CN105907679A - Composition or composite bacterium agent for treating waste well drilling mud - Google Patents

Composition or composite bacterium agent for treating waste well drilling mud Download PDF

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Publication number
CN105907679A
CN105907679A CN201610307356.9A CN201610307356A CN105907679A CN 105907679 A CN105907679 A CN 105907679A CN 201610307356 A CN201610307356 A CN 201610307356A CN 105907679 A CN105907679 A CN 105907679A
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culture
days
bacillus subtilis
pseudomonas stutzeri
mud
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CN105907679B (en
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陈五岭
于烽
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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

Abstract

The invention provides a composition or composite bacterium agent for treating waste well drilling mud. The invention provides a composition, a composite bacterium agent and application thereof. The composition or the composite bacterium agent comprises bacillus subtilis, pseudomonas stutzeri and acinetobacter johnsonii and is used for treating the waste well drilling mud.

Description

For processing compositions or the composite bacteria agent capable of waste drilling mud
Technical field
The application belongs to oil gas field environmental protection technical field.Specifically, the application relates to place The compositions of reason waste drilling mud or composite bacteria agent capable.
Background technology
The discarded slurry discharged in shale gas drilling process is to be added by crude oil, oil-base mud, mud Add the mixture of agent composition.As water-base mud, mud system comprises mineral oil, oil sulphur Change acid, Calx, alkyl benzene calcium sulfonate, calcium chloride, sodium dichromate, Caustic soda, sodium hexameta phosphate, Sulfonated-methyl-tannin, ferrochrome lignosulfonate, oxidized asphalt, chromium brown coal, sulfonated-pheno-formoldehyde resin, Organic and inorganic compound, petroleum pollution and the heavy metal ion such as sodium polyacrylate, sulfonation tannin Deng harmful substance.Be different from water-base mud with water as disperse medium, oil-base mud using oil as disperseing Medium, is a kind of complicated heterogeneous system containing mineral oil, phenolic compound etc., major pollutants Coming from the base oil of oil-base mud and major-minor breast mud additive base, the most conventional oil base includes bavin Oil base, white oil base and synthesis base oil base mesh.Aromatic hydrocarbon in base oil diesel oil has bigger biological poison Property, region environment will be caused significant impact, than water base mud if the most effectively processed Starch the most difficult.
Under field conditions (factors), above-mentioned all kinds of harmful substances are difficult to degrade, serious harm soil ecology ring Border.Later stage process will be carried out according to environmental requirement, discarded oil-base mud and oil-base mud drilling cuttings. Process discarded oil-base mud traditionally and oil-based drill cuttings mode has burning disposal, cracking to process, biology Degraded etc..The processing method of waste drilling mud mainly has at present: simple process landfill, re-injection peace Full stratum, solidification, soil cultivating etc., the most most widely used is first to enter waste drilling mud After row cured, more progressive comprehensive utilization.
Along with the exploitation upsurge of Unconventional forage-shale gas that China starts, the thing followed a few days ago It is exactly exploration and the exploitation in a large amount of shale gas field, owing to shale gas exploitation must use oil-base mud, This will certainly increase the generation amount of discarded oil-base mud.To this end, shale gas field Discarded Drilling Materials oil base mud The economical and effective of slurry processes the individual hot subject becoming the most urgently to be resolved hurrily.
Summary of the invention
On the one hand, the application provides a kind of compositions, comprises bacillus subtilis (Bacillus Subtilis), Pseudomonas stutzeri (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii)。
On the other hand, the application provides a kind of composite bacteria agent capable, and it comprises bacillus subtilis (Bacillus Subtilis), Pseudomonas stutzeri (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii)。
In one embodiment, described composite bacteria agent capable is made up of above-mentioned three kinds of bacterial strains.
In one embodiment, described composite bacteria agent capable comprises the culture of above-mentioned three kinds of bacterial strains.
On the other hand, the application further relates to the preparation method of a kind of composite bacteria agent capable, including:
By bacillus subtilis (Bacillus subtilis), Pseudomonas stutzeri (Pseudomonas Stutzeri) cultivate respectively with Acinetobacter johnsonii (Acinetobacter johnsonii)；And
Obtain the respective culture of described bacterial strain, and described culture is carried out according to combinations thereof Mixing, obtains described composite bacteria agent capable.
Optionally, above-mentioned cultivation is solid culture, Semi-solid cell culture or liquid culture.
On the other hand, the application further relates to a kind of process or the method for aid in treatment waste drilling mud, Including:
Described compositions or described composite bacteria agent capable are put in pending waste drilling mud and processes At least 20 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, extremely Few 50 days, at least 55 days, at least 60 days, at least 65 days, at least 70 days, at least 75 days, at least 80 days, at least 85 days, at least 90 days, at least 95 days, at least 100 days, at least 105 days, at least 110 days, at least 115 days, at least 120 days, at least 125 days, at least 130 days, at least 135 days, At least 140 days, at least 145 days, at least 150 days, at least 155 days, at least 160 days, at least 165 My god, at least 170 days, at least 175 days or at least 180 days.
In one embodiment, described compositions or described composite bacteria agent capable are put into pending discarding Drilling mud is carried out process at least 30 days or at least 35 days.
In one embodiment, described waste drilling mud is discarded oil-base mud or waste water-base Mud.
In one embodiment, described discarded oil-base mud is to discharge in shale gas drilling process Discarded oil-base mud.
In some embodiments, ambient temperature during process be at least 10 DEG C, at least 15 DEG C, extremely Few 20 DEG C, at least 25 DEG C, at least 30 DEG C or at least 35 DEG C, it is therefore preferable at least 12～30 DEG C, 16～28 DEG C or 16～30 DEG C.During process, the temperature in described pending waste drilling mud compares environment Temperature is low at least 5 DEG C, at least 8 DEG C, at least 10 DEG C, at least 12 DEG C, at least 13 DEG C, at least 14 DEG C Or at least 15 DEG C.
In some embodiments, ambient temperature is the highest, and the temperature in waste drilling mud is the highest, Strain growth is the rapidest, processes the time the shortest.The north process time, typically in May～JIUYUE, works as ring When border temperature is at least 20 DEG C, the time of process is at least 40～50 days；When ambient temperature is at least When 25 DEG C, the time of process is at least 30～40 days；When ambient temperature is at least 30 DEG C, process Time is at least 20～30 days.
In some embodiments, the pH value of described pending waste drilling mud be at least 7, extremely Few 8, at least 9, at least 10, at least 11, at least 12 or at least 13.
In some embodiments, the optimum pH of the growth of bacterial strain described in composite bacteria agent capable 7～ Between 9.
In some embodiments, described compositions or described composite bacteria agent capable are pending relative to described The inoculum concentration of waste drilling mud is at least 0.5%, at least 0.7%, at least 1%, at least 3%, at least 5% or at least 7%w/v.
In one embodiment, described compositions or described composite bacteria agent capable are pending relative to described The inoculum concentration of waste drilling mud is at least 0.5% or at least 0.7%w/v.
On the other hand, the application obtains after further relating to process waste drilling mud by method disclosed herein To solid content waste drilling mud repair in application.
Accompanying drawing explanation
Fig. 1 is the photo of shale gas well waste drilling mud pond processing procedure, before wherein A is for processing, B terminates for stirring after adding composite bacteria agent capable for whipping process after addition composite bacteria agent capable, C, and D is for adding Composite bacteria agent capable post processing the 7th day, E is for adding composite bacteria agent capable post processing the 18th day, and F is for adding compound bacteria Agent post processing the 30th day.
Fig. 2 is the photo of oil well waste drilling mud pond processing procedure, and before wherein A is for processing, B is Adding whipping process after composite bacteria agent capable, C terminates for stirring after adding composite bacteria agent capable, and D is compound for adding Microbial inoculum post processing the 10th day, E is for adding composite bacteria agent capable post processing the 18th day, and F is for adding composite bacteria agent capable Post processing the 35th day.
Fig. 3 is the photo of gas well waste drilling mud pond processing procedure, and before wherein A is for processing, B is Adding whipping process after composite bacteria agent capable, C terminates for stirring after adding composite bacteria agent capable, and D is compound for adding Microbial inoculum post processing the 5th day, E is for adding composite bacteria agent capable post processing the 15th day, after F is for adding composite bacteria agent capable Process the 30th day.
