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Both bottles contain 25 ml of tryptic soy broth, 0.05% sodium polyanetholesulfonate, and mixed resins.
The new formulation, designated BACTEC Plus, contains 25 ml of tryptic soy broth, 0.05% sodium polyanetholesulfonate, and mixed resins.
The isolates of S. epidermidis were inoculated in 10 mL of tryptic soy broth with 0.25% glucose and incubated overnight with shaking at 37°C.
The hydroxyapatite discs and the bacterial suspension were added into sterile test tubes containing 2 ml of tryptic soy broth to make the final concentration of the bacterial cells approximately 2 × 107 CFU/ml.
For each experiment, bacteria were subcultured on tryptic soy agar (TSA, Merck, Darmstadt, Germany) for about 36 h at 37°C and then grown for 24 h in 15 mL of tryptic soy broth (TSB, Merck), at 37°C under a constant agitation of 120 rpm (SI50; Stuart Scientific, Redhill, UK).
Biofilms were grown, in 24-well plates made of polystyrene plastic (Orange Scientific, Braine-l'Alleud, Belgium), by inoculating 1.5 µL of an overnight culture into 1 mL of Tryptic Soy Broth (Liofilchem, Teramo, Italy) supplemented with glucose to a final concentration of 0.65 % (v/v) (TSB0.65 %G), and by incubating at 37 °C with agitation at 120 rpm.
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The colony was moved to 3 ml of sterile tryptic soy broth (TSB) in a culture tube and through an overnight shaking at 225 rpm and 37 °C.
For the mid-log phase culture, fifty microliters of this culture was transferred to 3 ml of sterile tryptic soy broth (TSB), at 225 rpm and 37 °C, and then diluted with sterile 1% TSB to OD600 = 0.01 for inoculation into the flow cells.
Pulsed cells were immediately diluted with 1 ml of either Tryptic Soy Broth (TSB) medium or TSB supplemented with 5 mM MgCl2, 5 mM MgSO4, and 250 mM sucrose (TSB-plus medium) and incubated during recovery period with shaking (120 rpm) for 2 or 3 h before spreading on LB medium agar plate including tetracycline (10 μg ml-1) or kanamycin (5 μg ml-1) as indicated.
One day prior to testing, 100 mL of TSB (Tryptic Soy) were inoculated with lyophilized challenge species and incubated at 36°C ± 1°C.
Following growth, a single isolated colony was selected and used to inoculate 3 mL of 20% tryptic soy broth (TSB) medium.
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