EA000660B1 - Biomass reduction and bioremediation processes and products - Google Patents

Abstract

1. A process for bioremediating a media containing hydrocarbon contaminants, said process comprising: A. exposing the hydrocarbon contaminants in said media in-situ to a viable culture of aerobic bacteria obtained by -mixing said waste mass with manure and secondary sewage sludge to form a compost biomass pile, said manure and sewage sludge having sufficient moisture and indigenous bacteria, including aerobic bacteria, to carry out composting of said pile; -allowing said compost pile to autothermally react for an initial time period, said initial time period being terminated upon said compost pile reaching a temperature of from about 46 degree C (115 degrees F) to about 60 degree C (140 degrees F); -adding to and distributing an alkaline material throughout said compost pile to quickly raise the pH of said pile to at least 10; -allowing said compost pile to autothermally react for a second period of time, said second period being terminated upon said compost pile reaching a temperature of at least 82 degree C (180 degrees F); -adding nutrients selected from the group comprising inorganic compounds of nitrogen, phosphorus, sulfur and potassium to said compost pile to support continuing metabolic reactions within said compost pile; and -allowing said compost pile to further autothermally react for a third period of time, while adding water sufficient to maintain a moisture level in said pile of about 60 to about 82 percent throughout said third time period, said third time period terminating upon said biomass degrading to produce a non-pathogenic compost product; -said reaction being carried out during said third period of time in the absence of mechanical aeration of the compost pile or the addition of gaseous oxygen into said compost pile obtaining a viable culture of aerobic bacteria, which may be capable of bioremediating chlorinated hydrocarbons and long chain hydrocarbons, including paraffin and asphalt, in the presence of nutrients and water to bioremediate said hydrocarbons by enzymatically catalyzed hydrolysis of the hydrocarbons; B. assuring the presence of sufficient nutrients and moisture to support bioremediation of said hydrocarbons; C. carrying out an enzymatically catalyzed hydrolysis of said hydrocarbon contaminants; D. periodically extracting a water solution containing bioremediation products from the situs of the in-situ bioremediation process; and E. adding fresh water to said situs in an amount approximately equal to the amount extracted in step D. above; F. said water extracting step D and said water addition step E being repeated in alternating steps until said site is sufficiently bioremediated. 2. A process according to Claim 9. characterized in that said media is a subsurface earth material or said media is located in a pit. 3. A system for in-situ subterranean bioremediation of a horizontally extending stratum in an earthen area (12) contaminated with organic or hydrocarbon material, said system characterized by: A. at least two small diameter (18, 34) wells extending vertically from the surface (14) of said earthen area (12) to points at least immediately below said stratum (16) of earth to be bioremediated; B. each said well (18, 34} having an outer tube (20, 36) enclosing a vertically extending electrode (26, 42) extending to the level of said stratum (16); C. a direct current power source (30); D. means (4, 6) for connecting a first electrode (42) in a first one of said wells (34) to a negative terminal of said power source (30); E. means (32) for connecting a second electrode (26) in a second one of said wells (18) to a positive terminal of said power source (30); F. at least the outer tube (36) of said one well (34) being perforated at the level of said stratum (16); G. means (48, 50) for supplying to said one well (34) sufficient aqueous mixture of bioremediating bacteria to allow a bioremediation reaction to take place; H. said bioremediation reaction being stimulated by said electrodes (26,42), said electrodes being energized by said power source (30): and I. means to activate said power source (30) to provide an electric field between said electrodes (26, 42), whereby said water supported bioremediation reaction is caused to migrate from said first well (34) to said second well (18) within said stratum (16) to progressively bioremediate said stratum in-situ. 4. A system for in-situ subterranean bioremediation in accordance with Claim 3, characterized in that at least said first electrode (42) in said first well (34) is an electrically conducting tube which has perforations (44) at the level of said stratum (16) and wherein at least part of said aqueous mixture is supplied through said conducting tube. 5. A system for in-situ subterranean bioremediation in accordance with Claim 3, characterized by multiple pairs of oppositely charged electrodes (26. 42) located in multiple respective wells (18,34) for driving such aqueous mixtures across differing respective contaminated stratum portions. 6. A system for in-situ subterranean bioremediation in accordance with Claim 3. characterized by a tank (48) at the surface of said site for controlled gravity feed therefrom of a portion of said mixture to said first well (34). 7. A method for in-situ subterranean bioremediation of a horizontally extending stratum (16) in an earthen area (12) contaminated with organic or hydrocarbon material, said method characterized by the steps of: A. introducing a mixture of water and bioremediating bacteria to a first location within said stratum (16); B. subjecting said stratum (16) to a direct current electric field having a positive pole and a negative pole, said positive pole being in the vicinity of said first location within said stratum; and C. allowing a bioremediation reaction to proceed at a reaction site in the vicinity of said first location, said reaction site being caused to migrate across said stratum (16) from said first location to a second location in the vicinity of said negative pole under the influence of said electric field. 8. The method for in-situ subterranean bioremediation of Claim 7, characterized in that said bioremediating bacteria comprise a viable culture of aerobic bacteria according to Claim 5 and said bioremediation reaction occurs principally by enzymatically catalyzed hydrolysis of the organic or hydrocarbon contaminant material. 9. The method for in-situ subterranean bioremediation of Claim 19, characterized in that water is added at said first location over the period of said bioremediation reaction.

Description

The present invention relates to the field of biomass processing or composting methods for obtaining a useful microbiological product, intended either for subsequent biological treatment of various substances, or for direct application to the soil as an agricultural fertilizer. The method includes a thermophilic alkalinephilic reaction with the predominant participation of aerobic microbes, which activates the microbial population in high humidity and high pH at the first stage and the subsequent reaction at high humidity, low pH and adding nutrients that ensure the preserved composting operation in a new form when aerobic microbes decompose or compost waste materials without the usual mixing, turning over or yachivaniya compostable mass to aerate it, as well as without introducing air or oxygen using conventional mechanical devices, in particular, pumps or blowers. The present invention also relates to the field of biological processing of contaminated soils or other industrial wastes and to the production of valuable fatty acids and amino acids.

Environmental pollution is considered one of the most serious problems for government agencies, as well as for private owners of land or real estate and for users. The main problem of all public utilities is the provision of safe sanitary conditions for the disposal of ordinary household waste. Even after the selection of recyclable glass, metal, paper and plastics, there remains a huge amount of mixed harmful and useful materials that are sent to landfills, septic tanks, etc. and that require treatment for various reasons, in particular, to eliminate harmful materials and pathogens, as well as other materials that pose a risk of contamination by leaks or seeps into aquatic systems, soil or groundwater, followed by unwanted movement or migration to other uncontaminated and often inaccessible zone. Biological processing of such polluting products and waste is currently considered as a possible way to solve pollution problems. However, the methods and products used prior to the present invention have never provided ideal parameters in terms of safety, low cost, volume reduction, and ease of achieving the results that natural bacteria produce when biologically recycling ordinary household waste. Contamination of the environment at a landfill or elsewhere is a problem, even if places of such contamination are accessible, in particular, if the contaminants are discharged or drained onto the ground surface, however, the difficulty of accessing the contaminants due to deep underground leakage or penetration of contaminants ground beneath buildings, pavements, driveways, under impermeable rocks, or soil formations exacerbates processing problems. Such pollution caused by seepage is not at all limited to the disposal of household waste and often occurs due to unwanted release and leakage of hazardous materials from refineries, chemical plants, industrial plants that use or get various chemicals and paints as by-products, and are often associated with spills of harmful materials during their transportation. Although federal, state, and municipal laws and decrees attempt to regulate environmental pollution, there are thousands of places where pollution has been the result of an accident or mistake, or occurred in the past when environmental problems were not recognized as much as they exist today, when environmental protection has become both a public and private matter.

