Swimming Cells Research Articles

A new mode of swimming in singly flagellated Pseudomonas aeruginosa.

SignificanceThe monotrichous Pseudomonas aeruginosa was usually thought to swim in a pattern of "run and reverse" (possibly with pauses in between), where straight runs alternated with reverses with angular changes of swimming direction near 180°. Here, by simultaneously tracking the cell swimming and the morphology of its flagellum, we discovered a swimming mode in P. aeruginosa-the wrap mode, during which the flagellar filament wrapped around the cell body and induced large fluctuation of the body orientation. The wrap mode randomized swimming direction, resulting in a broad distribution of angular changes over 0 to 180° with a peak near 90°. This allowed the bacterium to explore the environment more efficiently, which we confirmed by stochastic simulations of P. aeruginosa chemotaxis.

Transport of complex and active fluids in porous media

Complex and active fluids find broad applications in flows through porous materials. Nontrivial rheology can couple to porous microstructure leading to surprising flow patterns and associated transport properties in geophysical, biological, and industrial systems. Viscoelastic instabilities are highly sensitive to pore geometry and can give rise to chaotic velocity fluctuations. A number of recent studies have begun to untangle how the pore-scale geometry influences the sample-scale flow topology and the resulting dispersive transport properties of these complex systems. Beyond classical rheological properties, active colloids and swimming cells exhibit a range of unique properties, including reduced effective viscosity, collective motion, and random walks, that present novel challenges to understanding their mechanics and transport in porous media flows. This review article aims to provide a brief overview of essential, fundamental concepts followed by an in-depth summary of recent developments in this rapidly evolving field. The chosen topics are motivated by applications, and new opportunities for discovery are highlighted.

Metformin Alters the Chemotaxis and Flagellar Motility of Escherichia coli.

Metformin is a biguanide molecule that is widely prescribed to treat type 2 diabetes and metabolic syndrome. Although it is known that metformin promotes the lifespan by altering intestinal microorganism metabolism, how metformin influences and alters the physiological behavior of microorganisms remains unclear. Here we studied the effect of metformin on the behavior alterations of the model organism Escherichia coli (E. coli), including changes in chemotaxis and flagellar motility that plays an important role in bacterial life. It was found that metformin was sensed as a repellent to E. coli by tsr chemoreceptors. Moreover, we investigated the chemotactic response of E. coli cultured with metformin to two typical attractants, glucose and α-methyl-DL-aspartate (MeAsp), finding that metformin prolonged the chemotactic recovery time to the attractants, followed by the recovery time increasing with the concentration of stimulus. Metformin also inhibited the flagellar motility of E. coli including the flagellar motor rotation and cell swimming. The inhibition was due to the reduction of torque generated by the flagellar motor. Our discovery that metformin alters the behavior of chemotaxis and flagellar motility of E. coli could provide potential implications for the effect of metformin on other microorganisms.

Coordinated motion of active filaments on spherical surfaces

The coordinated motion of active filaments, most notably cilia and flagella, are used by numerous micoorganisms and swimming cells to facilitate swimming. This Letter explores how the coordinated states of active filaments depend on the topology of the surface to which they are attached, as well as whether the surface is held fixed or free to move. Most notably, the Letter demonstrates that for spherical surfaces, the coordinated state changes dramatically when the sphere is released due to defect-induced alterations in filament motion.

Differential Expression Pattern of Pathogenicity-Related Genes of Ralstonia pseudosolanacearum YQ Responding to Tissue Debris of Casuarina equisetifolia

Ralstonia solanacearum species complex (RSSC) contains a group of destructive plant pathogenic bacteria, causing bacterial wilt of >200 species of crops and trees, such as Casuarina equisetifolia, worldwide. RSSC can survive in the soil environment for a long time and start infection after activation by host plants. This study conducted a transcriptome analysis on the expression pattern of the pathogenicity-related genes of a new isolated RSSC strain YQ (Ralstonia pseudosolanacearum phylotype I-16) in response to C. equisetifolia cladophyll (a branch of a stem that resembles and functions as a leaf) and root debris under invitro culture. The cladophyll debris induced more genes up-regulated than the root debris, including pathogenicity-related genes involved in motility, effectors, type III secretion systems, quorum sensing, and plant cell wall degradation. Besides, many differentially expressed genes were related to transcriptional regulator such as cyclic dimeric guanosine monophosphate. Moreover, the cultures with cladophyll debris induced a faster wilting in bioassays, and the cell swimming was enhanced by cladophyll exudate. C. equisetifolia cladophylls could activate the expression of pathogenicity-related genes of strain YQ and accelerate infection. Our findings suggest that litterfall management in C. equisetifolia forests, or even other plantations, should receive attention to prevent the induction of bacterial wilt disease caused by RSSC.

