“…As described by Alfa et al ( 20 , 21 ) BBF was formed over eight days at room temperature inside PTFE channels (Endoscopy Development Company, Maryland Heights, MO, USA) using ATS-2015 (Healthmark, Fraser, MI, USA) containing 8 Log 10 /mL (day 1) of E. faecalis and P. aeruginosa . On days 3, 4, and 5, the PTFE channels were rinsed and exposed to glutaraldehyde partial fixation (1:50 dilution of glutaraldehyde) and then repeat biofilm formation allowed to develop overnight ( 13 ).…”