Evaluation of in situ biosurfactant production by inoculum of P. putida and nutrient addition for the removal of polycyclic aromatic hydrocarbons from aged oil-polluted soil.

This work aimed to conduct a laboratory study to evaluate the use of Pseudomonas putida CB-100 and nutrient addition for the removal of PAHs from an aged oil-polluted soil of Veracruz, Mexico. Pseudomonas putida is a biosurfactant-producing bacterium capable of metabolizing polycyclic aromatic hydrocarbons (PAHs), which are toxic compounds with low water solubility, high melting, and boiling points, and low vapor pressure; characteristics that increase as their molecular weight increases and make them more recalcitrant. The methodology consisted in sampling the long-term oil-polluted soil and testing the use of Gamma irradiation (25 kGy) for the sterilization of the soil for abiotic control. We evaluated serological bottles containing 20 g of 35% moist soil (irradiated and non-irradiated) with the following treatments: the addition of nutrients (NH 4 Cl, NaNO 3 , KH 2 PO 4, and K 2 HPO 4 ), an inoculum of P. putida, and both P. putida and nutrients. The parameters assessed were pH, organic matter, humidity, available phosphorus, total nitrogen, cultivable heterotrophic microorganisms, CO 2 production, rhamnolipids, surface tension, and the removal of eleven PAHs. The non-irradiated soil added with P. putida was the most efficient in the removal of PAHs; the pattern was: Benzo(a)anthracene > Phenanthrene > Fluoranthene > Benzo(k)fluoranthene > Chrysene > Pyrene > Anthracene > Acenaphthylene > Benzo(b)fluoranthene. In conclusion, P. putida in the non-irradiated soil produced in situ biosurfactants (1.55 mg/kg of rhamnolipids and an 11.9 mN/m decrease in surface tension) and removed PAHs in 10 days.