Apparently, one of the easiest ways to treat industrial waste is their usual stacking in heaps and composting, in which chemical decomposition of the waste occurs under the action of natural bacteria. Well known are such processes where aerobic thermophilic reactions take place with occasional overturning or stirring of the mass in order to aerate it, providing oxygen to support the aerobic process. Other known methods of aeration include maintaining the porosity of the mass by introducing wood chips and forcing air or oxygen into the compostable mass through perforated pipes or supporting grids. The availability of mechanical equipment and operators for management and maintenance is a significant factor in the cost of such composting operations. If waste composting areas are limited or absent, removal of such products is a major problem.

Another waste disposal problem arises in connection with sewage sludge, which creates both physiological and physical problems of its use and / or disposal in safe for health conditions.

To further substantiate the present invention in this description, the following articles are cited for reference:

Abelson, Reed, Bugs clean their act, p. 144, Forbes, Sept. 28, 1992;

Bouwer, E.J. Bioremediation of Organic contaminants in the Subsurface in the Subsurface, Chapter 11, pp. 287-318 of Enviromental Microbiology Edited by Ralph Mitchel, Wiley-Liss 1992;

Feinstein, Melvin S., Composting in the Context of Municipal Solid Waste Management, Chapter 14, pp. 355-374 of Environmental Microbiology Edited by Ralph Mitchel, Wiley-Liss 1992;

Gillis, Anna Maria, Shrinking the trash heap, p. 90 (4 pp.), BioScience, V. 42, N. 2, Feb., 1992;

Glass, David J., Waste Management, p. 5 (4 pp.), Environment, V. 33, N. 9, Nov. 1991;

Madsen, E.L, Sinclair, J.L, Ghiorse, Wm.C. In Situ Biodegradation: Microbiological Patterns in Contaminated Aquifer Stover, Dawn, TOXIC AVENGERS p. 70, (6 pp.) Popular Science, July, 1992.

The advantages of composting using household sludge mixed with waste are described in the Gillis article cited. This article indicates that, despite significant improvements in the cultivation of various plants and vegetables, it may be necessary to evaluate the use of basic products of composting, taking into account the type of plants grown, irrigation and geographical regions to determine the optimal rate of compost material to the soil surface. It is said that improved methods similar to those described in the present invention can overcome the problems associated with the safe disposal of harmful materials that may be prohibited by a regulation of the Environmental Protection Agency (EPA) or other federal, state or local laws. or regulations that may limit the disposal of hazardous waste in landfills or in composting areas.

Although it is known that natural organisms can perform biological processing of, for example, household waste landfills where composting occurs under aerobic conditions, natural organisms are used in the present invention, however, the processing operations proposed in the invention occur when reacting with enzyme catalysis, which not only reduces the volume of overturned compaction mass for aeration of the heap or the need for an expensive injection process into the air or oxygen mass, but also provides a very fast yōkan reaction in which the composting operation may be nearly completed during a period somewhat more than one month, usually inaccessible to conventional composting methods.

The present invention, both in terms of processing technology and the use of products from the main composting operation, is useful for solving many of the problems described in the above articles, including (but not only) decomposition and disposal of solid household waste, biological processing of hazardous materials, safe decomposition and disposal waste in production areas, biological processing of surface soil contamination, biological processing of subsurface soil and liquids, decomposition of carbon compounds with shorter chain carbon chains, cheap production of useful amino acids and fatty acids as biological processing by-products, and non-pathogenic fertilizer as a by-product of composting with increased nutrient retention for plants in combination with increased moisture retention upon application into the soil for agrotechnical purposes. If the scope of a biological processing project is small enough, practical and convenient is the recovery in barrel-shaped containers, in particular to transform the processed materials into more useful products in the industrial field.

Another important aspect of the present invention relates to improved methods for processing subsoil pollution. Microbiological products obtained according to the invention are used for biological processing of contaminants at their location, and the use of direct electric current causes bacteria to pass through the soil, which can be clay or other soils that have poor permeability under normal conditions. In the process of applying electrical voltage add nutrients.

The characteristics of the invention and its technical advantages are evident from the following description of a preferred embodiment of the invention, together with the claims and the accompanying drawings, which show:

in fig. 1 to 4 are diagrams of test results of the effect of bacterial strains according to the present invention on trichlorethylene (TCE);

in fig. 5 - two electrically interconnected underground wells connected to a direct current source and a bacteria and nutrient supply tank;

in fig. 6 is an enlarged vertical section of the negatively charged well shown in FIG. five;

in fig. 7a is a layout diagram of several electrically connected subsurface wells shown in FIG. 5 and located in the center and around the circumference of the zone of the subsoil biological processing;

in fig. 7b is a diagram similar to FIG. 7a, with the location of the wells on opposite sides of the rectangle;

in fig. 8 is a vertical section of a tank used for biological processing of hazardous waste of toxaphen;

in fig. 9 is a schematic sectional view of a pit containing soil contaminated with a hydrocarbon, with a central tube for extracting biological processing products;

in fig. 10 is a schematic section of a biological reactor for processing industrial waste or for obtaining useful products by bioconversion.

Without being tied to the theory outlined below, explaining why the methods and products described here proceed in unexpected ways and give unexpected results, it is assumed that the initial stage of activating the bacterial population at high temperature, high pH and high humidity in the compost heap leads in about two days to the formation of a practically non-pathogenic population of bacteria capable of continuing reproduction in this heap and at the same time decomposing wastes within one to two months after the initial and activation. During this decomposition period, nutrients and water are added to the mass to maintain the reaction with enzyme catalysis at the interface of bacteria, water, nutrients and waste substrates so that the oxygen required for the process, which is considered aerobic, is obtained from water that is continuously added. in mass. Since this elemental or molecular oxygen in the composting process is not fed into the mass by aeration or inflation, aerobic bacteria in the heap act in an anaerobic environment. Oxygen in this case is not initially supplied in gaseous form, and it is formed in sufficient quantity at the active phase boundary, where the hydrolysis of waste occurs in the presence of water due to the reaction of the aerobic type. Water is also regenerated in the compost heap due to the decomposition of organic compounds whose carbon interacts with oxygen to form carbon dioxide, and hydrogen reacts with oxygen and forms water. Decomposition of harmful hydrocarbon can be performed as part of a composting operation using the present method, or, in the case of a specific substrate material, by acting on it in a similar way using bacteria that are formed during the composting process. The use of bacteria resulting from composting is preferable to ensure a high degree of metabolic activity supported by the formation of oxygen with enzymatic catalysis in order to accelerate any process of biological processing. The method of obtaining these bacteria formed during composting, if desired, can be repeated using readily available waste, collection sludge, horse manure, water and protozoa bacteria in the combinations indicated in this description.