Effect of inoculum size and antibiotics on bacterial traveling bands in a thin microchannel defined by optical adhesive

Phenotypic diversity in bacterial flagella-induced motility leads to complex collective swimming patterns, appearing as traveling bands with transient locally enhanced cell densities. Traveling bands are known to be a bacterial chemotactic response to self-generated nutrient gradients during growth in resource-limited microenvironments. In this work, we studied different parameters of Escherichia coli (E. coli) collective migration, in particular the quantity of bacteria introduced initially in a microfluidic chip (inoculum size) and their exposure to antibiotics (ampicillin, ciprofloxacin, and gentamicin). We developed a hybrid polymer-glass chip with an intermediate optical adhesive layer featuring the microfluidic channel, enabling high-content imaging of the migration dynamics in a single bacterial layer, i.e., bacteria are confined in a quasi-2D space that is fully observable with a high-magnification microscope objective. On-chip bacterial motility and traveling band analysis was performed based on individual bacterial trajectories by means of custom-developed algorithms. Quantifications of swimming speed, tumble bias and effective diffusion properties allowed the assessment of phenotypic heterogeneity, resulting in variations in transient cell density distributions and swimming performance. We found that incubation of isogeneic E. coli with different inoculum sizes eventually generated different swimming phenotype distributions. Interestingly, incubation with antimicrobials promoted bacterial chemotaxis in specific cases, despite growth inhibition. Moreover, E. coli filamentation in the presence of antibiotics was assessed, and the impact on motility was evaluated. We propose that the observation of traveling bands can be explored as an alternative for fast antimicrobial susceptibility testing.

Helical trajectories of swimming cells with a flexible flagellar hook

The flexibility of the bacterial flagellar hook is believed to have substantial consequences for microorganism locomotion. Using a simplified model of a rigid flagellum and a flexible hook, we show that the paths of axisymmetric cell bodies driven by a single flagellum in Stokes flow are generically helical. Phase-averaged resistance and mobility tensors are produced to describe the flagellar hydrodynamics, and a helical rod model which retains a coupling between translation and rotation is identified as a distinguished asymptotic limit. A supercritical Hopf bifurcation in the flagellar orientation beyond a critical ratio of flagellar motor torque to hook bending stiffness, which is set by the spontaneous curvature of the flexible hook, the shape of the cell body, and the flagellum geometry, can have a dramatic effect on the cell's trajectory through the fluid. Although the equilibrium hook angle can result in a wide variance in the trajectory's helical pitch, we find a very consistent prediction for the trajectory's helical amplitude using parameters relevant to swimming P. aeruginosa cells.

Purely viscous acoustic propulsion of bimetallic rods

Synthetic microswimmers offer models for cell motility and their tunability makes them promising candidates for biomedical applications. Here we measure the acoustic propulsion of bimetallic micro-rods that, when trapped at the nodal plane of a MHz acoustic resonator, swim with speeds of up to 300 microns per second. While past acoustic streaming models predict speeds that are more than one order of magnitude smaller than our measurements, we demonstrate that the acoustic locomotion of the rods is driven by a viscous, non-reciprocal mechanism relying on shape anisotropy akin to that used by swimming cells and that reproduces our data with no adjustable parameters.

Prospects for the Mechanism of Spiroplasma Swimming.

Spiroplasma are helical bacteria that lack a peptidoglycan layer. They are widespread globally as parasites of arthropods and plants. Their infectious processes and survival are most likely supported by their unique swimming system, which is unrelated to well-known bacterial motility systems such as flagella and pili. Spiroplasma swims by switching the left- and right-handed helical cell body alternately from the cell front. The kinks generated by the helicity shift travel down along the cell axis and rotate the cell body posterior to the kink position like a screw, pushing the water backward and propelling the cell body forward. An internal structure called the “ribbon” has been focused to elucidate the mechanisms for the cell helicity formation and swimming. The ribbon is composed of Spiroplasma-specific fibril protein and a bacterial actin, MreB. Here, we propose a model for helicity-switching swimming focusing on the ribbon, in which MreBs generate a force like a bimetallic strip based on ATP energy and switch the handedness of helical fibril filaments. Cooperative changes of these filaments cause helicity to shift down the cell axis. Interestingly, unlike other motility systems, the fibril protein and Spiroplasma MreBs can be traced back to their ancestors. The fibril protein has evolved from methylthioadenosine/S-adenosylhomocysteine (MTA/SAH) nucleosidase, which is essential for growth, and MreBs, which function as a scaffold for peptidoglycan synthesis in walled bacteria.

