Attenuation of petroleum hydrocarbon fractions using rhizobacterial isolates possessing alkB, C23O, and nahR genes for degradation of n-alkane and aromatics.

This work assessed the catabolic versatility of functional genes in hydrocarbon-utilizing bacteria obtained from the rhizosphere of plants harvested in aged polluted soil sites in Ogoni and their attenuation efficacy in a bioremediation study. Rhizosphere soil was enumerated for its hydrocarbon-utilizing bacteria. The bacteria were in-vitro screened and selected through the quantification of their total protein and specific intermediate pathway enzyme (catechol 2,3-dioxygenase) activity in the metabolism of hydrocarbon. Thereafter, agarose gel electrophoresis technique was deployed to profile the genome of the selected strains for catechol 2,3-dioxygenase (C23O), 1,2-alkane monooxygenase (alkB), and naphthalene dioxygenase (nahR). Four rhizobacterial isolates namely Pseudomonas fluorescens (A3), Achromobacter agilis (A4), Bacillus thuringiensis (D2), and Staphylococcus lentus (L1) were selected based on the presence of C23O, alkB, and nahR genes. The gel electrophoresis results showed an approximate molecular weight of 200 bp for alkB, 300 bp for C23O, and 400 bp for nahR. The gas chromatogram for residual total petroleum hydrocarbon (TPH) revealed mineralization of fractions C8-C17, phytane, C18-C30. TPH for in-vitro bioremediation of crude oil-polluted soil was observed to have an optimal reduction/loss of 97% within the 56th day of the investigation. This study has further revealed that the microbiome of plants pre-exposed to crude oil pollution could serve as a reservoir for mining group of bacterial with broad catabolic potentials for eco-recovery and waste treatment purposes.

Optimization of bioremediation-cocktail for application in the eco-recovery of crude oil polluted soil.

Background: Environmental sustainability is the driver for finding the optimal bioremediation cocktail with the combination of highly potent hydrocarbonoclastic strains and the nutrient additives that significantly enhance mineralization of crude oil in polluted soil in order to mitigate its deleterious effects on the environment. In this study, four hydrocarbon-degrading bacterial strains were pre-selected from mined rhizobacterial isolates in aged crude oil-contaminated soil.  Method: Agrowaste residues of poultry-droppings, corn chaff, and plantain peel were selected among others for their ability to support high biomass of selected bacterial strains. Baseline proximate analysis was performed on the agrowaste residues. Simplified, one variable at a time (OVAT) was employed in the validation of the variables for optimization using the Multivariate analysis tool of Response Surface Methodology (RSM). To test the significant formulation variables, the Box-Behnken approach using 15 runs design was adopted. Results:  The rate of contaminant removal was observed to fit into a quadratic function. For optimal rate or contaminant removal, the fitted model predicted the optimal formulation cocktail condition to be within 0.54 mg/kg (Corn steep liquor), phosphate 137.49 mg/kg (poultry droppings) and 6.4% inocula for initial TPH of 9744 mg kg -1 and THC of 9641 mg kg -1 contaminant level. The model for the application of the bioremediation product and the variables evaluated had a significant p-value < 0.005 for the attainment of 85 to 96 % of TPH and THC removal after 56 days of treatment. Conclusions:  This study has shown the need to harness the abundant agrowaste nutrients in supporting high throughput rhizobacteria in the formulation of a bioremediation agent suitable for use in the reclamation of oil spill sites in the Niger Delta oil-producing region.