RU2114071C1 - Method of biological preparation-assisted cleaning of soil and natural and waste waters polluted by petroleum and petroleum products - Google Patents

Abstract

FIELD: oil pollution removal. SUBSTANCE: new form of biological preparations is provided. They are stored and transported to pollution spot in the form of suspension of biomass with microorganism density 1×10¹¹-5×10¹² col/ml, NaCl concentration 10% and additives of biologically active substances (vitamins) and osmosis protector (betaine). Temperature of suspension is maintained at 10 ± 1 C. Preparations include both separate stains and consortiums of microorganisms. Prior to be used, preparations are diluted. They are characterized by a high vitality of microorganisms. EFFECT: reduced preparation cost and facilitated storage and application. 5 cl, 4 tbl

Description

 The invention relates to the technological use of biological products for cleaning the soil, drive and wastewater from pollution by oil and oil products, and, in particular, a new form of biological products.

 In the course of oil production and subsequent oil refining, rather serious environmental pollution occurs. The annual loss of oil as a result of accidents at oil pipelines in Russia, according to various estimates, ranges from 15 to 50 million tons of crude oil [1]. Annually, accidents occur on the sea routes for transporting oil and oil products, some of which are catastrophic.

 Nature has a certain potential for self-purification, which is based on microbiological processes. However, the speed of these processes is low, the time taken to clean up contaminated territories is estimated for decades. The use of individual strains or consortia of specialized microorganisms can significantly intensify the process of cleaning polluted ecosystems.

 Known biological product "Putidoyl" [2, 3], preparations of the series "Biodestructor" [4], used to clean water and soil from oil pollution.

 The usual form of biological products is dry biomass, dehydrated using freeze-drying or thermal vacuum drying. When dehydrating a biological product, energy costs increase sharply (more than 90% of the prime cost falls on the energy sector, and drying is the most energy-intensive process). In addition, in the final product - a dry powder, the survival rate of microorganism cells in the best case reaches 1 - 10% with strict adherence to the drying regimes, the real result is usually even worse. Thus, with a sharp increase in the cost of the drug during drying, the product is of the worst quality. In practice, this means that when using dry powder instead of a live culture, to obtain the desired effect, it is necessary to spend a larger amount of a more expensive drug.

 A dry preparation requires preparation for work, as microorganisms must be revitalized and activated. In practice, this requires special equipment (for mixing and aeration of the drug) and time. In addition, a specialist microbiologist is required to assess the quality of training.

A living culture of microorganisms is preferable to work, because it has a higher specific activity (1 g of crude biomass - 10 ¹³ living cells, whereas in a powder of 1 g at best 10 ¹⁰ - 10 ¹¹ cells), it does not require preparation and is much cheaper.

 However, the bulk of this biological product is water (99.9%), in addition, it is characterized by a very short time to maintain quality, since microorganisms require air oxygen to maintain viability. In the absence of active aeration, microorganisms are lysed. Biomass can be condensed on the separators, but at the same time microorganisms die from a lack of oxygen even faster.

A known method of treating water and soil contaminated with oil and oil products using biological products, including storing or delivering manufactured dry biological products to a contaminated area or water area, activating dry biological products for use (first preparing a basic suspension of a biological product with a density of at least 2 • 10 ¹³ for microorganisms cells / ml, then the working suspension immediately before use - diluting the base suspension with water 100 times). The introduction of biological products into the cleaned environment is carried out by spraying the working suspension over the contaminated area at positive ambient temperatures [5].

The main disadvantages of this method are:
- for the dry form of the drug - low survival of microorganisms, the drug is quite expensive in terms of the cell of a living microorganism;
- for the liquid form of the drug, the impossibility of long-term storage and difficulties in delivery (transportation);
- the need for preliminary activation of a dry biological product in the preparation of the base suspension.

 The objective of the invention is to obtain a technical result, expressed in increasing the efficiency of cleaning contaminated with oil or oil products of territories and water areas due to the high number of viable microorganisms in biological products, ensuring the storage or delivery of biological products in a form convenient for use, preservation for a sufficiently long time and simplified preparation for use .

The problem is solved in that in the known method, which includes the storage or delivery of manufactured biological products to a contaminated area or water area, the preparation of biological products for use and their introduction into the cleaned environment at positive temperatures, the storage or delivery of manufactured biological products is carried out in the form of a biomass suspension with a density of microorganisms 1 • 10 ¹¹ - 5 • 10 ¹² cells / ml, NaCl concentration of 10% and additives of biologically active substances and osmoprotectant, while maintaining the temperature of the biomass sys in the range (-10) - (- 1) ^o C, and preparation of biological products for use is carried out by diluting the biomass suspension to the required concentration.

 The achievement of the technical result is facilitated by the fact that individual strains or consortia of microorganisms, for example, the consortium of hydrocarbon-oxidizing microorganisms Devoroil (Rhodococcus maris, Rhodococcus erythropolis, Pseudomonas stutzeri, Yarrowia lipolytica), are used as biological preparations.

In one of the preferred embodiments of the method, vitamins A, B ₁ , B ₂ , B ₆ , C, B ₁₂ , D ₂ , D ₃ , E, H, K ₁ , biotin, folic acid, nicotinamide are used as additives of biologically active substances and pantothenic acid, and betaine as an osmoprotector, with the content of each of vitamins B ₁₂ , D ₂ , H, biotin and folic acid 1.5 times, and each of vitamins C and D ₃ twice the content of any other vitamins and betaine mentioned.