Describe in detail
There is provided defined below and method in order to preferably to define the application and to put into practice middle finger in the application Lead those of ordinary skill in the art.Unless otherwise mentioned, term is according to association area ordinary skill people The common usage of member understands.All patent documentations, scientific paper and other disclosures cited herein Publication, full content therein is incorporated herein by reference.
Definition
Term as used herein " compositions ", refers to comprise two or more microorganism fungus kind Mixture, or be made up of described microorganism fungus kind.If it is required, described compositions can also be wrapped Include other be conducive to described bacterial strain deposit or the material of activity, such as, culture medium, trace element, Vitamin, aminoacid, meat soup etc..
Term as used herein " composite bacteria agent capable ", refers to comprise two or more and mutual the most not antagonism The microorganism formulation made of microorganism fungus kind, described composite bacteria agent capable becomes with described strain for activity Point, it is also possible to additionally comprise carrier or excipient, it is also possible to comprise other is conducive to described bacterium Strain is deposited or the material of activity, such as, and culture medium, trace element, vitamin, aminoacid, meat Soup etc..In one embodiment, the general compatibility of described microbial inoculum is reasonable.
Term as used herein " culture ", refers under specific process conditions control by microbial bacteria Plant the microbial product formed the most afterwards in specific culture medium, after it mainly comprises fermentation Microorganism fungus kind cell mass, microorganism fungus kind extracellular metabolic product and culture medium.
Term as used herein " drilling mud ", i.e. drilling fluid, refer in drilling process, be used for clear At the bottom of well-flushing, cutting carring to bottom surface, maintain the circulation flushing fluid that is normally carried out of drill-well operation, one As for oil and natural gas probing in.
Term as used herein " oil-base mud ", refers to comprise crude oil, oil-base mud, mud interpolation The mixture of agent, using oil as disperse medium, mud system comprises mineral oil, oil sulfonated acid, Calx, alkyl benzene calcium sulfonate, calcium chloride, sodium dichromate, Caustic soda, sodium hexameta phosphate, sulphur methyl Tannin, ferrochrome lignosulfonate, oxidized asphalt, chromium brown coal, sulfonated-pheno-formoldehyde resin, poly-third The organic and inorganic compound such as olefin(e) acid sodium, sulfonation tannin, petroleum pollution and heavy metal ion etc. have Pest matter；Major pollutants come from base oil and major-minor breast mud additive base, the mesh of oil-base mud Front conventional oil base includes diesel oil base, white oil base and synthesis base oil base mesh.
Term as used herein " water-base mud ", refers to water as disperse medium, comprises basicity regulation Agent, fluid loss additive, flocculant, coating agent, inhibitor, lubricant, anti-sloughing agent, flow pattern regulation Material, leak stopping material, soil powder or weighting material etc. are protected in agent, oil.
Term as used herein " pH ", also known as hydrogen ionexponent, pH-value, refer in solution The ratio of the amount of hydrionic sum and total material, represents solution acidic or the numerical value of degree of alkalinity, i.e. The negative value of the common logarithm of contained hydrogen ion concentration.
Term as used herein " five-day BOD (BOD5) ", refer to that microorganism is at 20 DEG C Time, under, the condition determination that do not stirs sufficient at oxygen, during using standard as mensuration BOD on the 5th Between, some in the microbial decomposition certain volume water of mensuration can oxidized material, particularly have Machine material, the quantity of the dissolved oxygen consumed, represent with mg/litre or percentage rate, ppm.
Term as used herein " COD (COD) ", refers under certain condition, with certain The amount of oxidant that consumed when processing water sample of strong oxidizer, represent with the mg/litre of oxygen； Utilize chemical oxidizing agent, the reducing substances in water sample is aoxidized, then from remaining oxidation The gauge of agent calculates the consumption of oxygen.
Term as used herein " sulfide ", refers to the organic compound containing divalent sulfur, such as Hydrogen sulfide (H2S), thioether (R-S-R), thiophenol/mercaptan (Ar/R-SH), thioaldehydes (R-CSH), Thiocarboxylic acid and disulphide (R-S-S-R) etc..
Can term as used herein " volatile phenol ", steam according to phenols together with steam, can Being divided into volatile phenol and non-volatile phenol, volatile phenol refers to boiling point phenols below 230 DEG C.
Term as used herein " ammonia nitrogen ", refers in water with free ammonia (NH3) and ammonium ion (NH4+) form exist nitrogen.
Term as used herein " cyanide ", refers to the compound with cyano group (CN), wherein Carbon atom be connected by three key with nitrogen-atoms, be divided into inorganic cyanide and organic cyanide； Inorganic cyanide refers to include the inorganic salt of cyanide ion (CN-), common are potassium cyanide And Cyanogran.；Organic cyanide, is to be combined into other carbon atom by singly-bound by cyano group, Depending on the difference of combination, organic cyanide can be categorized as nitrile (C-CN) and isonitrile (C-NC), Accordingly, cyano group is referred to alternatively as itrile group (-CN) or isonitrile base (-NC).
Term as used herein " petroleum-type ", refers to various hydro carbons (alkane, alkene, alkynes, ring Alkane, aromatic hydrocarbon) mixture, waste water can be present in the dissolved, emulsifying state and dispersed In, refer to during detection by carbon tetrachloride extraction and not by the material of adsorption of magnesium silicate.
Term as used herein " solid content ", refers to destroy the colloform texture of drilling mud, converts Petroleum hydrocarbon in mud and heavy metal, the material of mummification after making the moisture in mud evaporate.
Detailed description of the invention
On the one hand, the application provides a kind of compositions, comprises bacillus subtilis (Bacillus Subtilis), Pseudomonas stutzeri (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii)。
On the other hand, the application provides a kind of composite bacteria agent capable, and it comprises bacillus subtilis (Bacillus Subtilis), Pseudomonas stutzeri (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii)。
In one embodiment, described composite bacteria agent capable is made up of above-mentioned three kinds of bacterial strains.
In one embodiment, described composite bacteria agent capable comprises the culture of above-mentioned three kinds of bacterial strains.
In some embodiments, total living bacteria count in described compositions or in described composite bacteria agent capable ≥1×108cfu/g。
In some embodiments, in terms of the weight of described strain culture, wherein said compositions In, or bacillus subtilis culture described in described composite bacteria agent capable, Pseudomonas stutzeri culture With the weight portion of Acinetobacter johnsonii culture it is:
Bacillus subtilis culture 1～3
Pseudomonas stutzeri culture 1～6
Acinetobacter johnsonii culture 1～6,
Optionally, described strain culture comprises bacterial strain and culture medium；
Optionally, effective viable bacteria of bacillus subtilis described in described bacillus subtilis culture Number is for >=1 × 108cfu/g；Described in described Pseudomonas stutzeri culture, Pseudomonas stutzeri is effective Viable count is >=1 × 108cfu/g；Acinetobacter johnsonii described in described Acinetobacter johnsonii culture Living bacteria count is >=1 × 108cfu/g；
Optionally, described culture medium is solid medium, semisolid culturemedium or fluid medium.
In one embodiment, in described compositions or hay spore described in described composite bacteria agent capable The weight ratio of alphacterium culture, Pseudomonas stutzeri culture and Acinetobacter johnsonii culture is 1:2:2,1:1.5:2 or 1:1:1.
On the other hand, the application further relates to the preparation method of a kind of composite bacteria agent capable, including:
By bacillus subtilis (Bacillus subtilis), Pseudomonas stutzeri (Pseudomonas Stutzeri) cultivate respectively with Acinetobacter johnsonii (Acinetobacter johnsonii)；And Obtain the respective culture of described bacterial strain, and described culture is carried out according to combinations thereof Mixing, obtains described composite bacteria agent capable.
Optionally, above-mentioned cultivation is solid culture, Semi-solid cell culture or liquid culture.
