Remediation technologies for oil-polluted soil:

• The first soil remediation technology was developed and practically used in the Klaipėda production base. It includes scattering of the soil over a polygon in a 0.35–0.45 cm thick layer, mellowing twice a week with a special device, insertion of biopreparation four times a year (on May 5–10, June –10, July 5–10, and August 5–10) and fertilization with nitrogen fertilizers twice a month, and phosphorus and potassium fertilizers once a month.
• The second soil remediation technology is applied in the Jonava production base. It includes formation of soil stacks on permeable draining ground following an established order, installation of aeration system, watering with a special solution and insertion of biogenic elements.
• The third remediation technology is based on the use of higher plants. The aim is to select the plants resistant to oil products (fuel-oil), determine the limit values of fuel-oil in the soil (at which higher plants are unable to grow), and to evaluate the functional capacity of the rhizosphere of the selected plants to disintegrate fuel-oil. The plants used were: wheat-grass, reed, lupine, white clover, fescue grass, flax, meadow grasses, oats, tares and rye.

In most cases, there occurs pollution with oil products containing high concentrations of polyaromatic compounds and pitch. These hydrocarbons are especially resistant to biological disintegration.

In order to have a normal course of the process of biological remediation it is necessary to ensure optimal temperature, moisture, aeration and pH conditions and the necessary content of N, P, K and trace elements. Under these conditions, the oil-decomposing microorganisms Pseudomonas fluorescens, Bacillus cereus, Penicillium decumbens Thom, Serratia, Acinetobacter, Trichoderma, and others intensively propagate, and the degradation of oil products is optimal.

The technological control of remediation process is implemented by the technological service of the enterprise in conformity with the regulations:

• visiting the pollution source;
• analysis of the causes of pollution;
• estimation of the levels and scope of pollution;
• selection of remediation technologies and creation of normal medium for oil-decomposing microorganisms (pH, ratio N-P-K, abstergent to be or not to be used);
• regular control of remediation process, pH, N-P-K ratio and colonies of oil-decomposing microorganisms.

The end of remediation process (cleaning to the established standards depending on the further use of the soil) is committed by the Regional Department of Environment after the evaluation of the content of oil products in the soil implemented by the National Department of Analytical Control. In our opinion, other organizations implementing the in situ and ex situ soil remediation works should establish a similar control system.

In cooperation with the specialists of scientific organizations we upgraded and patented the effective technologies for liquidation of pollution with oil products:

