CN113016800A - Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method - Google Patents
Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method Download PDFInfo
- Publication number
- CN113016800A CN113016800A CN202110262680.4A CN202110262680A CN113016800A CN 113016800 A CN113016800 A CN 113016800A CN 202110262680 A CN202110262680 A CN 202110262680A CN 113016800 A CN113016800 A CN 113016800A
- Authority
- CN
- China
- Prior art keywords
- reducing bacteria
- sulfate
- sulfate reducing
- oil field
- inhibitor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/30—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests characterised by the surfactants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N33/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds
- A01N33/16—Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds containing nitrogen-to-oxygen bonds
- A01N33/18—Nitro compounds
- A01N33/20—Nitro compounds containing oxygen or sulfur attached to the carbon skeleton containing the nitro group
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/18—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds
- A01N57/20—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-carbon bonds containing acyclic or cycloaliphatic radicals
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/52—Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning
- C09K8/528—Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning inorganic depositions, e.g. sulfates or carbonates
- C09K8/532—Sulfur
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2208/00—Aspects relating to compositions of drilling or well treatment fluids
- C09K2208/20—Hydrogen sulfide elimination
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Environmental Sciences (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Agronomy & Crop Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pest Control & Pesticides (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- General Life Sciences & Earth Sciences (AREA)
- Materials Engineering (AREA)
- Toxicology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cosmetics (AREA)
Abstract
The invention belongs to the technical field of environmental microbiology, and particularly relates to an oilfield sulfate reducing bacteria activity inhibitor, wherein the oilfield sulfate reducing bacteria enrichment culture inhibitor is obtained by compounding three chemical bactericides, namely glutaraldehyde, bronopol and tetrakis hydroxymethyl phosphonium sulfate, with a biosurfactant rhamnolipid. The method solves the problems that the existing method for inhibiting the amount of hydrogen sulfide produced by the sulfate reducing bacteria in the oil field has low bacteriostasis efficiency, complex bacteriostasis method and difficult implementation, and the oil field microorganisms have drug resistance and other defects in the prior art. Compared with the single use of chemical bactericides or biological surfactants, the inhibitor has higher efficiency of inhibiting the activity of sulfate reducing bacteria for generating hydrogen sulfide, can reduce the potential environmental pollution risk caused by the excessive use of the bactericides, and can be applied to the prevention and control of the harm of the sulfate reducing bacteria in the development process of oil fields.
Description
Technical Field
The invention relates to the technical field of environmental microbiology, in particular to an inhibition method for an oil field sulfate reducing bacteria enrichment culture and application thereof.
Background
At present, most of oil exploitation in China enters a water injection and oil displacement stage of a later development period, and the extremely high water content in produced liquid at the stage causes serious Microbial Corrosion (MIC), so that the increasing cost of oil field exploitation is finally caused. With the development of anaerobic technology, and the further understanding of anaerobic bacteria, it was found that sulfate-reducing bacteria (SRB) are the most important flora causing microbial corrosion. Sulfate-reducing bacteria refer to microorganisms that produce hydrogen sulfide gas using sulfur or sulfur-containing compounds as substrates. SPB, while oxidizing nutrients to reduce sulfate, produces a large amount of sulfides, which are responsible for microbial corrosion. In an oil field system, hydrogen sulfide can not only accelerate corrosion, pollute crude oil, reduce the quality of the crude oil and increase the subsequent treatment cost, but also seriously threaten the life of workers due to higher toxicity. Meanwhile, the generated ferrous sulfate precipitate can cause oil layer blockage so as to reduce the oil layer leakage rate, and the exploitation difficulty is greatly increased. Losses from sulfate-reducing bacteria are in the hundreds of millions of dollars per year. Therefore, eliminating or reducing the content of hydrogen sulfide produced by reservoir sulfate-reducing bacteria is an urgent problem to be solved.
