Roseanne M. Ford

Reversible and Irreversible Adhesion of Motile Escherichia coli Cells Analyzed by Total Internal Reflection Aqueous Fluorescence Microscopy

ABSTRACT 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.

Reversal of Flagellar Rotation Is Important in Initial Attachment of Escherichia coli to Glass in a Dynamic System with High- and Low-Ionic-Strength Buffers

ABSTRACT The attachment rates of wild-type, smooth-swimming, tumbly, and paralyzed Escherichia coli to glass was measured at fluid velocities of 0.0044 and 0.044 cms−1 (corresponding to shear rates of 0.34 and 3.4 s−1, respectively), in 0.02 and 0.2 M buffer solutions. At the highest ionic strength, we did not observe a significant difference in the attachment rate of wild-type and paralyzed cells at either fluid velocity. However, when the ionic strength was reduced, paralyzed bacteria attached at rates 4 and 10 times lower than that of the wild type under fluid velocities of 0.0044 and 0.044 cms−1, respectively. This suggested that the rotation of the flagella assisted in attachment. We then compared the attachment rates of smooth-swimming (counterclockwise rotation only) and tumbly (clockwise rotation only) cells to the wild type to determine whether the direction of rotation was important to cell attachment. At 0.0044 cms−1, the smooth-swimming cells attached at rates similar to that of the wild type in both buffer solutions but significantly less at the higher fluid velocity. Tumbly cells attached at much lower rates under all conditions. Thus, the combination of clockwise and counterclockwise flagellar rotation and their coupling appeared to be important in cell attachment. We considered a number of hypotheses to interpret these observations, including a residence time analysis and a comparison of traditional Derjaguin-Landau-Verwey-Overbeek (DLVO) theory to soft-particle theory.

Analysis of chemotactic bacterial distributions in population migration assays using a mathematical model applicable to steep or shallow attractant gradients

The mathematical model developed by Rivero et al. (1989, Chem. Engng Sci. 44, 2881-2897) is applied to literature data measuring chemotactic bacterial population distributions in response to steep as well as shallow attractant gradients. This model is based on a fundamental picture of the sensing and response mechanisms of individual bacterial cells, and thus related individual cell properties such as swimming speed and tumbling frequency to population parameters such as the random motility coefficient and the chemotactic sensitivity coefficient. Numerical solution of the model equations generates predicted bacterial density and attractant concentration profiles for any given experimental assay. We have previously validated the mathematical model from experimental work involving a step change in the attractant gradient (Ford et al., 1991 Biotechnol. Bioengng, 37, 647-660; Ford and Lauffenburger, 1991, Biotechnol. Bioengng, 37, 661-672). Within the context of this experimental assay, effects of attractant diffusion and consumption, random motility, and chemotactic sensitivity on the shape of the profiles are explored to enhance our understanding of this complex phenomenon. We have applied this model to various other types of gradients with successful interpretation of data reported by Dalquist et al. (1972, Nature New Biol. 236, 120-123) for Salmonella typhimurium validating the mathematical model and supporting the involvement of high and low affinity receptors for serine chemotaxis by these cells.

Random Walk Calculations for Bacterial Migration in Porous Media

Bacterial migration is important in understanding many practical problems ranging from disease pathogenesis to the bioremediation of hazardous waste in the environment. Our laboratory has been successful in quantifying bacterial migration in fluid media through experiment and the use of population balance equations and cellular level simulations that incorporate parameters based on a fundamental description of the microscopic motion of bacteria. The present work is part of an effort to extend these results to bacterial migration in porous media. Random walk algorithms have been used successfully to date in nonbiological contexts to obtain the diffusion coefficient for disordered continuum problems. This approach has been used here to describe bacterial motility. We have generated model porous media using molecular dynamics simulations applied to a fluid with equal sized spheres. The porosity is varied by allowing different degrees of sphere overlap. A random walk algorithm is applied to simulate bacterial migration, and the Einstein relation is used to calculate the effective bacterial diffusion coefficient. The tortuosity as a function of particle size is calculated and compared with available experimental results of migration of Pseudomonas putida in sand columns. Tortuosity increases with decreasing obstacle diameter, which is in agreement with the experimental results.