In some embodiments, in the preparation method of described composite bacteria agent capable, each with described bacterial strain From the weight meter of culture mix, described bacillus subtilis culture, Amur during mixing The weight portion of pseudomonas culture and Acinetobacter johnsonii culture is:
Bacillus subtilis culture 1～3
Pseudomonas stutzeri culture 1～6
Acinetobacter johnsonii culture 1～6；
Optionally, described strain culture comprises bacterial strain and culture medium；
Optionally, effective viable bacteria of bacillus subtilis described in described bacillus subtilis culture Number is for >=1 × 108cfu/g；Described in described Pseudomonas stutzeri culture, Pseudomonas stutzeri is effective Viable count is >=1 × 108cfu/g；Acinetobacter johnsonii described in described Acinetobacter johnsonii culture Living bacteria count is >=1 × 108cfu/g；
Optionally, total living bacteria count >=1 × 10 in described composite bacteria agent capable8cfu/g；
Optionally, described culture medium is solid medium, semisolid culturemedium or fluid medium.
In one embodiment, in the preparation method of described composite bacteria agent capable, each with described bacterial strain From the weight meter of culture mix, described bacillus subtilis culture, Amur during mixing The weight ratio of pseudomonas culture and Acinetobacter johnsonii culture is 1:2:2,1:1.5:2 or 1:1:1.
Strain used herein is known strain, can by routine screening, commercial means or other Approach obtains.
In one embodiment, can be the most withered by being either commercially available strain as herein described Grass bacillus cereus can be bacillus subtilis (Bacillus subtilis), selectively, and can be from Xi'an Zi Rui bio tech ltd (address: eastern section, Mei Wu main road, new district, riverfront, Mei County, Shaanxi Province, postcode: 722300) above-mentioned bacterial strains is obtained.
In one embodiment, such as can execute by being either commercially available strain as herein described Family name pseudomonas can be Pseudomonas stutzeri (Pseudomonas stutzeri), selectively, and can be from Zi Rui bio tech ltd, Xi'an (address: eastern section, Mei Wu main road, new district, riverfront, Mei County, Shaanxi Province, Postcode: 722300) obtain above-mentioned bacterial strains.
In one embodiment, can be by being either commercially available strain as herein described, e.g., from about Family name's acinetobacter calcoaceticus Ke Yishiyueshi acinetobacter calcoaceticus (Acinetobacter johnsonii), selectively, can From Zi Rui bio tech ltd, Xi'an (address: eastern section, Mei Wu main road, new district, riverfront, Mei County, Shaanxi Province, Postcode: 722300) obtain above-mentioned bacterial strains.
In some embodiments, the solid medium of the solid culture for cultivating described strain can For bran mass, potato culture, Testa oryzae culture medium, soybean cake powder culture medium or peanut cake powder Culture medium.
In some embodiments, the fluid medium of the liquid culture for cultivating described strain can For beef extract-peptone fluid medium, seed culture medium (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) or LB liquid Culture medium.
In some embodiments, for cultivating the Semi-solid cell culture of the Semi-solid cell culture of described strain Base can be liquid medium within adds the preparation of a small amount of coagulator (agar such as 0.2%～0.5%) and Become.
Culture mainly (such as enzyme, polysaccharide, lipid, is had by microorganism fungus kind extracellular metabolic product Machine acid etc.), fermentation after culture medium and microorganism fungus kind cell mass constitute.
In one embodiment, the preparation method of described composite bacteria agent capable includes at least one side following Method or step:
1) shake-flask culture: by bacillus subtilis, Pseudomonas stutzeri and Acinetobacter johnsonii respectively It is inoculated in beef extract-peptone fluid medium and carries out shake-flask culture, all under the conditions of 25～35 DEG C Cultivate 24～48 hours, obtain bacillus subtilis shaking flask strain, Pseudomonas stutzeri shaking flask respectively Strain and Acinetobacter johnsonii shaking flask strain.
2) seed tank culture: by step 1) in bacillus subtilis shaking flask strain, Amur vacation unit cell Bacterium shaking flask strain, Acinetobacter johnsonii shaking flask strain be inoculated into respectively seed culture medium (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out seed tank culture, under the conditions of 25～35 DEG C, all cultivate 24～48 little Time, obtain bacillus subtilis seed liquor, Pseudomonas stutzeri seed liquor and about family name's not lever respectively Bacterium seed liquor.
3) fermentor cultivation: by step 2) in bacillus subtilis seed liquor, Pseudomonas stutzeri kind Sub-liquid and Acinetobacter johnsonii seed liquor in 3～10% ratio (percent by volume) inoculum concentration inoculate respectively (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) to seed culture medium Potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out fermentor cultivation, all under the conditions of 25～35 DEG C Cultivate 24 hours or 48 hours, obtain bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacterium respectively Suspension and Acinetobacter johnsonii bacteria suspension；Respectively bacteria suspension is carried out microscopy, bacterial strain living bacteria count It is >=1 × 109cfu/mL。
4) solid culture: by step 3) in bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacterium hang Liquid and Acinetobacter johnsonii bacteria suspension are inoculated into wheat bran training respectively in 5～10% (percentage by weight) ratio Support base (consisting of: 10% (w) rice husk, 5% (w) bean cake powder and 85% (w) wheat bran (such as Testa Tritici)) In carry out solid culture, cultivation temperature is 25～35 DEG C, and incubation time is 24 hours, 48 hours Or 72 hours, obtain bacillus subtilis solid culture, Pseudomonas stutzeri solid culture respectively Thing and Acinetobacter johnsonii solid culture；Respectively solid culture is carried out microscopy, hay spore In bacillus solid culture, the living bacteria count of bacillus subtilis is >=1 × 108cfu/g；Amur vacation list In born of the same parents' bacterium solid culture, the living bacteria count of Pseudomonas stutzeri is >=1 × 108cfu/g；Yue Shi is motionless In bacillus solid culture, the living bacteria count of Acinetobacter johnsonii is >=1 × 108cfu/g；
Containing B. subtilis cell group, bacillus subtilis in bacillus subtilis solid culture Bacterium metabolite (such as lipopeptid, protease, lipase, cellulase, α-amylase, lactic acid etc.) With the bran mass after fermentation；Containing Pseudomonas stutzeri in Pseudomonas stutzeri solid culture Wheat bran training after cell mass, Pseudomonas stutzeri metabolite (such as lipase, oxidase) and fermentation Support base；In Acinetobacter johnsonii solid culture motionless containing Acinetobacter johnsonii cell mass, Yue Shi Bran mass after bacillus metabolite (such as lipase) and fermentation.
5) by above-mentioned bacillus subtilis solid culture, Pseudomonas stutzeri solid culture peace treaty Family name's acinetobacter calcoaceticus solid culture is dried respectively, be dried can be dust dry, be vacuum dried or Fluid bed drying, respectively obtains bacillus subtilis solid fungicide, Pseudomonas stutzeri solid fungicide With Acinetobacter johnsonii solid fungicide；In bacillus subtilis solid fungicide, bacillus subtilis has Effect viable count is >=1 × 108cfu/g；In Pseudomonas stutzeri solid fungicide, Pseudomonas stutzeri is effective Viable count is >=1 × 108cfu/g；In Acinetobacter johnsonii solid fungicide, the effective of Acinetobacter johnsonii lives Bacterium number is >=1 × 108cfu/g。
6) by bacillus subtilis solid fungicide, Amur acinetobacter calcoaceticus solid fungicide and about family name's not lever Bacterium solid fungicide mixes according to the percentage by weight of 1:2:2,1:1.5:2 or 1:1:1, is combined Microbial inoculum；Total living bacteria count >=1 × 10 in composite bacteria agent capable8cfu/g。
On the other hand, the application further relates to a kind of process or the method for aid in treatment waste drilling mud, Including:
Described compositions or described composite bacteria agent capable are put in pending waste drilling mud and processes At least 20 days, at least 25 days, at least 30 days, at least 35 days, at least 40 days, at least 45 days, extremely Few 50 days, at least 55 days, at least 60 days, at least 65 days, at least 70 days, at least 75 days, at least 80 days, at least 85 days, at least 90 days, at least 95 days, at least 100 days, at least 105 days, at least 110 days, at least 115 days, at least 120 days, at least 125 days, at least 130 days, at least 135 days, At least 140 days, at least 145 days, at least 150 days, at least 155 days, at least 160 days, at least 165 My god, at least 170 days, at least 175 days or at least 180 days.
In one embodiment, described compositions or described composite bacteria agent capable are put into pending discarding Drilling mud is carried out process at least 30 days or at least 35 days.