There are many methods for inhibiting sulfate-reducing bacteria in China, and the methods are mainly divided into physical methods, chemical methods and biological methods. The physical methods mainly comprise technical methods such as cathodic protection, ultraviolet irradiation, ultrasonic treatment, high-voltage pulse electric field sterilization and the like, but in practical application, the physical methods are found to be difficult to effectively inhibit sulfate reducing bacteria for a long time, and have no obvious effect on the generated biological membrane. The biological method is to use the same habit as SPB but does not produce H2The microorganisms of S achieve the aim of inhibiting SPB through the relations of competition, antagonism or symbiosis and the like. Or using some microbes capable of producing metabolites inhibiting the growth of SPB to prevent the growth and propagation of SPB in the system. But because the biological method is relatively complex, the application in oil field systems in China is less. The chemical method, namely adding a certain amount of bactericide into the reinjection water of the oil field so as to achieve the purpose of killing or inhibiting SPB, is relatively simple and easy to operate, so that the method is widely used in China in the eighties of the last century. Although the purpose of killing SPB can be achieved by destroying cell walls, cell membranes, enzymes or substrates of SRB cells, the bactericide is the most direct and effective method for inhibiting SRB at present. However, long-term use of chemical fungicides not only causes environmental pollution, but also causes SPB drug resistance.
Disclosure of Invention
The invention provides an efficient bacteriostatic inhibitor and an inhibition method, which solve the problems of low bacteriostatic efficiency, complex bacteriostatic method, difficult implementation, drug resistance of oilfield microorganisms and other defects of the prior art in the existing method for inhibiting the hydrogen sulfide production amount of oilfield sulfate reducing bacteria. .
In order to achieve the purpose, the invention adopts the technical scheme that: the oil field sulfate reducing bacteria enrichment culture inhibitor is obtained by compounding three chemical bactericides, namely glutaraldehyde, bronopol and tetrakis hydroxymethyl phosphonium sulfate, with a biosurfactant rhamnolipid.
The oil field sulfate reducing bacteria enrichment culture inhibitor is characterized in that three chemical bactericide mother solutions of glutaraldehyde, bronopol and tetrakis hydroxymethyl phosphonium sulfate are prepared respectively by using sterile water in a super clean bench, the mother solutions are stored in a refrigerator, activated pseudomonas aeruginosa is inoculated in a sterilized culture medium for producing rhamnolipid by metabolism according to 5 percent in the super clean bench, and the rhamnolipid is obtained by fermentation for 7d at 37 ℃ and 180 r/min.
In the oil field sulfate reducing bacteria enrichment culture inhibitor, the culture medium for metabolizing to produce rhamnolipid is 3-4% of glycerol and NaNO3 0.2-0.4％，K2HPO4·3H2O 0.35-0.45％，KH2PO4 0.4-0.5％，MgSO4·7H2O 0.003-0.005％，CaC120.01-0.02%, KCl 0.08-0.14%, NaCl 0.08-0.14%, yeast powder 0.1-0.2%, and water in balance.
The inhibitor for enrichment culture of sulfate reducing bacteria in oil fields comprises (by mass concentration ratio) glutaraldehyde, bronopol, tetrakis hydroxymethyl phosphonium sulfate and rhamnolipid, (10-100 mg/L) and (0.1-10 mg/L).
The method for inhibiting the oil field sulfate reducing bacteria enrichment culture inhibitor from producing hydrogen sulfide comprises the following steps,
1) inoculating the oilfield produced water into a sterilized sulfate reducing bacteria culture medium, and standing and culturing in a biochemical incubator at 45 ℃ until the enrichment liquid in the anaerobic tube becomes black to obtain an oilfield sulfate reducing bacteria enrichment culture;
2) inoculating the oil field sulfate reducing bacteria enrichment culture into an oil field sulfate reducing bacteria enrichment culture inhibitor according to the inoculation amount of 1-20%, detecting the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture inhibitor by using a methylene blue spectrophotometry, then placing the oil field sulfate reducing bacteria enrichment culture inhibitor into a biochemical incubator at 37 ℃ for standing culture for 2-60 days, and detecting the hydrogen sulfide content again.
In the method for inhibiting any of the oilfield sulfate reducing bacteria enrichment culture inhibitors, in the step 1), the components of the sulfate reducing bacteria culture medium are sodium lactate (50 percent) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0.