Perturbation expansion of Alt's cell balance equations reduces to Segel's one-dimensional equations for shallow chemoattractant gradients

The cell balance equations of Alt are rigorously studied and perturbatively expanded into forms similar to Segels one-dimensional phenomenological cell balance equations by considering the simplifying case of bacterial density possessing symmetry in the x and y directions responding to an attractant gradient present only in the z direction. We prove that for shallow attractant gradients the lumped integrals involving the tumbling probability frequency distribution and bacterial density distribution in the

Bacterial chemotaxis transverse to axial flow in a microfluidic channel

\theta

Mathematical model for characterization of bacterial migration through sand cores.

direction can be explicitly expressed as a product of three quantities: the mean tumbling frequency, the bacterial subpopulation density, and a reversal probability. We also derive expressions for the bacterial net flux in the Fickian form from which two macroscopic transport parameters, the random motility coefficient and the chemotactic velocity, are explicitly related to individual cell properties and chemical gradients.

On the relationship between cell balance equations for chemotactic cell populations

Swimming bacteria sense and respond to chemical signals in their environment. Chemotaxis is the directed migration of a bacterial population toward increasing concentrations of a chemical that they perceive to be beneficial to their survival. Bacteria that are indigenous to groundwater environments exhibit chemotaxis toward chemical contaminants such as hydrocarbons, which they are also able to degrade. This phenomenon may facilitate bioremediation processes by bringing bacteria into closer proximity to these contaminants. A microfluidic device was assembled to study chemotaxis transverse to advective flow. Using a T‐shaped channel design (T‐sensor), two fluid streams were brought into contact by impinging flow. They then flowed adjacent to each other along a transparent channel. An advantage to this design is that it allows real‐time visualization of bacterial distributions within the channel. Under laminar flow conditions a chemotactic driving force was created perpendicular to the direction of flow by diffusion of the chemical attractant from one input stream to the other. A comparison of the chemotactic band behavior in the absence and presence of flow showed that fluid velocity did not significantly impede chemotactic migration in the transverse direction. Biotechnol. Bioeng. 2008;100: 653–663.

Analysis of biodegradation and bacterial transport: Comparison of models with kinetic and equilibrium bacterial adsorption

The migration of chemotactic bacteria in liquid media has previously been characterized in terms of two fundamental transport coefficients-the random motility coefficient and the chemotactic sensitivity coefficient. For modeling migration in porous media, we have shown that these coefficients which appear in macroscopic balance equations can be replaced by effective values that reflect the impact of the porous media on the swimming behavior of individual bacteria. Explicit relationships between values of the coefficients in porous and liquid media were derived. This type of quantitative analysis of bacterial migration is necessary for predicting bacterial population distributions in subsurface environments for applications such as in situ bioremediation in which bacteria respond chemotactically to the pollutants that they degrade.We analyzed bacterial penetration times through sand columns from two different experimental studies reported in the literature within the context of our mathematical model to evaluate the effective transport coefficients. Our results indicated that the presence of the porous medium reduced the random motility of the bacterial population by a factor comparable to the theoretical prediction. We were unable to determine the effect of the porous medium on the chemotactic sensitivity coefficient because no chemotactic response was observed in the experimental studies. However, the mathematical model was instrumental in developing a plausible explanation for why no chemotactic response was observed. The chemical gradients may have been too shallow over most of the sand core to elicit a measurable response. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 487-496, 1997.

CELLULAR DYNAMICS SIMULATIONS OF BACTERIAL CHEMOTAXIS

This paper considers several cell balance equations describing the time and spatial evolution of the number density of chemotactic cell populations. The conditions are determined under which the three-dimensional cell balance equations of Alt reduce to the simpler, one-dimensional equations of Segel that have been used by Lauffenberger and colleagues to describe chemotaxis of flagellar bacteria and polymorphonuclear leukocytes, and to interpret experimental measurements on such systems.