The overlay agar method referred to Abudoleh & Mahasneh [22] was used with modifications for this screening. Each Actinomycetes isolate inoculated onto YMEA with a straight streak method and incubated at 28 °C for 2 days. C. violaceum was grown separately in 50 mL of Luria-Bertani Broth (LB) (Oxoid) and incubated at 28 °C 24 h. Indicator bacteria then diluted until OD600 = 0.132, then a total of 100 μL culture were taken and mixed with semisolid agar (0.75% LA) of 5 mL volume for an overlay on top of the selected Actinomycetes bacteria in the YMEA before, later incubated at 28 °C for 2 days. The positive results were indicated by the absence of violacein pigmentation around the Actinomycetes isolates.

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