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Detection of Biofilm-Grown Aspergillus fumigatus by Means of Atomic Force Spectroscopy: Ultrastructural Effects of Alginate Lyase

Published online by Cambridge University Press:  02 October 2012

M. Papi ,
A. Maiorana ,
F. Bugli ,
R. Torelli ,
B. Posteraro ,
G. Maulucci ,
M. De Spirito  and
M. Sanguinetti
M. Papi
Affiliation:
Istituto di Fisica, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
A. Maiorana
Affiliation:
Istituto di Fisica, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
F. Bugli
Affiliation:
Istituto di Istituto di Microbiologia, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
R. Torelli
Affiliation:
Istituto di Istituto di Microbiologia, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
B. Posteraro
Affiliation:
Istituto di Istituto di Microbiologia, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
G. Maulucci
Affiliation:
Istituto di Fisica, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
M. De Spirito*
Affiliation:
Istituto di Fisica, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
M. Sanguinetti
Affiliation:
Istituto di Istituto di Microbiologia, Università Cattolica del S. Cuore, L. go F. Vito 1, 00168 Roma, Italy
*
*Corresponding author. E-mail: [email protected]
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Abstract

Aspergillus fumigatus has become a leading cause of fungal morbidity and mortality, especially in immunocompromised patients. This fungus is able to grow as a multicellular community and produce a hydrophobic extracellular matrix (ECM), mainly composed of galactomannan and α-1,3 glucans, to protect itself from host defenses and antimicrobial drugs. This matrix envelops the fungus hyphae, binding them into a contiguous sheath on the colony surface, forming a biofilm and increasing the fungal resistance to adverse environmental factors. Adherence to host cells and resistance to physical removal play a key role in fungal colonization and invasion of the host and in a wide range of infections. Here we show that, by using atomic force spectroscopy, it is possible to exploit the peculiar hydrophobicity of the biofilm components (i.e., cell walls, ECM) to detect the biofilm spread, its growth, and lysis on rough surfaces. By means of this approach, we demonstrate that alginate lyase, an enzyme known to reduce negatively charged alginate levels in microbial biofilms, reduces the biofilm adhesion forces suggesting a loss of ECM from the biofilm, which could be used to enhance pharmacological treatments.

Keywords

biofilmfungiatomic force microscopyadhesionAspergillus fumigatusatomic force spectroscopyextracellular matrixhydrophobicityalginate
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 18 , Issue 5 , October 2012 , pp. 1088 - 1094
Copyright
Copyright © Microscopy Society of America 2012