The invention has the beneficial effects that:
the invention provides a mixed bacteriostatic agent for inhibiting oil field sulfate reducing bacteria from producing hydrogen sulfide, which is prepared by mixing a chemical bactericide and a biological surfactant in proportion, and can improve the capability of killing or inhibiting the oil field sulfate reducing bacteria.
The chemical bactericide and the biosurfactant are compounded for use, compared with the use of a single chemical bactericide as the oilfield sulfate reducing bacteria inhibitor, the addition amount is reduced to a certain extent, the pollution to the oilfield and the surrounding environment is reduced, and the drug resistance of the oilfield sulfate reducing bacteria can be resisted. Compared with the single use of the biosurfactant, the method has the advantages of rapid, efficient and long-term sterilization and bacteriostasis effects.
The invention has wide application range, can be used for most oil fields in China and has simple operation.
The invention can be used for a long time, is environment-friendly, has high sterilization and bacteriostasis efficiency and long duration, and is beneficial to field popularization and application.
The specific implementation mode is as follows:
the present invention will be further described with reference to the following examples, but the present invention is not limited to the following examples.
Material preparation
1. Chemical germicide and biological surfactant
Three chemical bactericides, namely glutaraldehyde, bronopol and THPS, are purchased from the market, and after mother liquor is prepared, the mother liquor is stored in a refrigerator at 4 ℃ for later use. The biosurfactant is prepared by inoculating activated pseudomonas aeruginosa at 5% into sterilized culture medium capable of metabolizing to produce rhamnolipid, and fermenting at 37 deg.C for 7d at 180 r/min.
The medium for metabolizing to produce rhamnolipid is 3-4% of glycerol and NaNO3 0.2-0.4％，K2HPO4·3H2O 0.35-0.45％，KH2PO4 0.4-0.5％，MgSO4·7H2O 0.003-0.005％，CaC120.01-0.02%, KCl 0.08-0.14%, NaCl 0.08-0.14%, yeast powder 0.1-0.2%, and water in balance.
2. Culture medium
The culture medium of the sulfate reducing bacteria comprises the following components: lactic acidSodium (50%) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO41.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0.
3. Laboratory apparatus
Constant temperature incubator
Super clean bench
Vertical pressure steam sterilizer
Miniature ultraviolet spectrophotometer
Laboratory pH meter
Series vacuum suction filtration device
Example 1 sulfate-reducing bacteria inhibition experiment in certain northeast oilfield
Collecting produced liquid from the oil production well port end of a certain oil field in northeast of China by using a presterilized polyethylene plastic barrel (10.0L), closing a sampling port valve after the polyethylene plastic barrel is fully taken, rapidly covering and sealing the polyethylene plastic barrel, and transporting the polyethylene plastic barrel to a laboratory by a logistics company after marking. And then carrying out oil-water separation on the produced liquid of each oil well. And respectively filtering the produced liquid of 10 oil wells by using medium-speed qualitative filter paper to complete oil-water separation. Introducing 99.99 percent high-purity nitrogen into the filtered produced water for a period of time, removing air, and storing the produced water in a laboratory. Used for preparing enriched culture of sulfate reducing bacteria.
In a super clean bench, the treated oilfield produced water is inoculated in a sterilized sulfate reducing bacteria culture medium by a sterile injector in a proportion of 5%. Sulfate reducing bacteria culture medium with sodium lactate (50%) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0. Then placing the mixture in a biochemical incubator at 45 ℃ for static culture. Observing the change condition of the enrichment liquid at regular time every day until the enrichment liquid in the anaerobic tube becomes black (the formed FeS black precipitate indicates the growth of the strain), thus obtaining the enrichment culture of sulfate reducing bacteria in the somebody's northeast oilfield.
