CN113862198A - Compound microbial agent for degrading oil pollutants in oily sludge and preparation method and application thereof - Google Patents
Compound microbial agent for degrading oil pollutants in oily sludge and preparation method and application thereof Download PDFInfo
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- 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
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
Abstract
The invention discloses a compound microbial agent for degrading oil pollutants in oily sludge and a preparation method and application thereof, wherein the compound microbial agent is prepared by mixing a Pseudomonas aeruginosa (Pseudomonas aeruginosa) microbial agent, a Bifidobacterium (Bifidobacterium) microbial agent, a Bacillus subtilis microbial agent and a Bacillus halodenitrificans (Virgibacillus halodendrons) microbial agent according to a volume ratio of (1-4) to (1-4); the invention also relates to application of the compound microbial agent for degrading oil pollutants in the oily sludge in degrading petroleum substances. The composite microbial agent provided by the invention has a good degradation effect on petroleum hydrocarbon substances in the oily sludge, so that oil substances in the oily sludge can be removed, the oil concentration in the oil sludge is reduced, and the oil sludge treatment work is greatly promoted.
Description
Technical Field
The invention relates to microbial degradation and treatment of oil pollutants, in particular to a compound microbial agent for degrading oil pollutants in oily sludge and a preparation method and application thereof.
Background
The crude oil is a mixture composed of a plurality of simple or complex hydrocarbon compounds, generally consists of saturated hydrocarbon, aromatic hydrocarbon, colloid, asphaltene, a small amount of nitrogen compounds, sulfur compounds and other non-hydrocarbon compounds, has important application in various industries in China, and plays a very important role in the economic development of China. The oily sludge is an oily pollution mixture formed after crude oil is mixed with soil, water, garbage and the like in the processes of crude oil exploitation, refining and transportation, wherein the oily pollution mixture contains a large amount of pathogenic bacteria, parasites (eggs), heavy metals and a large amount of benzene series, phenols, polychlorinated biphenyl and other various refractory toxic and harmful substances, and if the oily pollution mixture is directly discharged without treatment, the oily sludge occupies a large amount of cultivated land, and pollutes surrounding soil, water and air. The volume of the oily sludge is huge, the statistics shows that the volume of the oily sludge is only 2020, 1.95 million tons of crude oil are produced in China, the increment of the oily sludge in China in 2020 is nearly 600 million tons calculated according to the amount of the produced oily sludge as 3 percent of the yield of the crude oil, and the historical residual oily sludge in China exceeds 1.4 million tons due to the immature treatment technology and incomplete legal regulations in the past. Therefore, the treatment of the oil sludge becomes the most important part of the treatment work of the hazardous waste in China and also becomes one of the factors for limiting the crude oil exploitation in China. At present, the methods for treating the oil sludge are mainly divided into physical, chemical and microbial methods, the physical and chemical methods are gradually replaced because of high treatment cost and secondary pollution problems, and the microbial treatment technology is gradually the most widely applied treatment method in multiple tests and applications because of low cost investment and good environmental benefit. The microbial treatment of the oily sludge is mainly to degrade crude oil into CO through the self metabolism by taking the crude oil as a nutrient substance required by the growth and the propagation of the crude oil through the growth metabolism of a crude oil degrading flora2And H2O is discharged out of the body, thereby reducing the petroleum content in the oily sludge and achieving the aim of safe treatment.
The microbial treatment of oily sludge is divided into biostimulation and bioaugmentation technologies. The biostimulation technology is characterized in that original indigenous bacteria in soil are activated under an artificially optimized environmental condition, so that the indigenous bacteria can perform a better growth and metabolism process, and petroleum pollutants can be degraded more easily; the biological strengthening technology is a technology of domesticating high-efficiency degrading bacteria and then inoculating the bacteria into oil sludge, and can overcome the limitation of the biological stimulation technology, so that the exogenous microorganisms are generally added in a biological strengthening way to achieve a better treatment effect. However, in practical application, the effect of treating the oil-containing sludge by using a single strain is not ideal, the single strain has specificity, and the single degradation strain is difficult to achieve good degradation effect on various oil sludges due to differences of oil sludge properties and components caused by different crude oil concentrations and extraction processes in various regions.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a compound microbial agent for degrading oil pollutants in oily sludge, which can effectively degrade petroleum substances in soil, and a preparation method and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
the compound microbial agent is prepared by mixing a Pseudomonas aeruginosa (Pseudomonas aeruginosa) microbial agent, a Bifidobacterium (Bifidobacterium) microbial agent, a Bacillus subtilis (Bacillus subtilis) microbial agent and a mycobacterium denitrificans (Virgibacillus halodendron) microbial agent in a volume ratio of (1-4) to (1-4).
