CN109234201B - Microbial oil recovery bacterium W-Y10 and application thereof - Google Patents
Microbial oil recovery bacterium W-Y10 and application thereof Download PDFInfo
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- E21B—EARTH OR ROCK DRILLING; OBTAINING OIL, GAS, WATER, SOLUBLE OR MELTABLE MATERIALS OR A SLURRY OF MINERALS FROM WELLS
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Abstract
The invention discloses a microbial oil recovery bacterium W-Y10 and application thereof. The microbial oil recovery bacterium W-Y10 of the invention has been preserved in China general microbiological culture Collection center (CGMCC) in 2018, 9 and 26, and the preservation number is CGMCC No. 16533. Experiments prove that: the microbial oil recovery bacterium W-Y10 can emulsify crude oil, can be used for microbial oil recovery and oil recovery rate improvement, can be used for treatment and restoration of oil pollution, and has good application prospect.
Description
Technical Field
The invention relates to the technical field of microbial oil recovery, in particular to a microbial oil recovery bacterium W-Y10 and application thereof.
Background
The petroleum pollution is widely distributed, and relates to various industries such as oil extraction, oil refining, chemical engineering, machinery and the like, and various polluted environments such as petroleum leakage and the like. The petroleum hydrocarbon components are complex and diverse, and have strong hydrophobicity, so that the petroleum hydrocarbon components are difficult to be contacted, degraded and utilized by common microorganisms. The microbial oil recovery technology is to inject nutrients or microbes into the stratum and to utilize the growth and metabolism of microbes in oil deposit to raise the yield and recovery rate of crude oil. The traditional petroleum functional bacterial strain is separated from water and soil. The existing research shows that the microorganisms are widely distributed in the petroleum oil phase besides water and soil respectively, so that the separation of related functional strains from the petroleum oil phase has important significance in the microbial oil recovery technology.
A new strain named Aquamiumaestivarii was isolated by Hyun Mijin et al by enrichment at the beach contaminated with crude oil, but it was not examined whether it has a function of degrading crude oil (Aquamiumaestivarii sp. nov., a marine bacterium isolated from a tidaflat). A strain of benzo (a) pyrene degrading bacteria named Aquamiumlusatiense is separated by Ferying and the like (separation and identification of the benzo (a) pyrene degrading bacteria in soil of a coal polluted area). A chlorohexadecane degrading bacterium named Aquamicrobiumaerosum is separated from Wangjianing and the like (diversity analysis of the chlorohexadecane degrading bacterium in surface seawater of the arctic pole and research on a genome and polycyclic aromatic hydrocarbon degrading mechanism of Alteromonas sp.P127). Androenia et al isolated a non-degrading strain of bacteria (Bacterial communities and enzymologicalvisions of PAHs poluted soils) named Aquamibium defluvium. Wang X et al isolated a strain of a petroleum hydrocarbon (phenanthrene and pyrene) degrading bacterium named Aquamicrobium defluvium (Draft genome sequence of Aquamicrobium defluvium strain w13Z1, a polysaccharide microorganism strain hydro-degrading bacterium and a Draft genome sequence of a polysaccharide microorganism strain hydrocarbon-degrading strain W13Z 2).
Disclosure of Invention
One purpose of the invention is to provide an Aquamibium sp.W-Y10 strain.
The Aquamibium sp.W-Y10 provided by the invention has a preservation number of CGMCC No.16533, is classified and named as Aquamibium sp, and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms (CGMCC for short, the address: No. 3 of West Lu 1 of the morning district of Beijing, China academy of sciences, postal code 100101) in 26.9.2018.
Another purpose of the invention is to provide a new application of Aquamibium sp.W-Y10 or a bacterial suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial inoculum containing the same.
The invention provides application of aquamicrobioum sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum recovery.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of petroleum products.
The invention also provides application of aquamicrobioum sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in microbial oil recovery.
The invention also provides application of aquamicrobioum sp.W-Y10 or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in preparation of microbial oil recovery products.
The invention also provides application of Aquamicrobium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in crude oil emulsification.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of crude oil emulsified products.
The invention also provides application of Aquamicrobium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum degradation.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of petroleum degradation products.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum pollution treatment.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of products for treating petroleum pollution.