In a super clean bench, three kinds of mother liquid of the chemical germicide, glutaraldehyde, bronopol and THPS, are prepared separately with sterile water. And (4) after the mother liquor is prepared, storing the mother liquor in a refrigerator at 4 ℃ for later use. Inoculating activated pseudomonas aeruginosa in a sterilized culture medium which is metabolized to produce rhamnolipid according to 5 percent in a super-bacterium platform, and fermenting at 37 ℃ for 7d at 180r/min to obtain the rhamnolipid for later use. Then, the mother liquor of the chemical bactericide and the biological surfactant are inoculated into a culture medium of the sterilized sulfate reducing bacteria according to the mass ratio of glutaraldehyde to bronopol to THPS to rhamnolipid of 10mg/L to 5mg/L to 0.5 mg/L. Then, the oil field sulfate reducing bacteria enrichment culture is inoculated into a culture medium of the sulfate reducing bacteria according to the inoculation amount of 5 percent, and the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture is detected by a methylene blue spectrophotometry. And then placing the mixture in a biochemical incubator at 45 ℃ for standing culture for 45 days, and detecting that the increment of hydrogen sulfide is zero, so that the mixed inhibitor has good inhibition effect on sulfate reducing bacteria enrichment culture in certain oil fields in northeast China.
Example 2 Xinjiang oil field sulfate reducing bacteria inhibition experiment
Collecting produced liquid from the oil production well port end of a certain oil field in Xinjiang by using a presterilized polyethylene plastic barrel (10.0L), closing a sampling port valve after the polyethylene plastic barrel is fully taken, rapidly covering and sealing the polyethylene plastic barrel, and transporting the polyethylene plastic barrel to a laboratory by a logistics company after marking. And then carrying out oil-water separation on the produced liquid of each oil well. And respectively filtering the produced liquid of 10 oil wells by using medium-speed qualitative filter paper to complete oil-water separation. Introducing 99.99 percent high-purity nitrogen into the filtered produced water for a period of time, removing air, and storing the produced water in a laboratory. Used for preparing enriched culture of sulfate reducing bacteria.
In a super clean bench, the treated oilfield produced water is inoculated in a sterilized sulfate reducing bacteria culture medium by a sterile injector in a proportion of 10%. The culture medium is sulfate-reducing bacteria culture medium, and contains sodium lactate (50%) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0. Then placing the mixture in a biochemical incubator at 37 ℃ for static culture. Observing the change condition of the enrichment liquid every day at regular time until the enrichment liquid in the anaerobic tube becomes black (the formed FeS black precipitate shows the growth of the strain), thus obtaining the enrichment culture of the sulfate reducing bacteria in the Xinjiang oil field.
In a super clean bench, three kinds of mother liquid of the chemical germicide, glutaraldehyde, bronopol and THPS, are prepared separately with sterile water. And (4) after the mother liquor is prepared, storing the mother liquor in a refrigerator at 4 ℃ for later use. Inoculating activated pseudomonas aeruginosa in a sterilized culture medium which is metabolized to produce rhamnolipid according to 5 percent in a super-bacterium platform, and fermenting at 37 ℃ for 7d at 180r/min to obtain the rhamnolipid for later use. Then, the mother liquor of the chemical bactericide and the biological surfactant are inoculated into a culture medium of the sterilized sulfate reducing bacteria according to the mass ratio of glutaraldehyde to bronopol to THPS to rhamnolipid of 10mg/L to 0.5 mg/L. Then, the oil field sulfate reducing bacteria enrichment culture is inoculated into a culture medium of the sulfate reducing bacteria according to the inoculation amount of 5 percent, and the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture is detected by a methylene blue spectrophotometry. And then placing the mixture in a biochemical incubator at 37 ℃ for standing culture for 45 days, and detecting that the increment of hydrogen sulfide is zero, so that the mixed inhibitor has good inhibition effect on the sulfate reducing bacteria enrichment culture of the Xinjiang oil field.
Example 3 sulfate-reducing bacteria inhibition experiment in certain oil field in sea area
Collecting produced liquid from the oil production well end of a certain oil field in the sea area by using a presterilized polyethylene plastic barrel (10.0L), closing a sampling port valve after the polyethylene plastic barrel is fully taken by 10.0L, rapidly covering and sealing the polyethylene plastic barrel, and transporting the polyethylene plastic barrel to a laboratory by a logistics company after marking. And then carrying out oil-water separation on the produced liquid of each oil well. And respectively filtering the produced liquid of 10 oil wells by using medium-speed qualitative filter paper to complete oil-water separation. Introducing 99.99 percent high-purity nitrogen into the filtered produced water for a period of time, removing air, and storing the produced water in a laboratory. Used for preparing enriched culture of sulfate reducing bacteria.
