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 (NH4Cl, NaNO3, KH2PO4, and K2HPO4), 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, CO2 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.
