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Surface-associated lifestyles dominate in the bacterial world. Large multicellular assemblies, called biofilms, are essential to the survival of bacteria in harsh environments and are closely linked to antibiotic resistance in pathogenic strains. Biofilms stem from the surface colonization of a wide variety of substrates encountered by bacteria, from living tissues to inert materials. Here, we demonstrate experimentally that the promiscuous opportunistic pathogen Pseudomonas aeruginosa explores substrates differently based on their rigidity, leading to striking variations in biofilm structure, exopolysaccharides (EPS) distribution, strain mixing during co-colonization and phenotypic expression. Using simple kinetic models, we show that these phenotypes arise through a mechanical interaction between the elasticity of the substrate and the type IV pilus (T4P) machinery, that mediates the surface-based motility called twitching. Together, our findings reveal a new role for substrate softness in the spatial organization of bacteria in complex microenvironments, with far-reaching consequences on efficient biofilm formation.

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Astronauts are spending longer periods locked up in ships or stations for scientific and exploration spatial missions. The International Space Station (ISS) has been inhabited continuously for more than 20 years and the duration of space stays by crews could lengthen with the objectives of human presence on the moon and Mars. If the environment of these space habitats is designed for the comfort of astronauts, it is also conducive to other forms of life such as embarked microorganisms. The latter, most often associated with surfaces in the form of biofilm, have been implicated in significant degradation of the functionality of pieces of equipment in space habitats. The most recent research suggests that microgravity could increase the persistence, resistance and virulence of pathogenic microorganisms detected in these communities, endangering the health of astronauts and potentially jeopardizing long-duration manned missions. In this review, we describe the mechanisms and dynamics of installation and propagation of these microbial communities associated with surfaces (spatial migration), as well as long-term processes of adaptation and evolution in these extreme environments (phenotypic and genetic migration), with special reference to human health. We also discuss the means of control envisaged to allow a lasting cohabitation between these vibrant microscopic passengers and the astronauts.

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Surface colonization underpins microbial ecology on terrestrial environments. Although factors that mediate bacteria-substrate adhesion have been extensively studied, their spatiotemporal dynamics during the establishment of microcolonies remains largely unexplored. Here, we use laser ablation and force microscopy to monitor single-cell adhesion during the course of microcolony formation. We find that adhesion forces of the rod-shaped bacteria Escherichia coli and Pseudomonas aeruginosa are polar. This asymmetry induces mechanical tension, and drives daughter cell rearrangements, which eventually determine the shape of the microcolonies. Informed by experimental data, we develop a quantitative model of microcolony morphogenesis that enables the prediction of bacterial adhesion strength from simple time-lapse measurements. Our results demonstrate how patterns of surface colonization derive from the spatial distribution of adhesive factors on the cell envelope.

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We introduce and analyze a model for osmotically spreading bacterial colonies at solid-air interfaces that includes wetting phenomena, i.e., surface forces. The model is based on a hydrodynamic description for liquid suspensions which is supplemented by bioactive processes. We show that surface forces determine whether a biofilm can expand laterally over a substrate and provide experimental evidence for the existence of a transition between continuous and arrested spreading for Bacillus subtilis biofilms. In the case of arrested spreading, the lateral expansion of the biofilm is confined, albeit the colony is biologically active. However, a small reduction in the surface tension of the biofilm is sufficient to induce spreading. The incorporation of surface forces into our hydrodynamic model allows us to capture this transition in biofilm spreading behavior.

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Understanding interactions between membranes requires measurements on well-controlled systems close to natural conditions, in which fluctuations play an important role. We have determined, by grazing incidence X-ray scattering, the interaction potential between two lipid bilayers, one adsorbed on a solid surface and the other floating close by. We find that interactions in this highly hydrated model system are two orders of magnitude softer than in previously reported work on multilayer stacks. This is attributed to the weak electrostatic repulsion due to the small fraction of ionized lipids in supported bilayers with a lower number of defects. Our data are consistent with the Poisson-Boltzmann theory, in the regime where repulsion is dominated by the entropy of counter ions. We also have unique access to very weak entropic repulsion potentials, which allowed us to discriminate between the various models proposed in the literature. We further demonstrate that the interaction potential between supported bilayers can be tuned at will by applying osmotic pressure, providing a way to manipulate these model membranes, thus considerably enlarging the range of biological or physical problems that can be addressed.

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Bacteria inhabit a wide variety of environments in which fluid flow is present, including healthcare and food processing settings and the vasculature of animals and plants. The motility of bacteria on surfaces in the presence of flow has not been well characterized. Here we focus on Pseudomonas aeruginosa, an opportunistic human pathogen that thrives in flow conditions such as in catheters and respiratory tracts. We investigate the effects of flow on P. aeruginosa cells and describe a mechanism in which surface shear stress orients surface-attached P. aeruginosa cells along the flow direction, causing cells to migrate against the flow direction while pivoting in a zig-zag motion. This upstream movement is due to the retraction of type IV pili by the ATPase motors PilT and PilU and results from the effects of flow on the polar localization of type IV pili. This directed upstream motility could be beneficial in environments where flow is present, allowing bacteria to colonize environments that cannot be reached by other surface-attached bacteria.

