In-vitro assessment of staphylococci biofilms formed under biologically-relevant conditions and correlation to the biofilm genotype

Abstract

Staphylococci have been implicated in chronic device-related infections due to their ability to form resistant biofilms on implanted medical devices. For a long time, two different mechanisms of biofilm formation were known in Staphylococcus spp., the ica-dependent biofilms in MSSA and CoNS and the ica-independent biofilms in MRSA. Recently, a new fibrin-based biofilm phenotype was identified when S. aureus isolates were allowed to construct biofilms in biologically-relevant conditions using plasma-coated surfaces and RPMI-1640 for biofilm development (RPMI-1640/Pl). In this study, 140 staphylococci clinical isolates (91 MRSA, 27 MSSA and 22 CoNS) were tested for biofilm formation, biofilm formers were selected and used to scrutinize the ability of RPMI-1640/Pl to support staphylococci biofilm formation. Results showed that, in RPMI-1640/Pl, the biofilm formation abilities of MRSA and MSSA isolates were non-significantly different compared to those formed in TSB and BHI, (Kruskal Wallis test, P = 0.3275 and 0.466 for MRSA and MSSA isolates, respectively). However, a significantly different biofilm formation ability was observed regarding the tested CoNS isolates (ANOVA test, P = 0.0006). Furthermore, biofilm formation in RPMI-1640/Pl under different incubation conditions was tested, and among the tested conditions, 48h of static incubation showed significantly elevated biofilm for both MRSA and MSSA. Finally, PCR was used to detect genes implicated in biofilm formation, and the genotypes were correlated to the biofilm formation ability in different tested conditions. In contrast to ordinary media, biofilm formation by staphylococci in RPMI-1640/Pl was positively correlated to coa, fnbA, fnbB and clfB.