CN109482638A - Pseudomonas oil-reducing bacterium L-1, mixed microbial inoculum thereof and in-situ remediation method for petroleum-polluted soil - Google Patents
Pseudomonas oil-reducing bacterium L-1, mixed microbial inoculum thereof and in-situ remediation method for petroleum-polluted soil Download PDFInfo
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C2101/00—In situ
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- Engineering & Computer Science (AREA)
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- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Soil Sciences (AREA)
- Environmental & Geological Engineering (AREA)
- Processing Of Solid Wastes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides pseudomonas oil-reducing bacteria, a mixed microbial inoculum thereof and an in-situ remediation method of petroleum-polluted soil, wherein the pseudomonas oil-reducing bacteria; the preservation number is CGMCC NO. 16557. The mixed microbial inoculum contains pseudomonas oil-reducing bacteria and indigenous bacteria with the preservation number of CGMCC NO. 16557. The method can effectively reduce the content of petroleum hydrocarbon and polycyclic aromatic hydrocarbon in the oily sludge, realize the harmless treatment of the oily sludge, eliminate the harm of the polluted soil to human bodies and recover the ecological function of the soil. Therefore, the invention takes the degradation of petroleum hydrocarbon as a starting point, separates and cultures microorganisms with obvious degradation effect on soil petroleum, and further provides a concrete implementation measure of the in-situ remediation technology of the microorganisms in the soil polluted by oil on site. Can degrade the oil soil with 3 percent of concentration to 77 percent in 30 days.
Description
Technical field
The present invention provides a kind of pseudomonas drop oil bacterium L-1 and its microorganism mix bacterium agent and oil-polluted soils original position
Restorative procedure belongs to Biotechnology in energy production and field of environmental biotechnology, which can effectively degrade stone in soil and greasy filth
Petroleum hydrocarbon substance can be effectively applied to oil-polluted soils biodegrade processing.
Background technique
With the development of petroleum industry, petroleum is increasingly severe to the pollution of environment.Especially buried tank and petroleum pipeline
Line corrosion leakage contaminated soil and underground water source generate a large amount of oily sludge and oily wastewater, cause the salinization of soil, poison
Phenomena such as, human body is caused greatly to damage eventually by food chain accumulation.
The method for administering oily sludge at present mainly has physical method, chemical method and biological restoration.In practical applications,
Peripheral doses and chemical remediation all have certain limitation, should not use and be also easy to produce under partial picture secondary pollution,
The problems such as repairing effect is undesirable, energy consumption is high, comprehensive utilization degree is low low with resource recovery.It is biological prosthetic because of repair time
Long, larger by environmental factor interference, pollutant initial concentration is more low to limit its popularization and application, but because its green is without secondary dirt
Dye, low power consuming, do not destroy soil texture, can ira situ degradation pollutant the advantages that, be increasingly becoming renovation of organic pollution soil
Emerging technology.Bioremediation technology be considered as it is a kind of it is environmentally protective, without secondary pollution, efficient, can thorough degradation of contaminant
Oil pollution restorative procedure with development prospect, the key problem in technology are to utilize functional microorganism efficient degradation different kinds of petroleum hydrocarbon
Component.
Soil Microorganism is resourceful, microbial metabolism and inactive in natural situation.When these microorganisms encounter conjunction
Recovery starts metabolic process when suitable environmental stimulus, by stimulated difference, under different environmental stimulus, and recovery generation
The microbe species the thanked dominant bacteria generated that is also not quite similar is not also identical.In the soil for a long time by oil pollution, micro- life
The condition that itself can be altered in steps in object deacclimatizes environment, carries out selective enrichment and hereditary variation occurs, therefore can be this
The bacterial strain of efficient degradation petroleum is isolated in screening in soil.
Summary of the invention
The present invention provides a kind of pseudomonas drop oil bacterium L-1 and its micro- primarily directed to the status of oil-polluted soils
Biological mix bacterium agent and oil-polluted soils in-situ remediation method.