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In most environments, such as natural aquatic systems, bacteria are found predominantly in self-organized sessile communities known as biofilms. In the presence of a significant flow, mature multispecies biofilms often develop into long filamentous structures called streamers, which can greatly influence ecosystem processes by increasing transient storage and cycling of nutrients. However, the interplay between hydrodynamic stresses and streamer formation is still unclear. Here, we show that suspended thread-like biofilms steadily develop in zigzag microchannels with different radii of curvature. Numerical simulations of a low-Reynolds-number flow around these corners indicate the presence of a secondary vortical motion whose intensity is related to the bending angle of the turn. We demonstrate that the formation of streamers is directly proportional to the intensity of the secondary flow around the corners. In addition, we show that a model of an elastic filament in a two-dimensional corner flow is able to explain how the streamers can cross fluid streamlines and connect corners located at the opposite sides of the channel.

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Although ubiquitous, the processes by which bacteria colonize surfaces remain poorly understood. Here we report results for the influence of the wall shear stress on the early-stage adhesion of Pseudomonas aeruginosa PA14 on glass and polydimethylsiloxane surfaces. We use image analysis to measure the residence time of each adhering bacterium under flow. Our main finding is that, on either surface, the characteristic residence time of bacteria increases approximately linearly as the shear stress increases (∼0-3.5 Pa). To investigate this phenomenon, we used mutant strains defective in surface organelles (type I pili, type IV pili, or the flagellum) or extracellular matrix production. Our results show that, although these bacterial surface features influence the frequency of adhesion events and the early-stage detachment probability, none of them is responsible for the trend in the shear-enhanced adhesion time. These observations bring what we believe are new insights into the mechanism of bacterial attachment in shear flows, and suggest a role for other intrinsic features of the cell surface, or a dynamic cell response to shear stress.

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We investigate a new type of behavior whereby small vesicles orbiting around a larger vesicle in a toroidal electrohydrodynamic flow undergo dynamic angular segregation. Application of a low frequency (approximately 50 Hz) electric field induces aggregation of adjacent unilamellar vesicles near the electrode, in a manner similar to that observed with rigid colloidal particles. For polydisperse vesicle suspensions, however, small vesicles (<10 microm) are often observed to "orbit" around larger vesicles (>20 microm) in a toroidal electrohydrodynamic flow field. While orbiting, the smaller vesicles gradually segregate into well-defined angular cross sections. Viewed from above, the vesicles appear to form dynamic "bands" at prescribed angles, separated by regions devoid of vesicles. We interpret the angular segregation in terms of induced dipolar interactions, and we propose a model based on point dipoles rotating in a cellular flow field. We demonstrate that the model yields a surprisingly diverse range of vesicle trajectories, including many that are qualitatively consistent with the experimental observations.

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Bacterial biofilms have an enormous impact on medicine, industry and ecology. These microbial communities are generally considered to adhere to surfaces or interfaces. Nevertheless, suspended filamentous biofilms, or streamers, are frequently observed in natural ecosystems where they play crucial roles by enhancing transport of nutrients and retention of suspended particles. Recent studies in streamside flumes and laboratory flow cells have hypothesized a link with a turbulent flow environment. However, the coupling between the hydrodynamics and complex biofilm structures remains poorly understood. Here, we report the formation of biofilm streamers suspended in the middle plane of curved microchannels under conditions of laminar flow. Experiments with different mutant strains allow us to identify a link between the accumulation of extracellular matrix and the development of these structures. Numerical simulations of the flow in curved channels highlight the presence of a secondary vortical motion in the proximity of the corners, which suggests an underlying hydrodynamic mechanism responsible for the formation of the streamers. Our findings should be relevant to the design of all liquid-carrying systems where biofilms are potentially present and provide new insights on the origins of microbial streamers in natural and industrial environments.

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Proteins in bacteria often deploy to particular places within the cell, but the cues for localization are frequently mysterious. We found that the peripheral membrane protein SpoVM (VM) recognizes a geometric cue when localizing to a particular membrane during sporulation in Bacillus subtilis. Sporulation involves an inner cell maturing into a spore and an outer cell nurturing the developing spore. VM is produced in the outer cell, where it embeds in the membrane that surrounds the inner cell but not in the cytoplasmic membrane of the outer cell. We found that VM localized by discriminating between the positive curvature of the membrane surrounding the inner cell and the negative curvature of the cytoplasmic membrane. Membrane curvature could be a general cue for protein localization in bacteria.

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Poly( L-lysine) (PLL)/hyaluronic acid (HA) multilayers are films whose thickness increases exponentially with the number of deposition steps. Such a growth process was attributed to the diffusion, in and out of the whole film, of at least one of the polyelectrolytes constituting the film. In the case of PLL/HA, PLL is known to be the diffusing species. In order to better understand the growth mechanism of such films, it is of primary importance to well characterize the diffusion process of the polyelectrolytes in the multilayer. This process is studied here by fluorescence recovery after pattern photobleaching. We show that the diffusion behavior is different when we consider either PLL chains that are deposited on top of the film or PLL chains embedded in the film, even below only one HA layer. For chains that are embedded, we find two populations: a mobile one with a diffusion coefficient, D, of the order of 0.1 microm(2) x s(-1) and a population that appears immobile ( D < 0.001 microm(2) x s(-1)). For chains deposited on top of the multilayer, a third population appears which is rapidly diffusing ( D congruent with 1 microm(2) x s(-1)). These results confirm the validity of the model generally accepted for the exponential growth process and in particular the existence of up to three subgroups of PLL chains from the point of view of their diffusion coefficient.

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