Rhamnolipid assessment
Rhamnolipid production was quantified using the Blue Agar Plate (Bap) method, following a previously established procedure (Bhat et al., 2015). The detection of rhamnolipid was conducted using a mineral salt agar medium supplemented with 2% glucose, 0.05% cetyltrimethylammonium bromide, and 0.02% methylene blue. Each treatment utilized a 0.5 McFarland’s suspension prepared from 24-hour bacterial culture, to which sterile chitosan was added in concentrations of 10 and 100 μg/L. Cork borers were used to create 4 mm diameter wells on methylene blue agar plates, which were then loaded with 30 μL of fresh culture from individual isolates. The plates were incubated at 37°C for 48-72 hours. Positive results were observed as a dark blue halo zone around the culture and assessed using a transilluminator at a wavelength of 365 nm.