Although bacterium of the bacillus genus is one of the most studied microbial species, the present invention has demonstrated the unique ability of strains of this genus, which are the final product of the present invention, not only for extreme fecundity and rapid colonization (see FIG. 8), but also the properties of their enzymes have a catalytic effect on the reaction involving water, nutrients and contaminated substrates, providing biomass decomposition and biological processing that can turn t pichnye waste in fruitful and beneficial ingredients, including the chemical by-products or agricultural fertilizers. In addition, the finished product is also capable of exerting an enzymatic catalytic effect on the biological processing of chlorinated hydrocarbons and long-chain hydrocarbons, especially those contained in petroleum. The method according to the present invention provides a biological processing of materials on site, in a manner that is not possible with known analogs. These methods have great economic dignity, since they do not require the usual plowing or cultivation of huge bands of waste characteristic of typical composting operations. The invention also provides biological processing of deep and inaccessible contaminated soil layers without excavating or removing soil to gain access to contaminants, as is the case in some enterprises using external reactor tanks for processing toxic chemical compounds in the presence of microorganisms that react with these compounds.

The present invention provides a low-cost method for producing large amounts of enzymes that are an enzyme catalyst for biological waste treatment and hydrocarbon materials that do not require the use of mechanical equipment for supplying air or oxygen, usually used to maintain aerobic reactions of biological processing.

The unique property of microbes and enzymes according to the invention is that the biological processing of contaminated materials occurs with the use of bacteria, which are usually considered aerobic, however, these aerobic bacteria act in an environment that is usually considered anaerobic. Thus, air or oxygen is not supplied to the reaction, while nutrients, contaminated materials and water interact with microbes, and the enzyme catalysis of the reaction ensures efficient biological processing of contaminated substances. For the case when the nutrient contains oxygen, such as, for example, nitrates ΝΘ ₃ , it should be noted that this method is assumed to be independent of any oxygen in the specified nutrient.

The percentage of strains of aerobic and anaerobic types of bacteria in the compostable product according to the invention is shown below in table. one.

Table 1

The percentage of strain types in the compost sample

Strain Compost Aerobic one ten% 2 50% 3 ten% four ten% five 20% Anaerobic 6 40% 7 40% eight 20%

The ability of aerobic bacteria, producing compost according to the present invention, to produce biological processing in an anaerobic environment is confirmed by the results of laboratory studies presented below in table. 2, and FIG. 1-4, which show that aerobic strains grow better in trichlorethylene (TCE) under anaerobic conditions and are practically inhibited in TCE under aerobic conditions.

table 2

Growth of aerobic strains in the presence of TCA under aerobic and anaerobic conditions

Strain Aerobic 24 hours Aerobic 48 h Anaerobic 48 h one Suppressed Suppressed Average 2 Suppressed Suppressed Suppressed 3 Suppressed Suppressed Good four Suppressed Suppressed Good five Suppressed Suppressed Average 6 Average Suppressed Suppressed 7 Suppressed Suppressed Average eight Suppressed Suppressed Minimum

The present invention is capable of obtaining a biological processing product in the areas where common household waste is located, as well as material that is useful either as a fertilizer or for biological processing of other materials or products, and has a long shelf life during storage. Tests that were carried out with the use of such a product obtained as a result of composting operations showed a shelf life of more than one year.

Composting method

In a preferred embodiment of the invention, which is initially used for composting household waste, the initial biomass contains mixed garbage, horse manure and secondary sewage sludge. The sludge is preferably free from oil contamination and contains about 18 percent of the solid phase. Quicklime or fly ash is added to raise the pH to 1 2 or more and to achieve a rapid autocatalytic temperature rise in a few seconds to about 93 ° C (200 ° F). Water is added to maintain the solid phase in the compost heap at about 18%. In this way, a moisture content of about 82% is maintained, which is sufficient saturation to cause aerobic bacteria to react in an almost anaerobic environment. High thermal and alkaline conditions contribute to the activation of bacteria, therefore, after 24-36 hours, the bacteria are brought to the active state, i.e. The bacteria population is stable and is ready for the subsequent addition of nutrients to the compost heap, which cools the heap and prepares it for one to two months to complete the biomass recovery and biological decomposition of all organic materials in the compost heap.

After the first addition of nutrients, the compost heap can be aged for 7-10 days with only pure water added to maintain moisture levels. After that, a bunch of turn so that the outer areas were inside. Then the nutrients are added again, and then the heap can be kept for two weeks. After that, a bunch of overturned once so that the outer areas were inside. Then the nutrients are added again, and then the heap can be kept for another two weeks. After that, a bunch of turn over for the third time and again add nutrients. Then, reactions with enzymatic catalysis continue in the compost heap, the completion signal of which is a gradual decrease in temperature to 27-32 ° C (80-90 ° F) at the end of the decomposition process. This temperature is one of the indicators of the end of the process. Another indicator is the presence of finely granulated or sludge consistency of the obtained organic product.

It should also be noted that high values of temperature and pH during the first two days of activation of bacteria effectively destroy pathogens in the compost heap.

In general, the preferred embodiment of the composting method is implemented according to the following example.

Example 1. A compost mixture consisting of garbage, horse manure, secondary sewage sludge, cellulose, carbohydrates and other typical household organic wastes was collected into a critical mass, i.e. 75 cubic yards (about 57.3 m ³ ) with an initial pH of 7 or 8, to ensure the onset of an autocatalytic exothermic reaction due to the activity of microbes in the presence of moisture. Such a compost heap usually contains natural (endogenous) microorganisms involved in the aerobic thermophilic and alkaline-phase reaction, which is a fundamental aspect of the present method.

The mixture contains a sufficient amount of organic and inorganic nutrients, including inorganic compounds of nitrogen, phosphorus, sulfur and potassium, necessary to maintain the reaction, which proceeds with increasing temperature. After reaching a temperature of 46-60 ° C (115-140 ° F) after about 10 hours, the activity of thermophilic alkaline-acidic microorganisms increases or accelerates due to additional mixing with the basic or alkaline material, in particular, with CO, CaO (lime), NaO, MgO or fly ash, in sufficient quantity to raise the pH of the initial mixture to about 12 or higher. It is advisable to wait for the temperature to rise before adding alkaline material, as otherwise there is a danger of destroying the beneficial microbes.

After adding an alkaline material that raises the pH, the exothermic reaction continues with a significant increase in temperature. In addition to aerobic organisms that require molecular oxygen, additional organisms are forcibly becoming aerobic. Reactions are maintained by dissolved oxygen, which is contained in sufficient quantities (ie, at least 1 part per million) to ensure the survival of aerobic organisms. The combination of high pH and temperature is supposed to destroy pathogenic bacteria, while the primary aerobic thermophilic microbial strains, including aerobic bacteria of the genus Bacillus, and their high-energy enzymes continue to be reproduced or produced. It is assumed that these remaining living microorganisms and enzymes pass through an evolutionary process, as a result of which the final composition of living bacteria and enzymes is formed and useful products such as amino acids and fatty acids are produced.

The primary enzymes that are produced during the decomposition process are lipase and protease, but lignase is also produced. The exact nature of the intended mutation or recombination has not been established.

Raising the temperature of a new mixture after changing the pH can reach the boiling point of water. However, it has been found that beneficial surviving microorganisms can withstand significantly higher temperatures in the presence of some moisture.

In addition to maintaining high temperatures, surviving microorganisms have been found to tolerate temperatures close to the freezing point of water.

