Role of Lipopolysaccharides in the Adhesion, Retention, and Transport of Escherichia coli JM109

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Abstract

The role of lipopolysaccharides (LPS) in bacterial adhesion was investigated via atomic force microscopy (AFM). Adhesion between a silicon nitride tip and Escherichia coli JM109 was measured in water and 0.01 M phosphate-buffered saline (PBS) on untreated cells and on a sample of E. coli treated with 100 mM ethylenediaminetetraacetic acid (EDTA), which removes approximately 80% of the LPS molecules. LPS removal decreased the adhesion affinity between the bacterial cells and the AFM tip from -2.1 +/- 1.8 to -0.40 +/- 0.36 nN in water and from -0.74 +/- 0.44 to -0.46 +/- 0.23 nN in 0.01 M PBS (statistically different, Mann-Whitney rank sum test, P < 0.01). The distributions of adhesion affinities between E. coli LPS macromolecules and the AFM tip could be described by gamma distribution functions. Direct measurements of the adhesive force between E. coil and a surface were compared with adhesion in batch and column experiments, and agreement was observed between the influences of LPS on adhesion in each system. Bacterial batch retention to glass or in packed beds to quartz sand decreased after LPS removal. When interaction forces were measured during the approach of the AFM tip to a bacterium, steric repulsive forces were seen for both treated and untreated cells, but the repulsion was greater when the LPS was intact A model for steric repulsion predicted a reduction of the equilibrium length of the surface polymers from 242 to 64 nm in water and from 175 to 81 nm in buffer, after removal of a portion of the LPS. DLVO calculations based on conventional and soft-particle DLVO theories predicted higher energy barriers to adhesion for all surfaces after LPS removal, consistent with experimental findings. Adhesion forces between the AFM tip and bacterial polymers were correlated with bacterial attachment and retention, while measurements of interaction forces during the approach of the AFM tip to the bacterium did not correlate with subsequent adhesion behavior to glass or quartz sand.

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