The technical solution of the invention is as follows: a kind of oily bacterium L-1 of pseudomonas drop is isolated from Oilfield Oil-Contained Silt,
Purpose bacterial strain is obtained from petroleum aromatic contaminated soil suspension culture, isolates and purifies to obtain in bacterial strain that select 1 plant of degradation capability most strong
Growing Strain Designation the most vigorous is that oily bacterium L-1 drops in pseudomonas, which belongs to pseudomonas
(Pseudomonadaceae), deposit number is CGMCC NO.16557, and DNA sequence dna is as shown in SEQ NO<400>1.
The microorganism mix bacterium agent that oily bacterium L-1 is dropped comprising a kind of pseudomonas is containing indigenous bacterium and deposit number
Oily bacterium L-1 drops in the pseudomonas of CGMCC NO.16557, and it is 1 that oil bacterium L-1 and indigenous bacterium solution drop in pseudomonas in mass ratio:
Bacteria suspension is made in 3~1:6.
A kind of oil-polluted soils in-situ remediation method using the microorganism mix bacterium agent,
(1) required microorganism mix bacterium agent bacterial strain is first brought back to life, then expands culture, fermenting and producing is spare；Indigenous bacterium
For bacillus (Bacillus encimensis strain encimensis), Bacillus foecalis alkaligenes (Alcaligenes
Faecalis strain), yellow class Nocard's bacillus (Nocardioides luteus), acinetobacter (Acinetobacter
Sp), penicillium oxalicum (Penicillium oxalicum), by 6 plants of microbial inoculums in microorganism mix bacterium agent according to pseudomonas
Oily bacterium L-1: bacillus: Bacillus foecalis alkaligenes: yellow class Nocard's bacillus: acinetobacter: penicillium oxalicum bacterium solution volume drops
25~35 DEG C are put into fermentor than the ratio for 1:1:1:1:1:1~1.1:1.1:1.1:1.1:1.1:0.5,170~230r/
Min fermentation is spare；
(2) microorganism mix bacterium agent and microbial inoculum nutrient solution NS-1 are added according to 4% (wt)~6% (wt), make C in soil:
N:P=100:10:1~100:10:5；Dig ventilation and watering are carried out daily, increases oxygen-supplying amount, keep moisture 40-50%.
(3) periodic detection oil content, oil component and microbial activities situation, enhance process are monitored and controlled.
The microorganism mix bacterium agent to the condition of oil degradation be 25-35 DEG C of temperature, pH between 7-8, TDS is in 0-
Between 800ppm.
Microbial inoculum nutrient solution NS-1 is added in soil to cultivate the pseudomonas drop oil bacterium L-1 and microorganism mixing
Microbial inoculum, the nutrient solution NS-1 formula:
Ammonium nitrate (NH3NO3): 1.5-2.5g/L
Dipotassium hydrogen phosphate (K2HPO4): 1-2g/L
Potassium dihydrogen phosphate (KH2PO4): 2.5-3.5g/L
Magnesium sulfate (MgSO4): 0.01-0.1g/L
Calcium chloride (CaCl2): 0.01-0.02g/L
Sodium chloride (NaCl): 0.3-0.6g/L
Ferrous sulfate (FeSO4): 0.01-0.02g/L
Sodium ethylene diamine tetracetate (Na2EDTA): 0.01-0.02g/L.
Applicant isolates a kind of high efficient petroleum degrading bacteria in Changqing oilfields oil-containing mud sand, and has studied bacterium
Strain is to the degradation capability of petroleum hydrocarbon, to be used for oil-containing mud sand bioremediation technology.
The content of petroleum hydrocarbon in oily sludge, polycyclic aromatic hydrocarbon can be effectively reduced using method of the invention, realize oil-containing
The harmlessness disposing of sludge eliminates harm of the contaminated soil to human body, restores the ecological functions of soil.
