Microfluidics Expanding the Frontiers of Microbial Ecology

Microfluidics has significantly contributed to the expansion of the frontiers of microbial ecology over the past decade by allowing researchers to observe the behaviors of microbes in highly controlled microenvironments, across scales from a single cell to mixed communities. Spatially and temporally varying distributions of organisms and chemical cues that mimic natural microbial habitats can now be established by exploiting physics at the micrometer scale and by incorporating structures with specific geometries and materials. In this article, we review applications of microfluidics that have resulted in insightful discoveries on fundamental aspects of microbial life, ranging from growth and sensing to cell-cell interactions and population dynamics. We anticipate that this flexible multidisciplinary technology will continue to facilitate discoveries regarding the ecology of microorganisms and help uncover strategies to control microbial processes such as biofilm formation and antibiotic resistance.National Science Foundation (U.S.) (Grant OCE-0744641-CAREER)National Science Foundation (U.S.) (Grant IOS-1120200)National Science Foundation (U.S.) (Grant CBET-1066566)National Science Foundation (U.S.) (Grant CBET-0966000)National Institutes of Health (U.S.) (NIH grant 1R01GM100473-0)Human Frontier Science Program (Strasbourg, France)Human Frontier Science Program (Strasbourg, France) (award RGY0089)Gordon and Betty Moore Foundation (Microbial Initiative Investigator Award

The Classical Nature of Thermal Conduction in Nanofluids,

We show that a large set of nanofluid thermal conductivity data falls within the upper and lower Maxwell bounds for homogeneous systems. This indicates that the thermal conductivity of nanofluids is largely dependent on whether the nanoparticles stay dispersed in the base fluid, form large aggregates, or assume a percolating fractal configuration. The experimental data, which are strikingly analogous to those in most solid composites and liquid mixtures, provide strong evidence for the classical nature of thermal conduction in nanofluid

Transport of Pseudomonas aeruginosa in Polymer Solutions

Bacteria often live surrounded by polymer solutions, such as in animal respiratory, gastrointestinal, and reproductive tracts. In these systems, polymer solutions are often exposed to fluid flow, and their complex rheology can affect the transport of chemical compounds and microorganisms. Recent studies have focused on the effect of polymer solutions on the motility of bacteria in the absence of fluid flow. However, flow can be a game-changer on bacterial transport, as demonstrated by the depletion of motile bacteria from the low-shear regions and trapping in the high-shear regions in simple fluids, even for flows as simple as the Poiseuille one. Despite the relevance of polymer solutions in many bacterial habitats, the effect of their complex rheology on shear-induced trapping and bacterial transport in flow has remained unexplored. Using microfluidic experiments and numerical modeling, we studied how the shear rate and the rheological behavior of Newtonian and non-Newtonian polymer solutions affect the transport of motile, wild-type Pseudomonas aeruginosa in a Poiseuille flow. Our results show that, in Newtonian solutions, an increase in viscosity reduces bacterial depletion in the low-shear regions at the microchannel center, due to a reduction in the bacterial swimming velocity. Conversely, in the non-Newtonian solution, we observed a depletion comparable to the buffer case, despite its zero-shear viscosity being two orders of magnitude higher. In both cases, bacterial swimming and polymer fluid rheology control the magnitude of bacterial depletion and its shear-rate dependence. Our observations underscore the importance of the rheological behavior of the carrier fluid in controlling bacterial transport, in particular, close to surfaces giving rise to velocity gradients, with potential consequences on surface colonization and biofilm formation in many naturally relevant microbial habitats

The structural role of bacterial eDNA in the formation of biofilm streamers

Across diverse habitats, bacteria are mainly found as biofilms, surface-attached communities embedded in a self-secreted matrix of extracellular polymeric substances (EPS), which enhance bacterial recalcitrance to antimicrobial treatment and mechanical stresses. In the presence of flow and geometric constraints such as corners or constrictions, biofilms can take the form of long, suspended filaments (streamers), which bear important consequences in industrial and clinical settings by causing clogging and fouling. The formation of streamers is thought to be driven by the viscoelastic nature of the biofilm matrix. Yet, little is known about the structural composition of streamers and how it affects their mechanical properties. Here, using a microfluidic platform that allows growing and precisely examining biofilm streamers, we show that extracellular DNA (eDNA) constitutes the backbone and is essential for the mechanical stability of Pseudomonas aeruginosa streamers. This finding is supported by the observations that DNA-degrading enzymes prevent the formation of streamers and clear already formed ones and that the antibiotic ciprofloxacin promotes their formation by increasing the release of eDNA. Furthermore, using mutants for the production of the exopolysaccharide Pel, an important component of P. aeruginosa EPS, we reveal an concurring role of Pel in tuning the mechanical properties of the streamers. Taken together, these results highlight the importance of eDNA and of its interplay with Pel in determining the mechanical properties of P. aeruginosa streamers and suggest that targeting the composition of streamers can be an effective approach to control the formation of these biofilm structures. Significance: Streamers, filamentous bacterial biofilms formed in flowing systems, are ubiquitous in natural and artificial environments, where they cause clogging of devices and spreading of infections. Despite their impact, little is known about the nature and properties of streamers and their response to fluid flow. Here, we uncover the specific contribution of bacterial secreted extracellular DNA and exopolysaccharide Pel, two important components in Pseudomonas aeruginosa biofilms, to the formation and the mechanical properties of the streamers. We then show how this knowledge can be used to control biofilm streamer formation, both to inhibit or to promote it

