The method of cleaning water from oil pollution
The invention relates to applied Microbiology and can be used for cleaning the surface of the water from spilled oil and petroleum products. For implementing the method in contaminated by oil and water make a bacterial culture, representing a consortium of microorganism cells of Bacillus brevis IB DT 5-1 and Arthrobacter species IB DT 5-3 in the form of the culture fluid with a total titer of not less than 108CFU/ml Aeration in the pond contaminated by oil or oil products, to maintain the activity of aerobic microorganisms-oil degrading carry out a forced intake of contaminated water and its distribution by the method of sprinkling on the surface of the water body. The method allows to effectively clean the water surface at high oil content or oil in the water. 1 C.p. f-crystals, 1 table.
The invention relates to applied Microbiology and can be used for cleaning the surface of the water from spilled oil and petroleum products.
Known bacterial preparation “Poutical designed for cleaning oil-contaminated soil and water, made on the basis of bacterialike “Poutical” is a dry bacterial mass, and prior to his “recovery” necessary labour-intensive set of activities performed in the field: heating large quantities of water (5 m3) to a temperature of 28° C, providing aeration for 12 to 16 hours, stirring.
Known bioreagent for purification of water and soil from oil pollution, representing bacterial culture immobilized on gidrofobizirovannym peat /3/. This gidrofobizirovannym peat is an oil adsorbent, due to this there is the destruction of the oil film and increases the access of oxygen that activates the process of biodegradation of petroleum products.
A disadvantage of the known technical solutions adopted for the prototype, is the low efficiency of its use in stagnant and parinamah reservoirs with a high degree of oil pollution.
The technical task of the invention is to increase the efficiency of water purification from oil pollution.
The problem is solved by the use of process water treatment consortium of natural aerobic oil-oxidizing microorganisms, consisting of Bacillus brevis IB DT 5-1 and Arthrobacter species IB DT 5-3, priceville from the gray forest soil, artificially contaminated with diesel fuel, and deposited in the Collection of microorganisms, Institute of biology, Ufa scientific center of RAS.
The characteristic strain of Bacillus brevis IB DT 5-1.
Morphological and cultural characteristics. Straight bacilli with rounded ends. Gram-positive. Movable. The spherical endospores. On agar media, form round, with smooth edges, dense in the center of the colony for the matte, opaque, luminous. Saprophyte.
Physiological and biochemical properties. Aerobes. Chemoorganotrophic. Catalase and oxidizability. Well metabolizes sucrose, xylose, fructose, maltose, lactose, lures, mannose, arabinose, L-rhamnose, glycerol. Absorbs asparagine, arginine, alanine; weakly absorbs valine, lysine. The reaction Wages-Proskauer negative. Gas from glucose does not form. Indole is not formed. Growth under anaerobic agar negative. Produces acid from glucose. Starch is not decompose. Able to oxidize the hydrocarbon oil. Non-pathogenic. Characteristics of the strain of Arthrobacter species IB DT 5-3.
Morphological and cultural characteristics. Short sticks with age are shortened to cocci. Gram-positive. On solid nutrient media form round, definition properties. Aerobe. Catalanopolonesa, oxidization. The oxidative metabolism type. Well metabolizes sucrose, fructose, maltose, lactose, xylose, mannitol, mannose, arabinose, L-rhamnose, glycerol. When grown on glucose forms a gas. Absorbs asparagine, arginine, alanine; weakly absorbs valine, lysine. Consumes citrate as the sole carbon source. Gelatin is not hydrolyses. Starch is not decompose. Indole is not formed. The reaction Wages-Proskauer negative. The lack of growth on medium containing 6% NaCl. Able to oxidize the hydrocarbon oil. Non-pathogenic.
Oxidative activity of the consortium of microorganisms was determined by the final product of oxidation, i.e. the allocation of CO2according to the method described in /4/. In a flask with a capacity of 500 ml contribute absorber of carbon dioxide BA(OH)2, 250 ml of culture medium Raymond, 2.5 ml of diesel fuel and 5 ml three-day culture of the consortium with the contents of the cells 1,0· 108CFU/ml, as well as individual pure cultures (5 ml) of microorganisms Bacillus brevis IB DT 5-1 and Arthrobacter species IB DT 5-3 with a titer of 1.0-108CFU/ml, obtained by selecting them from the specified consortium. Oxidative activity of the consortium in the experiment duration 30 days costino, 750 mg CO2and 870 mg CO2. Thus, the type of the specified consortium is a natural symbiotic Association of two strains of microorganisms.