The process continues for a sufficiently long time, approximately 24 hours, in order to ensure that the compost pile reaches the desired recycled, uncontaminated state. After adding nutrients, ammonium sulphate 21-0-0, phosphate 0-45-0 and potassium sulphate 0-0-60 in equal quantities by weight, the temperature decreases, as well as the pH value to a level of 7-7.5, close to neutral. Microorganisms can continue to multiply in the presence of moisture and nutrients, while the final product includes high-energy enzymes with an increased ability to break down for the biological purification of soil from hydrocarbon contaminants to produce harmless water and CO2.

Throughout the process described, waste materials and additives remain in a solid state, which facilitates work with these materials.

Metal ions in the reaction mixture do not interfere with the formation of useful enzymes that can be isolated as useful products for the biological purification of hydrocarbons and for the regeneration of biologically contaminated activated carbon used in other processes.

In a similar alternative implementation, garbage and horse manure can be added to the compost mixture after the activation period ends. It is preferable to first add horse manure, since in this case the bacteria are exposed to high temperature and high pH before adding garbage that has a high paper content and a tendency to a rapid decrease in pH.

In both of these embodiments, the compost heaps are continuously sprayed with water to maintain high humidity. Similarly, in both embodiments of the invention, nutrients are added approximately 48 hours after the start of the activation period with a high moisture content in the compost heap before the end of the composting.

For these methods, as well as for the others described here, a gradual decrease in temperature to about 27-32 ° C (80-90 ° F) at the end of the decomposition process is typical. This temperature is one of the indicators of the end of the process. Another indicator is the presence of finely granulated or silty consistency of the resulting organic product.

Although the molecular structure of microbes and their enzymes resulting from individual processing operations according to the present invention is not precisely known, it has been established that these microbes and their enzymes are exposed to extreme temperatures and pH values during the passage of the aerobic metabolic process. At least, in the definition of aerobic strains and the short-term repetition of about 24 hours, microbiological products are subject to a very high load, which, as expected, only high-thermophilic microbes can withstand. It is also assumed that pathogenic microbes are destroyed or permanently change their natural properties, and those thermophilic microbes that can change their properties at temperatures of the order of 100 ° C can restore them during cooling, which occurs approximately 48 hours after the addition of nutrients. After the addition of nutrients, not only temperature decreases, but also pH values from about 12-14 to about 7-8.

Product Composting Analysis Figure 3 shows the results of laboratory analysis — a total plate count for determining the types of bacteria in the compost material to identify strains.

Table 3: Bacteria in the composting product

Name strain Primary identification GCFAME Coeff. similarities Coeff. differences Primary identification of Biolog TM Type of plates Coeff. similarities Coeff. differences 3238-1 Bacillus circulans CLIN 0.489 3,864 closest view of Bacillus insolitus GP 0.921 0.962 3238-2 Bacillus coagulans 0.143 6.376 Bacillus pasteurii GP 0.791 2,976 3238-3 Bacillus latersporus 0.530 3,642 Bacillus insolitus GP 0,900 1,260 3238-4 Staphylococcus aureus 0,607 3.232 Bacillus pasteurii GP 0.440 6,126 3238-5 Micrococcus varians 0.350 6,684 closest view of Bacillus pasteurii GP 0.169 11,916 AN 3238-6 Prevotella veroralis 0.001 10,900 AN 3238-7 Rothia denticariosa 0.016 8.516 AN 3238-8 Staphylococcus epidermidis 0,005 9,753

These results were obtained using the standard method for counting bacterial plates for isolating strains. Five aerobic and three anaerobic strains were found in a sample of a composting product. After isolation, the strains were individually placed on TSA. After incubation for 24 hours, TSA plates were processed according to [Method 1 of GS-FAME] and using clinical aerobic (CLIN [rev.3.80]) or anaerobic databases [ANER1] GCFAME. Then, a suspension of aerobic strains was prepared in sterile saline and loaded to prepare the Bilog (TM) assay in appropriate microtiter plates (gram-negative and gram-positive). The plates were incubated for 24 hours and then analyzed according to version 3.5 of the Biolog database (TM) using an automatic microplate reader. The coefficients of similarity and differences for each strain are presented in table. 3 indicate the similarity and difference with respect to the hypothetical average organism in the database. An organism in a database has a coefficient of similarity equal to one, and a coefficient of difference equal to zero. The closer the corresponding coefficients of the strain to one and zero, the more it approaches the average organism in the database. A good match occurs when the similarity coefficient is greater than 0.5 and the difference coefficient is less than 7.

In tab. 4 shows the growth rates of aerobic and anaerobic bacteria according to the invention.

Table 4

Total heterotro count ( ) tablets 24 h 48 h Types Aerobic compost 5.06 x 10 ⁵ 8.15x10 ⁹ five Anaerobic compost 7.20x10 ⁴ 1.18x10 ⁵ 3

Biological processing and fertilizers

The secondary aspect of this method is mixing a concentrated portion of solid material obtained from the process described above and containing preserved living or active organisms with water and using a liquid mixture for inoculation and biological treatment of soil contaminated with hydrocarbon compounds (for example, when fuel is spilled or leaked tanks) and chlorinated hydrocarbons (pesticides, polychlorinated diphenyl, etc.).

Fertilizers

The present methods of composting and biological processing combine about 50% of microbiological metabolic processes and about 50% of synthetic or enzymatic processes, especially extracellular processes. If the method according to a preferred embodiment of the present invention is used to obtain a by-product containing gypsum for use as an agricultural fertilizer, then nutrients such as ammonium sulphate 21-0-0, ternary superphosphate 0-45-0 and potassium 0- are used. 0-60 in equal amounts. These additives reduce the pH to neutral and ensure the formation of gypsum (CaSO ₄ 2H ₂ O) as the reaction proceeds. Gypsum is a valuable fertilizer that persists in the soil for much longer than other fertilizers, and also helps retain moisture in the soil. Depending on the gypsum requirements for specific types of crops and soil, the percentage of gypsum in the final composting product may vary in the range of about 3 to 12% due to changes in the amount of added nutrients. At the same time, phosphogypsum will also be present in a significant amount, and its relative content can be changed depending on the relative amount of the applied ternary superphosphate (0-45-0).

Biological processing

As indicated above, the final composting product obtained by the preferred composting method, instead of being used as an organic fertilizer, can be used for the biological processing of various organic and hydrocarbon materials. The production and isolation of useful products such as amino acids and fatty acids gives a significant economic effect due to these methods.

An important additional feature of the present invention is the universality of bacteria appearing in the first composting operation. This means that they can be used for biological processing of many different hazardous or polluting materials and, therefore, can provide greater opportunities for general application of this method without the need for extensive analytical preparation to determine the location of contaminants, such as bacteria, necessary for the biological processing of contaminants, and also make it possible to convert most of the impurities processed by the products according to the present invention into such p Leznov or innocuous materials such as water and carbon dioxide, as well as potentially useful acid.

If the final product of composting in accordance with a preferred embodiment of the invention is to be used as an agent for the subsequent biological processing of organic or hydrocarbon materials, urea (46-0-0), ammonium nitrate (32-0-0) are the preferred nutrients for this method of subsequent processing. , phosphate (0-450) and potash (0-0-30), and the mass ratio of nitrogen to the amount of phosphorus and potassium is 10 to 1. This method of subsequent processing can be of the type described gave her and the illustrated fig. 5-7 and 9-10. For biological processing of the soil at its location, the moisture content can be reduced to about 20 wt.% If the necessary conditions are met for the biological reaction to take place with enzyme catalysis.

