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International audienceBacteria living on surfaces form heterogeneous three-dimensionalconsortia known as biofilms, where they exhibit many specific properties oneof which is an increased tolerance to antibiotics. Biofilms are maintained by apolymeric network and display physical properties similar to that of complexfluids. In this work, we address the question of the impact of antibiotic treatmenton the physical properties of biofilms based on recently developed tools enablingthe in situ mapping of biofilm local mechanical properties at the micron scale.This approach takes into account the material heterogeneity and reveals thespatial distribution of all the small changes that may occur in the structure. Withan Escherichia coli biofilm, we demonstrate using in situ fluorescent labelingthat the two antibiotics ofloxacin and ticarcillin—targeting DNA replicationand membrane assembly, respectively—induced no detectable alteration of thebiofilm mechanical properties while they killed the vast majority of the cells. Inparallel, we show that a proteolytic enzyme that cleaves extracellular proteinsinto short peptides, but does not alter bacterial viability in the biofilm, clearlyaffects the mechanical properties of the biofilm structure, inducing a significantincrease of the material compliance. We conclude that conventional biofilm control strategy relying on the use of biocides targeting cells is missing a key target since biofilm structural integrity is preserved. This is expected toefficiently promote biofilm resilience, especially in the presence of persistercells. In contrast, the targeting of polymer network cross-links—among whichextracellular proteins emerge as major players—offers a promising route for thedevelopment of rational multi-target strategies to fight against biofilms

Bacterial biofilm mechanical properties persist upon antibiotic treatment and survive cell death