The quantitative content of each of the strains of microorganisms belonging to the consortium, constituted in culture liquid of 108CFU/ml and did not change during prolonged storage in appropriate conditions. The method of mathematical analysis established that this consortium of Bacillus brevis IB DT 5-1 refers to the dominant form (51%) of all the studied microorganisms in the population. Bacteria Arthrobacter species IB DT 5-3 are codominant culture (49%).
The consortium used in the form of the culture fluid obtained during joint fermentation of these strains of microorganisms, under the General title of microorganisms not less than 1 to 108CFU/ml, and aeration in the pond is achieved by forced intake of contaminated water and its distribution by the method of sprinkling on the surface of the water object.
The method is as follows.
To obtain cell biomass consortium of microorganisms Bacillus brevis IB DT 5-1 and Arthrobacter species IB DT 5-3 them together grown in liquid nutrient medium followed by 7H2O - 0,02; NaH2RHO4TO 1.5; K2NRA4- 1,0; MgSO4·7H2O - 0,2; NH4NO3- 2,0; water water 1000 ml.
As the only carbon source use diesel fuel.
The cultivation process is conducted continuously or intermittently at a temperature of 18-20° C in aerobic conditions until titer of microorganisms in the culture fluid of value not less than 108CFU/ml (working title biopreparation “Lenoil”).
Example 1. In reservoirs, representing aquariums with a volume of 10 l, bring tap water to a volume of 6 l, crude oil Pravdinskaya field (or commodity diesel fuel) in a volume of 0.5 l and the suspension of microorganisms biological product “Lenoil” with a titer of 1.2· 108CFU/ml in a volume of 200 ml. of Aeration in the tank is provided laboratory peristaltic pump, with the inlet of the pump to force water intake is below the level of the oil film, and the outlet with a rubber tube, provided with perforated holes for distributing pumped from a reservoir of water above the surface of the oil film.
In the control tank, in which onmicroorganisms biological product “Lenoil”, immobilized on gidrofobizirovannym peat according to known technical solution.
Experiments carried out at a temperature of 20° C, the residual oil content in water is determined by gravimetric method. The results of the experiments are given in the table.
Example 2. In stagnant swamp area of 1.0 hectares intensity crude oil contamination of the Pravdinskaya field is 1 kg/m2(the total number of oil pollution around 10 tons). Processing biological product “Lenoil” held under the title of microorganisms in the culture fluid of 1.8· 108CFU/ml Flow rate of a biological product amounted to 4.0 m3. Day temperatures ranged 14-20° C. the Aeration of the water object was achieved by the forced diversion of water by means of a pump brand VXm 10/50 and water supply and distribution through pipelines to the surface contaminated by the oil film of water.
24 days later analyses showed that the total number of oil pollution decreased by 93%.
Presents the results showed the effectiveness of the proposed method, allowing to clean the water surface from oil pollution in the high degree of this sealing water bodies does not cause any difficulties and is determined only by the presence of a biological product and pumping equipment.
1. USSR author's certificate No. 1076446, 02 F 3/34, 1984.
2. USSR author's certificate No. 1428809, 02 F 3/34, 1988.
3. RU patent No. 2081854, 02 F 3/34, 1997.
RU patent No. 2115629, 02 F 3/34, In 09 With 1/10, 1998.
1. The method of purification of the water surface from oil and oil products, including the introduction of a bacterial culture, characterized in that as a bacterial culture using a consortium of natural strains of aerobic oil-oxidizing microorganisms Bacillus brevis IB DT 5-1 and Arthrobacter species IB DT 5-3 in the form of the culture fluid with a total titer of microorganisms, not less than 108CFU/ml of culture fluid, and the cleaning process is carried out with the help of aeration of the water object.
2. The method according to p. 1, characterized in that the aeration in the pond contaminated by oil or oil products, by abstraction of contaminated water and its distribution by the method of sprinkling on the surface of the water object.