The capacity of fungi to serve as vectors for the dispersion of pollutant-degrading bacteria was analyzed in laboratory model systems mimicking water-saturated (agar surfaces) and unsaturated soil environments (glass-bead-filled columns). Two common soil fungi (Fusarium oxysporum and Rhexocercosporidium sp.) forming hydrophilic and hydrophobic mycelia, respectively, and three polycyclic aromatic hydrocarbon degrading bacteria (Achromobacter sp. SK1, Mycobacterium frederiksbergense LB501TG, and Sphingomonas sp. L138) were selected based on the absence of mutual antagonistic effects. It was shown that fungal hyphae act as vectors for bacterial transport with mobilization strongly depending on the specific microorganisms chosen: The motile strain Achromobacter sp. SK1 was most efficiently spread along hyphae of hydrophilic F. oxysporum in both model systems with transport velocities of up to 1 cm d(-1), whereas no dispersion of the two nonmotile strains was observed in the presence of F. oxysporum. By contrast, none of the bacteria was mobilized along the hydrophobic mycelia of Rhexocercosporidium sp. growing on agar surfaces. In column experiments however, strain SK1 was mobilized by Rhexocercosporidium sp. It is hypothesized that bacteria may move by their intrinsic motilitythrough continuous (physiological) liquid films forming around fungal hyphae. The results of this study suggest that the specific stimulation of indigenous fungi may be a strategy to mobilize pollutant-degrading bacteria leading to their homogenization in polluted soil thereby improving bioremediation.

Knowing how motile bacteria move near and along a solid surface is crucial to understanding such diverse phenomena as the migration of infectious bacteria along a catheter, biofilm growth, and the movement of bacteria through the pore spaces of saturated soil, a critical step in the in situ bioremediation of contaminated aquifers. In this study, a tracking microscope is used to record the three-dimensional motion of Escherichia coli near a planar glass surface. Data from the tracking microscope are analyzed to quantify the effects of bacteria-surface interactions on the swimming behavior of bacteria. The speed of cells approaching the surface is found to decrease in agreement with the mathematical model of Ramia et al. [Ramia, M., Tullock, D. L. & Phan-Tien, N. (1993) BiophysJ. 65,755-778], which represents the bacteria as spheres with a single polar flagellum rotating at a constant rate. The tendency of cells to swim adjacent to the surface is shown in computer-generated reproductions of cell traces. The attractive interaction potential between the cells and the solid surface is offered as one of several possible explanations for this tendency.In a homogeneous fluid medium, peritrichously flagellated bacteria such as Escherichia coli and Salmonella typhimurium execute random walks as they alternate between two phases of motion: running (motion in essentially straight paths) and tumbling (changes in direction while remaining in place) (1, 2).This behavior is similar to molecular diffusion except that changes in direction are due to reversal of flagellar rotation and not molecular collisions. In many natural systems, the characteristic motion of swimming bacteria is modified by the presence of solid surfaces. Examples of such systems include the motion of bacteria in saturated soil (3-5), the migration of bacteria through small-diameter capillary tubes (6), and the migration of infectious bacteria along medically implanted surfaces, such as prostheses and catheters (7).Bacterial transport rates in the presence of solid boundaries are different from transport rates in the absence of such boundaries. Depending on the system studied, the migration of bacteria can either be enhanced (6, 7) or attenuated (5, 8) by solid surfaces. In this study we have quantified the change in the characteristic motion of bacteria when cells approach solid surfaces by measuring the speed as a function of the separation distance. The experimental results were compared with solutions of the mathematical model of Ramia et al. (9) based on hydrodynamic forces to determine the validity of the application of this model. We also observed bacteria swimming in circles parallel to solid surfaces, as previously seen in other studies (6, 10). We illustrate the motion exhibited by both wild-type cells (cells that change direction by tumbling) and smooth-swimming cells (cells that do not tumble) through three-dimensional projections of cell traces near solid surfaces.The publication costs of this article were defrayed in part by page charge...