Clostridioides difficile Single Cell Swimming Strategy: A Novel Motility Pattern Regulated by Viscoelastic Properties of the Environment.

Flagellar motility is important for the pathogenesis of many intestinal pathogens, allowing bacteria to move to their preferred ecological niche. Clostridioides difficile is currently the major cause for bacterial health care-associated intestinal infections in the western world. Most clinical strains produce peritrichous flagella and are motile in soft-agar. However, little knowledge exists on the C. difficile swimming behaviour and its regulation at the level of individual cells. We report here on the swimming strategy of C. difficile at the single cell level and its dependency on environmental parameters. A comprehensive analysis of motility parameters from several thousand bacteria was achieved with the aid of a recently developed bacterial tracking programme. C. difficile motility was found to be strongly dependent on the matrix elasticity of the medium. Long run phases of all four motile C. difficile clades were only observed in the presence of high molecular weight molecules such as polyvinylpyrrolidone (PVP) and mucin, which suggests an adaptation of the motility apparatus to the mucin-rich intestinal environment. Increasing mucin or PVP concentrations lead to longer and straighter runs with increased travelled distance per run and fewer turnarounds that result in a higher net displacement of the bacteria. The observed C. difficile swimming pattern under these conditions is characterised by bidirectional, alternating back and forth run phases, interrupted by a short stop without an apparent reorientation or tumbling phase. This motility type was not described before for peritrichous bacteria and is more similar to some previously described polar monotrichous bacteria.

High-speed, three-dimensional imaging reveals chemotactic behaviour specific to human-infective Leishmania parasites

Cellular motility is an ancient eukaryotic trait, ubiquitous across phyla with roles in predator avoidance, resource access, and competition. Flagellar motility is seen in various parasitic protozoans, and morphological changes in flagella during the parasite life cycle have been observed. We studied the impact of these changes on motility across life cycle stages, and how such changes might serve to facilitate human infection. We used holographic microscopy to image swimming cells of different Leishmania mexicana life cycle stages in three dimensions. We find that the human-infective (metacyclic promastigote) forms display 'run and tumble' behaviour in the absence of stimulus, reminiscent of bacterial motion, and that they specifically modify swimming direction and speed to target host immune cells in response to a macrophage-derived stimulus. Non-infective (procyclic promastigote) cells swim more slowly, along meandering helical paths. These findings demonstrate adaptation of swimming phenotype and chemotaxis towards human cells.

Retraction of the Research Article: “Human sperm uses asymmetric and anisotropic flagellar controls to regulate swimming symmetry and cell steering”

In our recent Research Article “Human sperm uses asymmetric and anisotropic flagellar controls to regulate swimming symmetry and cell steering” ( 1 ), we …

Mathematical modeling of chemotaxis guided amoeboid cell swimming

Cells and microorganisms adopt various strategies to migrate in response to different environmental stimuli. To date, many modeling research has focused on the crawling-based Dictyostelium discoideum (Dd) cells migration induced by chemotaxis, yet recent experimental results reveal that even without adhesion or contact to a substrate, Dd cells can still swim to follow chemoattractant signals. In this paper, we develop a modeling framework to investigate the chemotaxis induced amoeboid cell swimming dynamics. A minimal swimming system consists of one deformable Dd amoeboid cell and a dilute suspension of bacteria, and the bacteria produce chemoattractant signals that attract the Dd cell. We use the mathematical amoeba model to generate Dd cell deformation and solve the resulting low Reynolds number flows, and use a moving mesh based finite volume method to solve the reaction–diffusion–convection equation. Using the computational model, we show that chemotaxis guides a swimming Dd cell to follow and catch bacteria, while on the other hand, bacterial rheotaxis may help the bacteria to escape from the predator Dd cell.

Hopping trajectories due to long-range interactions determine surface accumulation of microalgae

The accumulation of motile cells at solid interfaces increases the rate of surface encounters and the likelihood of surface attachment, leading to surface colonization and biofilm formation. The cell density distribution in the vicinity of a physical boundary is influenced by the interactions between the microswimmers and their physical environment, including hydrodynamic and steric interactions, as well as by stochastic effects. Disentangling the contributions of these effects remains an experimental challenge. Here, we use a custom-made four-camera view microscope to track a population of motile puller-type Chlamydomonas reinhardtii in a relatively unconstrained three-dimensional (3D) domain. Our experiments yield an extensive sample of 3D trajectories including cell-surface encounters with a planar wall, from which we extract a full description of the dynamics and the stochasticity of swimming cells. We use this large data sample and combine it with Monte Carlo simulations to determine the link between the dynamics at the single-cell level and the cell density. Our experiments demonstrate that the near-wall scattering is bimodal, corresponding to steric and hydrodynamic effects. We find, however, that this near-wall dynamics has little influence on the cell accumulation at the surface. On the other hand, we present evidence of a cell-induced surface-directed rotation leading to a vertical orbiting behavior and hopping trajectories, consistent with long-range hydrodynamic effects. We identify this long-range effect to be at the origin of the significant surface accumulation of cells.

