The emergence of new infectious diseases, the resurgence of several infections that appeared to have been controlled and the increase in bacterial resistance have created the necessity for studies directed towards the development of new antimicrobials. Considering the failure to acquire new molecules with antimicrobial properties from microorganisms, the optimization for screening methods used for the identification of antimicrobials from other natural sources is of great importance. The objective of this study was to evaluate technical variants used in screening methods to determine antibacterial activity of natural products. Thus, a varied range of natural products of plant, fungi and lichen origin were tested against two bacterial species, Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922, by two variants of the agar diffusion method (well and disc), two variants of the bioautographic method (direct and indirect) and by microdilution assay. We concluded that the well-variant of the diffusion method was more sensitive than the disc-variant, whilst the direct-variant of the bioautographic method exhibited a greater sensitivity if compared to indirect-variant. Bioautographic and diffusion techniques were found to have similar sensitivity; however the latter technique provided more suitable conditions for the microbial growth. In this study, we also discussed the best conditions for the determination of minimal inhibitory concentration.
agar diffusion; antimicrobial activity; bioautographic methods; minimum inhibitory concentration; minimum bactericidal concentration; natural products
