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Supplemental Materials: Effects of Medium Components on the Bulk Rheology and on the Formation of Ferning Patterns for Biofilm of Pseudomonas aeruginosa

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  • Pseudomonas aeruginosa is a ubiquitous bacterium that has an incredible ability to survive in various extreme environments and to persist despite bombardment by all types of antimicrobial agents. Its virulence and success within a broad range of hosts are largely thanks to the strength of its biofilms. The rheology of biofilm of P. aeruginosa was measured to investigate the bacterial response to nutritional conditions (medium that was modified with glycerol, glucose, sucrose, sodium chloride, and silver nitrate). The elastic modulus and the yield stress of the biofilm of P. aeruginosa increased in response to increases in glycerol, glucose, and sodium chloride. Alternatively, silver nitrate and glycerol inhibited biofilm formation at concentrations that were greater than 0.1 mM and 10 v/v%, respectively. While the addition of sucrose did not affect the biofilm rheology, it did alter the ferning pattern of the biofilm. Ferning patterns form as a result of diffusion-limited desiccation of the salt-macromolecule solution. Ferning coverage of about 50% and a ferning complexity score of 5 were found for most of the biofilm samples. The complexity increased with modifications that caused strengthening of the biofilm, while the coverage and complexity dropped to zero when no biofilm growth was observed due either to inhibition or to incomplete drying. The birefringent bundles of liquid crystals in the biofilm gained a new level of complexity and order within the ferning pattern that correlates with the biofilm robustness as characterized by its rheology, and these properties are heavily influenced by the nutritional environment of P. aeruginosa.
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