Resilience of bacterial quorum sensing against fluid flow

Quorum sensing (QS) is a population-density dependent chemical process that enables bacteria to communicate based on the production, secretion and sensing of small inducer molecules. While recombinant constructs have been widely used to decipher the molecular details of QS, how those findings translate to natural QS systems has remained an open question. Here, we compare the activation of natural and synthetic Pseudomonas aeruginosa LasI/R QS systems in bacteria exposed to quiescent conditions and controlled flows. Quantification of QS-dependent GFP expression in suspended cultures and in surface-attached microcolonies revealed that QS onset in both systems was similar under quiescent conditions but markedly differed under flow. Moderate flow (Pe > 25) was sufficient to suppress LasI/R QS recombinantly expressed in Escherichia coli, whereas only high flow (Pe > 102) suppressed QS in wild-type P. aeruginosa. We suggest that this difference stems from the differential production of extracellular matrix and that the matrix confers resilience against moderate flow to QS in wild-type organisms. These results suggest that the expression of a biofilm matrix extends the environmental conditions under which QS-based cell-cell communication is effective and that findings from synthetic QS circuits cannot be directly translated to natural systems

The effect of flow on swimming bacteria controls the initial colonization of curved surfaces

The colonization of surfaces by bacteria is a widespread phenomenon with consequences on environmental processes and human health. While much is known about the molecular mechanisms of surface colonization, the influence of the physical environment remains poorly understood. Here we show that the colonization of non-planar surfaces by motile bacteria is largely controlled by flow. Using microfluidic experiments with Pseudomonas aeruginosa and Escherichia coli, we demonstrate that the velocity gradients created by a curved surface drive preferential attachment to specific regions of the collecting surface, namely the leeward side of cylinders and immediately downstream of apexes on corrugated surfaces, in stark contrast to where nonmotile cells attach. Attachment location and rate depend on the local hydrodynamics and, as revealed by a mathematical model benchmarked on the observations, on cell morphology and swimming traits. These results highlight the importance of flow on the magnitude and location of bacterial colonization of surfaces

The effect of flow on swimming bacteria controls the initial colonization of curved surfaces

The colonization of surfaces by bacteria is a widespread phenomenon with consequences on environmental processes and human health. While much is known about the molecular mechanisms of surface colonization, the influence of the physical environment remains poorly understood. Here we show that the colonization of non-planar surfaces by motile bacteria is largely controlled by flow. Using microfluidic experiments with Pseudomonas aeruginosa and Escherichia coli, we demonstrate that the velocity gradients created by a curved surface drive preferential attachment to specific regions of the collecting surface, namely the leeward side of cylinders and immediately downstream of apexes on corrugated surfaces, in stark contrast to where nonmotile cells attach. Attachment location and rate depend on the local hydrodynamics and, as revealed by a mathematical model benchmarked on the observations, on cell morphology and swimming traits. These results highlight the importance of flow on the magnitude and location of bacterial colonization of surfaces.Fil: Secchi, Eleonora. Eidgenössische Technische Hochschule Zurich. Institute of Environmental Engineering; SuizaFil: Vitale, Alessandra. Universitat Zurich; SuizaFil: Miño, Gastón Leonardo. Universidad Nacional de Entre Ríos. Facultad de Ingeniería. Laboratorio de Microscopía Aplicada a Estudios Moleculares y Celulares; Argentina. Universidad Nacional de Entre Ríos. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática; ArgentinaFil: Kantsler, Vasily. University of Warwick; Reino UnidoFil: Eberl, Leo. Universitat Zurich; SuizaFil: Rusconi, Roberto. Humanitas University. Department of Biomedical Sciences; Italia. Humanitas Clinical and Research Center; ItaliaFil: Stocker, Roman. Eidgenössische Technische Hochschule Zurich. Institute of Environmental Engineering; Suiz