Control of Bacterial Biofilms by Microparticle Spacers

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Sergent, Olivia Gabrielle
Allison, Kyle
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Bacteria can be found in two distinct states: planktonic or as part of a sessile biofilm. While planktonic, free-floating bacteria are moderately easy to eradicate with antibiotics, bacteria in biofilms are not. Bacterial biofilms are tightly packed together, making it difficult for antibiotics to pass through the film and eliminate the bacteria; this antibiotic resistance makes it difficult to treat many bacterial infections. To try to decrease the antibiotic resistance of bacterial biofilms, this study utilized a microparticle spacer to create distance between cells in biofilms. The microparticle spacer was made by annealing Dynabead magnetic beads to Cholesterol-containing DNA; the Cholesterol on the spacer could bind to the lipid bilayer of bacteria. The spacer was mixed with various proportions of Escherichia coli K-12 wild-type and analyzed with fluorescence microscopy to determine binding affinity between the spacer and bacteria. Binding was quantified both visually and computationally, and the two methods were compared to determine accuracy. The spacers and bacteria showed high levels of binding, with over 84.8% of all spacers bound to bacteria according to the visual method. 13.3% difference was determined between the two methods; therefore, the computational method still needs improved precision and accuracy before it is used consistently. Due to a demonstrated high binding affinity, we hypothesize that this microparticle spacer could successfully add space between bacteria in biofilms.
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