Biological processing of a mixture of crude paraffin oil and water at the site in an open pit

Another embodiment of the invention is the biological processing of crude paraffin oil in a mixture with other compost materials. Such paraffinic raw materials are usually found in a pit of earth and mixed with water. As a first operation, secondary cooking sludge containing about 3-5% by weight of moisture is mixed with horse manure in a compost heap, using the sludge as a wetting agent. After mixing, the total moisture content is reduced by about 50% by weight. Then all the garbage is added to the pit, mainly paper products, in particular, magazines, cardboard boxes, brochures and newspapers, to absorb raw oil and water to form an oily trash mixture that has an average carbon content of 30% (300,000 million). h) the total content of petroleum hydrocarbons (SNU) according to infrared spectrometric analysis 418.1. Then both mixtures are mixed together to obtain a mixture with a jelly-like consistency. About ten hours after the start of the biological reaction, the temperature of the resulting mixture rises to about 54 ° C (130 ° F). After 48 hours, the temperature reaches about 71 ° C (160 ° F).

The number of ingredients in the mixture is 110 cubic yards (about 84 m ³ ) of horse manure, about 2000 gallons (about 7.6 m ³ ) of sludge, 50 cubic yards (about 38 m ³ ) of garbage and 25 cubic yards (about 19 m ³ ) crude paraffin oil.

After 10 days, the oily consistency disappears and the SNU drops to about 5,000 ppm. At this time, the compost pile can be flipped so that the outer layers are inside the pile. With the same high moisture content of at least 60-75%, the entire mixture is biologically decomposed, while the hydrocarbons are subjected to biological processing due to constant microbiological exposure with enzyme catalysis. After 30 days, the temperature of the compost pile is set to about 54 ° C (130 ° F), and the pile is turned over again so that the outer layers are inside again for better effect. At this time, SNU is reduced to 2000-3000 parts per million. After 60-90 days, the value of SNU becomes less than 100 million hours, which is an acceptable level of satisfactory processing of hydrocarbons.

Biological processing of subsoil layers at their location

Another embodiment of the invention relates to the use of an aerobic microbiological product for the biological treatment of subsurface or subsurface pollution, such as spills or leaks near buried tanks, commonly used to store gasoline and other liquid products that are considered harmful pollutants because of their ability to migrate through soil or other soil materials, mix with groundwater and pollute them, making them unsuitable for drinking or, necks least, less useful. In the United States, there are many places where biological treatment is required before the migration of contaminant fluid goes too far. In many of these places, a number of small diameter pilot wells have already been made according to a previously developed scheme for taking control samples from each well in order to periodically monitor and determine the extent and change of contaminants. Contaminated areas in such places are often difficult to access because they are at considerable depths, perhaps measured in hundreds of feet, underground, or because they are under pavements, buildings, or other structures that cannot be moved for various reasons.

Despite the fact that electrodes with applied voltage were previously used in similar wells for sampling, the present invention assumes that the guaranteed biological processing in such zones is successfully and expediently implemented in the presence of suitable microbes and enzymes with essential nutrients and sufficient water to ensure aerobic reactions of a microbiological product with nutrients and subsoil pollution that need to be turned into useful effects. ucts.

As shown in FIG. 5-7, this embodiment of the invention involves the use of at least two separate vertically located wells or drilled small-diameter bore holes extending to a depth at least equal to the depth of the lowest level of the contaminated subsoil layer.

According to FIG. 5, the biological soil treatment system 10 is installed in the ground 1 2 having a surface 1 4 and a contaminated zone 1 6 located at a depth of about 5 to 10 feet (about 1.5 to 3.0 m) from the surface 1 4. Positive charged well 18 is located vertically, passing from the surface 1 4 down through the contaminated zone 1 6 in this case to a depth of about 1 0 feet (approximately 3.0 m). The liner 20 passes along the entire length of the well 18, tightly adjacent to the inner surface of the hole, and has an end cap at the lower embedded end 22. The liner 20 has holes 24 located in its lower part within the contaminated zone 16, i.e. lower five feet (about 1.5 m). The liner 20 may be made of a polyvinyl chloride (PVC) tube. The holes 24 have a diameter of about 1/4 inch (about 6.3 mm) and pass through the wall of the liner 20. The perforated portion of the PVC tube may be a conventional indicator PVC tube well with many small arcuate, extending to the periphery of the slots, passing along the length of the perforated tube . The positively charged coaxial electrode 26 is placed inside the well 18 concentrically with the liner 20 and extends almost throughout its depth, while the lower part of the electrode 5 feet long (about 1.5 m) communicates with the contaminated soil section 1 6 of the soil through the holes 24 in the liner 20. The coaxial electrode 26 is made of a standard copper tube with a diameter of 3/4 to 1 inch (about 2.0 to 2.5 cm) and is perforated with holes 28 having a diameter of about 1/8 inch (about 0.3 cm) with an interval of 3 inches (about 7.6 cm) on the length of the electrode 26, corresponding to the performance The injected area of the liner 20, preferably at least 2 feet (about 61 cm) of the perforated section of the liner 20. Positive charge is applied to the electrode 26 by connecting to a DC electric generator 30 using a conductor 32.

The negatively charged well 34 is located at some distance from the positively charged well 18 and is preferably located in that part of the contaminated zone 16 that is remote from the positively charged well 18. The negatively charged well 34 may have a configuration identical to that of the positively charged well 18. The well 34 contains an insert 36 with an end cap 38, the lower part of which has holes 40 on it. The negatively charged concentric electrode 42 may be identical in configuration the positively charged electrode 26 and is located inside the well 34, passing through almost its entire depth, while the lower part of the electrode 42 with a length of 5 feet (about 1.5 m) communicates with the contaminated soil section 1 6 through the holes 40 in the liner 36. Electrode 42 made of a standard copper tube similar to electrode 26 and perforated with holes 44 arranged similarly to holes 28 in electrode 26. Negative charge is applied to electrode 42 by connecting to an electric DC generator 30 using conductor 46. The upper part of the negatively charged electrode 42 is connected to the tank 48 by means of a hose 50, through which liquid flows from the tank 48 to the electrode 42.

FIG. 6 shows the negatively charged well 34, the negatively charged electrode 42 and its connection to the hose 50 on an enlarged scale.

FIG. 7a is a top view of a typical well pattern of FIG. 5 for the case of a circular contaminated zone 16, while the negatively charged well 34 is located in the center of the contaminated zone 1 6, and several positively charged electrodes are placed at regular intervals near the perimeter of the contaminated zone 1 6 and are at the same distance from the central negatively charged well 18. Although relative charges can be reversed; it is most convenient to supply liquid from tank 48 (see FIG. 5) to one negative electrode. For the case shown in the figure, six pairs of wells are formed by six positively charged wells 1 8 and one negatively charged well 34.

FIG. 7b is a top view of a typical well pattern of FIG. 5 for the case of a rectangular contamination zone 1 6, with several negatively charged wells 34 placed along one side of the contaminated zone 1 6, and the corresponding number of positively charged wells 1 8 are located along the opposite side of the contaminated zone 1 6. For the case shown in the figure, three pairs of wells they are formed by three positively charged wells 1 8 and three negatively charged wells 34.