The initial events in bacterial adhesion are often explained as resulting from electrostatic and van der Waals forces between the cell and the surface, as described by DLVO theory (developed by Derjaguin, Landau, Verwey, and Overbeek). Such a theory predicts that negatively charged bacteria will experience greater attraction toward a negatively charged surface as the ionic strength of the medium is increased. In the present study we observed both smooth-swimming and nonmotile Escherichia coli bacteria close to plain, positively, and hydrophobically coated quartz surfaces in high-and low-ionic-strength media by using total internal reflection aqueous fluorescence microscopy. We found that reversibly adhering cells (cells which continue to swim along the surface for extended periods) are too distant from the surface for this behavior to be explained by DLVO-type forces. However, cells which had become immobilized on the surface did seem to be affected by electrostatic interactions. We propose that the "force" holding swimming cells near the surface is actually the result of a hydrodynamic effect, causing the cells to swim at an angle along the glass, and that DLVO-type forces are responsible only for the observed immobilization of irreversibly adhering cells. We explain our observations within the context of a conceptual model in which bacteria that are interacting with the surface may be thought of as occupying one of three compartments: bulk fluid, near-surface bulk, and near-surface constrained. A cell in these compartments feels either no effect of the surface, only the hydrodynamic effect of the surface, or both the hydrodynamic and the physicochemical effects of the surface, respectively.The goal of this work was to determine the force or forces controlling reversible adhesion of motile bacterial cells to surfaces. Reversible bacterial adhesion is operationally defined here as a situation in which a bacterium remains very close (within the same plane of focus for a light microscope) to a surface for a period of several minutes. Reversibly adhering bacteria are presumed to retain their ability to move laterally along the surface (37), by swimming or Brownian motion, and these cells may also eventually leave the vicinity of the surface. Cells behaving in this manner have been observed in numerous experiments and will often spend long times (Ͼ1 min) swimming near the surface (5,12,22,25,41). In irreversible adhesion, by contrast, bacteria adhering to the surface do not move, either by swimming or Brownian motion, for the duration of observation (36). In general, bacteria that have become immobilized on the surface are described as irreversibly adhered to the surface, while cells that can still swim along the surface are described as reversibly adhered. Cells may also become tethered to the surface, when a flagellum adheres to the surface but the cell body still rotates freely. Figure 1 illustrates the definitions of these terms.Adhesion of individual cells to a surface is the first step in the formation of biof...

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Subsurface bioremediation is often hindered by the inability to achieve good mixing between injected bacteria and residual contaminants. Chemotaxis, which is the ability of bacteria to migrate preferentially toward higher concentrations of certain chemical attractants, could potentially increase bacterial transport into the contaminated zone. To observe and quantify this chemotactic enhancement to bacterial dispersion transverse to groundwater flow, a microfluidic device--a porous T-sensor-was created. It allowed two streams of equal flow rate to enter side-by-side into a porous channel; the transverse mixing of the two streams was controlled primarily by dispersion. When a suspension of the chemotactic bacteria Escherichia coli HCB1 and a solution of chemical attractant alpha-methylaspartate were injected as the two incoming streams, enhanced bacterial migration into the attractant stream was observed relative to a control experiment with dispersion alone. Chemotaxis was observed under lower flow rates comparable to natural groundwaterflow. The chemotactic response was greater than that predicted by an advection-dispersion equation model using a chemotactic coefficient derived under quiescent experimental conditions, which suggests that flow in porous media may further enhance transverse migration for chemotactic bacteria. This study provided direct evidence of the significance of bacterial chemotactic transverse migration at groundwater flow rates.