The invention also provides a preparation method of the compound microbial agent for degrading oil pollutants in oily sludge, which comprises the following steps:
1) solid slant culture: respectively inoculating pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis into a beef extract peptone solid culture medium, and culturing at constant temperature to fully activate the strains;
2) first-order seed culture: respectively inoculating the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the bacillus thuringiensis activated in the step 1) into a beef extract peptone liquid culture medium, and culturing at constant temperature until OD is obtained600Stopping culturing when the value reaches 0.8, and respectively obtaining primary seed solutions of pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis;
3) secondary seed culture: respectively inoculating the primary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 2) into a fermentation culture medium, and culturing for 2d at 37 ℃ and 130r/min to respectively obtain secondary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium;
4) inoculating the secondary seed solutions of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 3) into a fermentation culture medium respectively in an inoculation amount of 1% of the fermentation culture medium by volume ratio, and performing fermentation culture for 1d at 37 ℃ and 130r/min to respectively obtain a pseudomonas aeruginosa microbial inoculum, a bifidobacterium microbial inoculum, a bacillus subtilis microbial inoculum and a salt denitrogenation mycobacterium microbial inoculum;
5) and mixing the pseudomonas aeruginosa microbial inoculum, the bifidobacterium microbial inoculum, the bacillus subtilis microbial inoculum and the bacillus halodenitrificans microbial inoculum according to the volume ratio of (1-4) to obtain the compound microbial inoculum for degrading the oil pollutants in the oily sludge.
Further, the fermentation medium in the step 3) and the fermentation medium in the step 4) comprise, by mass percent: 0.3 to 0.5 percent of ammonium chloride, 0.3 to 0.5 percent of monopotassium phosphate, 0.1 to 0.3 percent of dipotassium phosphate, 0.001 to 0.02 percent of anhydrous calcium chloride, 0.3 to 1 percent of sodium chloride, 0.01 to 0.02 percent of magnesium sulfate heptahydrate, 1 to 1.5 percent of molasses and the balance of water, and the pH value is 7.0 to 7.5.
The invention also relates to application of the compound microbial agent for degrading oil pollutants in oily sludge in degrading petroleum substances.
Further, when the composite microbial agent for degrading oil pollutants in the oily sludge is used for treating the oily sludge, the adding amount of the composite microbial agent is 10% of the volume of the oil sludge degradation liquid.
Further, the oil sludge degradation liquid comprises oil-containing sludge with the mass fraction of 2% and a degradation culture medium with the mass fraction of 98%, wherein the degradation culture medium comprises, by mass: 0.3% -0.5% of ammonium chloride, 0.3% -0.5% of monopotassium phosphate, 0.1% -0.3% of dipotassium phosphate, 0.001% -0.02% of anhydrous calcium chloride, 0.3% -1% of sodium chloride, 0.01% -0.02% of magnesium sulfate heptahydrate, and the balance of water, wherein the pH value of the degradation culture medium is 7.0-7.5.
Compared with the prior art, the invention has the following technical effects:
the pseudomonas aeruginosa and the bacillus subtilis in the composite microbial agent for degrading the oil pollutants in the oily sludge can both generate surfactants and reduce the interfacial tension, so that the emulsification is better promoted to improve the degradation effect of crude oil; the bifidobacterium and the bacillus halodenitrificans have certain specificity on the degradation of crude oil components, and the degradation performance of the composite microbial inoculum on petroleum hydrocarbon substances is greatly improved by combining the use, so that the current situations of difficult treatment and high cost of the existing oily sludge can be well solved, and the application prospect is good.
