Graduate Thesis Or Dissertation
 

Interactions between microbial dynamics, water flow, and solute transport in unsaturated porous media

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  • Bioremediation in the vadose zone is unpredictable because of poor understanding of factors influencing microbial growth in this environment. A lab-scale experimental system was developed to examine, noninvasively, interactions between microbial growth, water flow, and solute transport in unsaturated porous media. Measurements of microbial colonization, and its impact on hydrology, were facilitated by using the luxCDABE-containing reporter bacterium Pseudomonas fluorescens HK44 and digital CCD imaging. Experiments were conducted in glass-walled two-dimensional flow cells (45 x 50 x 1 cm) packed with silica sand. Several bioengineering problems associated with chamber design and function required solution before microbial experiments were successful. These included: choice of materials for chamber components; development of sterilization, packing, and inoculation protocols; and development of procedures for data collection and chamber maintenance during experiments lasting several days. Bacterial growth was mapped daily by quantifying development of salicylate-induced bioluminescence. A model relating the rate of increase in light emission after induction successfully predicted microbial densities over four orders of magnitude (R²=0.95) provided that sufficient oxygen for the bioluminescence reaction was available. Total model-predicted growth during a one-week experiment agreed with potential growth calculated from the mass-balance of the system and previously established kinetic parameters (predicted, 1.2x10¹² cells; calculated, 1.7x10¹² cells). Although the rate of expansion of the colonized zone (and predicted populations in newly colonized regions) remained relatively constant, the proportion of the daily potential growth remaining within the chamber declined over time. Monitoring of bioluminescence revealed the development of an (hypothesized) anaerobic zone associated with microbial growth in the unsaturated porous media. Water content and flow streams were measured using light transmission. Accumulation of microbial growth modified the hydrologic properties of the sand causing up to 50% decrease in saturation within the colonized zone, diversion of flow around the colonized zone, and lowering (5 cm) of the capillary fringe height. Apparent solute velocity through the colonized region was reduced from 0.39 cm min⁻¹ (R²=0.99) to 0.25 cm min⁻¹ (R²=0.99). These experiments provide proof-of-concept for combining light transmission and bioluminescence technologies to study interactions between microbial growth and hydrology in unsaturated porous media.
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