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Swimming plates were prepared with either Bromfield medium (0.04% tryptone, 0.01% yeast extract, and 0.01% CaCl2.2H2O) containing 0.3% Bacto agar or with MM containing 0.3% purified agar.
After carefully placing 3-μl suspension droplets in the centre of soft swimming plates (0.03% peptone, 0.03% yeast extract, 0.3% agar), plates were incubated at 25°C and evaluated after either 4 or 7 days by measuring the diameter of the swimming zone.
Briefly, bacterial stabs were placed on swimming plates (5 g/L NaCl, 10 g/L tryptone, and 0.03% (w/v) agarose), and swarming plates (8 g/L nutrient broth, 5 g/L glucose, with 0.5% (wt/vol) agar and after incubation at 30°C the diameter of bacterial growth was measured.
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The swimming motility of P. flavipulchra JG1 observed by swimming plate method showed a distinct motility halo, while the swarming ability was not detected.
Non-swarming (vegetative) cells were analyzed on either solid (NBG, 1.5% agar) or swim plates.
Follow the instructions provided by the manufacturer to interpret the MIC results on solid and swim plates.
Briefly, swim plates prepared by using of 1% tryptone, 0.5% NaCl and 0.3% (wt/vol) agarose, were inoculated with bacteria using a sterile toothpick.
In order to measure motility, 2-µl samples were spotted onto swimming agar plates (0.3% Difco agar, 0.4% glucose in LB broth) or swarming agar plates (0.45% Eiken agar, 0.4% glucose in LB broth), and incubated for 8 and 18 hours, respectively before inspection.
Quantitation of this phenotype showed that on average the zone of growth of K56-2 in swimming motility plates with the vehicle control was more than twice the size of the growth in plates with polymyxin B. Assays were also conducted to determine if swarming motility is impaired by polymyxin B. K56-2 wableble to swarm across plates containing the vehicle control.
The swimming motility plates were prepared with 0.3% "Difco" agar, 1.0% "Difco" Bacto-tryptone, 0.5% "Difco" Yeast Extract and 0.5% NaCl [ 7].
To measure bacterial motility we inoculated swimming media plates (triptone 10 g L−1, NaCl 5 g L−1, thymine 50 μg ml−1 and agar 0.25%) with 5 μL of cells in exponential phase.
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