In some embodiments, ambient temperature during process be at least 10 DEG C, at least 15 DEG C, extremely Few 20 DEG C, at least 25 DEG C, at least 30 DEG C or at least 35 DEG C, it is therefore preferable at least 12～30 DEG C, 16～28 DEG C or 16～30 DEG C.During process, the temperature in described pending waste drilling mud compares environment Temperature is low at least 5 DEG C, at least 8 DEG C, at least 10 DEG C, at least 12 DEG C, at least 13 DEG C, at least 14 DEG C Or at least 15 DEG C.
In some embodiments, ambient temperature is the highest, and the temperature in waste drilling mud is the highest, Strain growth is the rapidest, processes the time the shortest.The north process time, typically in May～JIUYUE, works as ring When border temperature is at least 20 DEG C, the time of process is at least 40～50 days；When ambient temperature is at least When 25 DEG C, the time of process is at least 30～40 days；When ambient temperature is at least 30 DEG C, process Time is at least 20～30 days.
In some embodiments, the pH value of described pending waste drilling mud be at least 7, at least 8, at least 9, at least 10, at least 11, at least 12 or at least 13.
In some embodiments, the optimum pH of the growth of bacterial strain described in composite bacteria agent capable is 7～9 Between.
In some embodiments, described compositions or described composite bacteria agent capable are pending relative to described The inoculum concentration of waste drilling mud is at least 0.5%, at least 0.7%, at least 1%, at least 3%, at least 5% or at least 7%w/v.
In one embodiment, described compositions or described composite bacteria agent capable are pending relative to described The inoculum concentration of waste drilling mud is at least 0.5% or at least 0.7%w/v.
In one embodiment, described waste drilling mud is discarded oil-base mud or water-base mud.
In one embodiment, described discarded oil-base mud is to discharge in shale gas drilling process Discarded oil-base mud.
In some embodiments, the water content of described waste drilling mud be at least 50%, at least 60%, at least 70%, at least 80% or at least 90%.
In one embodiment, the water content of described waste drilling mud is at least 80% or at least 90%.
Mud can be divided into oil-base mud and water-base mud.Oil-base mud mainly has two big class: Yi Zhongshi Oil phase drilling fluid, is oxidized asphalt, Organic Acid and Base, stabilizer and the mixture of high-flash diesel oil, The water of the most mixed 3%～5%；Another kind is water in oil emulsion drilling fluid (anti-phase drilling fluid), has Various additives are used to make water emulsification and stablize, and this system High water cut is up to 50%.Oil base mud Slurry be mainly used in bore complicated and troublesome shale, bore high temperature deep well, production layer creep into coring, Brill salt deposit, anhydrite formation, carnallite layer, the drilling fluid of directional well, the drilling fluid of slim-hole, brill contain H2S and CO2Stratum, perforation and completion fluid, the soak of unfreezing, packer fluids, workover fluid, anti- Corrosion control liquid, casing packing fluid etc..The oil-base mud that above-mentioned operation is discharged is discarded oil base mud Slurry, can be shale gas drilling well, shale oil drilling well, sandstone pneumatic drill well, sandstone oil drilling well, Fractured The discarded oil base discharged in carbonate buried hill pneumatic drill well, fractured carbonate rock buried hill oil drilling process Mud.
In some embodiments, dumping mud treatment terminates to be based on the concrete physical and chemical index of mud Carry out judging, be typically reduced to a certain degree table at the bottom of (such as less than 45%), pond with mud moisture content Face occurs that dry and cracked, people station will not be sunk etc. as apparent criterion up, it is also possible in water content Dropping to and process with regard to stopping after exposing pond basal surface, follow-up employing additive method processes.Owing to not having Unification of the motherland standard, processes qualified specific targets and is specified by examination side.
In one embodiment, ambient temperature is 12～30 DEG C, 16～28 DEG C or 16～30 DEG C, The outdoor process in pending waste drilling mud original place, the water content of pending waste drilling mud is 80%～90% or at least 90%, the pH value of pending waste drilling mud is 13 or 11～12.Throwing The plant equipment such as high-front shovel are used waste drilling mud to be stirred, so before adding composite bacteria agent capable After by composite bacteria agent capable and pending waste drilling mud according at least 0.5% or at least 0.7% w/v Ratio is thrown in pending waste drilling mud, uses the plant equipment such as high-front shovel to stir Mixing, process 30 days or 35 days, the mud water content after processing at the end of process is down to less than 45%, place In mud pit after reason, solid phase solid comprcssive strength reaches 1MPa, and earth's surface pollutant reach harmless safety Landfill disposal requirement.
On the other hand, the application obtains after further relating to process waste drilling mud by method disclosed herein To solid content waste drilling mud repair in application.
Testing index and assay method in the solid content obtained after process and leachate thereof are as follows:
PH value measures: the mensuration glass electrode method of GB 6920-1986 water pH value；
Five-day BOD (BOD5) measure: HJ 505-2009 water quality five-day BOD (BOD5) mensuration dilution and inocalation method；
COD (COD) measures: the mensuration weight of GB 11914-1989 water chemical oxygen demand Chromate process；
Sulfide content measures: the mensuration iodimetric titration of HJ/T 60-2000 water quality sulfide；
Volatile phenol content measures: the mensuration 4-AA of HJ 503-2009 water quality volatile phenol Spectrophotography；
Ammonia-nitrogen content measures: the mensuration distillation-neutralization titration of HJ 537-2009 Water quality ammonia nitrogen；
Total cyanide assay: the mensuration volumetric method of HJ 484-2009 water quality cyanide and light splitting Photometry；
Petroleum content measures: the mensuration of HJ 637-2012 water-quality petroleum and animals and plants oils is red Outer spectrophotography；
Copper, zinc-content determination: GB/T 17138-1997 soil quality copper, Direct spectrophotometry flame are former Sub-absorptiometry；
Cadmium content measures: GB/T 17141-1997 soil quality lead, Cadmium detrmination sampling Graphite Furnace Atomic Absorptiometry；
Chrome determination: the mensuration Flame Atomic Absorption Spectrometry of the total chromium of GB/T 17137-1997 soil quality Spectrophotography；
Mercury Determination: GB/T 22105.1-2008 soil quality total mercury, total arsenic, the survey of total lead Determine atomic fluorescence method part 1: the mensuration of Mercury In Soil；
Arsenic content: GB/T 22105.2-2008 soil quality total mercury, total arsenic, the survey of total lead Determine atomic fluorescence method part 2: the mensuration of total Arsenic In Soil；
Determination of Pb: GB/T 22105.3-2008 soil quality total mercury, total arsenic, the survey of total lead Determine atomic fluorescence method third portion: the mensuration of total lead in soil；
Nickel assay: GBT 17139-1997 soil quality nickel-silicon nitride Flame Atomic Absorption Spectrometry divides Light photometry；
Boron contents measures: NY/T 149-1990 Effective Boron of Soil assay method.
Barium assay: the mensuration atomic absorption spectrphotometry of GB/T 15506-1995 water quality barium Method；
Determination of Selenium: the mensuration sampling Graphite Furnace Atomic Absorption of GB/T 15505-1995 water quality selenium is divided Light photometry.
The reference standard of the innoxious security landfill that waste drilling mud processes is soil environment quality mark Quasi-GB15618-2008, three grades of Soil standards.
Therefore, compositions or the composite bacteria agent capable for processing waste drilling mud that the application provides, There is at least one advantage following:
(1), in the compositions of the application or composite bacteria agent capable, each bacterial strain adapts to natural environment, is inoculated into In waste drilling mud, institute's selection-breeding bacterial strain all can be survived, and carries out metabolic activity and Discarded Drilling Materials mud Slurry processes.
(2), in the compositions of the application or composite bacteria agent capable, each bacterial strain is dominant strain, adapts to Waste drilling mud environment, its metabolite, such as lipopeptid, protease, lipase etc. are conducive to Process waste drilling mud.
(3) in the compositions of the application or composite bacteria agent capable, each bacterial strain reasonable mixture ratio, it is possible to play each Degraded advantage.
(4) the application uses composite bacteria agent capable in-situ treatment in discarded drilling mud pond so that process Low cost, handling process is simple, and harmless treatment is effective, and economic benefit and social benefit are notable.
(5) the discarded oil base mud of discharge during the applicable object of the application can be shale gas wellbore construction Slurry, it is also possible to be oil-base mud and the water-base mud of the discharge of general oilfield drilling, therefore can solve The environmental pollution produced during oil-gas mining and ecological disruption problem, have important to pollution amelioration Meaning.