Therefore, for the present invention using decomposing petroleum hydrocarbon as starting point, separating and having cultivated has obvious degradation to soil petroleum-type
The microorganism of effect has studied the growth of this quasi-microorganism and carries out biological prosthetic nutrient solution prescription, while to degradation kinetics
Probed into organic evolution parameter, it is further provided live oily sludge edaphon based technique for in-situ remediation it is specific
Implementing measure.It can make to degrade for petroleum-contaminated soil 30 days of 3% concentration and reach 77%.
Detailed description of the invention
Fig. 1 is the phylogenetic tree schematic diagram that oily bacterium L-1 bacterial strain drops in pseudomonas.
Preservation proves
The classification naming of biomaterial: Pseudomonas stutzeri Pseudomonas stutzeri strain
Depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on November 6th, 2018
Deposit number: CGMCC No 16557
Specific embodiment
Possessed by the specific embodiment bacterial strain that present invention be described in more detail and its feature and application
Technical effect, but the present invention is not therefore subject to any restriction.
The separation and identification of 1 pseudomonas Pseudomonadaceae bacterial strain of embodiment
The present invention provides a kind of pseudomonas to drop oily bacterium L-1, is isolated from Changqing oilfields oil-containing mud sand, by it
Morphological feature, Physiology and biochemistry and 16S rDNA gene sequencing Preliminary Identification belong to pseudomonas
(Pseudomonadaceae).The bacterial strain can be using petroleum hydrocarbon as the sole carbon source of growth, in 25-35 DEG C of temperature, pH in 7-8
Between, TDS between 0-800ppm under the conditions of simulation test to show that the pseudomonad removes petroleum hydrocarbon obvious, removal rate is reachable
77%.
It is sole carbon source that oil-polluted soils suspension, which is inoculated in the inoculum concentration of mass ratio 15% with crude oil (6g/L),
In nutrient solution NS-1,150r/min 3~4d of shaking table culture under 28 DEG C of aerobic conditions transfers 1 time under the same terms.It dilutes, be applied to
Using crude oil as the solid medium of sole carbon source, (solid is made in addition 20g agar after 6g crude oil is added in 1000mL nutrient solution NS-1
Culture medium) in, 28 DEG C of 4~5d of constant temperature incubation form single colonie.Picking grows rapid, neat in edge bacterium colony and accesses with crude oil
(6g/L) is in the nutrient solution NS-1 of sole carbon source, and shaking flask secondary screening obtains purpose bacterial strain.
From isolating and purifying to obtain, 1 plant of degradation capability most Johnson & Johnson head Strain Designation the most vigorous is selected in bacterial strain is false unit cell
Oily bacterium L-1 drops in Pseudomonas, is further identified.
The identification of molecular biology level
16s rDNA sequence is as shown in sequence table SEQ No<400>1.
The analysis of 16s rDNA phylogenetic tree
With the neighbouring phase connection (Neighbor mono- of Molecular Evolutionary Genetics credit analysis and sequence alignment program MEGA5.2
JoiningMethod phylogenetic analysis) is carried out to clustalx multiple alignment result, using Boot strap method 500
Repeat certificate authenticity.By studying position of the resulting 16SrDNA sequence in GenBank database, such as Fig. 1 institute
Show.
Oily bacterium L-1 bacterial strain drops in pseudomonas and west figure Ze Shi pseudomonad is alike, and 16s rDNA sequence and homology
With west figure Ze Shi pseudomonad (Pseudomonas stutzeri strain) 16s rDNA sequence have 100.0% it is homologous
Property, and show it and Xi Tu Ze Shi pseudomonad (Pseudomonas stutzeri strain) tool on systematic evolution tree
Have compared with into genetic distance.Therefore according to result above, the oily bacterium L-1 bacterial strain of pseudomonas drop is belonged into the false list of west figure Ze Shi
Its preliminary designation is Pseudomonas stutzeri by born of the same parents bacterium (Pseudomonas stutzeri strain) Pseudomonas
strain。
The preparation of 2 microorganism mix bacterium agent of embodiment
1. microbial strains inoculation activation
Aseptically, by pseudomonas drop oil bacterium L-1 and indigenous bacterium, [indigenous bacterium is bacillus
(Bacillus encimensis strain encimensis), Bacillus foecalis alkaligenes (Alcaligenes faecalis
Strain) yellow class Nocard's bacillus (Nocardioides luteus) acinetobacter (Acinetobacter sp) oxalic acid is green
Mould (Penicillium oxalicum)] according to the ratio of 1:1:1:1:1:1 it is inoculated into the triangle equipped with nutrient broth respectively
In bottle (after sterilizing), the above triangular flask is put into 30 DEG C respectively, cultivates 48h in the shaking table of 200r/min, for use.
