Bioavailability enhanced rhizosphere remediation of petroleum hydrocarbon contaminated soil

Aliphatic, aromatic and polycyclic aromatic oil hydrocarbons are structurally complicated man-caused pollutants that are constantly brought into biosphere. Oil production in Russia, so as all over the world, is connected with pollution of biotopes, ecosystems and agro-landscapes. Presently large funds are allocated either for oil leak prevention or for discharged oil gathering. At the same time, in spite of large necessity in technologies for efficient reconstruction of soil bio-productivity, reliable regional systems of their remediation in situ have not been developed yet. One such method is rhizosphere remediation, a biotechnology, based on the functioning of plant-microbial complexes. Little is known about bioavailability in phyto-remediation systems. Specific bioavailability-promoting mechanisms, operating in soil with hydrocarbon-degrading populations, may be responsible for increased rates of pollutant transformation (increased bacterial adherence to the pollutants, production of bio-surfactants by bacteria or by plants, possible role of chemotaxis). In the course of work collection of 42 chemo-tactically active bio-surfactant producing strain-degraders of petroleum hydrocarbons including polycyclic aromatic hydrocarbons (PAHs) was created. Two representative strains were selected for detailed chemotaxis studies with PAHs (naphthalene, phenanthrene, anthracene, and pyrene), bacterial lipopolysaccharide and root exudates from seven different plants. These strains are produce the bio-surfactants (rhamno-lipid). The chemotactic response was quantified with a capillary and densitometric chemotaxis assay. Surface tension of cultural liquid was measured after cultivation of strains in the presence of hexadecane or phenanthrene with the use of a ring tensiometer. Before measuring of surface tension microbial cells were collected from liquid culture by centrifugation. Total petroleum Hydrocarbons (TPH) in soil were analyzed by infra-red spectroscopy method. PAHs contain were analyzed by gas chromatography method. Four bioassays were used to measure toxicity during bio-remediation of soil contaminated by petroleum hydrocarbons: Microtox(R) test, SOSchromotest, lettuce seed germination and sheep red blood cell (RBS) hemolysis assay. Rhizosphere remediation was found to be effective for removal of polycyclic aromatic hydrocarbons (PAHs), total petroleum hydrocarbons (TPHs) from soil with the use of alfalfa inoculated by the Pseudomonas stutzeri MEV-S1 strain (RU 2228952 patent) and oats inoculated by the Pseudomonas alcaligenes MEV strain (RU 2228953 patent) in vegetation and field experiments. The reduction of the TPH and PAH concentrations in soil was accompanied by the reduction of integral toxicity and genotoxicity, evaluated by bio-testing. It is conceivable, therefore, that a possible way to optimize petroleum hydrocarbons phyto-remediation is the use of selected plants and microbial inoculants with specific chemotactic affinities and bio-surfactant production. The proposed technology for soil bio-remediation with the use of integrated plant-microbial system is ecologically and toxicologically safe and economically attractive