The invention also provides application of Aquamibium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum pollution remediation.
The invention also provides application of Aquamicrobium sp.W-Y10 or bacterial suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of products for repairing petroleum pollution.
It is another object of the present invention to provide a product.
The active ingredient of the product provided by the invention is Aquamibium sp.W-Y10 or bacterial suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same;
the product has any one of the following functions 1) to 6):
1) oil recovery;
2) microbial oil recovery;
3) emulsifying crude oil;
4) degrading petroleum;
5) treating petroleum pollution;
6) and (4) repairing petroleum pollution.
It is yet another object of the present invention to provide a method for microbial oil recovery.
The microbial oil recovery method provided by the invention comprises the following steps: in the oil extraction process, the aquamicrobioum sp.W-Y10 or the bacterial suspension thereof, the culture solution thereof, the fermentation product thereof or the microbial inoculum containing the same is added.
It is a final object of the present invention to provide a method for remediation and/or rehabilitation of petroleum pollution.
The method for treating and/or repairing petroleum pollution provided by the invention comprises the following steps: the aquamicrobioum sp.W-Y10 or its bacterial suspension, its culture solution, its fermentation product or its bacteria agent containing it is used to treat petroleum pollutant.
In the above application or product or method, the preparation method of the microbial inoculum is as follows: the Aquamicrobium sp.W-Y10 CGMCC No.16533 is inoculated into a culture medium for culture to obtain the microbial inoculum. The concentration of the bacteria in the microbial inoculum is not less than 108cfu/mL。
The invention provides a microbial oil-producing strain W-Y10, which is classified and named as Aquamibium sp.A strain is preserved in China general microbiological culture Collection center (CGMCC) in 2018, 9 and 26, and the preservation number is CGMCC No. 16533. Experiments prove that: the microbial oil recovery bacterium W-Y10 can emulsify crude oil, can be used for microbial oil recovery and oil recovery rate improvement, can be used for treatment and restoration of oil pollution, and has good application prospect.
Drawings
FIG. 1 is a W-Y10 phylogenetic tree.
FIG. 2 is a phylogenetic tree of strains with similarity above 97%.
FIG. 3 shows W-Y10 emulsified thick oil. The experimental group W-Y10 is on the left, and the control group is on the right.
FIG. 4 is a graph of the displacement results of the model.
Deposit description
Latin name: aquamicrobioum sp.
The strain number is as follows: W-Y10
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)
Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: 9/26/2018
Registration number of the preservation center: CGMCC No.16533
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Example 1 isolation, identification and preservation of the W-Y10 Strain
Isolation of the first, W-Y10 Strain
In 5 months of 2017, oil phase petroleum is obtained from oil well produced liquid of a North China oil field of Erlianhaote, inner Mongolia, and a strain is obtained by separating the oil phase petroleum from the oil well produced liquid and named as a strain W-Y10.
Identification of the two, W-Y10 Strain
1. Morphological characterization of W-Y10 Strain
The W-Y10 strain has morphological characteristics that a round and regularly wet convex khaki colony with the diameter of 2mm is formed on a L B solid culture medium.
2. Physiological and biochemical identification of W-Y10 Strain
Physiological and biochemical characteristics of the W-Y10 strain: growth was possible at 30 ℃ with 1% sodium chloride, but not at 5% sodium chloride. Potassium gluconate and D-mannitol cannot be utilized.
3. Molecular characterization of the W-Y10 Strain
The activated W-Y10 strain is inoculated in a liquid L B culture medium (a solvent is water, solutes and the concentrations of the solutes are respectively 10 g/L NaCl, 10 g/L peptone, 5 g/L yeast powder and pH7-8), shaking culture is carried out at 35 ℃ and 150rpm for 24h, 1m L fresh culture solution is taken, the temperature is 4 ℃, 8000rpm is 5min, the thalli is collected in a 2m L centrifuge tube, DNA is extracted by a DNA extraction kit, after electrophoresis detection, PCR amplification is carried out by using a universal primer 8F/1492R, after electrophoresis detection of PCR products, a 16S rDNA gene sequence is determined, the specific sequence is shown as a sequence 1 in a sequence table, after B L AST search and comparison of the sequence 1 in NCBI, the W-Y10 strain is found to belong to Aquamobium, but has lower similarity with the existing strain, a software MEGA construction phylogenetic tree is shown as figure 1, the strain with the similarity of 97% is constructed again by using a software MEGA 2, wherein the strain is shown as a strain, and the highest physiological property of the strain W-Y-4838 strain is shown as a biological property shown in a biological growth tree shown in a software MEGA construction system shown in a picture 1.