The composite microbial agent provided by the invention has a good degradation effect on petroleum hydrocarbon substances in the oily sludge, so that oil substances in the oily sludge can be removed, the oil concentration in the oil sludge is reduced, and the oil sludge treatment work is greatly promoted.
After the oily sludge is degraded by the compound microbial agent, the oil-containing concentration of the oily sludge is obviously reduced, and the oily sludge can reach the industrial emission standard, so that the method is an environment-friendly treatment method. The microbial inoculum is applied to the hazardous waste treatment industry, and has good market prospect and economic benefit.
Drawings
FIG. 1 is a schematic diagram of the degradation of oily sludge in a certain oil production plant in northern Shaanxi by applying different microbial agent adding schemes related to example 1 in the invention;
FIG. 2 is a schematic diagram of the degradation of oily sludge of another oil production plant in northern Shaanxi by using different microbial agent adding schemes related to example 2 of the present invention.
Detailed Description
The present invention will be explained in further detail with reference to examples.
The invention provides a compound microbial agent for degrading oil pollutants in oily sludge, which is prepared by mixing a Pseudomonas aeruginosa (Pseudomonas aeruginosa) microbial agent, a Bifidobacterium (Bifidobacterium) microbial agent, a Bacillus subtilis (Bacillus subtilis) microbial agent and a mycobacterium salmonides (Virgibacillus halodenitriica) microbial agent in a volume ratio of (1-4) to (1-4); wherein the Bifidobacterium may be Bifidobacterium anti strain XD80, Bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium breve (Bifidobacterium breves), Bifidobacterium longum (Bifidobacterium longum), Bifidobacterium lactis (Bifidobacterium lactis), Bifidobacterium bifidum (Bifidobacterium bifidum) and Bifidobacterium pseudocatenulatum (Bifidobacterium eudiocin).
The invention also provides a preparation method of the compound microbial agent for degrading oil pollutants in oily sludge, which comprises the following steps:
1) solid slant culture: respectively inoculating pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis into a beef extract peptone solid culture medium, and culturing at constant temperature to fully activate the strains;
2) first-order seed culture: respectively inoculating the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the bacillus thuringiensis activated in the step 1) into a beef extract peptone liquid culture medium, and culturing at constant temperature until OD is obtained600Stopping culturing when the value reaches 0.8, and respectively obtaining primary seed solutions of pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis;
3) secondary seed culture: respectively inoculating the primary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 2) into a fermentation culture medium, and culturing for 2d at 37 ℃ and 130r/min to respectively obtain secondary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium;
4) inoculating the secondary seed solutions of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 3) into a fermentation culture medium respectively in an inoculation amount of 1% of the fermentation culture medium by volume ratio, and performing fermentation culture for 1d at 37 ℃ and 130r/min to respectively obtain a pseudomonas aeruginosa microbial inoculum, a bifidobacterium microbial inoculum, a bacillus subtilis microbial inoculum and a salt denitrogenation mycobacterium microbial inoculum;
5) and mixing the pseudomonas aeruginosa microbial inoculum, the bifidobacterium microbial inoculum, the bacillus subtilis microbial inoculum and the bacillus halodenitrificans microbial inoculum according to the volume ratio of (1-4) to obtain the compound microbial inoculum for degrading the oil pollutants in the oily sludge.
The invention also relates to application of the compound microbial agent for degrading oil pollutants in oily sludge in degrading petroleum substances.
To further illustrate the technical solution of the present invention, the following detailed description is given with reference to specific examples.
