Mesoscale tank experiments were performed to simulate bioremediation of saturated zone carbon tetrachloride (CCl4) originating from a vadose zone carbon tetrachloride source. The mesoscale tank is 2-m wide by 2-m high by 3-m long and was constructed of stainless steel, yielding a total volume of 12 m3. Simulated geology within the tank consisted of two unconsolidated sand layers separated by a clay layer containing variable-sized stainless steel tubes that represented fractures within a consolidated porous medium. The thickness of the upper sand layer was approximately 55 cm, the thickness of the virtual fracture layer was 25 cm, and the thickness of the lower sand layer was approximately 98 cm. The water table was located at an elevation of approximately 54 cm from the bottom of the tank. CCl4 was added to the sealed tank by pouring 500 ml of neat CCl4 into a beaker buried approximately 10 cm below the upper sand surface through a stainless steel tube. The CCl4 was then allowed to partition through the reactor over time, eventually coming to equilibrium. Once CCl4 equilibrium had occurred in the saturated zone (~500 ppb); the reactor was bioaugmented with a CCl4 degrading culture enriched from the Subsurface Disposal Area at the INEEL. The culture was grown to a cell density of ~ 1.0 x 108 cells/ml and injected into the simulated aquifer through a center sampling port. Following injection of the culture, an initial aliquot of lactate (1,000 g/L), nitrogen, and phosphorus were added to the reactor. Lactate was injected every 3 – 5 days for one month. After 1 month of operation, a continuous supply of lactate (1,000 g/L) was pumped into the reactor at an average rate of 50 mL/min. CCl4 concentrations in the unsaturated zone were measured using hollow fiber membrane samplers, while liquid samples were analyzed to monitor levels in the simulated aquifer zone. Samples were also taken for analysis of volatile organic acids and cell density. As would be expected, increases in cell density over the length of the cell correlated with the flow of the water through the cell. One week following injection microbes and lactate, cell numbers were in the range of 5.0 x 106 cells/mL, by the end of the experiment cell numbers had increased to approximately 1.94 x 107 cells/mL. Five days after lactate injection was initiated, chloroform appeared in liquid samples taken for chlorinated VOC analysis. CCl4 concentrations in the liquid phase dropped to approximately 180 ppb. At the conclusion of the batch lactate injection phase of the bioaugmentation, CCl4 levels averaged 40 ppb and chloroform levels averaged 44 ppb. Interestingly, once continuous lactate addition was initiated, CCl4 concentrations in the saturated zone increased with spikes as high as 3,000 ppb. Chloroform concentrations also increased following continuous addition of lactate. Since the CCl4 source in the breaker had been depleted, vadose zone concentrations of CCl4 dropped steadily during addition of lactate to the saturated zone. CCl4 levels of ~ 800 ppmv were noted at the beginning of the experiment, levels decreased to below 200 ppmv by the end of the bioaugmentation phase. No chloroform was noted in the vadose zone during testing