 For use, the biomass suspension is diluted, for example, with an aqueous solution of mineral salts of nitrogen and phosphorus in the range from 1000 to 10000 times.

To maintain biomass in an active state, it is necessary to reduce the respiratory activity of microorganisms by physiologically mild methods. This is achieved as follows. 12 hours before the end of the biomass growth process, NaCl in a final concentration of 3.5% is added to the nutrient medium. 6 hours before the end of the cultivation process, the NaCl content in the nutrient medium is raised to 7%, the biomass is separated to a density of microorganisms of 1 • 10 ¹¹ - 5 • 10 ¹² cells / ml. Next, NaCl is added to the resulting biomass to a final concentration of 10%, special additives of biologically active substances (vitamins) and osmoprotector (betaine). The biomass is cooled and stored or delivered to a contaminated area or water area in the temperature range (-10) - (- 1) ^o C in special thermal containers.

 The composition and content of special additives of biologically active substances (vitamins) and osmoprotectant (betaine) in a preservation medium in one example of the implementation of the method are given in table 1.

 Ice crystals do not form inside bacterial cells due to the high salinity of the medium. Respiratory activity is reduced to the maximum extent due to low temperatures and high salinity. A possible osmotic shock is relieved by the presence of an osmoprotector - betaine. The selected composition and content of vitamins allows cells to maintain a minimum level of intracellular metabolism.

 A decrease in the number of hydrocarbon-oxidizing microorganisms depending on temperature, medium composition and storage time can be traced on the basis of the studies (Tables 2, 3, 4).

From the data presented in table. 2-4, it is seen that the best results on the survival of microorganisms were obtained in variants with the addition of NaCl, betaine and vitamins in the temperature range (-10) - (- 1) ^o C.

The method is as follows. Biological products manufactured in accordance with the technology described in the present invention are stored in the form of a suspension of biomass at a temperature of (-10) - (- 1) ^o C in areas of potential contamination by oil or oil products. Storage is carried out for a period specified by regulatory and technical documents. During this period, biological products, if necessary, to clean the territories or water areas from pollution, are diluted with an aqueous solution of mineral salts of nitrogen and phosphorus to the required number of microorganisms in the range of ratios 1: 1000 1: 10000 and are introduced into the cleaned medium by spraying or sprinkling using any machine , mechanisms and assemblies designed for this.

 The proposed form of storage of biological products allows you to centrally produce a certain amount of them and quickly deliver to the place of contamination by any means of transport, suggests a high degree of their readiness for use and ease of preparation in work, which contributes to high efficiency, since it does not require specially trained personnel for this kind of works.

 Implementation of the proposed method allows the use of biological products manufactured with minimal energy costs, since their production does not require drying (more than 90% of the cost of biological products falls on the energy sector, and drying is the most energy-intensive process).

 In addition, in the biopreparation thus prepared, the number of living cells of microorganisms is several orders of magnitude higher than in dry powder. Separation reduces the cost of transporting the biological product, since without it, water accounts for more than 99% of the mass of the bacterial suspension. The storage of biological products in the form of the proposed biomass suspension is greatly simplified, since active aeration of the culture is not required in order to maintain the vital activity of microorganisms.

Sources of information:
1. The magazine. Oil and gas equipment market, M., 1996, N 1, p. 21-23.

 2. USSR author's certificate N 1076446, cl. C 02 F3 / 34, 1982.

 3. USSR author's certificate N 1428809, class C 02 F 3/34, 1988.

 4. The magazine. Oil industry, M., 1995, N 5/6, p. 83-85.

 5. PCT Application, W095 / 31408, CL C 02 F 3/34, 1995.

Claims (5)

1. The method of purification of soil, natural and wastewater contaminated with oil and oil products, using biological products, including the storage or delivery of manufactured biological products to a contaminated area or water area, preparing biological products for use and introducing them into the cleaned environment at positive temperatures, characterized in that that the storage or delivery of biologics manufactured performed as the biomass slurry with a density of microorganisms 1 • 10 ¹¹ - 5 • December ¹⁰ cells / ml, NaCl concentration of 10% and additives bIOL nical osmoprotectant active substances and, while maintaining the biomass slurry temperature in the range -10 ... -1 ^o C, and to the use of the preparation of biological products is performed by diluting the biomass to the desired concentration.  2. The method according to claim 1, characterized in that the individual strains or consortia of microorganisms are used as biological products.  3. The method according to claim 1, characterized in that as a consortium of microorganisms use a consortium of hydrocarbons - oxidizing microorganisms "Devoroil". 4. The method according to claim 1, characterized in that vitamins A, B ₁ , B ₂ , B ₆ , C, B ₁₂ , D ₂ , D ₃ , E, H, K ₁ , biotin are used as additives of biologically active substances , folic acid, nicotinamide and pantothenic acid, and betaine as the osmoprotector, with the content of each of vitamins B ₁₂ , D ₂ , H, biotin and folic acid 1.5 times, and each of vitamins C and D ₃ two times the content of any of the other vitamins and betaine mentioned.  5. The method according to claim 1, characterized in that the biomass suspension is diluted with water, and the necessary concentration of the biomass suspension when diluted with an aqueous solution of mineral salts of nitrogen and phosphorus is set in the range of ratios 1: 1000 - 10000.0

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