The biofilm formation was evaluated for all isolates on 96-well microtiter polystyrene plates TPP 92,096 (Techno Plastic Products; Trasadingen, Switzerland) as described by Ferreira et al. [21]. Twenty microliters of a bacterial suspension in sterile distilled water corresponding to the 0.5 McFarland standard were added in triplicate to the wells containing 180 µl of tryptic soy broth, TSB (Becton, Dickinson and Company; Sparks, MD, USA), supplemented with 1% glucose (Isofar; Duque de Caxias, RJ, Brazil). The plates were incubated at 37°C for 24 h without shaking. The biofilm was heat-fixed at 60°C for 1 h, stained with 0.1% (w/v) safranin for 15 min, and distained with 95% (v/v) ethanol for 30 min. The biofilm phenotype was categorized as absent, weak, moderate, or strong according to Stepanović et al [22]. Besides that, 13 isolates were randomly chosen in order to investigate biofilm production in wells coated with 50 µg/ml of human plasmatic fibrinogen (Sigma Chemical Company; St. Louis, MO, USA) [23]. All tests were performed in triplicate. Staphylococcus aureus ATCC 33591 was used as a control for biofilm formation.

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