2. prepared by the fermentation of liquid mix bacterium agent
The preparation of 2.1 bacterium seed liquors
By the above-mentioned pseudomonas drop oil bacterium L-1 for having cultivated 48h and indigenous bacterium, it is respectively connected to according to 5% access amount
After carrying out fermented and cultured in bacterial fermentation tank for 24 hours, bacterium seed liquor is obtained.
The preparation of 2.2 mix bacterium agents
It in mass ratio will be that 1:4 is made into bacterium with the west figure Ze Shi pseudomonas drop oil bacterium L-1 of top fermentation and indigenous bacterium solution
It is 20wt% that oil bacterium L-1 drops in suspension, i.e. pseudomonas.It places 4 DEG C of refrigerators to save, for use.
The processing of 3 oily sludge microorganism remediation of embodiment
The present invention also provides the oil-polluted soils in-situ remediation methods using microorganism mix bacterium agent.Vacation of the invention
In-situ immobilization can be used in combination with the oil-polluted soils original inhabitants bacterium after enrichment culture in zygosaccharomyces drop oil bacterium L-1, according to dirt
The crude content for contaminating soil typically constitutes from the 20wt% of microbial inoculum content after mixing.Oil-polluted soils in-situ remediation method, comprising:
(1) required microbial strains are first brought back to life, then expands culture, fermenting and producing is spare；According to a certain percentage
30 DEG C are put into fermentor, and 200r/min fermentation is spare；
(2) indigenous bacterium and microbial inoculum nutrient solution are added according to corresponding proportion, makes C:N:P=100:10:3 in soil；Daily into
Capable dig ventilation and watering, increase oxygen-supplying amount, keep moisture 40-50%.
(3) periodic detection oil content, oil component and microbial activities situation, enhance process are monitored and controlled.
In the present embodiment, microbial inoculum is made to oil-containing in the drop oil bacterium screened using embodiment 1, the method by embodiment 2
Sludge carries out biological prosthetic processing, repairs the oil of oil concentration 3% or so than 5% microorganism mix bacterium agent of addition by soil quality
Dirty soil.
Concrete operations are as follows:
(1) oily bacterium L-1 bacterium is dropped into the pseudomonas on the indigenous bacterium and storage medium of the oily sludge soil of reparation
Kind carries out activation culture, and large-scale production is spare；
Culture presevation culture medium (g/L): yeast extract 0.5, tryptone 0.25, peptone 0.75, glucose 0.5 can
Soluble starch 0.5, dipotassium hydrogen phosphate 0.3, magnesium sulfate 0.024, Sodium Pyruvate 0.3.
(2) petroleum-contaminated soil that oil concentration is 3% or so is manually configured in flowerpot.1kg is separately added into each flowerpot
The soil being sieved.3% standard oil is added in first group, sprays standard oil when digging sample, is uniformly mixed soil sample with standard oil.
Second group is added 50mL original inhabitants microbial inoculum on the basis of first group.50mL Mixed Microbes are added in third group on the basis of first group
Agent, west figure Ze Shi pseudomonad content is 20wt%.
