ALT Percentage of evaporation at 24 h in air and water-saturated atmospheres. 96-well plate with final volume of 100 µL and 384-well plate with final volume of 30 µL were incubated for 24 h at 37°C without agitation. Results are expressed in percentage of final volume at 24 h compared to initial volume. The figure shows a schematic representation of the top left-hand corner of a microtiter plate.

ALT Experimental design and workflow. (A) Serial passaging of planktonic and biofilms evolved independently by a daily aliquot/bead passage over 10 days. (B) Intermittent colistin experimental treatment conditions during evolution: 0 mg/L (no treatment, gray), 0.5 mg/L, (static, light blue), 1 mg/L, (static, blue), and 0.5–8 mg/L, (increasing, dark blue) at days 1, 3, 5, 7, and 9. (C) Daily population analysis profiling (PAP) tests were performed to determine the survival frequency in each evolved population. (D) Whole-genome sequencing analysis was performed to the ancestral PAO1 strain as well as representative resistant clones from the last day of evolution.