A polyphasic approach for assessing the suitability of bioremediation for the treatment of hydrocarbon-impacted soil

Bioremediation strategies, though widely used for treating hydrocarbon-contaminated soil, suffer from lack of biodegradation endpoint accountability. To address this limitation, molecular approaches of alkB gene analysis and pyrosequencing were combined with chemical approaches of bioaccessibility and nutrient assays to assess contaminant degrading capacity and develop a strategy for endpoint biodegradation predictions. In long-term hydrocarbon-contaminated soil containing 10.3g C₁₀-C₃₆ hydrocarbonskg^-1, 454 pyrosequencing detected the overrepresentation of potential hydrocarbon degrading genera such as Pseudomonas, Burkholderia, Mycobacterium and Gordonia whilst amplicons for PCR-DGGE were detected only with alkB primers targeting Pseudomonas. This indicated the presence of potential microbial hydrocarbon degradation capacity in the soil. Using non-exhaustive extraction methods of 1-propanol and HP-β-CD for hydrocarbon bioaccessibility assessment combined with biodegradation endpoint predictions with linear regression models, we estimated 33.7% and 46.7% hydrocarbon removal respectively. These predictions were validated in pilot scale studies using an enhanced natural attenuation strategy which resulted in a 46.4% reduction in soil hydrocarbon content after 320days. When predicted biodegradation endpoints were compared to measured values, there was no significant difference (P=0.80) when hydrocarbon bioaccessibility was assessed with HP-β-CD. These results indicate that a combination of molecular and chemical techniques that inform microbial diversity, functionality and chemical bioaccessibility can be valuable tools for assessing the suitability of bioremediation strategies for hydrocarbon-contaminated soil.