All targeted strains were grown at 37°C in LB broth under aerobic conditions (200 rpm), except for Clostridia that were grown in BHI-YH under anaerobic conditions (in a Trexler-type anaerobic chamber, without stirring). Peptides dissolved in sterile distilled water were sterilized 2 min under UV light and added to sterile F-bottom polypropylene 96-well microplates from 100 to 0.1 μM. Twofold series dilutions were performed in cell suspension of each bacterial target, including Cp, at 10−4 OD600 units. For other Clostridia, higher concentrations of bacteria (i.e., cell suspensions at 10−3 OD600 units) were used. MIC was defined as the lowest concentration of peptide that inhibited the visible growth of bacteria after 24 to 48 hours incubation at 37°C. Cell suspensions were then plated on appropriate solid medium devoid of any antimicrobial agent to evaluate bacterial growth. MBCs were defined as the lowest concentration of peptide that inhibited the visible growth of bacteria after 24 to 48 hours incubation at 37°C on a solid medium. MICs and MBCs were determined three times. Sterility and growth controls were prepared for each assay.

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