(6), in the compositions of the application or composite bacteria agent capable, harmful components are changed into plant by each bacterial strain can The materials such as the carbon source of utilization, nitrogen source, can supplement all kinds of nutrients in soil, promote plant Growth.
(7) in the compositions of the application or composite bacteria agent capable, each bacterial strain by discarded oil base mud by a huge sum of money Belong to and carry out adsorbing or converting, reduce the bio-toxicity in solid content.
In some embodiments, the compositions for processing waste drilling mud that the application provides Or composite bacteria agent capable, it is thus achieved that at least one effect below:
(1) compositions or the composite bacteria agent capable that use the application process after discarding oil-base mud, in solid phase Each content of beary metal all significantly reduces, and each content of beary metal reduces at least 15%, and concrete data see reality Execute example.Wherein, reducing at least 70% relative to cadmium content in solid phase before processing, lead content reduces at least 90%, arsenic content reduces at least 15%, is not detected by hydrargyrum.
(2) compositions or the composite bacteria agent capable that use the application process after discarding oil-base mud, and solid phase leaches PH value, BOD in liquid5, COD, sulfide content, volatile phenol content and petroleum content the most aobvious Writing and reduce, concrete data see embodiment.Wherein, relative to the BOD in solid phase leachate before processing5 Reducing at least 30%, COD reduces at least 20%, and petroleum content reduces at least 65%, does not detects To sulfide and volatile phenol.
(3) compositions or the composite bacteria agent capable that use the application process after discarding oil-base mud, and solid phase leaches In liquid, each content of beary metal all significantly reduces, and each content of beary metal reduces at least 8%, and concrete data are joined See embodiment.Wherein, at least 65% is reduced relative to mercury content during solid phase leaches before processing, nickel content Reduce at least 75%, be not detected by copper, zinc, cadmium, lead, chromium, arsenic and barium.
(4) compositions or the composite bacteria agent capable that use the application process after discarding oil-base mud, in mud pit Moisture is reduced to less than 45%, less than 40%, less than 35% or less than 30%, and preferably less than 45%.
(5) compositions or the composite bacteria agent capable that use the application process after discarding oil-base mud, in mud pit Solid phase firming body comprcssive strength can reach at least 1MPa, at least 1.5MPa or at least 2MPa, excellent Elect at least 1MPa as.
Following example are merely to illustrate and the purpose of unrestricted the application scope.
Embodiment
The process of embodiment 1 shale gas well abandoned oil base mud
The about 1000 cubic metres of discarded oil-base mud producing certain shale gas well carry out innoxious place Reason.
This shale gas well site is positioned at Mu us dese edge, and periphery geomorphological environment is sand dune and minority Farmland, own finishing drilling hands over nearly 1 year, the well-flushing waste liquid natural evaporation substantially of mud pit upper strata, pond Interior without the precipitation of obvious transudate, surface there are about the air dried coating of 2～3cm, and under air dried coating, mixing is discarded Thing water content is 80～90%, and pH value is 13, and physical state is viscous colloidal.Process front ring Border temperature is 30 DEG C, and in mud pit, temperature is 17 DEG C, is added by composite bacteria agent capable in waste sludge stock tank Processing after stirring, during process, ambient temperature is 12～30 DEG C, and the process monitoring time is 30 My god, there be twice rainfall period.
1) shake-flask culture: by bacillus subtilis, Pseudomonas stutzeri and Acinetobacter johnsonii respectively It is inoculated in beef extract-peptone fluid medium and carries out shake-flask culture, all under the conditions of 25～35 DEG C Cultivate 24～48 hours, obtain bacillus subtilis shaking flask strain, Pseudomonas stutzeri shaking flask respectively Strain and Acinetobacter johnsonii shaking flask strain.
2) seed tank culture: by step 1) in bacillus subtilis shaking flask strain, Amur vacation unit cell Bacterium shaking flask strain and Acinetobacter johnsonii shaking flask strain be inoculated into respectively seed culture medium (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out seed tank culture, under the conditions of 25～35 DEG C, all cultivate 24～48 little Time, obtain bacillus subtilis seed liquor, Pseudomonas stutzeri seed liquor and about family name's not lever respectively Bacterium seed liquor.
3) fermentor cultivation: by step 2) in bacillus subtilis seed liquor, Pseudomonas stutzeri kind Sub-liquid and Acinetobacter johnsonii seed liquor in 3～10% ratio (percent by volume) inoculum concentration inoculate respectively (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) to seed culture medium Potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out fermentor cultivation, all under the conditions of 25～35 DEG C Cultivate 48 hours, obtain bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacteria suspension peace treaty respectively Family name's acinetobacter calcoaceticus bacteria suspension；Bacteria suspension carries out microscopy respectively, and bacterial strain living bacteria count is ≥1×109cfu/mL。
4) solid culture: by step 3) in bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacterium hang Liquid and Acinetobacter johnsonii bacteria suspension are inoculated into wheat bran training respectively in 5～10% (percentage by weight) ratio Support in base (consisting of: 10% (w) rice husk, 5% (w) bean cake powder and 85% (w) wheat bran (such as Testa Tritici)) Carrying out solid culture, cultivation temperature is 25～35 DEG C, and incubation time is 72 hours, obtains respectively Bacillus subtilis solid culture, Pseudomonas stutzeri solid culture and Acinetobacter johnsonii are solid Body culture；Respectively solid culture is carried out microscopy, withered in bacillus subtilis solid culture The living bacteria count of grass bacillus cereus is >=1 × 108cfu/g；Pseudomonas stutzeri solid culture is executed The living bacteria count of family name pseudomonas is >=1 × 108cfu/g；In Acinetobacter johnsonii solid culture about The living bacteria count of family name's acinetobacter calcoaceticus is >=1 × 108cfu/g。
5) by step 4) in bacillus subtilis solid culture, Pseudomonas stutzeri solid culture Thing and Acinetobacter johnsonii solid culture carry out dusting dry, obtain bacillus subtilis respectively solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide, bacillus subtilis In bacterium solid fungicide, the living bacteria count of bacillus subtilis is >=1 × 108cfu/g；Pseudomonas stutzeri In solid fungicide, the living bacteria count of Pseudomonas stutzeri is >=1 × 108cfu/g；Acinetobacter johnsonii is solid In body microbial inoculum, the living bacteria count of Acinetobacter johnsonii is >=1 × 108cfu/g.Bacillus subtilis is solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide are in 1:2:2 ratio (weight Amount percentage ratio) mixing, it is thus achieved that composite bacteria agent capable, total living bacteria count >=1 × 10 in composite bacteria agent capable8cfu/g。
6) by step 5) composite bacteria agent capable prepared is transported to be positioned at the waste sludge stock tank near well site and uses In adding, use the plant equipment such as high-front shovel that waste sludge stock tank is uniformly stirred before adding Mix.Composite bacteria agent capable relative to the inoculum concentration of pending waste drilling mud be 0.5%w/v (ton/cube Rice), use the plant equipment such as high-front shovel by equal to composite bacteria agent capable and pending mud stirring after adding Even, harmless treatment 30 days after stirring.
After processing 30 days, to process drilling mud pond in solid phase in heavy metal (copper, zinc, cadmium, Lead, chromium, hydrargyrum, arsenic, nickel and boron) content is measured；Relevant physics and chemistry in solid phase leachate is referred to Mark (COD (COD), five-day BOD (BOD5), pH value, sulfide content, ammonia Nitrogen content, total cyanide content and petroleum content) and content of beary metal (copper, zinc, cadmium, lead, chromium, Hydrargyrum, arsenic, nickel, barium and selenium) it is measured.
The preparation method of solid phase leachate is with reference to HJ 557-2010 solid waste Leaching leaching side Prepared by method horizontal vibration method.
Processing procedure is as shown in Figure 1A to 1F, and in innocent treatment procedure, discarded slurry is through multiple Can destroy rapidly the colloidal state of mud after closing microbial inoculum effect, mud pit occurs that obvious solid-liquid divides From, greatly accelerate the evaporation rate of moisture in mud, reduce rapidly the pH value of aqueous phase, make water simultaneously The heavy metal ion, the chlorion that dissolve in mutually are retained in mud solid phase and continue by microbial action Degradation and Transformation.
