biofilm formation may be playing a role in protecting bacteria from the host immune system and antimicrobial treatments. A biofilm is characterized by aggregated cells surrounded by a self-produced matrix of hydrated extracellular polymeric substances (EPS) which contains polysaccharides, and other components such as extracellular DNA, proteins, and lipopolysaccharides. [3]In this work we present data obtained from Burkholderia contaminans LMG 23361 biofilms produced under batch conditions on Raman-compatible CaF2 windows. A multimodal analysis approach involving techniques like standard light microscopy, atomic force microscopy (AFM), FTIR hyperspectral imaging, confocal Raman microspectroscopy (CRM), confocal laser scan microscopy (cLSM) and scanning electron microscopy (SEM) has been developed and applied for studying structure and composition of Burkholderia contaminans biofilms at the micro- and nanoscale levels. Our novel correlative approach enabled us to get new insights into the spatial phenotypic heterogeneity of biofilms, to analyze the individual cell phenotypes and to characterize the biofilm's matrix composition at very high spatial resolution.References[1] P. Martina, M. Bettiol, C. Vescina, P. Montanaro, M.C. Mannino, C.I. Prieto, C. Vay, D. Naumann, J. Schmitt, O. Yantorno, A. Lagares, A. Bosch, Genetic diversity of Burkholderia contaminans isolates from cystic fibrosis patients in Argentina, Journal of clinical microbiology, 51 (2013) 339-344.[2] P. Martina, M. Leguizamon, C.I. Prieto, S.A. Sousa, P. Montanaro, W.O. Draghi, M. Stammler, M. Bettiol, C. de Carvalho, J. Palau, C. Figoli, F. Alvarez, S. Benetti, S. Lejona, C. Vescina, J. Ferreras, P. Lasch, A. Lagares, A. Zorreguieta, J.H. Leitao, O.M. Yantorno, A. Bosch, Burkholderia puraquae sp. nov., a novel species of the Burkholderia cepacia complex isolated from hospital settings and agricultural soils, International journal of systematic and evolutionary microbiology, DOI 10.1099/ijsem.0.002293(2017).[3] H.C. Flemming, J. Wingender, The biofilm matrix, Nature reviews. Microbiology, 8 (2010) 623-633.