Bacterial chemotaxis, the directed movement of a cell population in response to a chemical gradient, plays a critical role in the distribution and dynamic interaction of bacterial populations in nonmixed systems. Therefore, in order to make reliable predictions about the migratory behavior of bacteria within the environment, a quantitative characterization of the chemotactic response in terms of intrinsic cell properties is needed.The design of the stopped-flow diffusion chamber (SFDC) provides a well-characterized chemical gradient and reliable method for measuring bacterial migration behavior. During flow through the chamber, a step change in chemical concentration is imposed on a uniform suspension of bacteria. Once flow is stopped, diffusion causes a transient chemical gradient to develop, and bacteria respond by forming a band of high cell density which travels toward higher concentrations of the attractant. Changes in bacterial spatial distributions observed through light scattering are recorded on photomicrographs during a 10-min period. Computer-aided image analysis converts absorbance of the photographic negatives to a digital representation of bacterial density profiles. A mathematical model (part II) is used to quantitatively characterize these observations in terms of intrinsic cell parameters: a chemotactic sensitivity coefficient, chi(0), from the aggregate cell density accumulated in the band and a random motility coefficient, mu, from population dispersion in the absence of a chemical gradient.Using the SFDC assay and an individual-cell-based mathematical model, we successfully determined values for both of these population parameters for Escherichia coli K12 responding to fucose. The values obtained were mu = 1.1 +/- 0. 4 x 10(-5) cm(2)/s and chi(o) = 8 +/- 3 +/- 10(-5) cm(2)/s. We have demonstrated a method capable of determining these parameter values from the now validated mathematical model which will be useful for predicting bacterial migration in application systems.

A quantitative description of bacterial chemotaxis is necessary for making predictions about the migratory behavior of bacterial populations in applications such as biofilm development, release of genetically engineered bacteria into the environment, and in situ bioremediation technologies. The bacterial chemotactic response is characterized by a mathematical model which relates individual cell properties such as swimming speed and tumbling frequency to population parameters, specifically the random motility coefficient and the chemotactic sensitivity coefficient. Our model includes a nonlinear dependence of the chemotactic velocity on the attractant gradient as well as a dependence of the random motility coefficient on the temporal and spatial attractant gradients, both of which previous analyses have neglected. As we will show, these aspects are critical for interpreting the results from experiments like those performed in the stopped-flow diffusion chamber (SFDC) because the initial temporal and spatial gradients are very steep. Our analysis demonstrates that values for the random motility coefficient and chemotactic sensitivity coefficient can be obtained from experimental plots of net cell redistribution from initial conditions versus the square root of time. Values for these parameters are determined from experimental measurements of bacterial population distributions in the SFDC as described in the companion article. Using parameter values determined from independent experiments, mu = 1.1 +/- 0.4 +/- 10(-5) cm(2)/s and chi(0) = 8 +/- 3 +/- 10(-5) cm(2)/s, excellent agreement is found between theoretically predicted bacterial density profiles and actual experimental profiles for Escherichia coli K12 responding to fucose over two orders of magnitude in initial attractant concentration. Thus, our model captures the concentration dependence of this behavioral response satisfactorily in terms of cell population parameters which are derived from individual cell properties and will therefore be useful for making predictions about the migratory behavior of bacterial populations in the environment.

Bacterial chemotaxis has the potential to enhance biodegradation of organic contaminants in polluted groundwater systems. However, studies of bacterial chemotaxis in porous media are scarce. In this study we use magnetic resonance imaging (MRI) for the noninvasive measurement of changes in bacterial-density distributions in a packed column at a spatial resolution of 330 microm as a function of time. We analyze both the diffusive and the chemotactic behavior of Pseudomonas putida F1 in the presence of the chemical stimulus trichloroethylene (TCE). The migration of motile bacteria in experiments without TCE was described using an effective motility coefficient, whereas the presence of TCE required addition of a nonzero chemotactic sensitivity coefficient, indicating a significant response to TCE. The need for a chemotactic sensitivity term was justified by a test for statistical significance. This study represents the first quantification of bacterial chemotactic parameters within a packed column. For conditions under which chemotaxis occurs in porous media, it may potentially be exploited to significantly improve rates of in situ pollutant biodegradation in the subsurface environment, particularlyfor pollutants dissolved in water trapped in low-permeability formations or lenses.