Figure 3.

Biofilm eradication by RPMs FK20 and FdK. MRSA strain USA300 LAC (A) and P. aeruginosa strain PA01 (B) were cultured at 37 °C overnight in LB, then diluted 1:100 in M63 minimal media and aliquoted into a round-bottom 96-well plate. The plate was covered and incubated at 37 °C overnight. The liquid was shaken out from the plate, and it was rinsed with sterile H₂O. Fresh M63 media with RPMs, antibiotics (daptomycin 10 μg mL⁻¹, tobramycin 10 μg mL⁻¹), or PBS added was pipetted into the same wells. The plate was covered and again incubated at 37 °C overnight. The plate was dumped and rinsed again, and 125 μL of 0.1% crystal violet was added to each well. Wells were incubated at room temperature for 15 min, then dumped and rinsed 3 times with distilled H₂O. The plate was left to dry overnight at room temperature, uncovered. Acetic acid (125 μL of 30%) was added to each well, and the liquid from each of the wells was transferred to a new, flat-bottom 96-well dish. The plate was read at 550 nm, and absorbance was recorded. Error bars represent SD for n = 3 of a typical experiment performed independently three times. Significance was determined with ANOVA and Tukey HSD. *p < 0.05, **p < 0.01, and ***p < 0.001 significance compared to control; data without symbols are not significantly different.