Figure 1.

Confocal microscopical analysis of Pg biomass in 4-day old mixed-species biofilms under micro-aerobic conditions (6% O₂). (A) Pg was incubated under static conditions for biofilm growth (left bar) and under shaking conditions (100 rpm) to prevent biofilm formation and to maintain a planktonic state (right bar is absent due to lack of growth). After 4 days, the CFUs from both samples were tested. (B) Single and multiplex staining of Pg (green), Aa (red) and Ss (blue) with FISH probes specific to the conserved 16s ribosomal (rRNA) genes of these bacteria (scale bar shown in top left panel = 20 µm). (C) Pg biomass in (B) was calculated for single and multiplex biofilms. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, Student’s t-test. Means and standard deviations from triplicate experiments are shown.