RU2428471C1 - Eco-biopreparation centrum-mms for removing oil and oil products - Google Patents

Abstract

FIELD: chemistry. ^ SUBSTANCE: eco-biopreparation contains Pseudomonas fluorescens VKM V-6847 strain and Rhodococcus erythropolis IT VKM As-1769 strain in volume ratio 1:1, and additionally glycerine with the following ratio (wt %): consortium of the Pseudomonas fluorescens VKM V-6847 strain with content of bacterial cells of not less than (1.5-2.0)109-10 cl.cm-3 and the Rhodococcus erythropolis VKM As-1769 strain with cell content of (1.0-1.5)109-10 cl.cm-3 94-95; glycerine 5-6. ^ EFFECT: high efficiency of removing oil and oil products from environmental objects. ^ 1 tbl, 2 ex

Description

The invention relates to biotechnology for environmental protection and is intended for bioremediation measures for the purification of hydrocarbon pollutants from nature, primarily oil and fuels and lubricants. The invention can be used in the process of biological cleaning of the environment, local spills of crude oil, various fuels and lubricants, washing parts, fuel tanks, processing of industrial areas, gas stations, places of washing machines, etc.

A known strain of Rhodococcus erythropolis for the decomposition of oil and oil products (RF patent 2257409, MKI C12N 1/20, B09C 1/10, 2004). The strain is deposited as Rhodococcus erythropolis VKPM AC-1668 and has a relatively high rate of utilization of oil and oil products.

However, the disadvantages of the drug, created on the basis of the known strain, are, firstly, its monocomponent, that is, the drug contains one type of microorganism, which cannot provide a high potential for degradation activity, and therefore, the rate of utilization of oil products during soil remediation; secondly, the need to conduct additional activation of the drug; thirdly, there is no possibility to utilize "heavy" and aromatic hydrocarbons, as well as to use the strain for the biological decomposition of other types of toxicants (alcohols, aldehydes, industrial oils, fats, etc.).

Known bacterial preparation "Roder" for the purification of soils, oil sludge, fresh and mineralized waters from oil and oil products (RF patent 2295403, MKI B09C 1/10, C02F 3/04, C12N 1/20, 2007). The drug is based on the cultivation of the R-forms of strains of Rhodococcus erythropolis VKM AC-1514 and Rhodococcus rubber VKM AC-1513 D. The drug can be obtained in the form of a dry form and in the form of a concentrated suspension, therefore, it is characterized by long shelf life and ease of transportation.

However, a disadvantage of the known preparation is its composition, including strains from the same taxonomic group, which determines its insufficient degradation activity and limits the use in soil remediation, especially in case of technogenic pollution with oil products containing "heavy" and aromatic hydrocarbons. The drug is ineffective for the biological decomposition of other organic ecotoxicants (alcohols, aldehydes, industrial oils, fats), in addition, this drug has low activity at low temperatures and requires more than one treatment of the contaminated object.

Thus, the authors were faced with the task of developing a biological preparation characterized by high degradation activity with respect to crude oil and various types of hydrocarbon fuels, which is preserved at relatively low ambient temperatures, which makes it possible to use it for soil remediation, for example, in northern oil producing regions.

The problem is solved by using a new biological preparation for cleaning soil from oil and oil products based on biomass of a consortium of aerobic bacterial cultures containing microorganisms Rhodococcus erythropolis, characterized in that it contains a strain of Pseudomonas fluorescens VKM B-6847 and a strain of Rhodococcus erythropolis B KM Ac-1769 with a volume ratio of 1: 1, and additionally glycerin in the following ratio (wt.%):

consortium of strain Pseudomonas fluorescens VKM B-6847 when the cell content is not less than (1.5 ÷ 2.0) · 10 ^{9 ÷ 10} cells · cm ^-3 and strain Rhodococcus erythropolis VKM Ac-1769 when the cell content is not less than (1.0 ÷ 1.5) 10 ^{9 ÷ 10} cells · cm ^-3 94 ÷ 95 glycerol 5 ÷ 6

Currently, the biological product-destructor of oil and oil products is not known from the patent and scientific literature, the composition of which is the biomass of the consortium of aerobic bacterial cultures of the strain Pseudomonas fluorescens VKM B-6847 and strain Rhodococcus erythropolis VKM Ac-1769, taken in a certain mass ratio, and which further comprises glycerin.

