5.12. Time-Killing Assays

The time-killing assays were employed to determine the killing kinetics of the peptides on the selected bacteria strains. The experiment was performed as described previously [51]. In brief, the bacteria culture was prepared the same way as the antimicrobial activity assay. A bacteria suspension (OD₆₀₀ = 0.1) was mixed with the peptides at the concentration of their corresponding MICs and 4 × MICs. At different time intervals, 10 µL of aliquots of each sample was transferred into a clean tube and a series of 10-fold dilution was prepared in PBS. A volume of 100 µL of diluted bacteria suspension was spread on an MHA plate, and the number of the bacterial colonies that were formed after an overnight-incubation at 37 °C. The bacteria cultured without any treatment was employed as the negative (growth) control. Three experiments were run in triplicate.