TABLE 1 comparison of physiological and biochemical characteristics of the W-Y10 strain with the Aquamibium aestuarii strain
| Numbering | Color of colony | Potassium gluconate | D-mannitol |
| W-Y10 | Earthy yellow | - | - |
| Aquamicrobium aestuarii | Ivory color | + | + |
Note: -means no growth, + means growth
The comprehensive identification results show that the strain W-Y10 is a newly discovered Aquamiobium bacterium.
Deposit of the three, W-Y10 Strain
The W-Y10 strain is classified and named Aquamibium sp, and is preserved in China general microbiological culture Collection center (CGMCC for short, the address: No. 3 of West Lu 1 of Beijing Indoron area, Microbiol research institute of Chinese academy of sciences, postal code 100101) in 26.9.2018, and the preservation number is CGMCC No. 16533.
Example 2 emulsification of heavy oil by W-Y10 Strain
The oil in the embodiment is collected from North China oil field of Erlianhaote, inner Mongolia in 2017 and the viscosity of the oil at 35 ℃ is 570 mPa.s.
1. Inoculating the activated W-Y10 into liquid L B culture medium, and performing shaking culture at 35 deg.C and 150rpm for 24h to obtain W-Y10 bacterial liquid (bacterial concentration is not less than 10)8cfu/mL)。
2. Test group, 5 volume portions of bacterial liquid are inoculated to 95 volume portions of petroleum-inorganic salt culture medium (solvent is water, solute and its concentration are respectively 1 g/L petroleum, 5 g/L NaCl and 1 g/L NH)4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、3g/LKNO3pH7-8), 35 ℃ and 150rpm for 7 d.
Control group: 100 parts by volume of the petroleum-inorganic salt culture medium is subjected to shaking culture at 35 ℃ and 150rpm for 7 days.
3. After completion of step 2, the photographic results are shown in FIG. 3. In FIG. 3, the experimental group is shown on the left, and the control group is shown on the right. The strain W-Y10 has obvious emulsification effect on thick oil at 35 ℃.
4. And (3) measuring the residual petroleum amount in the system for completing the step (2) according to a method in the literature 'separation of crude oil degrading bacteria and degradation performance thereof', and calculating the petroleum degradation rate. Petroleum degradation rate (petroleum addition-petroleum residue)/petroleum addition. The calculation result shows that: the petroleum degradation rate is 50.1%.
Example 3 application of the W-Y10 Strain in microbial oil recovery technology
The oil in the embodiment is collected from North China oil field of Erlianhaote, inner Mongolia in 2017 and the viscosity of the oil at 35 ℃ is 570 mPa.s.
In the embodiment, the oil displacement effect of the microbial strains is evaluated through an indoor physical model oil displacement experiment. The specific experimental method is as follows:
1. using high pressure die-type pipe (specification)
Purchased from haian oil research instruments ltd) are filled with quartz sand with different meshes to form sand filling mold pipes. And measuring the permeability of the model pipe by adopting a permeability measuring device, and selecting the model pipe which meets the permeability. Vacuumizing saturated formation water, and calculating porosity; the parameters of the finally prepared model tubes for the subsequent steps are shown in table 2.
TABLE 2 model pipe Sand pack parameters
2. Continuously injecting petroleum from the inlet of the model pipe until the outlet of the model pipe detects the outflow of the petroleum, and measuring the original oil saturation; and aged at reservoir temperature (35 ℃) for 3 days.
3. And injecting water according to the injection quantity of the volume of the pore of the 1 model tube for displacing oil, stopping water drive when the outlet produced liquid reaches the limit water content (98%), and calculating the recovery ratio (namely the water drive recovery ratio).
4. Injecting bacteria-containing water according to the injection amount of the volume of the pore of the 1 model tube (after the injection is completed, calculating the recovery ratio, namely the recovery ratio in the process of injecting the microorganisms), and culturing for 15 days at the oil reservoir temperature (35 ℃).