In a super clean bench, the treated oilfield produced water is inoculated in a sterilized sulfate reducing bacteria culture medium by a sterile injector in a proportion of 10%. The culture medium is sulfate-reducing bacteria culture medium, and contains sodium lactate (50%) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0. Then placing the mixture in a biochemical incubator at 60 ℃ for static culture. Observing the change condition of the enrichment liquid at regular time every day until the enrichment liquid in the anaerobic tube becomes black (the formed FeS black precipitate indicates the growth of the bacterial strain), thus obtaining the enrichment culture of sulfate reducing bacteria in an oil field in the sea area.
In a super clean bench, three kinds of mother liquid of the chemical germicide, glutaraldehyde, bronopol and THPS, are prepared separately with sterile water. And (4) after the mother liquor is prepared, storing the mother liquor in a refrigerator at 4 ℃ for later use. Inoculating activated pseudomonas aeruginosa in a sterilized culture medium which is metabolized to produce rhamnolipid according to 5 percent in a super-bacterium platform, and fermenting at 37 ℃ for 7d at 180r/min to obtain the rhamnolipid for later use. Then, the mother liquor of the chemical bactericide and the biological surfactant are inoculated into a culture medium of the sterilized sulfate reducing bacteria according to the mass ratio of glutaraldehyde to bronopol to THPS to rhamnolipid of 10mg/L to 5mg/L to 0.5 mg/L. Then, the oil field sulfate reducing bacteria enrichment culture is inoculated into a sulfate reducing bacteria culture medium according to the inoculation amount of 5%, and the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture is detected by a methylene blue spectrophotometry. And then placing the mixture in a biochemical incubator at 60 ℃ for standing culture for 45 days, and detecting that the increment of hydrogen sulfide is zero, so that the mixed inhibitor has good inhibition effect on sulfate reducing bacteria enriched culture of certain oil field in the sea area.
Example 4 sulfate reducing bacteria inhibition experiment in certain oil field in northwest of China
Collecting produced liquid from the oil production well port end of a certain oil field in northwest by using a presterilized polyethylene plastic barrel (10.0L), closing a sampling port valve after the polyethylene plastic barrel is fully taken by 10.0L, rapidly covering and sealing the polyethylene plastic barrel, and transporting the polyethylene plastic barrel to a laboratory by a logistics company after marking. And then carrying out oil-water separation on the produced liquid of each oil well. And respectively filtering the produced liquid of 10 oil wells by using medium-speed qualitative filter paper to complete oil-water separation. Introducing 99.99 percent high-purity nitrogen into the filtered produced water for a period of time, removing air, and storing the produced water in a laboratory. Used for preparing enriched culture of sulfate reducing bacteria.
In a super clean bench, the treated oilfield produced water is inoculated in a sterilized sulfate reducing bacteria culture medium by a sterile injector in a proportion of 10%. The culture medium is sulfate-reducing bacteria culture medium, and contains sodium lactate (50%) 2.5 g.L-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0. Then placing the mixture in a biochemical incubator at 37 ℃ for static culture. Regularly observe rich every dayCollecting the liquid to change the situation until the enrichment liquid in the anaerobic tube becomes black (the formed FeS black precipitate indicates the growth of the strain), thus obtaining the enrichment culture of sulfate reducing bacteria in a certain oil field in northwest.
In a super clean bench, three kinds of mother liquid of the chemical germicide, glutaraldehyde, bronopol and THPS, are prepared separately with sterile water. And (4) after the mother liquor is prepared, storing the mother liquor in a refrigerator at 4 ℃ for later use. Inoculating activated pseudomonas aeruginosa in a sterilized culture medium which is metabolized to produce rhamnolipid according to 5 percent in a super-bacterium platform, and fermenting at 37 ℃ for 7d at 180r/min to obtain the rhamnolipid for later use. Then, the mother liquor of the chemical bactericide and the biological surfactant are inoculated into a culture medium of the sterilized sulfate reducing bacteria according to the mass ratio of glutaraldehyde to bronopol to THPS to rhamnolipid of 10mg/L to 0.5 mg/L. Then inoculating the oil field sulfate reducing bacteria enrichment culture into the oil field sulfate reducing bacteria enrichment culture according to the inoculation amount of 5%, and detecting the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture by using a methylene blue spectrophotometry method. And then placing the mixture in a biochemical incubator at 37 ℃ for standing culture for 45 days, and detecting that the increment of hydrogen sulfide is zero, so that the mixed inhibitor has good inhibition effect on the sulfate reducing bacteria enrichment culture of certain oil fields in northwest.
