Engineered in situ bioremediation of soil and groundwater polluted with weathered hydrocarbons

In this work, we present our experience in the engineered in situ biostimulation of a hydrocarbon-polluted subsoil of an old metal-working plant. The site had a long history of fuel, lubricant oil, and diesel spills and leakages that were initially treated by means of physico-chemical techniques. After one year of treatment, weathered hydrocarbons were firmly sorbed in the unsaturated zone, limiting the effectiveness of the physico-chemical methods being applied. Also, low nutrient and dissolved oxygen levels limited natural attenuation yields. Therefore, a bioremediation approach based on the injection of hydrogen peroxide, an oleophilic fertilizer, and a surfactant was applied. Total petroleum hydrocarbons and gas chromatography-mass spectrometry determinations were performed as a chemical means of monitoring the process. Microbial populations, including the presence of hydrocarbon-degrading bacteria, were simultaneously analyzed during the process, using enrichment techniques and confocal laser scanning microscopy observations with fluorescent indicators. Hydrocarbon-degrading bacteria were already present in the polluted subsoil prior to the implementation of this bioremediation technique and increased noticeably during the first 2 months of treatment. Hence, the suitability of the nutrient and oxygen amending approach was confirmed as further demonstrated by chemical determinations.