Example 1
A compound microbial agent for degrading oil pollutants in oily sludge is prepared by the following steps:
1) solid slant culture: respectively inoculating pseudomonas aeruginosa, Domibacillus anti strain XD80, bacillus subtilis and bacillus thuringiensis into a beef extract peptone solid culture medium, and culturing at the constant temperature of 37 ℃ for 2 days to fully activate the strains; the beef extract peptone solid medium comprises the following components in parts by weight: 0.5 wt% of beef extract, 1.0 wt% of peptone, 0.5 wt% of sodium chloride, 2 wt% of agar and the balance of water, wherein the pH value is 7.0-7.5;
2) first-order seed culture: respectively inoculating the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the bacillus thuringiensis activated in the step 1) into the beefCulturing in liquid peptone medium at 37 deg.C and 130r/min for 2d when OD is reached600Stopping culturing when the value reaches 0.8, and respectively obtaining primary seed solutions of pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis; the beef extract peptone liquid medium comprises the following components in percentage by weight: 0.5 wt% of beef extract, 1.0 wt% of peptone, 0.5 wt% of sodium chloride and the balance of water, wherein the pH value is 7.0-7.5;
3) secondary seed culture: respectively inoculating the primary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 2) into a fermentation culture medium, and culturing for 2d at 37 ℃ and 130r/min to respectively obtain secondary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium;
4) respectively inoculating the secondary seed solutions of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 3) into a fermentation culture medium in an inoculation amount of 1% (volume ratio), and performing fermentation culture for 1D at 37 ℃ and 130r/min to respectively obtain a pseudomonas aeruginosa microbial inoculum, a bifidobacterium microbial inoculum, a bacillus subtilis microbial inoculum and a salt denitrogenation mycobacterium microbial inoculum which are sequentially marked as a microbial inoculum A, a microbial inoculum B, a microbial inoculum C and a microbial inoculum D;
5) mixing a microbial agent A, a microbial agent B, a microbial agent C and a microbial agent D according to the volume ratio of 1:1:1:1 to obtain a compound microbial agent for degrading oil pollutants in oily sludge, and marking as the compound microbial agent;
in the present embodiment, the fermentation medium involved in step 3) and step 4) has a formulation of 1 wt% of molasses, 0.5 wt% of ammonium chloride, 0.3 wt% of monopotassium phosphate, 0.15 wt% of dipotassium phosphate, 0.001 wt% of anhydrous calcium chloride, 0.5 wt% of sodium chloride, 0.01 wt% of magnesium sulfate heptahydrate, and the balance of water, and the pH value of the fermentation medium is 7.0-7.5.
Application example 1
The compound microbial agent for degrading oil pollutants in oily sludge prepared in example 1 is applied to treatment of oily sludge in certain oil production plant in northern Shaanxi.
The microbial agent A, the microbial agent B, the microbial agent C, the microbial agent D and the compound microbial agent prepared in the example 1 are added into the oil sludge degradation liquid according to the adding amount of 10% of the volume of the oil sludge degradation liquid, oil-containing sludge in the oil sludge degradation liquid is degraded, and the scheme 1, the scheme 2, the scheme 3, the scheme 4 and the scheme 5 are marked in sequence; wherein the oil sludge degradation liquid comprises 2 wt% of oil-containing sludge and 98 wt% of degradation culture medium, and the formula of the degradation culture medium is as follows: 0.5% ammonium chloride, 0.3% monopotassium phosphate, 0.15% dipotassium phosphate, 0.001% anhydrous calcium chloride, 0.5% sodium chloride, 0.01% magnesium sulfate heptahydrate, and the balance water, wherein the pH is 7.2;
the test result is shown in fig. 1, the concentration of petroleum hydrocarbon in the initial oily sludge is 75.18mg/g, and after 7 days of degradation, the degradation of petroleum hydrocarbon substances in the oily sludge by adding different microbial inoculum is as follows: in the scheme 1, after being degraded by the microbial inoculum A, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 46.88mg/g, and the degradation rate is 37.65%; in the scheme 2, after degradation by the microbial inoculum B, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 47.62mg/g, and the degradation rate is 36.65%; according to the scheme 3, after degradation by the microbial inoculum C, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 46.57mg/g, and the degradation rate is 38.05%; scheme 4, after degradation by the microbial inoculum D, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 35.34mg/g, and the degradation rate is 53.00%; according to the scheme 5, after the composite microbial agent is degraded, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 25.36mg/g, and the degradation rate is 66.26%. Therefore, the microbial inoculum A, the microbial inoculum B, the microbial inoculum C and the microbial inoculum D have better treatment effect on petroleum hydrocarbon substances, and the degradation effect of the compound microbial inoculum is obviously better than that of a single strain.
