Bioavailability of weathered hydrocarbons in engine oil-contaminated soil: Impact of bioaugmentation mediated by Pseudomonas spp. on bioremediation.

Heavier fraction hydrocarbons (C15-C36) formed in soil after biotic and abiotic weatherings of engine oil are the continuing constraints in the bioremediation strategy, and their bioavailability remains a poorly quantified regulatory factor. In a microcosm study, we used two strains of Pseudomonas, P. putida TPHK-1 and P. aeruginosa TPHK-4, in strategies of bioremediation, viz., natural attenuation, biostimulation and bioaugmentation, for removal of weathered total petroleum hydrocarbons (TPHs) in soil contaminated long-term with high concentrations of engine oil (39,000-41,000 mg TPHs kg-1 soil). Both the bacterial strains exhibited a great potential in remediating weathered hydrocarbons of engine oil. Addition of inorganic fertilizers (NPK), at recommended levels for bioremediation, resulted in significant inhibition in biostimulation/enhanced natural attenuation as well as bioaugmentation. The data on dehydrogenase activity clearly confirmed those of bioremediation strategies used, indicating that this enzyme assay could serve as an indicator of bioremediation potential of oil-contaminated soil. Extraction of TPHs from engine oil-contaminated soil with hydroxypropyl-β-cyclodextrin (HPCD), but not 1-butanol, was found reliable in predicting the bioavailability of weathered hydrocarbons. Also, 454 pyrosequencing data were in accordance with those of bioremediation strategies used in the present microcosm study, suggesting the possible use of pyrosequencing in designing approaches for bioremediation.