Impact of Gba2 on neuronopathic Gaucher\u2019s disease and \u03b1-synuclein accumulation in medaka (Oryzias latipes)

Homozygous mutations in the lysosomal glucocerebrosidase gene, GBA1, cause Gaucher’s disease (GD), while heterozygous mutations in GBA1 are a strong risk factor for Parkinson’s disease (PD), whose pathological hallmark is intraneuronal α-synuclein (asyn) aggregates. We previously reported that gba1 knockout (KO) medaka exhibited glucosylceramide accumulation and neuronopathic GD phenotypes, including short lifespan, the dopaminergic and noradrenergic neuronal cell loss, microglial activation, and swimming abnormality, with asyn accumulation in the brains. A recent study reported that deletion of GBA2, non-lysosomal glucocerebrosidase, in a non-neuronopathic GD mouse model rescued its phenotypes. In the present study, we generated gba2 KO medaka and examined the effect of Gba2 deletion on the phenotypes of gba1 KO medaka. The Gba2 deletion in gba1 KO medaka resulted in the exacerbation of glucosylceramide accumulation and no improvement in neuronopathic GD pathological changes, asyn accumulation, or swimming abnormalities. Meanwhile, though gba2 KO medaka did not show any apparent phenotypes, biochemical analysis revealed asyn accumulation in the brains. gba2 KO medaka showed a trend towards an increase in sphingolipids in the brains, which is one of the possible causes of asyn accumulation. In conclusion, this study demonstrated that the deletion of Gba2 does not rescue the pathological changes or behavioral abnormalities of gba1 KO medaka, and GBA2 represents a novel factor affecting asyn accumulation in the brains.

Vertical distribution and longitudinal dispersion of gyrotactic microorganisms in a horizontal plane Poiseuille flow

A more concise and accurate generalized Taylor dispersion theory is applied to dispersion of active gyrotactic microorganisms in a plane Poiseuille flow. The joint effect of boundary conditions, cell shape anisotropy, swimming speed, and flow speed leads to the nonmonotonic variations of the phenomenological dispersion coefficients.

Escherichia coli Swimming back Toward Stiffer Polyetheylene Glycol Coatings, Increasing Contact in Flow.

Bacterial swimming in flow near surfaces is critical to the spread of infection and device colonization. Understanding how material properties affect flagella- and motility-dependent bacteria-surface interactions is a first step in designing new medical devices that mitigate the risk of infection. We report that, on biomaterial coatings such as polyethylene glycol (PEG) hydrogels and end-tethered layers that prevent adhesive bacteria accumulation, the coating mechanics and hydration control the near-surface travel and dynamic surface contact of E. coli cells in gentle shear flow (order 10 s-1). Along relatively stiff (order 1 MPa) PEG hydrogels or end-tethered layers of PEG chains of similar polymer correlation length, run-and-tumble E. coli travel nanometrically close to the coating's surface in the flow direction in distinguishable runs or "engagements" that persist for several seconds, after which cells leave the interface. The duration of these engagements was greater along stiff hydrogels and end-tethered layers compared with softer, more-hydrated hydrogels. Swimming cells that left stiff hydrogels or end-tethered layers proceeded out to distances of a few microns and then returned to engage the surface again and again, while cells engaging the soft hydrogel tended not to return after leaving. As a result of differences in the duration of engagements and tendency to return to stiff hydrogel and end-tethered layers, swimming E. coli experienced 3 times the integrated dynamic surface contact with stiff coatings compared with softer hydrogels. The striking similarity of swimming behaviors near 16-nm-thick end-tethered layers and 100-μm-thick stiff hydrogels argues that only the outermost several nanometers of a highly hydrated coating influence cell travel. The range of material stiffnesses, cell-surface distance during travel, and time scales of travel compared with run-and-tumble time scales suggests the influence of the coating derives from its interactions with flagella and its potential to alter flagellar bundling. Given that restriction of flagellar rotation is known to trigger increased virulence, bacteria influenced by surfaces in one region may become predisposed to form a biofilm downstream.

