Disk-diffusion assay bacteria

Another reason for careful interpretation of the disk-diffusion assay is that it is subject to false-positive results that can be misinterpreted as antibiotic activity. For example, physical characteristics of the extract (viscosity, pH, etc.) can generate small zones of growth inhibition when bacteria are inoculated directly onto the surface of the agar plate. In addition, we have observed that some primary metabolites can inhibit growth when tested at high concentrations. It is also possible that simple molecules, or extract degradation products, can exhibit mild antibiotic properties. For these reasons, it is important that replicate extracts are tested and that small zones of inhibition are interpreted with caution. It is also important to clearly state the concentrations tested, even if naturally occurring concentrations are not known, so that activities can be reproduced and evaluated at a later time. 

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