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References

Alkawash, M.A., Soothill, J.S. & Schiller, N.L. (2006). Alginate lyase enhances antibiotic killing of mucoid Pseudomonas aeruginosa in biofilms. APMIS 114, 131–138.Google Scholar
Azeredo, J. & Oliveira, R. (2003). The role of hydrophobicity and exopolymers in initial adhesion and biofilm formation. In Biofilms in Medicine, Industry and Environmental Biotechnology: Characteristics, Analysis and Control, Lens, P., O'Flaherty, V., Moran, A.P. & Stoodley, P. (Eds.), pp. 16–31. London: IWA Publishing.Google Scholar
Beauvais, A., Schmidt, C., Guadagnini, S., Roux, P., Perret, E., Henry, C., Paris, S., Mallet, A., Prévost, M.C. & Latgé, J.P. (2007). An extracellular matrix glues together the aerial-grown hyphae of Aspergillus fumigatus . Cell Microbiol 9, 1588–1600.Google Scholar
Beech, I.B., Smith, J.R., Steele, A.A., Penegar, I. & Campbell, S.A. (2002). The use of atomic force microscopy for studying interactions of bacterial biofilms with surfaces. Colloid Surface B 23, 231–247.Google Scholar
Bhushan, B. & Dandavate, C. (2000). Thin-film friction and adhesion studies using atomic force microscopy. J Appl Phys 87, 1201–1210.Google Scholar
Briandet, R., Herry, J. & Bellon-Fontaine, M. (2001). Determination of the van der Waals, electron donor and electron acceptor surface tension components of static Gram-positive microbial biofilms. Colloid Surface B 21, 299–310.Google Scholar
Brunelli, R., Papi, M., Arcovito, G., Bompiani, A., Castagnola, M., Parasassi, T., Sampaolese, B., Vincenzoni, F. & De Spirito, M. (2007). Globular structure of human ovulatory cervical mucus. FASEB J 21, 3872–3876.Google Scholar
Butta, H.J., Cappella, B. & Kappla, M. (2005). Force measurements with the atomic force microscope: Technique, interpretation and applications. Surf Sci Rep 59, 1–152.Google Scholar
Cerca, N., Pier, G.B., Vilanova, M., Oliveira, R. & Azeredo, J. (2005). Quantitative analysis of adhesion and biofilm formation on hydrophilic and hydrophobic surfaces of clinical isolates of Staphylococcus epidermidis . Res Microbiol 156, 506–514.Google Scholar
Doyle, R. (2000). Contribution of the hydrophobic effect to microbial infection. Microb Infect 2, 391–400.Google Scholar
Epstein, A.K., Pokroy, B., Seminara, A. & Aizenberg, J. (2011). Bacterial biofilm shows persistent resistance to liquid wetting and gas penetration. PNAS 108, 995–1000.Google Scholar
Fonseca, A., Granja, P., Nogueira, J. & Oliveira, R. (2001). Staphylococcus epidermidis RP62A adhesion to chemically modified cellulose derivatives. J Mater Sci-Mat Med 12, 543–548.Google Scholar
Jonas, K., Tomenius, H., Kader, A., Normark, S., Römling, U., Belova, L.M. & Melefors, Ö. (2007). Roles of curli, cellulose and BapA in Salmonella biofilm morphology studied by atomic force microscopy. BMC Microbiol 7, 70–78.Google Scholar
Loussert, C., Schmitt, C., Prevost, M.C., Balloy, V., Fadel, E., Philippe, B., Kauffmann-Lacroix, C., Latgé, J.P. & Beauvais, A. (2010). In vivo biofilm composition of Aspergillus fumigatus . Cell Microbiol 12, 405–410.CrossRefGoogle ScholarPubMed
Mowat, E., Butcher, J., Lang, S., Williams, C. & Ramage, G. (2007). Development of a simple model for studying the effects of antifungal agents on multicellular communities of Aspergillus fumigatus . J Med Microbiol 56, 1205–1212.Google Scholar
Müller, F.M.C., Seidler, M. & Beauvais, A. (2011). Aspergillus fumigatus biofilms in the clinical setting. Med Mycol 49, S96–S100.Google Scholar
Núnez, M.E., Martin, M.O., Chanb, P.H. & Spain, E.M. (2005). Predation, death, and survival in a biofilm: Bdellovibrio investigated by atomic force microscopy. Colloid Surface B 42, 263–271.Google Scholar
Oh, Y.J., Jo, W., Yang, Y. & Park, S. (2007a). Influence of culture conditions on Escherichia coli O157:H7 biofilm formation by atomic force microscopy. Ultramicroscopy 107, 869–874.Google Scholar
Oh, Y.J., Jo, W., Yang, Y. & Park, S. (2007b). Biofilm formation and local electrostatic force characteristics of Escherichia coli O157:H7 observed by electrostatic force microscopy. Appl Phys Lett 90, 143901.CrossRefGoogle Scholar
Papi, M., Sylla, L., Parasassi, T., Brunelli, R., Monaci, M., Maulucci, G., Missori, M., Arcovito, G., Ursini, F. & De Spirito, M. (2009). Evidence of elastic to plastic transition in the zona pellucida of oocytes using atomic force spectroscopy. Appl Phys Lett 94, 153902.Google Scholar
Philippe, B., Ibrahim-Granet, O., Prévost, M.C., Gougerot-Pocidalo, M.A., Sanchez Perez, M., Van der Meeren, A. & Latgé, J.P. (2003). Killing of Aspergillus fumigatus by alveolar macrophages is mediated by reactive oxidant intermediates. Infect Immun 71, 3034–3042.Google Scholar
Sedin, D.L. & Rowlen, K.L. (2000). Adhesion forces measured by atomic force microscopy in humid air. Anal Chem 72, 2183–2189.Google Scholar
Shunmugaperumal, T. (2010). Biofilm eradication and prevention. New York: John Wiley & Sons.Google Scholar
Teixeira, P. & Oliveira, R. (1999). Influence of surface characteristics on the adhesion of Alcaligenes denitrificans to polymeric substrates. J Adh Sci Technol 13, 1287–1294.Google Scholar
Thompson, G.R. & Patterson, T.F. (2008). Pulmonary aspergillosis. Semin Respir Crit Care Med 29, 103–110.Google Scholar
Thundat, T., Zheng, X-Y., Chen, G.Y., Sharp, S.L., Warmack, R.J. & Schowalter, L.J. (1993). Characterization of atomic force microscope tips by adhesion force measurements. Appl Phys Lett 63, 2150.Google Scholar
van Oss, C. (1995). Hydrophobicity of biosurfaces—Origin, quantitative determination and interaction energies. Colloid Surface B 5, 91–110.Google Scholar
Voung, C. & Otto, M. (2002). Staphylococcus epidermidis infections. Microb Infect 4, 481–489.Google Scholar
Yoon, E.S., Yang, S.H., Han, H.G. & Kong, H. (2003). An experimental study on the adhesion at a nano-contact. Wear 254, 974–980.Google Scholar