Inhibition of Staphylococcus aureus Biofilm Formation
S. aureus samples were incubated with MIC and sub-MIC of the examined agents in Tryptic soy broth with 2% glucose at 37 °C for 24 h. After incubation, the plate was washed twice with sterile PBS and fixed with methanol for 10 min. Methanol was removed by pipetting and the plate was air-dried. The biofilms were stained by using 0.1% crystal violet (Sigma Aldrich, Darmstadt, Germany) for 30 min, water washed and air dried, and the stain was dissolved in 96% ethanol (Zorka, Sabac, Serbia). The absorbance was read at 570 nm on a Multiskan™ FC microplate photometer (Thermo Scientific™, Waltham, MA, USA) and the results are presented as the inhibition of biofilm formation (%) [27 (link)].
Inhibition of the formed Staphylococcus aureus biofilm
The S. aureus inoculum was incubated in a microtiter plate with an adhesive bottom for 24 h at 37 °C. After incubation, the plate was washed 3× with saline and the adhered cells were treated with MBC of extracts for 30 s. After washing, addition of methanol, drying, staining with crystal violet dye and addition of ethanol, the absorbance was read at a wavelength of 570 nm on an automated Elisa reader (Multiskan™ FC microplate photometer, Thermo Scientific), and the percentage of biofilm diminishing was calculated [28 (link)].
S. aureus samples were incubated with MIC and sub-MIC of the examined agents in Tryptic soy broth with 2% glucose at 37 °C for 24 h. After incubation, the plate was washed twice with sterile PBS and fixed with methanol for 10 min. Methanol was removed by pipetting and the plate was air-dried. The biofilms were stained by using 0.1% crystal violet (Sigma Aldrich, Darmstadt, Germany) for 30 min, water washed and air dried, and the stain was dissolved in 96% ethanol (Zorka, Sabac, Serbia). The absorbance was read at 570 nm on a Multiskan™ FC microplate photometer (Thermo Scientific™, Waltham, MA, USA) and the results are presented as the inhibition of biofilm formation (%) [27 (link)].
Inhibition of the formed Staphylococcus aureus biofilm
The S. aureus inoculum was incubated in a microtiter plate with an adhesive bottom for 24 h at 37 °C. After incubation, the plate was washed 3× with saline and the adhered cells were treated with MBC of extracts for 30 s. After washing, addition of methanol, drying, staining with crystal violet dye and addition of ethanol, the absorbance was read at a wavelength of 570 nm on an automated Elisa reader (Multiskan™ FC microplate photometer, Thermo Scientific), and the percentage of biofilm diminishing was calculated [28 (link)].
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