Clonality, virulence and the occurrence of genes encoding antibiotic resistance among Staphylococcus warneri isolates...

Coagulase-negative Staphylococcuswarneri is an opportunistic pathogen capable of causing several infections, especially in patients with indwelling medical devices. We evaluated the virulence-associated properties of 23 clinical isolates recovered from blood specimens. In addition, the carriage of biofilm-associated genes, as well as antibiotic-resistant genes, was identified. S. warneri isolates appeared to be clonally unrelated and revealed a high degree of genetic diversity. All isolates revealed adhesion to epithelial cells, and 43.5 % of strains invaded the cells. Moreover, 52 % of isolates formed biofilm in vitro. PCR analysis demonstrated the presence of the ica operon, in two of the 12 biofilm-positive isolates. This indicated that biofilm formation, in this species, is not restricted to strains harbouring icaADBC genes, encoding polysaccharide intercellular adhesion. Analysis by confocal laser scanning microscopy revealed that biofilm-forming strains formed a three-dimensional structure, composed of mainly living cells. All strains revealed cell-contact cytotoxicity that was strongly associated with biofilm formation. Moreover, cell-free supernatants, of 95 % of the isolates, expressed a cytotoxic activity which caused the destruction of HeLa cells. S. warneri capable of forming biofilm carried significantly more genes encoding resistance to beta-lactams, aminoglicosides and macrolide-lincosamide streptogramin B antibiotics than biofilm-negative isolates. We have shown that tigecycline/rifampicin is effective against bacteria growing as a biofilm. The biofilm inhibitory concentration range of tigecycline/rifampicin was ≤1 µg ml-1. Results indicated that S. warneri have the ability to adhere, form biofilm, invade and destroy epithelial cells, which could be important mechanisms contributing to the development of diseases.