Application example 2
The compound microbial agent for degrading oil pollutants in oily sludge prepared in the example 1 is applied to treating oily sludge of another oil production plant in northern Shaanxi.
Different from the application example 1, the concentration of petroleum hydrocarbon in the initial oily sludge of the oil production plant is 231.70mg/g, and as shown in fig. 2, after 7 days of degradation, different microbial inoculum addition can degrade the petroleum hydrocarbon substances in the oily sludge as follows: in the scheme 1, after being degraded by the microbial inoculum A, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 160.82mg/g, and the degradation rate is 30.60%; in the scheme 2, after degradation by the microbial inoculum B, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 197.95mg/g, and the degradation rate is 14.56%; according to the scheme 3, after degradation by the microbial inoculum C, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 163.07mg/g, and the degradation rate is 29.62%; according to the scheme 4, after degradation by the microbial inoculum D, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 148.18mg/g, and the degradation rate is 36.05%; in the scheme 5, after the composite microbial agent is degraded, the concentration of petroleum hydrocarbon in the oil sludge is reduced to 138.18mg/g, and the degradation rate is 40.36%. The degradation effect of the obtained compound microbial agent is obviously better than that of a single strain.
Example 2
Different from the embodiment 1, the composite microbial agent is prepared by mixing a pseudomonas aeruginosa microbial agent, a bifidobacterium microbial agent, a bacillus subtilis microbial agent and a bacillus denitrificans microbial agent in a volume ratio of 1:2:3: 4; wherein the Bifidobacterium is Bifidobacterium bifidum;
the fermentation medium adopted for preparing the compound microbial agent comprises the following components in percentage by mass: 0.3 percent of ammonium chloride, 0.3 percent of monopotassium phosphate, 0.1 percent of dipotassium phosphate, 0.001 percent of anhydrous calcium chloride, 0.3 percent of sodium chloride, 0.01 percent of magnesium sulfate heptahydrate, 1 percent of molasses and the balance of water, and the pH value is 7.0.
Example 3
Different from the embodiment 1, the composite microbial agent is prepared by mixing a pseudomonas aeruginosa microbial agent, a bifidobacterium microbial agent, a bacillus subtilis microbial agent and a bacillus denitrificans microbial agent in a volume ratio of 1:2:4: 4; wherein the Bifidobacterium is specifically Bifidobacterium longum;
the fermentation medium adopted for preparing the compound microbial agent comprises the following components in percentage by mass: 0.5 percent of ammonium chloride, 0.5 percent of monopotassium phosphate, 0.3 percent of dipotassium phosphate, 0.02 percent of anhydrous calcium chloride, 1 percent of sodium chloride, 0.02 percent of magnesium sulfate heptahydrate, 1.5 percent of molasses and the balance of water, and the pH value is 7.2.
Example 4
Different from the embodiment 1, the composite microbial agent is prepared by mixing a pseudomonas aeruginosa microbial agent, a bifidobacterium microbial agent, a bacillus subtilis microbial agent and a bacillus denitrificans microbial agent in a volume ratio of 4:3:2: 1; wherein the Bifidobacterium is specifically Bifidobacterium breve;
the fermentation medium adopted for preparing the compound microbial agent comprises the following components in percentage by mass: 0.4% of ammonium chloride, 0.4% of monopotassium phosphate, 0.2% of dipotassium phosphate, 0.01% of anhydrous calcium chloride, 0.5% of sodium chloride, 0.015% of magnesium sulfate heptahydrate, 1.2% of molasses and the balance of water, wherein the pH value is 7.3.
Example 5
Different from the embodiment 1, the composite microbial agent is prepared by mixing a pseudomonas aeruginosa microbial agent, a bifidobacterium microbial agent, a bacillus subtilis microbial agent and a mycobacterium haloperi microbial agent in a volume ratio of 4:1:4: 1; wherein the Bifidobacterium is specifically Bifidobacterium adolescentis;
the fermentation medium adopted for preparing the compound microbial agent comprises the following components in percentage by mass: 0.3 percent of ammonium chloride, 0.4 percent of monopotassium phosphate, 0.2 percent of dipotassium phosphate, 0.015 percent of anhydrous calcium chloride, 0.8 percent of sodium chloride, 0.016 percent of magnesium sulfate heptahydrate, 1.4 percent of molasses and the balance of water, and the pH value is 7.5.