Process terminates such as table 1 institute of the content of beary metal measurement result in the solid phase in rear drilling mud pond Showing, compared with before process, various content of beary metal all significantly reduce, and reduce bio-toxicity.Its In, before processing, after process, in solid phase, copper content reduces 42.52%, and Zn content reduces 41.88%, cadmium content reduces 90.75%, and lead content reduces 96.53%, and chromium content reduces 83.26%, arsenic content reduces 18.71%, and nickel content reduces 40.48%, and Boron contents reduces 33.92%, solid phase does not detects hydrargyrum.
The testing result of solid phase content of beary metal before and after table 1 process
"/" represents and does not detects.
After process terminates, in solid phase leachate, each physical and chemical factor testing result is as shown in table 2, relatively Before processing, pH value, BOD5, COD, sulfide content, volatile phenol content and petroleum-type contain Amount significantly reduces, and ammonia-nitrogen content substantially increases.Wherein, before processing, solid phase leaching after process Go out BOD in liquid5Reducing 69.11%, COD reduces 64.75%, and petroleum content reduces 83.34%, it is not detected by sulfide and volatile phenol, ammonia-nitrogen content adds 50.88%.
The testing result of physical and chemical factor in solid phase leachate before and after table 2 process
"/" represents and does not detects.
In solid phase leachate, content of beary metal is significantly reduced before relatively processing, and result is as shown in table 3. Wherein, before processing, after process, in solid phase leachate, mercury content reduces 86.67%, and nickel contains Amount reduce 77.38%, Se content reduces 8.70%, be not detected by copper, zinc, cadmium, lead, chromium, Arsenic and barium.
The testing result of content of beary metal in solid phase leachate before and after table 3 process
"/" represents and does not detects.
After composite bacteria agent capable processes, in mud pit, moisture is reduced to less than 45%, and mud solid phase solidifies Body comprcssive strength can reach 1MPa, and earth's surface pollutant may finally reach at harmless security landfill The requirement of reason.The oxious component in mud after process is converted to all kinds of plant growing and can utilize Carbon source, the material such as nitrogen source, reduce bio-toxicity, all kinds of nutrients in soil can be supplemented, Promote plant absorption growth.Mud pit carries out smooth after being disposed, surface, smooth rear well site is clean and tidy And the foreign material such as noresidue mud, on the mud pit after process, thickness of earth covering is more than 30 centimetres.Mud pit After surfacing 30 days, it is sprinkled into the seed mixture of the plants such as stubborn bar, false indigo, Folium et Cacumen Artemisiae Halodendri, Herba Medicaginis, Carry out revegetation.
The process of oil-base mud discarded by embodiment 2 oil well
The about 800 cubic metres of discarded oil-base mud producing certain oil well carry out harmless treatment.
This oil well field is positioned at limit, village, and around geomorphological environment is in cultivated farmland, and firm finishing drilling is handed over, Bottom mud pH value is 13～14, and physical state is viscose glue body, and before processing, ambient temperature is 30 DEG C, mud pit TNE UPPER SEA TEMPERATURE is 18 DEG C, and bottom mud is 8～10 DEG C.Waste sludge after stirring Slurry water content is more than 90%, and pH value is 11～12, and temperature is 15～16 DEG C, physical state For dilute colloidal.Composite bacteria agent capable is added in waste sludge stock tank and process after stirring, process the phase Between ambient temperature be 16～28 DEG C, process the monitoring time be 35 days.
1) shake-flask culture: by bacillus subtilis, Pseudomonas stutzeri and Acinetobacter johnsonii respectively It is inoculated in beef extract-peptone fluid medium and carries out shake-flask culture, all under the conditions of 25～35 DEG C Cultivate 24～48 hours, obtain bacillus subtilis shaking flask strain, Pseudomonas stutzeri shaking flask respectively Strain and Acinetobacter johnsonii shaking flask strain.
2) seed tank culture: by step 1) in bacillus subtilis shaking flask strain, Amur vacation unit cell Bacterium shaking flask strain, Acinetobacter johnsonii shaking flask strain be inoculated into respectively seed culture medium (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out seed tank culture, under the conditions of 25～35 DEG C, all cultivate 24～48 little Time, obtain bacillus subtilis seed liquor, Pseudomonas stutzeri seed liquor and about family name's not lever respectively Bacterium seed liquor.
3) fermentor cultivation: by step 2) in bacillus subtilis seed liquor, Pseudomonas stutzeri kind Sub-liquid and Acinetobacter johnsonii seed liquor in 3～10% ratio (percent by volume) inoculum concentration inoculate respectively (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) to seed culture medium Potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out fermentor cultivation, all under the conditions of 25～35 DEG C Cultivate 48 hours, obtain bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacteria suspension peace treaty respectively Family name's acinetobacter calcoaceticus bacteria suspension；Respectively bacteria suspension is carried out microscopy, bacterial strain living bacteria count is >=1 × 109cfu/mL。
4) solid culture: by step 3) in bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacterium hang Liquid and Acinetobacter johnsonii bacteria suspension are inoculated into wheat bran training respectively in 5～10% (percentage by weight) ratio Support in base (consisting of: 10% (w) rice husk, 5% (w) bean cake powder and 85% (w) wheat bran (such as Testa Tritici)) Carrying out solid culture, cultivation temperature is 25～35 DEG C, and incubation time is 48 hours, obtains respectively Bacillus subtilis solid culture, Pseudomonas stutzeri solid culture and Acinetobacter johnsonii are solid Body culture；Respectively solid culture is carried out microscopy, withered in bacillus subtilis solid culture The living bacteria count of grass bacillus cereus is >=1 × 108cfu/g；Pseudomonas stutzeri solid culture is executed The living bacteria count of family name pseudomonas is >=1 × 108cfu/g；In Acinetobacter johnsonii solid culture about The living bacteria count of family name's acinetobacter calcoaceticus is >=1 × 108cfu/g。
5) by step 4) in bacillus subtilis solid culture, Pseudomonas stutzeri solid culture Thing and Acinetobacter johnsonii solid culture carry out dusting dry, obtain bacillus subtilis respectively solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide, bacillus subtilis In bacterium solid fungicide, the living bacteria count of bacillus subtilis is >=1 × 108cfu/g；Pseudomonas stutzeri In solid fungicide, the living bacteria count of Pseudomonas stutzeri is >=1 × 108cfu/g；Acinetobacter johnsonii is solid In body microbial inoculum, the living bacteria count of Acinetobacter johnsonii is >=1 × 108cfu/g.Bacillus subtilis is solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide are in 1:1.5:2 ratio (percentage by weight) mixes, it is thus achieved that composite bacteria agent capable, and total living bacteria count >=1 in composite bacteria agent capable × 108cfu/g。
6) by step 5) composite bacteria agent capable prepared is transported to be positioned at the waste sludge stock tank near well site and uses In adding, use the plant equipment such as high-front shovel that waste sludge stock tank is uniformly stirred before adding Mix.Composite bacteria agent capable relative to the inoculum concentration of pending waste drilling mud be 0.7%w/v (ton/cube Rice), use the plant equipment such as high-front shovel by equal to composite bacteria agent capable and pending mud stirring after adding Even, harmless treatment 35 days after stirring.
After processing 35 days, to process drilling mud pond in solid phase in heavy metal (copper, zinc, cadmium, Lead, chromium, hydrargyrum, arsenic, nickel and boron) content is measured；Relevant physics and chemistry in solid phase leachate is referred to Mark (COD (COD), five-day BOD (BOD5), pH value, sulfide content, ammonia Nitrogen content, total cyanide content and petroleum content) and content of beary metal (copper, zinc, cadmium, lead, chromium, Hydrargyrum, arsenic, nickel, barium and selenium) it is measured.
The preparation method of solid phase leachate is with reference to HJ 557-2010 solid waste Leaching leaching side Prepared by method horizontal vibration method.
Processing procedure is as shown in Fig. 2 A to 2F, and in innocent treatment procedure, discarded slurry is through multiple Can destroy rapidly the colloidal state of mud after closing microbial inoculum effect, mud pit occurs that obvious solid-liquid divides From, greatly accelerate the evaporation rate of moisture in mud, reduce rapidly the pH value of aqueous phase, make water simultaneously The heavy metal ion, the chlorion that dissolve in mutually are retained in mud solid phase and continue by microbial action Degradation and Transformation.
