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Macroscopic stab assay for twitching motility. Cells from an overnight LB agar (1.5%, wt/vol) plate were inoculated with a toothpick to the bottom of the LB agar (1%, wt/vol) plate and incubated for 24 h at 37°C. The diffuse interstitial zone is a measure of twitching motility. The denser and smaller zone represents surface colony growth. Strains: WT, PAO1; Δppk, PAOM5; ΔfliD, PAO-D; and ΔpilA, PAO-NP.
