Pore-scale analysis of anaerobic halorespiring bacterial growth along the transverse mixing zone of an etched silicon pore network.

The anaerobic halorespiring microorganism, Sulfurospirillum multivorans, was observed in the pore structure of an etched silicon wafer to determine how flow hydrodynamics and mass transfer limitations along a transverse mixing zone affect biomass growth. Tetrachloroethene (PCE, an electron acceptor, 0.2 mM) and lactate (an electron donor, 2 mM) were introduced as two separate and parallel streams that mixed along a reaction line in the pore structure. The first visible biomass occupied a single line of pores in the direction of flow, a few pore bodies from the micromodel centerline. This growth was initially present as small aggregates; over time, these grew and fused to form finger-like structures with one end attached to downgradient ends of the silicon posts and the other end extending into pore bodies in the direction of flow. Biomass did not grow in pore throats as expected, presumably because shear forces were not favorable. Over the next few weeks, the line of growth migrated upward into the PCE zone and extended over a width of up to five pore spaces. When the PCE concentration was increased to 0.5 mM, the microbial biomass increased and growth migrated down toward the lactate side of the micromodel. A new analytical model was developed and used to demonstrate that transverse hydrodynamic dispersion likely caused the biomass to move in the direction observed when the PCE concentration was changed. The model was unable, however, to explain why growth migrated upward when the PCE concentration was initially constant. We postulate that this occurred because PCE, not lactate, sorbed to biofilm components and that biomass on the lactate side of the micromodel was limited in PCE. A fluorescent tracer experiment showed that biomass growth changed the water flow paths, creating a higher velocity zone in the PCE half of the micromodel. These results contribute to our understanding of biofilm growth and will help in the development of new models to describe this complex process.