Correlation of biofilm expression types of Staphylococcus epidermidis with polysaccharide intercellular adhesin synthesis:: evidence for involvement of icaADBC genotype-independent factors

IcaADBC-encoded proteins mediate synthesis of the intercellular polysaccharide adhesin (PIA), which is essentially involved in Staphylococcus epidermidis biofilm formation. Seventy S. epidermidis isolates were investigated for their ability to form biofilm and synthesize PIA in different growth media including trypticase soy broth obtained from Becton Dickinson (TSBBBL), or Oxoid (TSBOXOID), and TSBOXOID supplemented with 0.5% N-acetylglucosamine, and for the presence of icaADBC. Dependent on the medium used (TSBBBL or TSBOXOID), the isolates exhibited a differential expression of PIA and biofilm formation, with 51 (72.85%) and 34 (48.57%) being biofilm positive, respectively. Using these growth media four different expression phenotypes were differentiated: similar quantities of biofilm formation in both TSBBBL and TSBOXOID (11 isolates, type A), significantly reduced biofilm expression in TSBOXOID compared to TSBBBL (23 isolates, type B), biofilm negative in TSBOXOID but biofilm producing in TSBBBL (17 isolates, type C) and biofilm negative in both media (19 isolates, type D). For all strains a biofilm-positive phenotype in a specific medium was closely linked to expression of PIA in that medium. All but one strain of expression type A-C and 7/19 expression type D strains were icaADBC positive. On the basis of restriction fragment length polymorphisms, the isolates were classified into two main icaADBC genotypes. There was no association between the observed biofilm-expression types and a defined icaADBC genotype. In the biofilm-negative S. epidermidis 5179, isolated from a ventriculo-atrial shunt infection, the insertion of IS257 interrupted the transcription of icaADBC, resulting in a PIA- and biofilm-negative phenotype. In all other icaADBC-positive, biofilm-negative isolates no major alterations of the icaADBC gene locus were identified. Obviously, expression of icaADBC, PIA synthesis and biofilm formation are integrated into a complex regulatory network involving other determinants independent of icaADBC genotype. Inactivation of icaADBC by IS elements is apparently a rare cause of a biofilm-negative phenotype in clinical S. epidermidis isolates.