Biofilm detection

Phenotypic assessment of biofilm production was done in micro-broth bioassay [20]. Briefly, overnight pure colonies were suspended in 200μl Brain Heart infusion (BHI) broth supplemented with 0.25% glucose and incubated at 37C for 24 hours. After incubation, the growth medium was aspirated away and the wells were washed thrice with Phosphate Buffer Saline (PBS). The wells were stained with 0.5% w/v Crystal Violet for 5 minutes and gently rinsed. Ethanol (70%) was added to dissolve the Crystal Violet and absorbance of the stained biofilm solution was measured with UV Spectrophotometer against absorbance of non-biofilm producing Staphylococcus aureus.