The antimicrobial activities of the peptides were measured by the two-fold dilution method [32 (link)]. Each strain was inoculated in the medium and cultivated to the logarithmic phase. A series of diluted peptides (concentrations of 3.1–100 μM, 50 μL) were prepared in a 96-well plate (Corning, New York, NY, USA), and the same volume of the bacterial suspension (the final concentration of 1 × 106 CFU/mL) was added. After incubation at 37 °C for 18–24 h, the absorbance at 600 nm was measured by using a microplate reader (Multiskan Spectrum, BioTek, Winuski, VT, USA). MIC was the lowest concentration of peptides that completely inhibited bacterial growth. All tests were performed in triplicate independent experiments.
From the MIC measuring plates, 50 μL of bacteria suspension in the presence of peptides when the concentrations were equal and up to MICs was spread on the BHI agar medium, and incubated at 37 °C for 24 h. The peptide concentration with no bacterial growth was defined as MBC [33 (link)].
From the MIC measuring plates, 50 μL of bacteria suspension in the presence of peptides when the concentrations were equal and up to MICs was spread on the BHI agar medium, and incubated at 37 °C for 24 h. The peptide concentration with no bacterial growth was defined as MBC [33 (link)].
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