Bacterial swarming reduces Proteus mirabilis and Vibrio parahaemolyticus cell stiffness and increases β-lactam susceptibility

Abstract

Swarmer cells of the gram-negative pathogenic bacteria Proteus mirabilis and Vibrio parahaemolyticus become long (>10-100 microns) and multinucleate during their growth and motility on polymer surfaces. We demonstrate increasing cell length is accompanied by a large increase in flexibility. Using a microfluidic assay to measure single-cell mechanics, we identified large differences in swarmer cell stiffness of (bending rigidity of P. mirabilis, 9.6 x 10^(-22) N m^2; V. parahaemolyticus, 9.7 x 10^(-23) N m^2) compared to vegetative cells (1.4 x 10^(-20) N m^2 and 3.2 x 10^(-22) N m^2, respectively). The reduction in bending rigidity (~3-15 fold) was accompanied by a decrease in the average polysaccharide strand length of the peptidoglycan layer of the cell wall from 28-30 to 19-22 disaccharides. Atomic force microscopy revealed a reduction in P. mirabilis peptidoglycan thickness from 1.5 nm (vegetative) to 1.0 nm (swarmer) and electron cryotomography indicated changes in swarmer cell wall morphology. P. mirabilis and V. parahaemolyticus swarmer cells became increasingly sensitive to osmotic pressure and susceptible to cell wall-modifying antibiotics (compared to vegetative cells)--they were ~30% more likely to die after 3 h of treatment with minimum inhibitory concentrations of the beta-lactams cephalexin and penicillin G. Long, flexible swarmer cells enables these pathogenic bacteria to form multicellular structures and promotes community motility. The adaptive cost of swarming is offset by a fitness cost in which cells are more susceptible to physical and chemical changes in their environment, thereby suggesting the development of new chemotherapies for bacteria that leverage swarming for survival

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