The proposed biological product is a mixture of two natural bacterial strains that actively oxidize hydrocarbons. Studies conducted by the authors revealed a synergistic effect in the degradation of hydrocarbons of oil and oil products. The strain Pseudomonas fluorescens VKM B-6847 is active in the utilization of "heavy" hydrocarbons, and the strain Rhodococcus erythropolis VKM As-1769 is primarily active in the utilization of aromatic hydrocarbons. But it was not obvious that a consortium of these strains would exhibit high biodegradation activity in relation to crude oil and various types of hydrocarbon fuels. The drug can also be used for the biological decomposition of other types of ecotoxicants (alcohols, aldehydes, industrial oils, fats, etc.). The advantage of the proposed drug is the possibility of its use for soil remediation at relatively low ambient temperatures (8-12 ° C), which is especially important for the northern oil producing regions.

The liquid bacterial preparation "CENTRUM-MMS" is a light cream suspension containing live microbial cells of two strains having hydrocarbon-oxidizing properties and a stabilizer. Glycerin is used as a stabilizer. Glycerin (Glycerin) - an organic compound related to polyols - alcohols containing several hydroxyl groups in the molecule.

Glycerin is widely used in medicine and pharmaceutical production. Glycerin is used to dissolve drugs, give moisture to tablets and pills, increase the viscosity of liquid preparations, protect against enzymatic changes during the fermentation of liquids and from drying ointments, pastes and creams. Glycerin is an excellent solvent for iodine, bromine, phenol, thymol, tannin, alkaloids and mercury chloride. Using glycerin instead of water, you can prepare highly concentrated medical solutions. Glycerin also has antiseptic properties, so it is used to prevent infection of wounds. The antiseptic and preservative properties of glycerol are associated with its hygroscopicity, due to which the dehydration of bacteria occurs, which contributes to their long-term preservation. Experimentally determined the optimal limits of the glycerol content in the proposed drug. Exceeding the amount of glycerol more than 6 wt.% Leads to its excess in relation to microorganisms, which can cause their excessive dehydration. When the content is less than 5 wt.%, There is a significant decrease in the conservation properties of the drug, and, accordingly, a reduction in its shelf life. To prepare the drug, a liquid culture of two bacterial strains grown together on a synthetic mineral medium is used (or separately).

Production strains isolated from the environment, belong to the group of saprophytes, are not pathogenic for humans and animals according to the classification of microorganisms given in the Sanitary Rules SP 1.2.731-99. Work with strains does not require special precautions.

Strains in the work are used in the form of agar and native liquid bacterial cultures.

Strain Pseudomonas fluorescens (VKM B-6847)

Cells are gram-negative rods. The colonies on nutrient agar are round shiny, at a low temperature form a diffusing non-fluorescent greenish pigment, in a liquid nutrient medium, the strain forms a uniform turbidity. Strict aerobic, oxidase- and catalase-positive, does not possess arginine hydrolase and denitrifying activity. Actively utilizes various organic substances and hydrocarbon compounds in the absence of other carbon sources. It grows well on synthetic liquid nutrient media, using hydrocarbon-containing substances as the sole source of energy and nutrition, and forms deep biomass during deep cultivation for 36-48 hours.

Strain Rhodococcus erythropolis (VKM Ac-1769)

Gram-positive motionless small coccopods. On nutrient agar forms round shiny colonies of a reddish color. In a liquid nutrient medium gives an abundant loose sediment. Respiration is of oxidizing type, oxidase- and catalase-positive, gelatin does not dilute, casein and starch do not hydrolyze, citrate utilizes slowly, actively assimilates hydrocarbon-containing substances, weakly glucose.