When performing the method, nutrients, microbes and water for maintaining the subsoil processing flow by gravity through the hose 50 from the tank 48 located on the surface 1 4, in selectively controlled quantities. Enough water is added to the wells to ensure the migration of bacteria into the soil material, if the contamination is not below the groundwater level. Water containing several ounces of aerobic bacteria and nutrients, including nitrates or urea and phosphates, flows by gravity into the well through a hollow negative electrode perforated with holes 44. Voltage is applied to the electrodes for several hours to stimulate or activate microbes and move the microbes and enzymes horizontally. and nutrients through the contaminated zone for their interaction with nutrients in order to carry out hydrolysis with enzyme catalysis of the substrate in the presence of water to provide enough oxygen to continue aerobic biological processing of pollutants even after disconnecting the power supply from the welding generator.

The presence of nutrient and microbial migration was demonstrated when the electrodes were separated by 50 feet (about 1–5 m) after four hours of applying voltage from a DC generator by reducing the nitrate content from 1 45 ppm to 5–8 ppm. negative electrode, while near the positive electrode, the nitrate content increased from less than one part per million to about 1 5 million hours. There was also a decrease in VTEX pollutants. Additional nutrients in an aqueous solution can be periodically supplied to a negative well or to both wells to maintain aerobic biological processing of pollutants. As in other embodiments of the present invention, hydrolysis of a substrate with enzymatic catalysis in the presence of moisture and microbiological effects on nutrients can continue under aerobic conditions without aeration or oxygen supply to the contaminated mass in any other way. Such hydrolytic reactions in the presence of water continue for a long time after cessation of the supply of electric voltage to water from the electrodes.

In another experiment, when removing electrodes about 430 feet (about 131 m), 80 gallo19 (about 303 l) of a solution with a high nutrient concentration, was fed through a negative electrode to a contaminated soil mass with nutrient deficiency at a depth of 7 to 12 feet ( from about 2.1 to 3.7 m), while the negative electrode was shielded at a depth of the first 7 feet (about 2.1 m) and opened in the zone of the contaminated layer 5 feet high (about 1.5 m) by perforation in PVC the tube.

Between the two vertical electrodes of opposite polarity, the DC electric field, as can be seen from the above, would have the shape of an oval or a ball for American football. When using multiple electrodes, these forms can be applied according to any desired pattern. One or more negative electrodes may be located in the center of the circular zone, as shown in FIG. 7a, either along one side or in the form of a grid, as shown in FIG. 7b to indicate a horizontally passing zone containing one or more fields of direct electric current.

Satisfactory tests were carried out using a Lincoln 200 DC welding generator having a 70 hp motor. and designed for a constant current of 200 A at a voltage of 40 V for operation as a voltage source. During the experiments, the generator was set to 110 A, although the current in the network during the experiments was approximately from 3 to 5 A. Biological processing of liquid waste in a container

In addition to the on-site biological processing method described above, when residual decomposed components can be left at their location, it is possible to apply this invention to the treatment of contaminants with their removal, i.e. placing a certain amount of liquid waste or contaminated solid material or soil in a container for biological treatment. The following categories are considered suitable for biological processing:

I. Petroleum

A. Asphalt

B. Paraffin

C. Pitch sand

D. Creosote

Ii. Chlorinated compounds

A. Trichlorethylene (THE)

B. Perchlorethylene (PCE)

C. Methylene chloride

D. All other chlorinated solvents

E. Pesticides

1. DDT

2. DDD

3. DDE

4. Toxaphen

5. Dieldrin

6. Chlordane

7. Dichloroethane

F. Phenolic compounds

G. Polychlorinated diphenyl

H. Herbicides

Iii. Glycols

A. Ethylene

B. Diethylene

C. Trimethylene

Iv. Amines

A. Ethanolamine

B. Alkalinamin

V. Paints

A. Enamel

B. Epoxy

C. Urethane

D. Xylene

E. Imronovye

Vi. Automotive fluids

A. Transmission Fluids

B. Brake Fluids

C. Engine oils

VII. Sediments in the tanks

A. Crude Oil Tanks

B. Diesel oil tanks

Viii. Refining slime

A. Styrene

B. Paraffin

C. Urethane

D. Asphalt

E. Anthracene

F. Pirin

G. Coal resins

Most, if not all, of these substances can be processed in containers of various sizes depending on the amount of material to be recycled.

Example 2. The method of such processing in accordance with one embodiment of the invention can be carried out in an inert plastic drum with a volume of 55 gallons (about 208 liters), made of PVC material (see Fig. 1 0) and intended for processing liquid substrates, in particular hydrocarbons. The drum is placed on the base and filled with sand, leaving about 10-12 inches (about 25-31 cm) free to the top. About 5-15 gallons (about 19-57 liters) of the liquid substrate are mixed with sand to cover the surface of the sand particles. Clean water is added to the drum to a level that exceeds a few inches of the mixture of sand and waste, as well as a small amount of seeding aerobic microbes and some nutrients. Hydrolysis of the substrate with enzymatic catalysis in the presence of water and the effect of microbes on nutrients occur under aerobic conditions without the need for aeration or the supply of oxygen to the mixture in any other way. The color of water at the top of the drum is a visual indicator that allows you to set the end of the process of biological processing. When the color of the water turns dark, indicating the end of this process, the liquid is removed from the drum, leaving about 6 inches (about 16 cm) at the bottom as a microbial culture for additional processing. Recycled waste can be removed through a drain valve located in the periphery of the lower part of the drum and having a manual or remote control. Separate drainage valves can be used to, on the one hand, maintain the substrate in the container at 6 inches (about 16 cm) for microbial culture, and, on the other hand, ensure the complete removal of the recycled substrate. After the end of the discharge into the sand, an additional amount of waste is added and preferably the mixture is kept for precipitation for about one hour to allow the substrate to migrate through the sand and cover the sand particles. The drum is then refilled with clean water, as described above, and nutrients are added for the next processing operation.

Repeated biological processing operations can be performed at regular intervals. This method can be used in an industrial enterprise or in a similar organization, where hazardous waste is constantly produced during a continuous process of manufacturing or processing products. If the substrate to be processed is any particular material, then when exposed to enzymes it is likely to produce one or more specific by-products in the form of amino acids or fatty acids. Products that are drained from the drum after processing can then be separated to isolate any desired product — amino acids or fatty acids. Such products usually have a significant market price, and their sale can be the basis for extensive recycling, as well as reduce the cost of disposal of hazardous waste.

Among the various types of biological processing byproducts using the present invention are fatty acids, which are produced as a result of microbiological effects on hydrocarbons, and amino acids, probably formed mainly as a result of decomposition or destruction (lysis) of microbes. It is obvious that the extraction of fatty acids as by-products of processing may have a certain commercial potential. They have a relatively high concentration - about one percent by weight of the samples.

When biological processing of contaminated solids or soil contaminated material can be introduced into the reactor alone or in a mixture with sand or other solid medium. After biological processing, the solid material should be periodically removed from the reactor. An example of the biological processing of contaminated soil is discussed below.

Thus, the cost of extracting these products should not be excessively high (the cost of production is assumed to be practically zero). Secondly, the market demand for fatty acids is very high, while in the mixture obtained during the experiments, the most valuable fatty acids with relatively long carbon chains (C10 and more) prevailed. In practice, it seems likely that some of the composition of the mixture of fatty acids can be controlled to some extent by changing the conditions of biological processing.

