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Abstract
Microtiter plates with 96 wells are routinely used in biofilm research mainly because they enable high-throughput assays. These platforms are used in a variety of conditions ranging from static to dynamic operation using different shaking frequencies and orbital diameters. The main goals of this work were to assess the influence of nutrient concentration and flow conditions on biofilm formation by Escherichia coli in microtiter plates and to define the operational conditions to be used in order to simulate relevant biomedical scenarios. Assays were performed in static mode and in incubators with distinct orbital diameters using different concentrations of glucose, peptone and yeast extract. Computational fluid dynamics (CFD) was used to simulate the flow inside the wells for shaking frequencies ranging from 50 to 200 rpm and orbital diameters from 25 to 100 mm. Higher glucose concentrations enhanced adhesion of E. coli in the first 24 h, but variation in peptone and yeast extract concentration had no significant impact on biofilm formation. Numerical simulations indicate that 96-well microtiter plates can be used to simulate a variety of biomedical scenarios if the operating conditions are carefully set.
Acknowledgements
The authors acknowledge financial support provided by the Operational Programme for Competitiveness Factors – COMPETE, the European Fund for Regional Development – FEDER and the Portuguese Foundation for Science and Technology – FCT, through Projects PTDC/EQU-FTT/105535/2008 and PTDC/EBB-BIO/104940/2008. Luciana Gomes acknowledges the receipt of a PhD grant from FCT (SFRH/BD/80400/2011) and José Araújo acknowledges financial support from FCT through Grant SFRH/BPD/64148/2009.