It grows well on synthetic nutrient media, using organic substances and hydrocarbon-containing compounds as the sole source of nutrition and energy, and when cultivated in depth for 48-52 hours, forms abundant biomass.

The proposed biological product is produced by the method of deep cultivation on a liquid synthetic medium of constant composition in a biological reactor.

The preparation of seed in flasks and cultivation in the apparatus is carried out on an environment of the following composition:

monosubstituted potassium phosphate, g 10.0 potassium phosphate disubstituted, g 4.0 ferrous sulfate, g 0.4 magnesium sulfate, g 0.1 drinking water, dm ³ 1,0

As a carbon source, glucose is used in an amount of not more than 1% for the entire preparation cycle of both inoculum and culture fluid.

The fermenter, filled into 2/3 of the volume with drinking tap water, is sequentially injected with the components of a synthetic nutrient medium. The medium is mixed and the required pH value is set (6.9 ± 0.1). The medium is sterilized at a temperature of (121 ± 2) ° C for (30 ± 1) min. The nutrient medium prepared in the apparatus should be considered a series. The shelf life of the nutrient medium is not more than 24 hours.

A sterile culture medium must meet the following requirements:

hydrogen indicator, units pH 6.5 ± 0.2 extraneous microflora not allowed

The same conditions must be met by the seed preparation medium.

Cultivation liquids of two production strains of Pseudomonas fluorescens (VKM B-6847) and Rhodococcus erythropolis (VKM Ac-1769) prepared in flasks are used as seed material for the fermenter; the seed dose of each component should be at least 1 · 10 ⁶ cells · cm ^-3 .

The seed for the fermenter is prepared in flasks placed in a thermostatically controlled apparatus for shaking flasks.

For cultivation, synthetic culture medium is added to the flasks in an amount of 1/5 of the volume of the flask. The inoculum is a bacterial suspension of strains grown separately on mowed nutrient agar. The sowing dose to the flask is at least 1 · 10 ⁶ cells · cm ^-3 .

Microbial cultures of the strains were grown at a temperature of (32 ± 1) ° C for (36 ± 4) h with constant mechanical stirring. The nutrient medium is introduced into flasks of various capacities, then the flasks are seeded. The concentration of cells in the initial stage of cultivation should be at least 1 · 10 ⁶ cells · cm ^-3 . Cultivation is carried out at (32 ± 1) ° C in the regime of 150-160 rpm ^-1 for (48 ± 4) hours.

The cultivation of a two-component microbial destructor is carried out by the method of deep cultivation in a fermenter

A glucose solution is introduced into the fermenter with a sterile nutrient medium in an amount of 2/3 of the apparatus volume at the rate of 0.4-0.6% of the total volume, the temperature is brought to (32 ± 1) ° C. The medium in the apparatus is stirred, the pH value is adjusted to a pH of 7.2 ± 0.1 with a solution of NH ₄ OH. After that, seed material of strains is introduced into the apparatus. The sowing dose in the device should be at least 1 · 10 ⁶ cells · cm ^-3 .

Co-cultivation of microbial cultures in the fermenter is carried out at a temperature of (32 ± 1) ° C with a constant supply of air for aeration and rotation of the mixer 340-360 rpm · ^-1 for 35 ± 2 hours. Control of temperature, air flow, vacuum in the cavity the fermenter and the pH value are carried out by rotameter, monovacuum meter, balanced bridge and pH meter. Cultivation is stopped when the desired pH value is reached (7.5 ± 0.5). Separate preparation of culture fluids of each strain is allowed, while using a synthetic nutrient medium that is identical in its component composition for both strain B-6847 and strain Ac-1769. In this case, both separately prepared culture liquids are mixed in a ratio of 1: 1 by volume.

