Bacterial Strains and Biofilm Inhibition for Pathogens: P. aeruginosa (ΔMexAB-OprM) and S. aureus (ATCC BAA-2313) were used in both the biofilm disruption and inhibition assays. Overnight cultures of the strains were grown in Luria-Bertani (LB) medium (Table ). The biofilm inhibition and disruption assays were performed as previously described by Hancock et al, except for the following variations. For the biofilm inhibition assay, 2 μL of each fraction (10 mg/mL in DMSO) was added to fresh 96-well plates in quadruplicates. Overnight cultures of each pathogen were diluted to a starting OD600 of 0.01 in M63 liquid broth (Table ) and 98 μL was transferred to each well, including solvent controls. Inoculated plates were incubated in a static incubator for 16 to 20 h at 37 °C. Bacterial growth was measured at 600 nm to ascertain whether the fractions exhibited antibacterial properties. Adhered biofilms were stained with 0.1% (wt/vol) crystal violet (aq) and placed on an orbital shaker at low speed for 30 min. Planktonic cells were washed with water, the plates were dried (30 -60 min) at rt and then the crystal violet bound biofilms were solubilized using 130 μL of 30% acetic acid (aq). The solubilized material was transferred to new 96well plates and absorbances (595 nm) measured. Raw absorbance readings were normalized to a vehicle control (negative control) and active fractions were described as those that did not negatively impact bacterial growth but inhibited >50% of the biofilm formation. All active fractions were then prioritized for biofilm disruptions assays.