Study of the formation of a biofilm by clinical strains of Staphylococcus aureus

Abstract

A study on biofilm formation was carried out using five methicillin-sensitive [MSSA] and five methicillin-resistant [MRSA] strains of S. aureus. In each group, there were four strains isolated from patients from Kinshasa (Democratic Republic of Congo, DRC) and one reference strain. All of the strains were hydrophobic. The adherence of the bacteria to an abiotic surface was studied with the Biofilm Ring Test (BFRT®) and the crystal violet staining method (CVSM). Both techniques showed that eight of the strains formed biofilms within 2–3 h. The extent of the biofilm formed by one strain could only be observed with the CVSM. Periodate prevented the formation of biofilms and, in separate experiments, destroyed the biofilm pre-formed by the MSSA reference, but not those pre-formed by the clinical strains. Proteinase K destroyed all pre-formed biofilms. Six of the strains were icaA⁺; the clinical MSSA strains were not. The results also indicated different mechanisms of biofilm development between MSSA and MRSA clinical strains. The BFRT® and the CVSM are complementary techniques to study the adhesion of bacteria and the development of biofilms.

Keywords:

• Biofilm Ring Test
• crystal violet
• periodate
• proteinase K

Acknowledgements

This work was supported by grant No. 3.4577.10 of the Fonds National de la Recherche Scientifique of Belgium and by a grant from the Coopération Technique Belge (CTB). J-M. Liesse-Iyamba is a recipient of a Research Fellowship from the CTB.