The bacteria-containing water is prepared by inoculating activated W-Y10 into liquid L B culture medium, and performing shake culture at 35 deg.C and 150rpm for 24 hr to obtain W-Y10 bacteria solution (bacteria concentration is not less than 10)8cfu/m L) in notesAdding W-Y10 bacterial liquid and NH into water4H2PO4、(NH4)2SO4、K2HPO4、KNO3Peptone and yeast powder, wherein the concentrations of the peptone and the yeast powder in the bacteria-containing water are respectively 5% (volume fraction) W-Y10 bacteria liquid and 1 g/L NH4H2PO4、1g/L(NH4)2SO4、1g/L K2HPO4、3g/L KNO32 g/L peptone and 1 g/L yeast extract powder.
5. Injecting water according to the injection quantity of the volume of the pore of the 1 model tube for displacing oil, stopping water drive until the outlet produced liquid reaches the limit water content (98%), and calculating the recovery ratio (namely the recovery ratio of the subsequent water drive under the action of the microorganisms). The physical model flooding result curve is shown in figure 4.
The recovery ratios in different stages of water flooding, microbial injection, microbial action follow-up water flooding and the like are compared, and the results are shown in table 3. Indoor physical model oil displacement shows that the W-Y10 strain can improve the recovery ratio by 8.38%.
TABLE 3 recovery ratio
| Stage of material model | Recovery ratio (%) | Enhanced recovery (%) |
| Water drive | 48.24 | / |
| Microbial injection process | 52.65 | 4.41 |
| Follow-up water drive of microbial action | 56.62 | 8.38 |
Sequence listing
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<120> microbial oil recovery bacterium W-Y10 and application thereof
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<400>1
tcgagcgccc cgcaagggga gcggcagacg ggtgagtaac gcgtgggaat ctacccagct 60
ctacggaata acccagggaa acttggacta ataccgtata cgtccttcgg gagaaagatt 120
tatcggagtt ggatgagccc gcgtcggatt agctagttgg tgaggtaatg gctcaccaag 180
gcgacgatcc gtagctggtc tgagaggatg atcagccaca ctgggactga gacacggccc 240
agactcctac gggaggcagc agtggggaat attggacaat gggcgaaagc ctgatccagc 300
catgccgcgt gagtgatgaa ggccctaggg ttgtaaagct ctttcaccgg ggaagataat 360
gacggtaccc ggagaagaag ccccggctaa cttcgtgcca gcagccgcgg taatacgaag 420
ggggctagcg ttgttcggaa ttactgggcg taaagcgcac gtaggcggac tattaagtca 480
ggggtgaaat cccggggctc aaccccggaa ctgcctttga tactggtagt ctagagttcg 540
agagaggtga gtggaattcc gagtgtagag gtgaaattcg tagatattcg gaggaacacc 600
agtggcgaag gcggctcact ggctcgatac tgacgctgag gtgcgaaagc gtggggagca 660
aacaggatta gataccctgg tagtccacgc cgtaaacgat ggaagctagc cgttggcaag 720
tttacttgtc ggtggcgcag ctaacgcatt aagcttcccg cctggggagt acggtcgcaa 780
gattaaaact caaaggaatt gacgggggcc cgcacaagcg gtggagcatg tggtttaatt 840
cgaagcaacg cgcagaacct taccagccct tgacatcccg atcgcggtta ccggagacgg 900
tttccttcag ttcggctgga tcggtgacag gtgctgcatg gctgtcgtca gctcgtgtcg 960
tgagatgttg ggttaagtcc cgcaacgagc gcaaccctcg cccttagttg cccttattaa 1020
gttaggcact ctaaggggac tgccggtgat aagccgagag gaaggtgggg atgacgtcaa 1080
gtcctcatgg cccttacggg ctgggctaca cacgtgctac aatggtggtg acagtgggca 1140
gcgagaccgc gaggtcgagc taatctccaa aagccatctc agttcggatt gcactctgca 1200
actcgagtgc atgaagttgg aatcgctagt aatcgcggat cagcatgccg cggtgaatac 1260
gttcccgggc cttgtacaca ccgcccgtca caccatggga gttggcttta cccgaaggcg 1320
ctgcgctaac cgca 1334
Claims (10)
1. The Aquamibium sp.W-Y10 has the preservation number of CGMCC No. 16533.
2. The application of aquamicrobioum sp.W-Y10 or bacterial suspension thereof or microbial inoculum containing the same in petroleum recovery as described in claim 1;
or, the use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the preparation of petroleum products.