As a result of biological processing of soil contaminated with oil, at its location or in a container with the help of microorganisms according to the present invention, mixtures of amino acids and fatty acids are obtained, which can potentially be extracted as valuable products. It is well known that the effect of microbes on long-chain hydrocarbons can lead to the formation of organic acids with different chain lengths. In addition, as cells age, microbes can break down, releasing the compounds they contain into the environment. However, other microorganisms usually capture these released compounds and use them. In this regard, we can expect the detection of both organic acids and amino acids, the products of protein decomposition in those areas where microbial populations actively affect long-chain hydrocarbons.

There is an extensive market for organic acids and amino acids. If it is economical to obtain these compounds from a mixture of biological products and sell them on this market, then it is possible to reduce or even eliminate the costs of recycling waste at their location.

Biological processing of soil at its location using an open pit

The method according to the present invention was applied to a pit (see FIG. 9) with dimensions of 12x18x2 feet deep (about 3.6 x5.5 x0.7 m) prior to the onset of the overall level of petroleum hydrocarbon in the soil, forming a hole about 20%. In the center of the pit, a vertical tube was installed, extending into the ground below the level of the pit to extract biological processing products. The treatment began by creating a basin by adding about 40 barrels (6.36 m ³ ) of water, fertilizer and about 10 ounces (about 283 g) of the final composting product according to the present invention to the pit. Every few days dark water was removed from the pit and then pure water and fertilizer or nutrient were added.

Samples representing the source material before the test, microbiologically treated sludge from the bottom of the treated pool, soil from a depth of several feet from the bottom of the pool and a sample from a very great depth — about 50 feet (about 15.2 m) from the bottom of the pool were selected. No fluid samples were taken.

Solid samples were analyzed by extraction with various solvents, followed by analysis of the extract using various means to determine individual amino acids, total fat, and individual fatty acids (saturated and unsaturated). In most cases, the results of the analysis are presented as a percentage of the non-extracted sample. See tab. 5 and 6 below.

Table 5

Amino acids produced by a single biological treatment at the site of waste crude paraffin oil in the pit

Analyzes results Total fat 7.41% Protease Not found Protein 56 mg / 100 g Lipase Not found P amino acid profile Aspartic acid 1.63 mg / 100 g Tyrosine 5.20 mg / 100 g Valin 4.42 mg / 100 g Methionine 4.87 mg / 100 g Cysteine 1.12 mg / 100 g Isoleucine 3.16 mg / 100 g Leucine 3.33 mg / 100 g Phenylalanine 4.51 mg / 100 g Lysine 5.01 mg / 100 g Glutamic acid 0.84 mg / 100 g Serine 2.20 mg / 100 g Glycine 2.33 mg / 100 g Histidine 3.19 mg / 100 g Arginine 3.64 mg / 100 g Threonine 2.12 mg / 100 g Alanine 1.92 mg / 100 g Proline 7.28 mg / 100 g Saturated fatty acid 1.48% Mole fatty fat 0.76% Polyunsaturated fat 0.22%

Table 6

Fatty acids produced by a single biological treatment at the site of waste crude paraffin oil in the pit

Composition Formula | Content% Fatty acid profile Caprilate C8: 0 0.74 Caprat C10: 0 6.94 Laurat C12: 0 5.43 Myristate C14: 0 4.68 Palmitate C16: 0 1.73 Palmitoleate C16: 1 1.33 Stearate C18: 0 1.21 Oleate C18: 1 4.86 Linoleate C18: 2 2.17

Linolenate C18: 3 0.47 Arahidat C20: 1 0.37 Eykozenat C22: 0 <0.1 Begenat C22: 1 2.88 Erukat C24: 0 2.85 Lignocerate 2.18 The total content of saturated fatty acids 3.34 The total content of monounsaturated fatty acids 1.15 The total content of polyunsaturated fatty acids 0.34 Total fat 12.70

The total fatty acid content in the solid phase is about 1% by weight. Fatty acids are volatile and at the same time poorly soluble, which allows their separation and purification using a relatively simple (and cheap) method of distillation and fractional crystallization.

The specific by-product obtained by the biological processing of the subsoil

It is assumed that the composting product obtained by biological processing according to the present invention is not specific to the processed and biologically processed substrates. However, as follows from the further description, all by-products obtained by this method of processing are specific for a particular substrate, i.e. the byproduct will depend on the composition of the decomposable substrate.

An example of the production of an amino acid from hazardous waste is to obtain tyrosine as a result of the biological decomposition of soil contaminated with toxaphene using microbiological products according to the present invention. Toxaphene is one of the compounds with a high resistance to biological effects due to the high chlorine content of about 70%. FIG. 8 shows a tank for biological treatment of soil contaminated with toxaphene. The technological operations used here are similar to those used for the biological processing of waste in the container described above. Clean water is added to the desired level (about 3 inches - about 7.6 cm) over contaminated soil. In addition, a small amount of seeding aerobic microbes obtained by composting using the method of the present invention is added, as well as some nutrients. Hydrolysis of the substrate with enzymatic catalysis in the presence of water and microbiological effects on pollution occur under aerobic conditions without aeration or oxygen supply to the soil in any other way. The color of the water on the surface of the tank becomes dark, and the liquid containing the product of biological processing - tyrosine - is decanted or otherwise drained to the level of the soil. Then add water again and repeat the process until the end of a sufficient biological processing. Toxaphene was widely used as a pesticide until it was banned by the EPA in the early 1980s. The results of the analysis of the product of biological processing toxaphene are presented below in table. 7

Table 7

Analyzes of amino acids in the product of biological processing toxaphene

Analysis results Amino acid profile Alanine Not found Alpartinic acid Not found Tyrosine 53,000 million hours Valin Not found Methionine Not found Cysteine Not found Isoleucine Not found Leucine Not found Phenylalanine Not found Lysine Not found Idroxyproline Not found Glutamic acid Not found Serine Not found Glycine Not found Histidine Not found Arginine Not found Threonine Not found Proline Not found Protein 5.38% Saturated fatty acid <0.1 Monounsaturated fat <0.1 Polyunsaturated fat <0.1 Total content fat 0.30%

These results determine the specificity of the biological processing reaction of the present invention when treating toxaphene to form the amino acid tyrosine as an exclusive amino acid product.

The procedure described above can be used for the biological processing of any polluted soils or other solids contaminated with products that can be biologically processed.

Theory of the process of biological processing

The hydrolysis reaction can be accelerated abiotically or biotically. Some sources confirm that hydrolysis occurs faster with the participation of microbes. Enzyme hydrolysis is accelerated if the enzymes hold the reacting elements in the correct position, increasing the speed of interaction. This ensures the elimination of adverse changes in the activation energy. The resulting compound and enzyme complexes are subsequently separated by the action of water. The end result is the same as if only water had a direct effect on the complexes. In operations using the unique composting products of the present invention for biological processing of soil and groundwater contaminated with oil and chlorinated compounds, the above data was collected, which show that hydrolysis with enzyme catalysis is the main decomposition mechanism of the treatment method of the present invention.

The moisture content was the limiting factor for both the rates (kinetics) and the degree of flow (thermodynamics) of the reactions, which was expected for the hydrolysis reaction. The processing results showed low substrate specificity. Dechlorination was usually accompanied by an increase in solubility, which was also expected during the hydrolysis reaction. For example, in the process of investigating the ability to process, the theoretical solubility of toxaphene increased 15 times (from 3 to 45 million hours). Nucleophilic dehalogenation by nucleophilic substitution of halogen groups is common in reactions with enzyme catalysis and leads to an increase in solubility due to hydroxylation.

