Abstract
Manual and automated methods were compared for routine screening of compounds for antimicrobial activity. Automation generally accelerated assays and required less user intervention while producing comparable results. Automated protocols were validated for planktonic, biofilm, and agar cultures of the oral microbe Streptococcus mutans that is commonly associated with tooth decay. Toxicity assays for the known antimicrobial compound cetylpyridinium chloride (CPC) were validated against planktonic, biofilm forming, and 24 h biofilm culture conditions, and several commonly reported toxicity/antimicrobial activity measures were evaluated: the 50 % inhibitory concentration (IC50), the minimum inhibitory concentration (MIC), and the minimum bactericidal concentration (MBC). Using automated methods, three halide salts of cetylpyridinium (CPC, CPB, CPI) were rapidly screened with no detectable effect of the counter ion on antimicrobial activity.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0191-2) contains supplementary material, which is available to authorized users.
Highlights
The automation of biological assays promises to improve reproducibility by minimizing ‘human error’ and to increase experimental throughput by tirelessly repeating standardized operations (Felder 1998; Tomar 1999; Sarkozi et al 2003)
We report here an evaluation of automated liquid handling instrumentation for routine, high-throughput screening of emerging antimicrobial compounds against S. mutans
Automated antimicrobial activity assay: planktonic culture The robotic liquid handler was used to automate a simple planktonic antimicrobial activity assay for the quaternary ammonium salt cetylpyridinium chloride (CPC), a known antimicrobial that is added to some commercially available mouthwashes and toothpastes
Summary
The automation of biological assays promises to improve reproducibility by minimizing ‘human error’ and to increase experimental throughput by tirelessly repeating standardized operations (Felder 1998; Tomar 1999; Sarkozi et al 2003). For both planktonic and biofilm cultures, 5 μL of suspended cells from each well of an antimicrobial activity assay plate were inoculated onto the agar surface using the liquid-level sensing capability of the automated liquid handler.
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