Fig. 6: iAMF and antibiotics can work on biofilm of various ages.

P. aeruginosa (PAO1) and S. aureus (UAMS1) biofilms were cultured until day 7 following the same protocol with media replenishment every 24 h. Then the biofilms were treated with iAMF doses at 0 and 12 h (T_max = 65 °C, Δt_exp = 5 min, 15 min per dose) and specified antibiotics. a 7-day P. aeruginosa (PAO1) biofilm log reduction (CFU) post 24 h with iAMF and 0.5 μg mL⁻¹ ciprofloxacin. CFU was counted at time points 0, 12 (pre- and post-AMF), and 24 h. b 7-day S. aureus (UAMS1) biofilm log reduction (CFU) post 24 h with iAMF and 2 μg mL⁻¹ linezolid. CFU was counted at time points 0, 12, and 24 h. c Comparison of prior 2-day (48 h) biofilm and 7-day P. aeruginosa (PAO1) biofilm under the same iAMF (T_max = 65 °C, Δt_exp = 5 min, 15 min per dose) treatment and 0.5 μg mL⁻¹ ciprofloxacin at time 0 and 24 h. d Comparison of 2-day (48 h) biofilm and 7-day S. aureus (UAMS1) biofilm under the same iAMF (T_max = 65 °C, Δt_exp = 5 min, 15 min per dose) treatment and 2 μg mL⁻¹ linezolid at time 0 and 24 h. n = 3. Error bars indicate SD. CFU limit of detection (LOD) = 0.78 log(CFU cm⁻²). Two-way ANOVA. Statistical significance: not significant (ns).