Effectiveness of Bioremediation on Oil-Contaminated Sand in Intertidal Zone

Bioremediation technologies were applied to experimental microcosms, simulating an oil spill in a lower intertidal area. Three treatments (oil only, oil plus nutrients, and oil plus nutrients and microbial inocula) were applied, and each microcosm was repeatedly filled and eluted with seawater every 12 h to simulate tidal cycles. To minimize washing-out of the inoculum by the tidal cycles, microbial cells were primarily immobilized on diatomaceous earth before they were applied to the oiled sand. Oil degradation was monitored by gravimetric measurements, thin layer chromatography/flame ionization detector (TLC/FID) analysis, and gas chromatography (GC) analysis, and the loss of oil content was normalized to sand mass or nor-hopane. When the data were normalized to sand mass, no consistent differences were detected between nutrient-amended and nutrient/inoculum-amended microcosms, although both differed from the oil-only microcosm in respect of oil removal rate by a factor of 4 to 14. However, the data relative to nor-hopane showed a significant treatment difference between the nutrient-amended and nutrient/inoculum-treated microcosms, especially in the early phase of the treatment. The accelerating effect of inoculum treatment has hardly been reported in studies of oil bioremediation in the Tower intertidal area. The inoculum immobilized on diatomaceous earth seemed to be a very effective formulation for retaining microbial cells in association with the sand. Results of this study also suggest that interpretation of the effectiveness of bioremediation could be dependent on the selection of monitoring methods, and consequently the application of various analytical methods in combination could be a solution to overcome the limitations of oil bioremediation monitoring.