Process terminates such as table 4 institute of the content of beary metal measurement result in the solid phase in rear drilling mud pond Showing, compared with before process, various content of beary metal all significantly reduce, and reduce bio-toxicity.Its In, before processing, after process, in solid phase, copper content reduces 48.49%, and Zn content reduces 61.20%, cadmium content reduces 71.60%, and lead content reduces 95.84%, and chromium content reduces 91.62%, arsenic content reduces 19.05%, and nickel content reduces 38.23%, and Boron contents reduces 59.84%, solid phase does not detects hydrargyrum.
The testing result of solid phase content of beary metal before and after table 4 process
"/" represents and does not detects.
After process terminates, in solid phase leachate, each physical and chemical factor testing result is as shown in table 5, relatively Before processing, pH value, BOD5, COD, sulfide content, volatile phenol content and petroleum-type contain Amount significantly reduces.Wherein, before processing, BOD in solid phase leachate after process5Reduce 45.65%, COD reduce 25.96%, and ammonia-nitrogen content reduces 6.61%, and petroleum content drops Low by 74.50%, it is not detected by sulfide and volatile phenol.
The testing result of physical and chemical factor in solid phase leachate before and after table 5 process
"/" represents and does not detects.
In solid phase leachate, content of beary metal is significantly reduced before relatively processing, and result is as shown in table 6. Wherein, before processing, after process, in solid phase leachate, mercury content reduces 66.67%, and nickel contains Amount reduce 84.62%, Se content reduces 12.12%, solid phase leachate is not detected by copper, zinc, Cadmium, lead, chromium, arsenic and barium.
The testing result of content of beary metal in solid phase leachate before and after table 6 process
"/" represents and does not detects.
After composite bacteria agent capable processes, in mud pit, moisture is reduced to less than 45%, and mud solid phase solidifies Body comprcssive strength can reach 1MPa, and earth's surface pollutant may finally reach at harmless security landfill The requirement of reason.The oxious component in mud after process is converted to all kinds of plant growing and can utilize Carbon source, the material such as nitrogen source, reduce bio-toxicity, all kinds of nutrients in soil can be supplemented, Promote plant absorption growth.Mud pit carries out smooth after being disposed, surface, smooth rear well site is clean and tidy And the foreign material such as noresidue mud, on the mud pit after process, thickness of earth covering is more than 30 centimetres.Mud pit After surfacing 30 days, it is sprinkled into the seed mixture of the plants such as stubborn bar, false indigo, Folium et Cacumen Artemisiae Halodendri, Herba Medicaginis, Carry out revegetation.
The process of oil-base mud discarded by embodiment 3 gas well
The about 200 cubic metres of discarded oil-base mud producing certain gas well carry out harmless treatment. This well site is positioned at loess hill area, and periphery geomorphological environment is farmland, and finishing drilling is handed over about partly Year, bottom mud pH value is 12, and physical state is viscose glue body, and upper strata about 100～150t is tried Well waste liquid, surface is thicker crude oil layer, and before processing, ambient temperature is 25 DEG C, temperature in mud pit It it is 13 DEG C.After stirring, discarded slurry water content is more than 90%, and pH value is 11～12, temperature Being 12 DEG C, physical state is dilute colloidal.Composite bacteria agent capable is added after waste sludge stock tank stirs Processing, during process, ambient temperature is 16～30 DEG C, and the process monitoring time is 30 days.
1) shake-flask culture: by bacillus subtilis, Pseudomonas stutzeri and Acinetobacter johnsonii respectively It is inoculated in beef extract-peptone fluid medium and carries out shake-flask culture, all under the conditions of 25～35 DEG C Cultivate 24～48 hours, obtain bacillus subtilis shaking flask strain, Pseudomonas stutzeri shaking flask respectively Strain and Acinetobacter johnsonii shaking flask strain.
2) seed tank culture: by step 1) in bacillus subtilis shaking flask strain, Amur vacation unit cell Bacterium shaking flask strain, Acinetobacter johnsonii shaking flask strain be inoculated into respectively seed culture medium (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out seed tank culture, under the conditions of 25～35 DEG C, all cultivate 24～48 little Time, obtain bacillus subtilis seed liquor, Pseudomonas stutzeri seed liquor and about family name's not lever respectively Bacterium seed liquor.
3) fermentor cultivation: by step 2) in bacillus subtilis seed liquor, Pseudomonas stutzeri kind Sub-liquid and Acinetobacter johnsonii seed liquor in 3～10% ratio (percent by volume) inoculum concentration inoculate respectively (consist of: 0.5% (w/v) yeast leaching powder, 1% (w/v) tryptone, 0.05% (w/v) to seed culture medium Potassium dihydrogen phosphate and 0.5% (w/v) sodium chloride) in carry out fermentor cultivation, all under the conditions of 25～35 DEG C Cultivate 24 hours, obtain bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacteria suspension peace treaty respectively Family name's acinetobacter calcoaceticus bacteria suspension；Respectively bacteria suspension is carried out microscopy, bacterial strain living bacteria count is >=1 × 109cfu/mL。
4) solid culture: by step 3) in bacillus subtilis bacteria suspension, Pseudomonas stutzeri bacterium hang Liquid and Acinetobacter johnsonii bacteria suspension are inoculated into wheat bran training respectively in 5～10% (percentage by weight) ratio Support in base (consisting of: 10% (w) rice husk, 5% (w) bean cake powder and 85% (w) wheat bran (such as Testa Tritici)) Carrying out solid culture, cultivation temperature is 25～35 DEG C, and incubation time is 24 hours, obtains respectively Bacillus subtilis solid culture, Pseudomonas stutzeri solid culture and Acinetobacter johnsonii are solid Body culture；Respectively solid culture is carried out microscopy, withered in bacillus subtilis solid culture The living bacteria count of grass bacillus cereus is >=1 × 108cfu/g；Pseudomonas stutzeri solid culture is executed The living bacteria count of family name pseudomonas is >=1 × 108cfu/g；In Acinetobacter johnsonii solid culture about The living bacteria count of family name's acinetobacter calcoaceticus is >=1 × 108cfu/g。
5) by step 4) in bacillus subtilis solid culture, Pseudomonas stutzeri solid culture Thing and Acinetobacter johnsonii solid culture carry out dusting dry, obtain bacillus subtilis respectively solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide, bacillus subtilis In bacterium solid fungicide, the living bacteria count of bacillus subtilis is >=1 × 108cfu/g；Pseudomonas stutzeri In solid fungicide, the living bacteria count of Pseudomonas stutzeri is >=1 × 108cfu/g；Acinetobacter johnsonii is solid In body microbial inoculum, the living bacteria count of Acinetobacter johnsonii is >=1 × 108cfu/g.Bacillus subtilis is solid Body microbial inoculum, Pseudomonas stutzeri solid fungicide and Acinetobacter johnsonii solid fungicide are in 1:1:1 ratio (weight Amount percentage ratio) mixing, it is thus achieved that composite bacteria agent capable, total living bacteria count >=1 × 10 in composite bacteria agent capable8cfu/g。
6) by step 5) composite bacteria agent capable prepared is transported to be positioned at the waste sludge stock tank near well site and uses In adding, use the plant equipment such as high-front shovel that waste sludge stock tank is uniformly stirred before adding Mix.Composite bacteria agent capable relative to the inoculum concentration of pending waste drilling mud be 0.5%w/v (ton/cube Rice), use the plant equipment such as high-front shovel by equal to composite bacteria agent capable and pending mud stirring after adding Even, harmless treatment 30 days after stirring.
After processing 30 days, to process drilling mud pond in solid phase in heavy metal (copper, zinc, cadmium, Lead, chromium, hydrargyrum, arsenic, nickel and boron) content is measured；Relevant physics and chemistry in solid phase leachate is referred to Mark (COD (COD), five-day BOD (BOD5), pH value, sulfide content, ammonia Nitrogen content, total cyanide content and petroleum content) and content of beary metal (copper, zinc, cadmium, lead, chromium, Hydrargyrum, arsenic, nickel, barium and selenium) it is measured.
The preparation method of solid phase leachate is with reference to HJ 557-2010 solid waste Leaching leaching side Prepared by method horizontal vibration method.