Transcriptomic analysis of the food spoilers Pseudomonas fluorescens reveals the antibiofilm of carvacrol by interference with intracellular signaling processes

Pseudomonas fluorescens as an important food spoilage organism, has strong biofilm forming-ability to resist common biocides, increasing persistence and contamination in the cold fresh food chain. This bacterial biofilm organization is regulated by luxI/luxR mediated quorum sensing (QS) signaling. Carvacrol with antimicrobial properties is well documented, however, the mechanisms of its antibiofilm activity targeting QS are still not well known. The aim of this study was to explore the inhibition of carvacrol on biofilm formation and spoilage potential of two P. fluorescens PFuk4 and PF07 strains, and their luxI mutants (ΔluxI). The minimum inhibitory concentration (MIC) of carvacrol against planktonic wild type (WT) strains and the corresponding ΔluxI was 1.6 and 1.4 mM, respectively, and maximum sublethal concentration (MSC) was 0.8 mM. The acyl-homoserine-lactones (AHLs) C4-HSL and C6-HSL, as major autoinducer molecules, were greatly decreased in PFuk4 and PF07 treated with 0.4 mM carvacrol, while those in the ΔluxI were almost abolished. Biofilm formation and exopolysaccharide production of the P. fluorescens WT and ΔluxI strains were significantly inhibited by carvacrol at the MSC, as well as the observed sparer and thinner biofilm structures. Bacterial cell swimming and swarming motilities were both decreased by carvacrol without affecting cell viability. Protease activity secreted by P. fluorescens was significantly reduced treated by MSC of carvacrol, in consistent with the repression of sensory attributes, especially in the WT strains. Transcriptomics profiling revealed that exposure to 0.4 mM carvacrol dramatically altered gene expression in P. fluorescens PF07 biofilm, as a total of 402 differentially expressed genes (DEGs) were upregulated and 308 DEGs were downregulated, compared with the untreated biofilms. Those DECs were mainly involved in pathways of ribosome, bacterial chemotaxis, citrate cycle. Furthermore, carvacrol also altered the transcription of P. fluorescens genes associated with flagellar assembly, chemotaxis, exopolysaccharide synthesis, signaling pathways of LuxR mediated QS and c-di-GMP, and amino acid degradation, which may contribute to biofilm development and spoilage potential. In addition, carvacrol was found to competitively bind with LuxR predicted protein to interrupt the interaction with C4-HSL, supported by molecular docking. Our results provide novel insights into the regulatory signaling networks of antibiofilm formation in P. fluorescens affected by carvacrol.

Controlling Cell Motion and Microscale Flow with Polarized Light Fields

We investigate how light polarization affects the motion of photoresponsive algae, Euglena gracilis. In a uniformly polarized field, cells swim approximately perpendicular to the polarization direction and form a nematic state with zero mean velocity. When light polarization varies spatially, cell motion is modulated by local polarization. In such light fields, cells exhibit complex spatial distribution and motion patterns which are controlled by topological properties of the underlying fields; we further show that ordered cell swimming can generate directed transporting fluid flow. Experimental results are quantitatively reproduced by an active Brownian particle model in which particle motion direction is nematically coupled to local light polarization.

Structure and Dynamics of the E. coli Chemotaxis Core Signalling Complex

Chemotactic bacteria, including numerous human and plant pathogens, navigate their habitats using large, multi-component protein arrays. Through the binding of environmental chemical ligands, these sophisticated arrays transduce highly-cooperative sensory signals over large distances, giving rise to an intracellular phosphorylation cascade that ultimately affects cell swimming pattern. The basic building block of the array, known as the core-signalling unit (CSU), comprises six transmembrane chemoreceptors, a dimeric histidine kinase, and two coupling proteins, which together span over 30 nm and have a molecular weight of more than 900 kDa. Despite considerable effort over the years, the large size and dynamic nature of the CSU have thwarted the determination of a high-resolution structure, rendering detailed molecular descriptions of array function difficult to resolve. Here, we present recent integrative modelling work, combining cryo-electron tomography structures of the E. coli CSU derived separately from reconstituted monolayer arrays and a newly-characterized minicell strain, to produce the first atomistic model of a complete, transmembrane CSU. This model, combined with all-atom molecular dynamics simulation, allows for a detailed investigation of kinase conformational dynamics, identifying multiple conformations of the critical catalytic and dimerization domains and highlighting residue-level features of their differential stabilization. In addition, Martini-based coarse-grained simulations provide insights into the previously-unresolved membrane-proximal regions of the CSU, characterizing key protein-lipid interactions as well as inter-receptor dynamics within the bilayer. These results provide a structural and methodological basis for testing and understanding molecular signalling mechanisms broadly within bacterial chemosensing