Broth Macrodilution Assay

Kinetic of inactivation was determined by macrodilution method, according to Klaus et al. (2015). The mushroom extract was diluted in 1 mL of bacterial culture in MHB to reach the concentrations ranging from the MIC to the MBC, previously obtained by the broth microdilution assay. Bacterial growth was followed by taking samples at 0, 3, 6, 9, and 24 h and plating on MHA after serial sample dilutions. The bacterial number was counted upon incubation under aerobic conditions, at 37°C for 24 h, and expressed as log₁₀ CFU mL^-1. Positive controls were performed in the same way, except without adding the mushroom extract.