The biofilm biomass was determined by staining the adherent bacteria on the discs with 0.1% safranin (Merck) for 30 min. The unabsorbed safranin and unattached bacteria were removed by washing in PBS. safranin was released from the biofilm by incubation in acetic acid (30%) for 10 min, and the amount of safranin was quantified by measuring the absorbance at 530 nm in a Cytation™ 3 Cell Imaging Multi-Mode Reader (BioTek, Santa Clara, CA, USA). The experiments were repeated three times.
Discs incubated in sterile modified BHI were used as a positive control, and discs with biofilm (no decontamination) were used as a negative control.
Discs incubated in sterile modified BHI were used as a positive control, and discs with biofilm (no decontamination) were used as a negative control.