To obtain a higher concentration of hydrocarbon-oxidizing bacteria than on a synthetic medium, cultivation can be carried out in a liquid nutrient medium based on 3-5% casein hydrolyzate under the above conditions.

The culture fluid, consisting of two strains, after a cycle of cultivation in a fermenter must meet the following requirements:

the total content of microbial cells, not less, class · cm ^-3 (2.5-3.5) · 10 ⁹ concentration of hydrogen ions, units pH 7.2 ± 0.2

The culture fluid obtained by culturing two strains of hydrocarbon-oxidizing bacteria, cooled to room temperature, determined by microscopic, colorimetric or other method, the total concentration of bacterial cells.

In the case of separate cultivation of the strains, the culture liquids are mixed in a volume ratio of 1: 1 to obtain the finished product. Then add sterile technical glycerin in an amount of 5-6 vol.% And mix thoroughly.

The shelf life of the finished liquid form of the drug at a temperature of (25 ± 4) ° C is at least 30 days, at (4 ± 2) ° C - at least 3 months.

The finished preparation "CENTRUM-MMS" according to its physicochemical and biological indicators must comply with the following requirements and standards:

Color light cream The concentration of hydrogen ions, units pH 7.2 ± 0.2 The total content of bacterial cells, C. · cm ^-3 no less than (2.5-3.5) · 10 ^{9 ÷ 10} The degree of utilization of petroleum products,% not less than 75

The proposed technical solution is illustrated by the following examples.

Example 1

The preparation of seed in flasks and subsequent cultivation is carried out on an environment of the following composition:

monosubstituted potassium phosphate, g 10.0 potassium phosphate disubstituted, g 4.0 ferrous sulfate, g 0.4 magnesium sulfate, g 0.1 drinking water, dm ³ 1,0

As a carbon source, glucose is used in an amount of not more than 1% for the entire preparation cycle of both seed and culture fluid in the fermenter.

The culture fluid is prepared by a BIOR-01 fermenter, which is a welded cylinder with a steel flat cap. Heating and cooling of the drug is done through the shirt. The apparatus is equipped with a three-tier mixer, a magnetic drive, a bubbler, a device for sampling and introducing additives. The frequency of rotation of the mixer is 0.4-10.0 rev · s ^-1 . In the apparatus, filled into 2/3 of the volume of drinking tap water, the components of the synthetic nutrient medium are sequentially introduced. The medium is stirred, the pH value is set to 6.9. The medium is sterilized at a temperature of (121 ± 2) ° C for 30 ± 1 min. The nutrient medium prepared in the apparatus should be considered a series. Shelf life of the medium is not more than 24 hours.

A sterile culture medium must meet the following requirements:

concentration of hydrogen ions, units pH 6.5 ± 0.2 extraneous microflora absent

The nutrient medium for the preparation of seeds should also meet the same requirements.

The seed material for the fermenter is the culture fluid of two production strains of Pseudomonas fluorescens (VKM B-6847) and Rhodococcus erythropolis (VKM Ac-1769), prepared in flasks, the inoculum dose of each component should be 1 · 10 ⁶ cells · cm ^-3 .

The seed for the apparatus BIOR-01 is prepared in flasks with a capacity of 250 or 750 cm ³ in thermostatic apparatus for shaking flasks.

For cultivation, synthetic culture medium is added to the flasks in an amount of 1/5 of the volume of the flask. The inoculum is a bacterial suspension of these strains grown separately in test tubes on mowed nutrient agar. The sowing dose to the flask is 1 · 10 ⁶ cells · cm ^-3 .

Cultivation in flasks is carried out at a temperature of (32 ± 1) ° C, in the regime of 150 rpm ^-1 for 48 hours.

