Antimicrobial Potency Evaluation via ATP

A. veronii ACCC61732 cells (1 × 10⁵ CFU/mL mid-log phase in MHB) were exposed to various concentrationds (1/2×, 2×, 4×, 8×, 16×, and 32× MIC) of N6NH₂ (MIC 4 μg/mL), DN6NH₂ (MIC 4 μg/mL), N6PNH₂ (MIC 16 μg/mL), V112N6NH₂ (MIC 16 μg/mL), Guo-N6NH₂ (MIC 8 μg/mL), and CIP (MIC 0.125 μg/mL) for 60 min. ATP was measured using the BacTiter-GloTM Microbial Cell Viability Assay (Promega) following the manufacturer’s instructions. The assay was performed for three biological replicates at 37 °C, with luminescence recorded using a Tecan Infinite M1000 Pro plate reader. The fold reduction of ATP was calculated as: Fold reduction ATP = 1 − (L_treat − L_media/L_control − L_media), where L_treat is the luminescence of treated cells, L_media is the luminescence of MHB without cells, and L_control is the luminescence of untreated cells. Resultant graphs show mean (n = 3) and SEM for each data point, prepared in Prism 7 [46 (link)].