Claims (6)
1. The oil field sulfate reducing bacteria enrichment culture inhibitor is characterized by being obtained by compounding three chemical bactericides, namely glutaraldehyde, bronopol and tetrakis hydroxymethyl phosphonium sulfate, with a biosurfactant rhamnolipid.
2. The oilfield sulfate reducing bacteria enrichment culture inhibitor as claimed in claim 1, wherein three chemical bactericide mother solutions of glutaral, bronopol and tetrakis hydroxymethyl phosphonium sulfate are prepared from sterile water in a super clean bench, stored in a refrigerator, and the activated pseudomonas aeruginosa is inoculated into a sterilized rhamnolipid-producing culture medium by metabolism at 5% in the super clean bench and fermented for 7d at 37 ℃ and 180r/min to obtain the rhamnolipid.
3. The oil field sulfate reducing bacteria enrichment culture inhibitor of claim 2The preparation is characterized in that the medium for metabolizing to produce rhamnolipid is 3-4% of glycerol and NaNO3 0.2-0.4％，K2HPO4·3H2O 0.35-0.45％，KH2PO40.4-0.5％，MgSO4·7H2O 0.003-0.005％，CaC120.01-0.02%, KCl 0.08-0.14%, NaCl 0.08-0.14%, yeast powder 0.1-0.2%, and water in balance.
4. The oilfield sulfate reducing bacteria enrichment culture inhibitor as defined in claim 3, wherein the weight concentration ratio of the glutaraldehyde, the bronopol, the tetrakis hydroxymethyl phosphonium sulfate and the rhamnolipid is (10-100 mg/L), (10-100 mg/L) and (0.1-10 mg/L).
5. The method for inhibiting the oilfield sulfate-reducing bacteria enrichment culture inhibitor according to any one of claims 1-4, wherein the inhibitor inhibits the oilfield sulfate-reducing bacteria from producing hydrogen sulfide, comprising the steps of,
1) inoculating the oilfield produced water into a sterilized sulfate reducing bacteria culture medium, and standing and culturing in a biochemical incubator at 45 ℃ until the enrichment liquid in the anaerobic tube becomes black to obtain an oilfield sulfate reducing bacteria enrichment culture;
2) inoculating the oil field sulfate reducing bacteria enrichment culture into an oil field sulfate reducing bacteria enrichment culture inhibitor according to the inoculation amount of 1-20%, detecting the hydrogen sulfide content of the oil field sulfate reducing bacteria enrichment culture inhibitor by using a methylene blue spectrophotometry, then placing the oil field sulfate reducing bacteria enrichment culture inhibitor into a biochemical incubator at 37 ℃ for standing culture for 2-60 days, and detecting the hydrogen sulfide content again.