Reactions involving bacterial cultures according to the present invention proceed faster than expected even when using advanced biotechnology. Long-chain aliphatic compounds decompose almost as easily as short-chain aliphatic compounds. High reaction rates, along with dechlorination, an increase in solubility and decomposition of various compounds (including such heavy compounds as paraffin and asphalt), indicate that these reactions take place at a reduced activation energy, which is a phenomenon characteristic of the action of the enzyme.

By-products of biological processing according to the present invention include amino acids that are in demand in the market. Saturation of treatment solutions with amines, amino acids and carboxylic acids, which are typical products of hydrolysis reactions, inhibited the decomposition of compounds. It turned out that the saturation of the treatment solutions with polar by-products neutralizes the active sites of water molecules, effectively reducing their reactivity. This option is similar to low moisture content. Decomposition of compounds is resumed after extraction of acids and replenishment with fresh water to continue decomposition. Salt water is supposed to be even more efficient.

It is assumed that enzymatic hydrolysis is the main decomposition mechanism used in the proposed method of treating contaminated soil. The enzymatic nature of the culture increases the rate of decomposition and reduces the specificity of the substrate for processing. Due to the increased energy levels during hydrolysis with enzymatic catalysis, such technological methods as air lapping (blowing) and soil extraction with steam (with fresh air displaced) may be absolutely unnecessary.

Despite the fact that when using the method according to the present invention, the metabolic and enzymatic processes, as a rule, occur in an anaerobic environment, the well-known enzymatic ammonia and unpleasant odors, often accompanying anaerobic processes, are minimized. This is particularly desirable for composting in landfills located near residential areas. It is also desirable to minimize unpleasant odors in industrial or industrial areas.

Other variations within the scope of the present invention are apparent from the examples given, it being understood that the present description illustrates the features of the invention included in the accompanying claims.

Claims (9)

CLAIM 1. The method of biological processing of the medium containing hydrocarbon pollution, in which: A. Carry out the impact on hydrocarbon pollution of the specified environment in-situ viable culture of aerobic bacteria obtained by - mixing the mass of household solid waste with manure and secondary sewage sludge to form a compost heap of biomass, while these manure and sewage sludge contain a sufficient amount of moisture and natural bacteria, including aerobic bacteria, for composting the specified mass; - keeping said compost heap to undergo an autothermal reaction during an initial period of time, wherein the duration of said initial period is determined by the time that said compost heap reaches a temperature of about 46 ° C (115 ° F) to about 60 ° C (140 ° F); - adding to said compost heap and distributing alkaline material therein to rapidly increase the pH in said compost heap to at least 1 0; - keeping said compost heap to undergo an autothermal reaction for a second period of time, wherein the duration of said second period is determined by the time the specified compost heap reaches a temperature of at least 82 ° C (180 ° F); - adding nutrients selected from the group comprising inorganic compounds of nitrogen, phosphorus, sulfur and potassium to said compost heap to maintain the continuity of metabolic reactions in said compost heap; and - maintaining the specified compost heap for further autothermal reaction during the third period of time with the addition of water in sufficient quantity to maintain humidity in the specified heap at a level of from about 60 to about 82% for the specified third period of time, while the specified third period of time ends after decomposition of the specified biomass with the formation of non-pathogenic compost product; - carrying out the specified reaction for the specified third period of time in the absence of mechanical aeration of the compost heap or adding gaseous oxygen to the specified compost heap to obtain a viable culture of aerobic bacteria that can biologically process chlorinated hydrocarbons and long-chain hydrocarbons, including paraffin and asphalt, in the presence of nutrients and water for the biological processing of these hydrocarbons by hydrolysis of hydrocarbons with enzymatic catalysis; B. Provide the presence of sufficient nutrients and moisture to support the biological processing of these hydrocarbons; C. Carry out the hydrolysis of these hydrocarbon contaminants with enzymatic catalysis; D. Periodically remove the aqueous solution containing biological products from the place of biological processing in place; E. Add fresh water to the indicated location in an amount approximately equal to the amount removed in the above step D; G. The indicated step D of removing the aqueous solution and the indicated step E of adding water are successively repeated until a sufficient degree of biological processing of the indicated site is achieved. 2. The method according to claim 1, characterized in that said medium is a subsurface soil layer or said medium is located in a pit. 3. The in-situ system of underground biological processing of the horizontal soil layer in the area (1 2) contaminated with organic or hydrocarbon material, characterized in that it includes: A. At least two small diameter wells (18, 34) extending vertically from the surface (14) of said soil section (12) to a level located at least directly below said soil layer (16) to be biologically processed ; B. Moreover, each specified well (18, 34) contains an outer pipe (20, 36), in which there is a vertically located electrode (26, 42), passing to the level of the specified layer (1 6); C. Source (30) of direct current electric power supply; D. Means (46) for connecting the first electrode (42) located in the first of these wells (34) to the negative pole of the specified power source (30); E. Means (32) for connecting the second electrode (26) located in the second of these wells (18) to the positive pole of the specified power source (30); F. In this case, at least the outer pipe (36) of one of said wells (34) is perforated at the level of said layer (1 6); G. Means (48, 50) for supplying to one specified well (34) a sufficient amount of an aqueous mixture containing bacteria that perform biological processing to ensure the course of the biological processing reaction; H. In this case, the specified biological processing reaction is stimulated by the indicated electrodes (26, 42), and the voltage from the specified power source (30) is supplied to the indicated electrodes; and I. Means for activating the specified source (30) of power supply in order to create an electric field between the indicated electrodes (26, 42), wherein the specified biological processing reaction is supported by water, moving from the specified first well (34) to the specified second well ( 18) within the specified layer (16) for the gradual biological processing of the specified layer in situ. 4. An in-situ underground biological processing system according to claim 3, characterized in that at least the first said electrode (42) in said first well (34) is an electrically conductive tube that has openings (44) at a level the specified layer (16), while at least part of the specified aqueous mixture is fed through the specified conductive tube. 5. An in-situ underground biological processing system according to claim 3, characterized in that several pairs of oppositely charged electrodes (26, 42) are located in several corresponding wells (18, 34) for passing said water mixtures through the corresponding different parts of the contaminated layer . 6. The system for in-situ underground biological processing according to claim 3, characterized in that it contains a tank (48) on the surface of the indicated section for controlled supply by gravity of a portion of the mixture to the specified first well (34). 7. An in-situ method for underground biological processing of a horizontal layer (16) in a section (12) of soil contaminated with organic or hydrocarbon material, characterized in that it includes operations in which: A. The mixture of water and bacteria performing biological processing is introduced into the first section of the specified layer (1 6); B. Affect said layer (16) with a constant electric field having a positive pole and a negative pole, wherein said positive pole is in close proximity to said first portion of said layer; and C. A biological treatment reaction is carried out in a reaction zone located in the immediate vicinity of said first section, wherein the influence of said electric field forces said reaction zone through said layer (1 6) from said first section to a second section in close proximity to specified negative pole. 8. The in-situ method of underground biological processing according to claim 7, characterized in that said bacteria that perform biological processing contain a viable aerobic bacteria culture obtained by the method according to claim 5, and said biological processing reaction proceeds mainly due to hydrolysis with enzymatic catalysis of material contaminated with organic or hydrocarbon compounds. 9. The in-situ method of underground biological processing according to claim 8, characterized in that water is added to the specified first section during the course of the specified biological processing reaction.