Further characterization of Staphylococcus epidermidis transposon mutants deficient in primary attachment or...

Biofilm formation is suggested to be the result of primary attachment of Staphylococcus epidermidis cells to a polymer surface followed by accumulation in multilayered cell clusters. Here we describe the further characterization of transposon (Tn917) mutants of Staphylococcus epidermidis O-47 having been biofilm-negative in a polystyrene microtiter plate adhesion assay. Among 5000 Tn917 insertion strains, we isolated four biofilm-negative mutants, each carrying one copy of Tn917. The mutants could be divided into two phenotypic classes: class A (mut1 and mut1a) and class B (mut2 and mut2a). Mutants of phenotype class A lacked four cell surface proteins and were affected in the primary attachment to polystyrene, but remained able to form multilayered cell clusters and to produce PIA. Mutants of phenotype class B were able to attach to polystyrene, but did not form multilayered cell clusters nor produce PIA. The cell surface protein pattern relative to the wild type was unchanged in class B mutants. On Congo red agar, the wild type and class A mutants formed black colonies (positive reaction on Congo red agar) while class B mutant colonies were red (negative reaction). The initial binding of cells to polystyrene and the ability to form multilayered cell clusters were found to be phenotypically and genetically distinct traits.