3. The use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in microbial oil recovery;
or, the use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the preparation of microbial oil recovery products.
4. Use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the emulsification of crude oil;
or, the use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the preparation of a crude oil emulsified product.
5. Use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in petroleum degradation;
or, the use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the preparation of petroleum degradation products.
6. Use of aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 in the treatment of petroleum pollution;
or, the application of aquamicrobioum sp.W-Y10 or the bacterial suspension thereof or the microbial inoculum containing the same in the preparation of products for treating petroleum pollution in claim 1.
7. The application of Aquamicrobium sp.W-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same in petroleum pollution remediation;
or, the application of aquamicrobioum sp.W-Y10 or the bacterial suspension thereof or the microbial inoculum containing the same in the preparation of products for repairing petroleum pollution in claim 1.
8. A product, wherein the active ingredient of the product is aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1;
the product has any one of the following functions 1) to 6):
1) oil recovery;
2) microbial oil recovery;
3) emulsifying crude oil;
4) degrading petroleum;
5) treating petroleum pollution;
6) and (4) repairing petroleum pollution.
9. A method of microbial oil recovery comprising the steps of: during oil recovery, aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing the same according to claim 1 is added.
10. A method for treating and/or remediating petroleum pollution, comprising the steps of: treating petroleum pollutants with aquamicrobioum sp.w-Y10 or a bacterial suspension thereof or a microbial inoculum containing same as defined in claim 1.
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| CN111961614B (en) * | 2020-08-04 | 2022-04-29 | 安徽科技学院 | Bacterium cd-1 for producing exopolysaccharide |
| CN115725432B (en) * | 2021-08-31 | 2023-11-28 | 中国石油天然气股份有限公司 | Acinetobacter capable of degrading crude oil heavy component to produce surfactant and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016122333A1 (en) * | 2015-01-28 | 2016-08-04 | Statoil Petroleum As | An automated method for selecting microbial strains which can degrade or emulsify oil |
| CN106635908A (en) * | 2016-12-29 | 2017-05-10 | 长安大学 | Advenella kashmirensis, microbial agent and application of advenella kashmirensis and microbial agent |
| KR101874956B1 (en) * | 2017-01-20 | 2018-07-05 | 신명옥 | Pseudomonas putida MZ3 bacteria for purifying oil-contaminated soil |
| CN108587987A (en) * | 2018-07-23 | 2018-09-28 | 北京润世能源技术有限公司 | One plant of microbe oil production bacterium W-Y6 and its application |
-
2018
- 2018-10-25 CN CN201811252377.0A patent/CN109234201B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016122333A1 (en) * | 2015-01-28 | 2016-08-04 | Statoil Petroleum As | An automated method for selecting microbial strains which can degrade or emulsify oil |
| CN106635908A (en) * | 2016-12-29 | 2017-05-10 | 长安大学 | Advenella kashmirensis, microbial agent and application of advenella kashmirensis and microbial agent |
| KR101874956B1 (en) * | 2017-01-20 | 2018-07-05 | 신명옥 | Pseudomonas putida MZ3 bacteria for purifying oil-contaminated soil |
| CN108587987A (en) * | 2018-07-23 | 2018-09-28 | 北京润世能源技术有限公司 | One plant of microbe oil production bacterium W-Y6 and its application |
Non-Patent Citations (3)
| Title |
|---|
| Draft Genome Sequence of Aquamicrobium defluvii Strain W13Z1，a Psychrotolerant Halotolerant Hydrocarbon Degrading Bacterium;Xinxin Wang等;《Genome Announcements》;20150831;第3卷(第4期);e00984-15 * |
| Petroleum Deradation： Promising Biotechnological Tools for Bioremediation;Maddela Naga Raju等;《Recent Insights in Petroleum Science and Engineering》;20180228;351-369 * |
| 中科院发现微生物采油菌株;郑宁来;《炼油技术与工程》;20140430;24 * |
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