(3) 3 flowerpots are successively put into KRQ-400 intelligent growth cabinet to cultivate.Adjust growth cabinet
35 DEG C of temperature, humidity 50%, 4 grades of illumination, pH is between 7-8 in flowerpot, and TDS content is between 0-800ppm.Growth cabinet
YC-D25 ultrasonic humidifier in connection, periodically adds distilled water to humidifier, and keeping the humidity of growth cabinet is 50%.In order to
Ensure that moisture content is 30%, soil one day in floral disc plus a water.In order to make the microorganism in soil connect oxygen area abundance, just
It is frequently grown breeding, grease uniformly mixes, and Soil ventilation is loose, keeps digging daily primary.Battalion is added in degradation process simultaneously
Nutrient solution keeps C:N:P=100:10:3 in soil.Required nutrient solution prescription are as follows:
Ammonium nitrate (NH3NO3): 2g/L
Dipotassium hydrogen phosphate (K2HPO4): 1.5g/L
Potassium dihydrogen phosphate (KH2PO4): 3g/L
Magnesium sulfate (MgSO4): 0.05g/L
Calcium chloride (CaCl2): 0.01g/L
Sodium chloride (NaCl): 0.5g/L
Ferrous sulfate (FeSO4): 0.01g/L
Sodium ethylene diamine tetracetate (Na2EDTA): 0.01g/L
(4) degradation kinetics is analyzed
By the resid amount in regression equation analysis soil.
The resid amount logarithm of the lower three groups of processing of 1 different time of table
In the identical situation of the contaminated degree of soil, different disposal can cause very big influence to degradation half life, the
One group of half-life period is 37.47 days, and second group of half-life period is 36.10 days, and the half-life period of third group is 14.82 days, explanation
The half-life period that west figure Ze Shi pseudomonad can be obviously shortened oil degradation is added.By degradation in 30 days, west figure Ze Shi is added
The processing group degradation rate highest of pseudomonad, has reached 77.2%, and first group with second group of degradation rate 45% or so.
(5) n-alkane organic evolution Parameter analysis
The ingredient of crude oil is divided into saturated hydrocarbons, aromatic hydrocarbon, nonhydrocarbon, asphalt colloid, and wherein the content of saturated hydrocarbons is up to
70% or so.Main component is alkane C12~C35 in saturated hydrocarbons, including normal alkane series, isoparaffin are serial and class is different
Pentadiene alkane series --- norpristane, pristane and phytane, they account for the 94.6% of saturated hydrocarbon fraction.Therefore can divide
The organic evolution parameter of n-alkane is analysed to explain degradation rule.Organic evolution parameter includes alkane main peak carbon, w (∑ C21-)/w
(∑ C22+), OEP, w (pr)/w (ph) etc..
Alkane main peak carbon and w (∑ C21-)/parameter of w (∑ C22+) 2 high carbon number when reflecting alkane by bacterial action
Trend of the alkane to low carbon number alkane transformations.The the Forward of main peak carbon the more, show that the high carbon number alkane ability of bacterial degradation is stronger.w
The ratio of n-alkane summation and n-alkane summation after C22, the parameter are got over before (∑ C21-)/w (∑ C22+) is C21
Greatly, then show that the ability of the high carbon number alkane of bacterial degradation is stronger.OEP value is the even odd carbon number mass ratio of high carbon number n-alkane,
OEP=(C25+6*C27+C29)/4 (C26+C28), the value reflect the energy of the high carbon number odd even carbon number n-alkane of bacterial degradation
Power, the ability for being worth smaller degradation odd number carbon alkane is stronger, and on the contrary then even number carbon alkane of degrading ability is stronger.Pristane to phytane ratio w (pr)/
W (ph) is to reflect sample Organic oxidation using the ratio of pr (pristane) and ph (phytane) common in isoprenoid hydrocarbon
The parameter of reducing degree.Following table is the organic evolution parameter value after degrading 30 days.
