Influence of Fluid Velocity and Cell Concentration on the Transport of Motile and Nonmotile Bacteria in Porous Media

Abstract

The effect of fluid velocity on the transport of motile and nonmotile bacteria was studied in saturated soil columns using radiolabeled cells. According to colloid filtration theory, decreasing the bulk fluid velocity in a porous medium increases the number of collisions of passive colloids with particles and, therefore, should result in increased colloid retention in porous media. However, for motile cells, there was a variation in cell retention significantly different from that predicted by filtration theory at low fluid velocities, leading to the conclusion that filtration theory is not applicable for this motile bacterial strain at low fluid velocities. As the pore velocity was decreased from 120 to 0.56 m/day, the fractional retention of motile cells (Pseudomonas florescens P17) decreased by 65%, and the collision efficiency (α) defined as the ratio of particles that attach to soil grains to particles that collide with the soil (calculated using a filtration equation) decreased from 0.37 (120 m/day) to 0.003 (0.56 m/day). For passive colloids, the fractional retention (if α is a constant equal to 0.01) would increase by more than 800% over this same velocity range. To support our conclusion that cell motility was the factor producing this change from filtration theory, we rendered P17 cells nonmotile and tested this strain and a second nonmotile strain [Burkholderia (Pseudomonas) cepacia G4] under the same conditions. Collision efficiencies for both nonmotile suspensions were constant. For nonmotile P17, α was equal to 0.018 ± 0.003 (0.56−590 m/day). Over a wide velocity range for nonmotile G4, α was equal to 0.22 ± 0.067 (11−560 m/day). Swimming cells were presumably able to avoid sticking to soil grains at low fluid velocities, but at high fluid velocities, cell motility did not reduce attachment. Two additional factors known to affect cell transport (solution ionic strength and cell concentration) were also examined with these two strains in porous media. Decreasing the ionic strength from 4.14 to 0.0011 mM (at a constant pH) decreased cell retention for motile P17 by 39 ± 12%, but this is less of a reduction than is typically observed for nonmotile strains. Increasing the cell concentrations of motile P17 increased the overall retention of cells, suggesting that previously deposited cells provided a more favorable surface for adhesion than the native soil (ripening). In contrast, increasing the cell concentrations of G4 resulted in lower retention, suggesting that deposited cells provided a less favorable collector surface (blocking). These results need to be further investigated with other motile and nonmotile species. However, our results do suggest that wider dispersal of cells during bioaugmentation than previously thought possible may be achieved by using a combination of motile cells, low pumping velocities, and low ionic strength solutions. Optimal cell concentrations to use for in situ bioaugmentation of contaminated soil will depend on the adhesion of the bacterial strains for soil grains and with each other, but in general blocking-type cells are capable of greater dispersal at higher concentration than ripening-type cells.