At the end of the cultivation process, biomass growth is estimated by determining the total number of microbial cells. Received seed should meet the following requirements:

the number of bacterial cells in terms of CFU: for strain Pseudomonas fluorescens (VKM B-6847), C. cm ^-3 , not less (1.5 ± 0.5) · 10 ¹⁰ for strain Rhodococcus erythropolis (VKM Ac-1769), C. cm ^-3 , not less (1.0 ± 0.5) · 10 ¹⁰ concentration of hydrogen ions, units pH 7.1 extraneous microflora absent

Ready seed can be stored at a temperature of (4 ± 2) ° C for up to 10 days. Microbial cultures obtained in one culture cycle in 6-12 flasks make up the batch.

In a fermenter with a capacity of 0.1 m ³ with a sterile nutrient medium in an amount of 2/3 of the volume of the apparatus, a glucose solution is introduced at the rate of 0.4% of the total volume, the temperature is brought to 32 ° C. The nutrient medium in the apparatus is stirred and the pH value is adjusted to a pH of 7.2 ± 0.1 with a solution of NH ₄ OH. After that, seed material of strains is introduced into the apparatus. The sowing dose in the apparatus is 1 · 10 ⁶ cells · cm ^-3 .

Co-cultivation of microbial cultures in a fermenter is carried out at a temperature of (32 ± 1) ° C with a constant supply of air for aeration and rotation of the mixer 340-360 rpm ^-1 for 35 hours. Control of temperature, air flow, vacuum in the fermenter cavity and pH value is carried out by rotameter, manovacuum meter, balanced bridge and pH meter. Cultivation is stopped when the pH reaches 7.5 ± 0.5.

The culture fluid, consisting of two strains, after a cycle of cultivation in a fermenter must meet the following requirements:

total content of microbial cells, cells · cm ^-3 , not less 2.510 ⁹ concentration of hydrogen ions, units pH 7.2

The culture fluid obtained by culturing two strains of hydrocarbon-oxidizing bacteria is cooled to room temperature. Microscopic or colorimetric methods determine the total concentration of bacterial cells. Then in the resulting culture fluid add sterile technical glycerin in an amount of 5 vol.% And mix thoroughly.

The shelf life of the finished liquid form of the drug at a temperature of (25 ± 4.0) ° C should be at least 30 days, at (4 ± 2) ° C - at least 3 months.

Get the biological product of the following composition, wt.%:

consortium of strain Pseudomonas fluorescens VKM B-6847 when the content of bacterial cells is 1.5 · 10 ⁹ cells · cm ^-3 and strain Rhodococcus erythropolis VKM Ac-1769 with cell content 1,0 · 10 ⁹ C. · cm ^-3 95 glycerol 5

Example 2

The preparation of seed in flasks and subsequent cultivation is carried out on an environment of the following composition:

monosubstituted potassium phosphate, g 10.0 potassium phosphate disubstituted, g 4.0 ferrous sulfate, g 0.4 magnesium sulfate, g 0.1 drinking water, dm ³ 1,0

As a carbon source, glucose is used in an amount of not more than 1% for the entire preparation cycle of both inoculum and culture fluid.

The culture fluid is prepared in a BIOR-01 fermenter, which is a welded cylinder with a steel flat cap. The device is heated and cooled through a jacket. The apparatus is equipped with a three-tier mixer, a magnetic drive, a bubbler, a device for sampling and introducing additives. The frequency of rotation of the mixer is 0.4-10.0 rev · s ^-1 . In the apparatus, filled into 2/3 of the volume of drinking tap water, the components of the synthetic nutrient medium are sequentially introduced. The medium is stirred, the pH value is set to 7.0. The medium is sterilized at a temperature of (121 ± 2) ° C for 30 ± 1 min. The nutrient medium prepared in the apparatus should be considered a series. The shelf life of the nutrient medium is not more than 24 hours.

A sterile culture medium must meet the following requirements:

concentration of hydrogen ions, units pH 6.5 ± 0.2 extraneous microflora absent

The nutrient medium for the preparation of seeds should also meet the same requirements.