2 n-alkane organic evolution parameter value of table
The OEP value of first group as can be seen from Table 2, second group and third group gradually rises, and illustrates that west figure Ze Shi is added
The ability for even carbon number alkane of degrading can be improved after pseudomonad.W (Σ C21-)/w (Σ C22+) parameter value 0.643 < 0.836 <
0.861, illustrate to embody low carbon number advantage in degradation process, shows that west figure Ze Shi pseudomonad, which is added, can be improved to high-carbon
The degradation of number alkane.W (Pr)/w (Ph) parameter value gradually decreases, and first group little with second group of gap, the variation of third group
It is very big, illustrate to promote isoprenoid alkenes that demethylation reaction has occurred after west figure Ze Shi pseudomonad is added, part is planted
Alkane molecule has sloughed a methyl and has been converted into pristane, and the ability of degradation isoprenoid alkenes can be improved.It is drilled by biology
Change Parameter analysis, can degrade high carbon number alkane difficult to degrade in n-alkane, even carbon number alkane is added after the figure Ze Shi pseudomonad of west
The components such as hydrocarbon and isoprenoid alkenes.Therefore degradation rate can be improved after west figure Ze Shi pseudomonad is added.
Sequence table
<110>Cnpc Changqing Drilling Engineering Co., Ltd. Changqing Downhole Technology Operating Company
<120>a kind of pseudomonas drop oil bacterium L-1 and its microorganism mix bacterium agent and oil-polluted soils in-situ remediation method
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1452
<212> DNA
<213>west figure Ze Shi pseudomonad (Pseudomonas stutzeri strain)
<400> 1
acgctgtgcg gcaggcctaa cacatgcaag tcgagcggat gagtggagct tgctccatga 60
ttcagcggcg gacgggtgag taatgcctag gaatctgcct ggtagtgggg gacaacgttt 120
cgaaaggaac gctaataccg catacgtcct acgggagaaa gtgggggatc ttcggacctc 180
acgctatcag atgagcctag gtcggattag ctagttggtg aggtaaaggc tcaccaaggc 240
gacgatccgt aactggtctg agaggatgat cagtcacact ggaactgaga cacggtccag 300
actcctacgg gaggcagcag tggggaatat tggacaatgg gcgaaagcct gatccagcca 360
tgccgcgtgt gtgaagaagg tcttcggatt gtaaagcact ttaagttggg aggaagggca 420
gtaagttaat accttgctgt tttgacgtta ccaacagaat aagcaccggc taacttcgtg 480
ccagcagccg cggtaatacg aagggtgcaa gcgttaatcg gaattactgg gcgtaaagcg 540
cgcgtaggtg gttcgttaag ttggatgtga aagccccggg ctcaacctgg gaactgcatc 600
caaaactggc gagctagagt atggcagagg gtggtggaat ttcctgtgta gcggtgaaat 660
gcgtagatat aggaaggaac accagtggcg aaggcgacca cctgggctaa tactgacact 720
gaggtgcgaa agcgtgggga gcaaacagga ttagataccc tggtagtcca cgccgtaaac 780
gatgtcgact agccgttggg atccttgaga tcttagtggc gcagctaacg cattaagtcg 840
accgcctggg gagtacggcc gcaaggttaa aactcaaatg aattgacggg ggcccgcaca 900
agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag gccttgacat 960
gcagagaact ttccagagat ggattggtgc cttcgggaac tctgacacag gtgctgcatg 1020
gctgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgtaacgagc gcaacccttg 1080
tccttagtta ccagcacgtt aaggtgggca ctctaaggag actgccggtg acaaaccgga 1140
ggaaggtggg gatgacgtca agtcatcatg gcccttacgg cctgggctac acacgtgcta 1200
caatggtcgg tacaaagggt tgccaagccg cgaggtggag ctaatcccat aaaaccgatc 1260
gtagtccgga tcgcagtctg caactcgact gcgtgaagtc ggaatcgcta gtaatcgtga 1320
atcagaatgt cacggtgaat acgttcccgg gccttgtaca caccgcccgt cacaccatgg 1380
gagtgggttg ctccagaagt agctagtcta accttcgggg ggacggttac cacggagtga 1440
ttcatgactg gg 1452
Claims (5)
1. oily bacterium L-1 drops in a kind of pseudomonas, it is characterised in that: be isolated from Oilfield Oil-Contained Silt, polluted from petroleum aromatic
Soil supension culture obtains purpose bacterial strain, isolates and purifies to obtain in bacterial strain that select 1 plant of degradation capability most Johnson & Johnson head the most vigorous
Strain Designation is L-1, which belongs to pseudomonas (Pseudomonadaceae), deposit number CGMCC
NO.16557, DNA sequence dna is as shown in SEQ NO<400>1.