1. Microbiological remediation of soil polluted with oil and oil products (LT patent 4559 B, issued 25 10 1999).
2. Bioremediation technology for soil polluted with oil and its products (LT patent 4620 B, issued. 25 02 2000).
3. Bacteria strain Serratia odorifera A3b GKM VIZR N 100, oxidizing hydrocarbons of oil and oil products (RU patent 2157839, issued 20 10 2000).
4. Bacteria strain Serratia odorifera A3p GKM VIZR N 99, oxidizing hydrocarbons of oil and oil products (RU patent 2157838, issued 20 10 2000).
5. Bacteria strain Serratia ficaria B3p GKM VIZR N 101, oxidizing hydrocarbons of oil and oil products (RU patent 2157840, issued 20 10 2000).
6. Bacteria strain Acetobacter pasterianum ABZ-2 GKM VIZR N 102, oxidizing hydrocarbons of oil and oil products (RU patent 2157841, issued 20 10 2000).
7. Bacteria strain Trichoderma lignorum L-1 GKM VIZR N 103, oxidizing hydrocarbons of oil and oil products (RU patent 2157842, issued 20 10 2000).
8. Bacteria strain Bacillus cereus B3b GKM VIZR N 98, oxidizing hydrocarbons of oil and oil products (RU patent 2157843, issued 20 10 2000).
9. Strains Penicillium decumbens Thom 4.5 VNB-AMFK and Penicillium frequentans Westling 6.4 PNB-AMFK, oxidizing hydrocarbons of oil and oil products (LT patent N 4791 B, issued 25 05 2001).
10. Strain Pseudomonas fluorescens IG N 57, oxidizing hydrocarbons of oil and oil products (LT patent N 4792 B, issued 25 05 2001).
11. Strain Candida lipolytica C.6.1-5, oxidizing hydrocarbons of oil and oil products (LT patent N 4793 B, issued 25 05 2001).
12. Strain Trichoderma harzianum VNB-16, decomposing fuel-oil; its isolation and use (LT patent N 4794 B, issued 25 05 2001).
13. Biological remediation technology for soil polluted with oil or oil products (LT patent N 4593 B, issued 27 12 1999).
14. DEVICE AND METHOD FOR VEGETATIVE WASTE WATER TREATMENT (LT patent N 5496 B, issued 25 04 2008).
15. PROCESS FOR PREPARING SAPROPEL FROM FRESHWATERS (LT patent N 5495 B, issued 25 04 2008).
16. PROCESS FOR BIODEGRADING INDUSTRIAL WASTE - MONOETHANOLAMINE (MEA) SOLUTION (LT patent N 5502 B, issued 26 05 2007).
17. THE METHOD OF TREATMENT MONOETANOLAMINE SOLUTION IDUSTRIAL WASTE (LT patent N 5373 B, issued 25 10 2006).
18. PROCESS AND DEVICE FOR REDUCING BIOGENES AMOUNT AND "FLOWERING" IN FRESH-WATER (LT patent N 5201 B, issued 25 03 2005).
19. BIOLOGICAL PREPARATION FOR REMOVING CRUDE OIL AND PETROLEUM PRODUCT CONTAMINANTS FROM SOIL AND WATER, METHOD FOR PRODUCTION AND USE THEREOF (LT patent N 5057 B, issued 25 09 2003).
20. PROCESS FOR BIODEGRADING INDUSTRIAL WASTE - MONOETHANOLAMINE (MEA) SOLUTION (LT patent N 5502, issued 2008-05-26).
21. PROCESS FOR BIOLOGICAL DECONTAMINATION OF SOIL CONTAMINATED WITH OIL OR PETROCHEMICALS (LT patent N 5579, issued 2009-06-25).
22. METHOD FOR PRODUCING OIL-OXIDISING MICROORGANISMS MEDIA AND MEDIA OBTAINED THEREBY (LT patent N 5587, issued 2009-08-25).
23. PROPHYLACTIC HYGIENIC COMPOSITION, PROCESS FOR PREPARING AND USES THEREOF (LT patent N 5632, issued 2010-02-25).
24. PHYTOCHEMICAL COMPOSITION, METHOD OF PREPARATION AND USE THEREOF (LT patent N 5701, issued 2010-12-27).
25. PHYTOCHEMICAL COMPOUNDS COMPOSITION AND METHOD OF ITS RECEIPT OF THE MALIGNANT TUMORS TREAT (LT paent N 5740, issued 2011-06-27).

Serratia                                                              Acinetobacter                                              Trichoderma

The SRS, using the patented strains of oil oxidizing microorganisms, produced test samples of original effective preparations for disintegrating oil products:

1. Biopreparation (Pseudomonas, Alcaligenes, Mucor, Scopulariopsis – the total of 7 strains) for disintegration of oil products (testing liophilisation method).
2. Biopreparation (Pseudomonas fluorescens, Candida lipolytica, Trichoderma harzianum – patented strains) for fuel-oil degradation.
3. Biopreparation (hydrophilous oil oxidizing microorganisms were used which aggregate in the water phase and lipophilous microorganisms which are present in the oil phase); hydrophilous bacteria – Pseudomonas sp., lipofilinės – Rhodococcus sp., Bacillus cereus and micromycetes – Candida sp., Trichoderma lignorum) designed for liquidation of different oil products.

The above mentioned patents are valuable because they contain full-scale descriptions of strain use (in other words, oil-polluted soil and water remediation) technologies.

It was determined that the roots of some higher plants (rhizosphere) effectively disintegrate the remains of oil products (6,000 mg/kg of soil). On this base, the technologies of cleaning territories polluted up to 0.5 m in depth in situ and restoration of the soil structure are in the process of improvement.

A close cooperation with the Biodestructor Research Laboratory, Institute of Botany of Nature Research Centre and the Department of Microbiology and Biotechnology, Faculty of Natural Science, Vilnius University took place during the time of described activities. Isolation of active oil decomposing mesophilous and thermophilous microorganisms from the natural sources, investigation of oil oxidizing bacteria and active micromycete strains and assessment of their physiological–biochemical properties were implemented by joint efforts.