Cultivation liquids of two production strains of Pseudomonas fluorescens (VKM B-6847) and Rhodococcus erythropolis (VKM Ac-1769) prepared in flasks are used as seed material for the fermenter; the inoculum dose of each component is 1 · 10 ⁶ cells · cm ^-3 _.

The seed for the apparatus BIOR-01 is prepared in flasks with a capacity of 250 or 750 cm ³ , which are then placed in a thermostatically controlled apparatus for shaking the flasks.

For cultivation, synthetic culture medium is added to the flasks in an amount of 1/5 of the volume of the flask. The inoculum is a bacterial suspension of these strains grown separately in test tubes on mowed nutrient agar. The sowing dose to the flask is 1 · 10 ⁶ cells · cm ^-3 .

Microbial cultures of the strains were grown at a temperature of (31 ± 2) ° C for 52 hours at 160 rpm.

At the end of the cultivation process, biomass growth is estimated by determining the total number of microbial cells. The resulting crop should meet the following requirements:

the number of bacterial cells in terms of CFU: for strain Pseudomonas fluorescens (VKM B-6847), C. cm ^-3 , not less 2.010 ¹⁰ for strain Rhodococcus erythropolis (VKM Ac-1769), C. cm ^-3 , not less 1.5 · 10 ¹⁰ concentration of hydrogen ions, units pH 7.3 extraneous microflora absent

Ready seed can be stored at a temperature of (4 ± 2) ° C for up to 10 days. Microbial cultures obtained in one culture cycle in 6-12 flasks make up the batch.

In a fermenter with a capacity of 0.1 m ³ with a sterile nutrient medium in an amount of 2/3 of the volume of the apparatus, a glucose solution is introduced at the rate of 0.6% of the total volume, the temperature is brought to (32 ± 1) ° C. The medium in the apparatus is stirred, the pH value is adjusted to a pH of 7.2 ± 0.1 with a solution of NH ₄ OH. After that, the seed material of the strains is introduced into the apparatus, the seed dose in the apparatus is 1 · 10 ⁶ cells · cm ^-3 .

Co-cultivation of microbial cultures in the fermenter is carried out at a temperature of (32 ± 1) ° C with a constant supply of air for aeration and rotation of the mixer 340-360 rpm · ^-1 for 35 hours. Control of temperature, air flow, vacuum in the cavity of the fermenter and pH value is carried out by rotameter, manovacuum meter, balanced bridge and pH meter. Cultivation is stopped when the pH reaches (7.5 ± 0.5).

The culture fluid, consisting of two strains, after a cycle of cultivation in a fermenter must meet the following requirements:

total content of microbial cells, cells · cm ^-3 , not less 3,510 ¹⁰ concentration of hydrogen ions, units pH 7.4

The culture fluid obtained by culturing two strains of hydrocarbon-oxidizing bacteria is cooled to room temperature. Check the microscopic or colorimetric method for the total concentration of bacterial cells. Then, sterile technical glycerin is added to the resulting culture fluid in an amount of 6 vol.% And mixed thoroughly.

The shelf life of the finished liquid form of the drug at a temperature of (25 ± 4) ° C should be at least 30 days, at (4 ± 2) ° C - at least 3 months.

Get the biological product of the following composition, wt.%:

consortium of strain Pseudomonas fluorescens VKM B-6847 when the content of bacterial cells 2.0 · 10 ¹⁰ cells · cm ^-3 and strain Rhodococcus erythropolis VKM Ac-1769 with a cell content of 1.5 · 10 ¹⁰ cells · cm ^-3 94 glycerol 6

In order to assess the effectiveness of the proposed drug was determined by its hydrocarbon-oxidizing activity.

The specific activity of the drug was evaluated by the degree of utilization of hydrocarbons of crude oil and petroleum products by IR spectroscopy in control and experimental samples.