2. including a kind of microorganism mix bacterium agent of the oily bacterium L-1 of pseudomonas drop described in claim 1, it is characterised in that:
Containing the pseudomonas that indigenous bacterium and deposit number are CGMCC NO.16557, oily bacterium L-1 drops, pseudomonas drop oily bacterium with
Indigenous bacterium solution is that bacteria suspension is made in 1:3~1:6 in mass ratio.
3. a kind of oil-polluted soils in-situ remediation method using microorganism mix bacterium agent described in claim 2, feature exist
In:
(1) required microorganism mix bacterium agent bacterial strain is first brought back to life, then expands culture, fermenting and producing is spare；Indigenous bacterium is bud
Spore Bacillus (Bacillus encimensis strain encimensis), Bacillus foecalis alkaligenes (Alcaligenes
Faecalis strain), yellow class Nocard's bacillus (Nocardioides luteus), acinetobacter (Acinetobacter
Sp), penicillium oxalicum (Penicillium oxalicum), by 6 plants of microbial inoculums in microorganism mix bacterium agent according to pseudomonas
Oily bacterium L-1: bacillus: Bacillus foecalis alkaligenes: yellow class Nocard's bacillus: acinetobacter: penicillium oxalicum bacterium solution volume drops
25~35 DEG C are put into fermentor than the ratio for 1:1:1:1:1:1~1.1:1.1:1.1:1.1:1.1:0.5,170~230r/
Min fermentation is spare；
(2) microorganism mix bacterium agent and microbial inoculum nutrient solution NS-1 are added according to 4% (wt)~6% (wt), makes C:N:P=in soil
100:10:1~100:10:5；Dig ventilation and watering are carried out daily, increases oxygen-supplying amount, keep moisture 40-50%；
(3) periodic detection oil content, oil component and microbial activities situation, enhance process are monitored and controlled.
4. oil-polluted soils in-situ remediation method as claimed in claim 3, it is characterised in that: the microorganism mix bacterium agent pair
The condition of oil degradation be 25-35 DEG C of temperature, pH between 7-8, total dissolved solidss TDS is between 0-800ppm.
5. a kind of oil-polluted soils in-situ remediation method as claimed in claim 3, it is characterised in that: microbial inoculum is added in soil
Nutrient solution NS-1 is matched with cultivating the pseudomonas drop oil bacterium L-1 and the microorganism mix bacterium agent, the nutrient solution NS-1
Side:
Ammonium nitrate (NH3NO3): 1.5-2.5g/L
Dipotassium hydrogen phosphate (K2HPO4): 1-2g/L
Potassium dihydrogen phosphate (KH2PO4): 2.5-3.5g/L
Magnesium sulfate (MgSO4): 0.01-0.1g/L
Calcium chloride (CaCl2): 0.01-0.02g/L
Sodium chloride (NaCl): 0.3-0.6g/L
Ferrous sulfate (FeSO4): 0.01-0.02g/L
Sodium ethylene diamine tetracetate (Na2EDTA): 0.01-0.02g/L.
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| CN109679871A (en) * | 2019-01-08 | 2019-04-26 | 西北民族大学 | A kind of method of PAM-SA immobilized microorganism degradation oily waste water |
| CN109929781A (en) * | 2019-03-26 | 2019-06-25 | 中国矿业大学 | One plant of degradation luxuriant and rich with fragrance bacterial strain and its application in soil remediation |
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| CN114214233A (en) * | 2021-12-22 | 2022-03-22 | 黄河三角洲京博化工研究院有限公司 | Styrene degrading bacterium and application thereof |
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