Processing procedure is as shown in Fig. 3 A to 3F, and in innocent treatment procedure, discarded slurry is through multiple Can destroy rapidly the colloidal state of mud after closing microbial inoculum effect, mud pit occurs that obvious solid-liquid divides From, greatly accelerate the evaporation rate of moisture in mud, reduce rapidly the pH value of aqueous phase, make water simultaneously The heavy metal ion, the chlorion that dissolve in mutually are retained in mud solid phase and continue by microbial action Degradation and Transformation.
Process terminates such as table 7 institute of the content of beary metal measurement result in the solid phase in rear drilling mud pond Showing, compared with before process, various content of beary metal all significantly reduce, and reduce bio-toxicity.Its In, before processing, after process, in solid phase, copper content reduces 51.19%, and Zn content reduces 59.14%, cadmium content reduces 75.00%, and lead content reduces 92.23%, and chromium content reduces 90.12%, arsenic content reduces 60.71%, and nickel content reduces 37.96%, and Boron contents reduces 59.60%, solid phase does not detects hydrargyrum.
The testing result of solid phase content of beary metal before and after table 7 process
"/" represents and does not detects.
After process terminates, in solid phase leachate, each physical and chemical factor testing result is as shown in table 8, relatively Before processing, pH value, BOD5, COD, sulfide content, volatile phenol content and petroleum-type contain Amount significantly reduces, and ammonia-nitrogen content substantially increases.Wherein, before processing, solid phase leaching after process Go out BOD in liquid5Reducing 31.76%, COD reduces 42.18%, and petroleum content reduces 68.75%, it is not detected by sulfide and volatile phenol, ammonia-nitrogen content adds 50.60%
The testing result of physical and chemical factor in solid phase leachate before and after table 8 process
"/" represents and does not detects.
In solid phase leachate, content of beary metal is significantly reduced before relatively processing, and result is as shown in table 9. Wherein, before processing, after process, in solid phase leachate, mercury content reduces 92.86%, and nickel contains Amount reduce 82.35%, Se content reduces 46.43%, solid phase leachate is not detected by copper, zinc, Cadmium, lead, chromium, arsenic and barium.
The testing result of content of beary metal in solid phase leachate before and after table 9 process
"/" represents and does not detects.
After composite bacteria agent capable processes, in mud pit, moisture is reduced to less than 45%, and mud solid phase solidifies Body comprcssive strength can reach 1MPa, and earth's surface pollutant may finally reach at harmless security landfill The requirement of reason.The oxious component in mud after process is converted to all kinds of plant growing and can utilize Carbon source, the material such as nitrogen source, reduce bio-toxicity, all kinds of nutrients in soil can be supplemented, Promote plant absorption growth.Mud pit carries out smooth after being disposed, surface, smooth rear well site is clean and tidy And the foreign material such as noresidue mud, on the mud pit after process, thickness of earth covering is more than 30 centimetres.Mud pit After surfacing 30 days, it is sprinkled into the seed mixture of the plants such as stubborn bar, false indigo, Folium et Cacumen Artemisiae Halodendri, Herba Medicaginis, Carry out revegetation.
Although, the most with a general description of the specific embodiments the present invention has been made detailed Description, but on the basis of the present invention, it can be made some modifications or improvements, this is to this area It is apparent from for technical staff.Therefore, done without departing from theon the basis of the spirit of the present invention These modifications or improvements, belong to the scope of protection of present invention.

Claims (9)

1. a compositions, comprises bacillus subtilis (Bacillus subtilis), Pseudomonas stutzeri (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii).
2. a composite bacteria agent capable, it comprises bacillus subtilis (Bacillus subtilis), Amur vacation list Born of the same parents bacterium (Pseudomonas stutzeri) and Acinetobacter johnsonii (Acinetobacter johnsonii), preferably Ground is made up of above-mentioned three kinds of bacterial strains, or comprises the culture of above-mentioned three kinds of bacterial strains.
3. as claimed in claim 2 composite bacteria agent capable, total living bacteria count in wherein said composite bacteria agent capable >= 1×108cfu/g；
Or, in terms of the weight of described strain culture, hay described in wherein said composite bacteria agent capable Bacillus culture, Pseudomonas stutzeri culture and the weight portion of Acinetobacter johnsonii culture For:
Bacillus subtilis culture 1～3
Pseudomonas stutzeri culture 1～6
Acinetobacter johnsonii culture 1～6,
Preferably, described bacillus subtilis culture, Pseudomonas stutzeri culture and about family name is not The weight ratio of dynamic alphacterium culture is 1:2:2,1:1.5:2 or 1:1:1；
Optionally, described strain culture comprises bacterial strain and culture medium；
Optionally, effective viable bacteria of bacillus subtilis described in described bacillus subtilis culture Number is for >=1 × 108cfu/g；Described in described Pseudomonas stutzeri culture, Pseudomonas stutzeri is effective Viable count is >=1 × 108cfu/g；Acinetobacter johnsonii described in described Acinetobacter johnsonii culture Living bacteria count is >=1 × 108cfu/g；
Optionally, described culture medium is solid medium, semisolid culturemedium or fluid medium.
4. a preparation method for composite bacteria agent capable, including:
By bacillus subtilis (Bacillus subtilis), Pseudomonas stutzeri (Pseudomonas Stutzeri) cultivate respectively with Acinetobacter johnsonii (Acinetobacter johnsonii)；And
Obtain the respective culture of described bacterial strain, and by the respective culture of described bacterial strain according to Combinations thereof mixes, and obtains described composite bacteria agent capable；
Optionally, above-mentioned cultivation is solid culture, Semi-solid cell culture or liquid culture.
5. method as claimed in claim 4, wherein with the weight of the respective culture of described bacterial strain Meter mixes, described bacillus subtilis culture during mixing, Pseudomonas stutzeri culture and The weight portion of Acinetobacter johnsonii culture is:
Bacillus subtilis culture 1～3
Pseudomonas stutzeri culture 1～6
Acinetobacter johnsonii culture 1～6,
Preferably, described bacillus subtilis culture during mixing, Pseudomonas stutzeri culture and The weight ratio of Acinetobacter johnsonii culture is 1:2:2,1:1.5:2 or 1:1:1；
Optionally, described strain culture comprises bacterial strain and culture medium；
Optionally, effective viable bacteria of bacillus subtilis described in described bacillus subtilis culture Number is for >=1 × 108cfu/g；Described in described Pseudomonas stutzeri culture, Pseudomonas stutzeri is effective Viable count is >=1 × 108cfu/g；Acinetobacter johnsonii described in described Acinetobacter johnsonii culture Living bacteria count is >=1 × 108cfu/g；
Optionally, total living bacteria count >=1 × 10 in described composite bacteria agent capable8cfu/g；
Optionally, described culture medium is solid medium, semisolid culturemedium or fluid medium.
6. process or a method for aid in treatment waste drilling mud, including:
By compositions described in claim 1, or composite bacteria agent capable described in Claims 2 or 3 puts into pending Waste drilling mud carries out processing at least 20 days, at least 25 days, at least 30 days, at least 35 days, At least 40 days, at least 45 days, at least 50 days, at least 55 days, at least 60 days, at least 65 days, extremely Few 70 days, at least 75 days, at least 80 days, at least 85 days, at least 90 days, at least 95 days, at least 100 days, at least 105 days, at least 110 days, at least 115 days, at least 120 days, at least 125 days, At least 130 days, at least 135 days, at least 140 days, at least 145 days, at least 150 days, at least 155 My god, at least 160 days, at least 165 days, at least 170 days, at least 175 days or at least 180 days；Preferably It it is at least 30 days or at least 35 days；
Preferably, described waste drilling mud is discarded oil-base mud or waste water-base mud；
It is highly preferred that described discarded oil-base mud is the discarded oil base discharged in shale gas drilling process Mud.
7. method as claimed in claim 6, wherein said compositions or described composite bacteria agent capable relative to The inoculum concentration of described pending waste drilling mud is at least 0.5%, at least 0.7%, at least 1%, extremely Few 3%, at least 5% or at least 7%w/v, preferably at least 0.5% or at least 0.7%w/v.
8. the solid content that method described in claim 6 or 7 obtains after processing waste drilling mud.
9. the solid content that method described in claim 6 or 7 obtains after processing waste drilling mud is discarded Application in drilling mud reparation.
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