The amount of fuel and lubricants and oil introduced was 50-100 grams per kilogram of soil. Soil samples were placed in tin trays with dimensions of 30 × 12 × 8 cm, and the proposed preparation and the comparison product Centrin were introduced into them. The concentration of microorganisms in all cases was (5-10) · 10 ⁹ . Ambient temperature (20-24) ° C. Observations were performed on 10, 20, and 30 days. Hydrocarbons in the experimental and control samples were extracted with carbon tetrachloride, then the samples for research were prepared according to the method of sample preparation for IR spectroscopy.

The calculation of the amount of hydrocarbons contained in the control and experimental samples was carried out using the FTIR-8201 software using a pre-built calibration dependence of the content of oil or oil products in the sample on the spectral area. The table shows comparative tests to evaluate the biodegradation activity of the drug "CENTRUM-MMS" and the widely used commercial drug "Centrin".

Efficiency of hydrocarbon biodegradation with the CENTRUM-MMS preparation Names of biological product The degree of degradation of hydrocarbons,% of the initial amount Raw oil Diesel fuel Fuel oil Duration of observation, days 10 twenty thirty 10 twenty thirty 10 twenty thirty "CENTRUM-MMS" 22.4 ± 2.7 62.8 ± 2.7 69.1 ± 8.4 15.3 ± 0.4 45.3 ± 5.1 84.5 ± 7.2 19.3 ± 1.3 46.8 ± 7.5 58.4 ± 5.2 Centrin (control) 6.0 ± 0.8 21.8 ± 3.3 28.2 ± 1.9 11.1 ± 0.3 24.8 ± 3.1 45.0 ± 2.3 4.8 ± 0.2 8.3 ± 0.8 12.5 ± 3.1

According to the data presented, the proposed two-component preparation "CENTRUM-MMS" has a higher biodegradation activity in contrast to the commercial comparison product "Centrin".

In the case of using CENTRUM-MMS, higher rates of biodegradation of oil products were also noted. So, diesel fuel was almost 80% utilized by the end of the observation period, the degree of degradation of oil and fuel oil exceeded the indicators obtained during the processing of Centrin samples. With the introduction of the biological product "CENTRUM-MMS" in contaminated soil, active growth of strains of introduced bacteria occurs, especially bacteria of strain AC-1769. This fact is especially valuable when treating poor sandy soils, which are the natural habitat of bacteria of the Rhodococcus group.

The results of the studies indicate that the developed liquid preparation based on two natural strains has high biodegradation activity and can be used to intensify the cleaning of soils contaminated with petroleum hydrocarbons. Activation of the degradation of petroleum products using the CENTRUM-MMS preparation was accompanied by a decrease in the inhibitory effect of hydrocarbons on groups of sensitive soil microorganisms and, consequently, an increase in the biological activity of soils.

Thus, the proposed biological preparation for environmental purposes "CENTRUM-MMS", which is characterized by:

the component composition represented by two types of microorganisms with a pronounced ability to utilize hydrocarbons when they are combined;

high biodegradation activity in relation to crude oil and various types of hydrocarbon fuels;

activity during utilization of "heavy" as well as aromatic hydrocarbons;

the possibility of application for soil remediation at relatively low ambient temperatures (8-12) ° C, which is especially important for the northern oil producing regions of the Russian Federation.

Claims (1)

Ecobiological product for cleaning oil and oil products based on biomass of a consortium of aerobic bacterial cultures containing microorganisms Rhodococcus erythropolis, characterized in that it contains a strain of Pseudomonas fluorescens BKM B-6847 and a strain of Rhodococcus erythropolis BKM Ac-1769
with a volume ratio of 1: 1, and additionally glycerin in the following ratio, wt.%:
consortium of strain Pseudomonas fluorescens BKM B-6847 when the content of bacterial cells is not less (1.5 ÷ 2.0) · 10 ^{9 ÷ 10} cells · cm ^-3 and strain Rhodococcus erythropolis BKM Ac-1769 with a cell content of (1.0-1.5) · 10 ^{9 ÷ 10} cells · cm ^-3 94-95 glycerol 5-6