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After 48 h, biofilms were washed once with 1 × PBS followed by treatment with different enzyme solutions prepared in 1 × PBS, pH 7.4, including: 5, 10, 20, and 40U DNase-I, 100, 200, 500 and 1000 μg/ml amylase and 50, 100, 200 and 500 μg/ml Proteinase K, for 24 h, at 37°C and 150 rpm, the control was treated with 1 × PBS alone