Claims (6)
1. The compound microbial agent for degrading oil pollutants in oily sludge is characterized by being prepared by mixing a Pseudomonas aeruginosa (Pseudomonas aeruginosa) microbial agent, a Bifidobacterium (Bifidobacterium) microbial agent, a Bacillus subtilis microbial agent and a mycobacterium salmonides (Virgibacter halodendrobii) microbial agent in a volume ratio of (1-4) to (1-4).
2. The preparation method of the compound microbial agent for degrading oil pollutants in oily sludge according to claim 1, which comprises the following steps:
1) solid slant culture: respectively inoculating pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis into a beef extract peptone solid culture medium, and culturing at constant temperature to fully activate the strains;
2) first-order seed culture: respectively inoculating the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the bacillus thuringiensis activated in the step 1) into a beef extract peptone liquid culture medium, and culturing at constant temperature until OD is obtained600Stopping culturing when the value reaches 0.8, and respectively obtaining primary seed solutions of pseudomonas aeruginosa, bifidobacterium, bacillus subtilis and bacillus thuringiensis;
3) secondary seed culture: respectively inoculating the primary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 2) into a fermentation culture medium, and culturing for 2d at 37 ℃ and 130r/min to respectively obtain secondary seed liquid of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium;
4) inoculating the secondary seed solutions of the pseudomonas aeruginosa, the bifidobacterium, the bacillus subtilis and the salt denitrogenation mycobacterium obtained in the step 3) into a fermentation culture medium respectively in an inoculation amount of 1% of the fermentation culture medium by volume ratio, and performing fermentation culture for 1d at 37 ℃ and 130r/min to respectively obtain a pseudomonas aeruginosa microbial inoculum, a bifidobacterium microbial inoculum, a bacillus subtilis microbial inoculum and a salt denitrogenation mycobacterium microbial inoculum;
5) and mixing the pseudomonas aeruginosa microbial inoculum, the bifidobacterium microbial inoculum, the bacillus subtilis microbial inoculum and the bacillus halodenitrificans microbial inoculum according to the volume ratio of (1-4) to obtain the compound microbial inoculum for degrading the oil pollutants in the oily sludge.
3. The method for preparing the compound microbial inoculant for degrading oil pollutants in oily sludge according to claim 2, wherein the fermentation medium in the step 3) and the fermentation medium in the step 4) comprise the following components in percentage by mass: 0.3 to 0.5 percent of ammonium chloride, 0.3 to 0.5 percent of monopotassium phosphate, 0.1 to 0.3 percent of dipotassium phosphate, 0.001 to 0.02 percent of anhydrous calcium chloride, 0.3 to 1 percent of sodium chloride, 0.01 to 0.02 percent of magnesium sulfate heptahydrate, 1 to 1.5 percent of molasses and the balance of water, and the pH value is 7.0 to 7.5.
4. The use of the complex microbial inoculant according to claim 1 for degrading oil pollutants in oily sludge in the degradation of petroleum substances.
5. The use of claim 4, wherein the amount of the compound microbial agent for degrading oil pollutants in oily sludge added is 10% of the volume of the degradation liquid of oily sludge when the oily sludge is treated by the compound microbial agent.
6. The application of claim 5, wherein the oil sludge degradation liquid comprises 2% of oil-containing sludge by mass and 98% of degradation medium by mass, and the degradation medium comprises the following components by mass percent: 0.3% -0.5% of ammonium chloride, 0.3% -0.5% of monopotassium phosphate, 0.1% -0.3% of dipotassium phosphate, 0.001% -0.02% of anhydrous calcium chloride, 0.3% -1% of sodium chloride, 0.01% -0.02% of magnesium sulfate heptahydrate, and the balance of water, wherein the pH value of the degradation culture medium is 7.0-7.5.
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