6. The method for inhibiting the oilfield sulfate-reducing bacteria enrichment culture inhibitor according to claim 5, wherein in the step 1), the sulfate-reducing bacteria culture medium comprises 2.5 g-L sodium lactate (50%) (sodium lactate)-1，NH4Cl 1.0g·L-1，Na2SO4 1.0g·L-11.0 g.L of sodium potassium tartrate-1，MgSO4·7H2O 0.5g·L-1，K2HPO4·3H2O 0.5g·L-10.5 g.L of yeast extract-1，Na2S2O3 0.2g·L-10.2 g.L of ferric ammonium citrate-1，NaCl 7.0g·L-1Ascorbic acid 0.1 g.L-1(ii) a Adding 1.7% agar strips as a solid culture medium, and adjusting the pH value to 7.0-8.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110262680.4A CN113016800B (en) | 2021-03-11 | 2021-03-11 | Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110262680.4A CN113016800B (en) | 2021-03-11 | 2021-03-11 | Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113016800A true CN113016800A (en) | 2021-06-25 |
| CN113016800B CN113016800B (en) | 2021-10-22 |
Family
ID=76469373
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110262680.4A Active CN113016800B (en) | 2021-03-11 | 2021-03-11 | Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113016800B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113956860A (en) * | 2021-09-16 | 2022-01-21 | 华东理工大学 | Construction method of microbial corrosion control system of oil field system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232708A (en) * | 2013-11-08 | 2014-12-24 | 中国科学院沈阳应用生态研究所 | Culture medium for metabolism production of rhamnolipid and application of culture medium |
| CN110697916A (en) * | 2019-10-17 | 2020-01-17 | 大庆石油管理局有限公司 | Multifunctional sulfide remover and preparation method thereof |
-
2021
- 2021-03-11 CN CN202110262680.4A patent/CN113016800B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232708A (en) * | 2013-11-08 | 2014-12-24 | 中国科学院沈阳应用生态研究所 | Culture medium for metabolism production of rhamnolipid and application of culture medium |
| CN110697916A (en) * | 2019-10-17 | 2020-01-17 | 大庆石油管理局有限公司 | Multifunctional sulfide remover and preparation method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113956860A (en) * | 2021-09-16 | 2022-01-21 | 华东理工大学 | Construction method of microbial corrosion control system of oil field system |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113016800B (en) | 2021-10-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zinder et al. | Methanogenesis in a thermophilic (58 C) anaerobic digestor: Methanothrix sp. as an important aceticlastic methanogen | |
| CN103666426B (en) | Oil field water biological treatment agent and preparation method thereof and using method | |
| DE3230197C2 (en) | Methane fermentation process | |
| CN104388348A (en) | Microbial preparation for purifying sewage and deodorizing garbage and preparation method thereof | |
| CN105660705A (en) | Composite sulfate reducing bacteria activity ecologic inhibitor and application thereof | |
| CN102586160A (en) | Stenotrophomonas maltophilia DS4 | |
| CN110408565A (en) | A kind of efficient denitrification composite bacteria agent, preparation and its application | |
| CN104845928A (en) | Method for processing hexavalent chromium pollution employing synergistic effects of mixed bacteria | |
| CN113016800B (en) | Oil field sulfate reducing bacteria enrichment culture inhibitor and inhibition method | |
| CN102776140B (en) | Cold-tolerant pseudomonas strain Den-05, and screening method and application thereof | |
| CN103708626A (en) | Method for treating solid-waste anaerobic fermentation biogas slurry | |
| CN103305443B (en) | Preparation and method for restoring ammonia nitrogen-containing industrial sewage | |
| CN109576189B (en) | Composite microbial inoculum for black and odorous water body and preparation method and application thereof | |
| CN101386822B (en) | Special effect phosphate accumulating organisms and waste water processing method using thereof | |
| CN105176868A (en) | Complex bactericide for efficiently purifying sewage anaerobic outlet water of pig farm and preparation method and application thereof | |
| CN102586111A (en) | Microorganism bacterium agent for subtracting organic sludge, production method of microorganism bacterium agent and complex culture medium | |
| CN104630112A (en) | Acinetobacter as well as preparation method and application of acinetobacter | |
| CN102168038A (en) | Xanthobacter sp. D7 capable of degrading dioxane and application thereof | |
| CN111997582A (en) | Biological treatment technology for hydraulic control sulfur extracted from oil field | |
| CN106635855B (en) | Microbacterium and its culture application are seen in a kind of north | |
| CN103381418B (en) | Method for processing tobacco waste or organic fluorine wastewater | |
| CN114262679B (en) | Autotrophic denitrifying bacteria and rapid expanding culture process thereof | |
| CN101613156A (en) | Adopt Klebsiella sp. DB-2 bacterium to repair the method that nitrate in groundwater is polluted | |
| CN106277308B (en) | A kind of well-drilling waste water anaerobic treatment method | |
| CN116254188A (en) | Strain SRB-6 and application thereof in degrading herbicide acetochlor under anaerobic condition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |