CN104357035A - Biological bactericide for preventing and controlling SRB (sulfate reducing bacteria) in high-temperature water body and SRB inhibition method of bactericide - Google Patents
Biological bactericide for preventing and controlling SRB (sulfate reducing bacteria) in high-temperature water body and SRB inhibition method of bactericide Download PDFInfo
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- CN104357035A CN104357035A CN201410569095.9A CN201410569095A CN104357035A CN 104357035 A CN104357035 A CN 104357035A CN 201410569095 A CN201410569095 A CN 201410569095A CN 104357035 A CN104357035 A CN 104357035A
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- 241000894006 Bacteria Species 0.000 title claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 15
- 230000005764 inhibitory process Effects 0.000 title abstract description 9
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 title abstract description 5
- 230000000844 anti-bacterial effect Effects 0.000 title abstract 7
- 239000003899 bactericide agent Substances 0.000 title abstract 7
- 239000007788 liquid Substances 0.000 claims abstract description 41
- 238000009655 industrial fermentation Methods 0.000 claims abstract description 15
- 238000010564 aerobic fermentation Methods 0.000 claims abstract description 9
- 230000001580 bacterial effect Effects 0.000 claims abstract description 8
- 241001663853 Parageobacillus toebii Species 0.000 claims abstract description 7
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 28
- 239000003129 oil well Substances 0.000 claims description 25
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 22
- 244000005700 microbiome Species 0.000 claims description 15
- 239000004317 sodium nitrate Substances 0.000 claims description 14
- 235000010344 sodium nitrate Nutrition 0.000 claims description 14
- 229940001516 sodium nitrate Drugs 0.000 claims description 14
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 13
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 239000002068 microbial inoculum Substances 0.000 claims description 12
- 229940045641 monobasic sodium phosphate Drugs 0.000 claims description 11
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 11
- 235000010288 sodium nitrite Nutrition 0.000 claims description 11
- 238000004321 preservation Methods 0.000 claims description 5
- 229960000819 sodium nitrite Drugs 0.000 claims description 4
- 241001037822 Bacillus bacterium Species 0.000 claims description 2
- 229910002651 NO3 Inorganic materials 0.000 abstract description 8
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 abstract description 8
- 238000005516 engineering process Methods 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 238000009629 microbiological culture Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 15
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 description 12
- 235000015097 nutrients Nutrition 0.000 description 12
- 239000002609 medium Substances 0.000 description 8
- 230000008859 change Effects 0.000 description 7
- 230000009467 reduction Effects 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 239000012530 fluid Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 239000000706 filtrate Substances 0.000 description 4
- 230000001629 suppression Effects 0.000 description 4
- 238000007400 DNA extraction Methods 0.000 description 3
- 230000007797 corrosion Effects 0.000 description 3
- 238000005260 corrosion Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 238000012408 PCR amplification Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 206010020718 hyperplasia Diseases 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 229910021653 sulphate ion Inorganic materials 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000186541 Desulfotomaculum Species 0.000 description 1
- 101001065501 Escherichia phage MS2 Lysis protein Proteins 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 241001226178 bacterium enrichment culture Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 244000144987 brood Species 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000001311 chemical methods and process Methods 0.000 description 1
- 238000005352 clarification Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000003398 denaturant Substances 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- IMBKASBLAKCLEM-UHFFFAOYSA-L ferrous ammonium sulfate (anhydrous) Chemical compound [NH4+].[NH4+].[Fe+2].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O IMBKASBLAKCLEM-UHFFFAOYSA-L 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 230000016507 interphase Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 230000003245 working effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/54—Compositions for in situ inhibition of corrosion in boreholes or wells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K8/00—Compositions for drilling of boreholes or wells; Compositions for treating boreholes or wells, e.g. for completion or for remedial operations
- C09K8/52—Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning
- C09K8/528—Compositions for preventing, limiting or eliminating depositions, e.g. for cleaning inorganic depositions, e.g. sulfates or carbonates
-
- E—FIXED CONSTRUCTIONS
- E21—EARTH OR ROCK DRILLING; MINING
- E21B—EARTH OR ROCK DRILLING; OBTAINING OIL, GAS, WATER, SOLUBLE OR MELTABLE MATERIALS OR A SLURRY OF MINERALS FROM WELLS
- E21B37/00—Methods or apparatus for cleaning boreholes or wells
- E21B37/06—Methods or apparatus for cleaning boreholes or wells using chemical means for preventing or limiting, e.g. eliminating, the deposition of paraffins or like substances
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- E—FIXED CONSTRUCTIONS
- E21—EARTH OR ROCK DRILLING; MINING
- E21B—EARTH OR ROCK DRILLING; OBTAINING OIL, GAS, WATER, SOLUBLE OR MELTABLE MATERIALS OR A SLURRY OF MINERALS FROM WELLS
- E21B41/00—Equipment or details not covered by groups E21B15/00 - E21B40/00
- E21B41/02—Equipment or details not covered by groups E21B15/00 - E21B40/00 in situ inhibition of corrosion in boreholes or wells
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2208/00—Aspects relating to compositions of drilling or well treatment fluids
- C09K2208/32—Anticorrosion additives
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Abstract
The invention relates to a technology for inhibiting SRB (sulfate reducing bacteria) in oilfield produced liquid, in particular to a biological bactericide for preventing and controlling the SRB in a high-temperature water body and an SRB inhibition method of the bactericide. The biological bactericide is JSHD-4 geobacillustoebii, has a sequence as shown in a sequence table SEQ ID NO:1, is collected in China General Microbiological Culture Collection Center (CCMCC) on March 5, 2013, and has a Latin literature name of Geobacillustoebii and a collection number of CGMCCNO7273. The JSHD-4 geobacillustoebii is subjected to microbial aerobic fermentation to prepare industrial fermentation liquid with the bacterial solution concentration of 1.0*10<6> to 1.3*10<10> cfu/mL, the tolerable temperature is 50-95 DEG C, and the optimal growth temperature is 60-70 DEG C. After the multifunctional biological bactericide prepared from the JSHD-4 geobacillustoebii is thrown into a well, nitrate reducing bacteria (DNB) are quickly proliferated in the high-temperature environment in the well, so that the biological bactericide is suitable for directly treating the SRB in the high-temperature environment in the well on line.
Description
Technical field
The present invention relates to the sulphate reducing bacteria suppression technology field in the water body of oil field, particularly a kind of bacteria agent preventing and treating SRB in high temperature water body and the method suppressing SRB thereof.
Background technology
Oil well etching problem long-standing problem oil exploration industry, is also the problem that oil well production and scientific research personnel pay close attention to always, is not solved preferably so far.Large quantity research shows, oil well biological corrosion mainly harmful microorganism sulphate reducing bacteria (SRB) is grown in a large number, causes sulfide concentration increase to cause.At present, oil well control SRB and sulfide mainly take chemistry and physical method, there is high, the easy secondary pollution of cost, form the deficiencies such as resistance.
Oil field sulfate reductive bacterium biological control collection is defeated to be risen in recent years gradually, the method that this technology mainly utilizes biological competition to eliminate, and by substituting of microbial population, harmful microorganism problem is become favorable factor; But the SRB that this technology main study subject is oilfield transportation system administers, and the research for this high temperature of oil well extreme environment (more than 50 DEG C) is less.
Multifunctional bio microbial inoculum in the present invention can use under the High Temperatures In Oil-well environment of 50 ~ 95 DEG C, make nitrate reduction flora (DNB) hyperplasia diffusion rapidly, and when competing living space and matrix with SRB, DNB prioritizing selection uses the matrix in oil field system, nutrition required for effective prevention SRB obtains, thus the metabolic activity of control SRB.
Summary of the invention
The object of the invention is the deficiency for SRB biological inhibition technology in prior art, by injecting thermophilic multifunctional bio microbial inoculum in oil field well hot environment, the quick growth of SRB in effective suppression hot environment, eliminate the sulfide produced, stimulate the growth of denitrifying bacterium simultaneously, change microorganisms in water structure of community, thus reach the object preventing biological corrosion.
An object of the present invention is, a kind of bacteria agent preventing and treating SRB in the high temperature water body environment of oil field is provided, described bacteria agent is JSHD-4 ground bacillus, there is the sequence shown in sequence table SEQ ID NO:1, be preserved in Chinese microorganism strain preservation management trust person and understand common micro-organisms preservation center, Latin name is Geobacillus
toebii, deposit number is CGMCC NO 7273 preservation date is on March 5th, 2013.
In the present invention, it is 1.0 × 10 that described JSHD-4 ground bacillus obtains bacterial concentration by microorganism aerobic fermentation
6~ 1.3 × 10
10the industrial fermentation liquid of cfu/mL, tolerable temperature is 50-95 DEG C, and optimum growth temp is 60-70 DEG C.JSHD-4 ground bacillus tolerable temperature in the present invention is high, in can adapting to high temperature subsurface environment in the suppression of SRB, prevent down-hole biological corrosion.
Another object of the present invention is, a kind of method adopting above-mentioned bacteria agent to suppress SRB in the high temperature water body environment of oil field is provided, adds the multifunctional bio microbial inoculum of following ratio in the amount of oil field high temperature water body: JSHD-4 ground bacillus: obtaining bacterial concentration by microorganism aerobic fermentation is 1.0 × 10
6~ 1.3 × 10
10the industrial fermentation liquid of cfu/mL, the ratio that adds of described industrial fermentation liquid is oil field high temperature water body volume 0.5 ~ 1%; SODIUMNITRATE 20 ~ 30 mg/L; Sodium Nitrite 50 ~ 150 mg/L; SODIUM PHOSPHATE, MONOBASIC 5 ~ 10 mg/L
As the further improvement of the inventive method, described JSHD-4 ground bacillus is 1.0 ~ 1.3 × 10 by the concentration of the industrial fermentation liquid of aerobic fermentation
8cfu/mL, the ratio of adding is oil field high temperature water body volume 0.8%; SODIUMNITRATE 25 mg/L; Sodium Nitrite 75 mg/L; SODIUM PHOSPHATE, MONOBASIC 8 mg/L.
Improve as a step more of the present invention, in method of the present invention, when adding described multifunctional bio microbial inoculum in target oil well, first well casing gas is pressed to casing pressure back to zero, JSHD-4 ground bacillus bacterium liquid directly adopts impact type to add to add in oil well casing, first be dissolved in the water of 25-30L after SODIUMNITRATE, Sodium Nitrite and SODIUM PHOSPHATE, MONOBASIC weigh in proportion, then adopt impact type to add and add in oil well casing, finally in oil well casing, add 50L water again.
The invention has the beneficial effects as follows: JSHD-4 ground bacillus of the present invention can use under the High Temperatures In Oil-well environment of 50 ~ 95 DEG C, it is 1.0 × 10 that bacterial classification can obtain bacterial concentration through biology aerobic fermentation
6~ 1.3 × 10
10the industrial fermentation liquid of cfu/mL gemma, after adopting the multifunctional bio microbial inoculum of JSHD-4 ground bacillus preparation to drop into down-hole, nitrate reduction flora (DNB) hyperplasia diffusion is rapidly made under the hot environment of down-hole, effectively can suppress the quick growth of SRB in hot environment, eliminate the sulfide produced, stimulate emulation the growth of microorganism denitrifying bacterium simultaneously, change microorganisms in water structure of community, its sulfide inhibiting rate is more than 93%, the work-ing life of effective prolongation pipeline, improve water quality, greatly reduce production cost.
biological deposits
Microorganism involved in the present invention submits biological deposits to:
Embodiment
Embodiment 1
JSHD-4 ground bacillus of the present invention is screened by following process:
(1) sample collecting
Gather the production fluid of 7 mouthfuls of oil wells at Jiangsu oilfield, sample ID is respectively: CH2-43, CH2-42, CH2-36, T83-1, T83-10, T83-7, H88-10.
(2) substratum configuration
Enrichment medium is mixed by solution A and B solution and forms, wherein:
Solution A: inorganic salt K1 2.0 g, l-asparagine 1.0 g, distilled water 500 ml, pH value 7.0 ~ 7.2;
B solution: Trisodium Citrate 8.5 g, sodium acetate 2 g, glucose 2 g, KH
2pO
40.5 g, K
2hPO
40.5 g, MgSO
47H
2o 1.0 g, FeCl
36H
2o 0.05 g, CaCl
20.1 g, yeast extract 0.5 g, distilled water 500 ml.PH value 7.0 ~ 7.2.
NRB liquid nutrient medium: A liquid and independent 121 DEG C of sterilizing 20 min of B liquid, equal proportion mixing after sterilizing.
NRB solid medium: respectively add the agar that mass ratio is 1.5% ~ 2% in A liquid and B liquid, equal proportion mixing after independent sterilizing.
(3) enrichment culture
By 7 mouthfuls of produced liquid in oil well sample equal proportion mixing in step (1), then add NRB liquid nutrient medium, be placed in 65 DEG C, 75 DEG C (whether should change 70 DEG C into) aerobic cultivations of incubator respectively; Meanwhile, with the Produced Liquid sterilizing sample processed equally as control sample.Be seeded in identical NRB liquid nutrient medium with the inoculum size of 5% after cultivating 6d under 65 DEG C of conditions and carry out switching cultivation, again transfer after cultivating 3 d, switching cultivation 3 times, carries out domestication and cultivates under being then forwarded to 70 DEG C of conditions.In cultivation rear mensuration nutrient solution, K1 ionic concn is in table 1.
Through three switchings, under 65 DEG C, 70 DEG C conditions, culture solution is muddy, and the clarification of control sample nutrient solution, show microorganism raised growth in experimental group nutrient solution, simultaneously, shown by table 1 nutrient solution K1 concentration determination, 70 DEG C of domestication cultivation 2 d, K1 concentration is reduced to 0.24 g/L by 2.0 g/L, and 65 DEG C of third time switching nutrient solution K1 concentration determination is 0, consume in a large number with K1 ion while that solution K1 ionic concn significantly reducing explanation microorganism growth after cultivating, show that experimental group nutrient solution contains nitrate reduction bacterium, and grow vigorous under experimental conditions.
(4) strains separation purifying
The sample of 65 DEG C of three switchings, 70 DEG C of domestications is applied to NRB solid medium slat chain conveyor, and from gained culture plate, picking list bacterium colony is cultivated in Erlenmeyer flask, cultivates 3 d and is placed on 4 DEG C of Refrigerator stores.
(5) nitrate reduction bacterium DNA extraction and qualification
(I) DNA extraction
By the nitrate reduction bacterium enrichment culture after separation and purification 3 days, with 2 mL centrifuge tubes centrifugal collecting cell under 12000 × g condition, adopt Axygen DNA extraction kit to extract nitrate reduction bacterium DNA, concrete operation step is as follows:
1) in the centrifuge tube that cell is housed, add 450 μ L lysates, vortex vibrates 15 s, 65 DEG C of water-bath 10 min;
2) continue to add 400 μ L protein denaturants and 1 mL to be separated liquid (4 DEG C of precoolings), firmly mixing, centrifugal 2 min of 12000 × g;
3) abandon phase, retain INTERPHASE CARBIDE PRECIPITATION and lower phase, add the liquid that is separated of 1 mL, 4 DEG C of precoolings, mixing, centrifugal 2 min of 12000 × g;
4) phase is abandoned, by lower phase transition to filter (filter is placed in 2 mL centrifuge tubes), centrifugal 1 min of 12000 × g;
5) abandon filter, in filtrate, add 400 μ L DNA in conjunction with liquid, mix;
6) being placed in 2 mL centrifuge tubes by preparing pipe, the mixed solution in step (5) being moved into and prepares in pipe, centrifugal 1 min of 12000 × g;
7) abandon filtrate, put get back to preparing pipe in former 2 mL centrifuge tubes, add 500 μ L washing fluid W1, centrifugal 1 min of 12000 × g;
8) abandon filtrate, put get back to preparing pipe in former 2 mL centrifuge tubes, add 700 μ L washing fluid W2, centrifugal 1 min of 12000 × g;
9) wash once with 700 μ L washing fluid W2 more in the same way;
10) abandon filtrate, put preparing pipe and get back in former 2 mL centrifuge tubes, centrifugal 1 min of 12000 × g;
11) will prepare pipe and be placed in 1.5 mL centrifuge tubes of another cleaning, add 100-200 μ L Eluent or deionized water in silica film central authorities, room temperature leaves standstill the centrifugal 1 min eluted dna of 1 min, 12000 × g, with for subsequent use at being kept at-20 DEG C.
(II) sample P CR amplification
By the DNA of the JSHD04 of extraction, 16S rDNA gene universal primer 1492R and 27F is adopted to carry out pcr amplification.
1) bacterial 16 S rDNA PCR reaction system
PCR reaction system composition is in table 2.
2) primer sequence
2) primer sequence
1492R：5‘-GGTTACCTTGTTACGACTT-3’
27F：5‘＞CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGAGAGTTTGATCTTGGCTCAG＜3’
3) PCR response procedures
1． 95 ℃ 3 min
2． 94 ℃ 40 sec
3． 52 ℃ 45 sec
4． Goto 2 35 times
5． 72 ℃ 10 min
(III) sequencing result
Jiangsu oilfield production fluid 65 DEG C of enriched sample isolate bacterium colony (numbering JSHD-4), picking list bacterium colony enrichment culture, and culturing cell extracts DNA, and pcr amplification product checks order, if the sequence of result as shown in SEQ ID NO:1.
By analysis comparison and comparison by analysis with
g. toebii strain R-32639similarity is 97%, is subspecies of ground bacillus.
Embodiment 2
Under System of Laboratories condition, JSHD-4 is to the inhibition of SRB
SRB culture medium prescription is improved according to table 3, be that solvent carries out substratum configuration with sterilized water, substratum is through 121 DEG C of sterilizing 20 min, and after cooling, often liter of substratum adds trace element solution (forming as shown in table 4) 1 mL, vitamin solution (forming as shown in table 5) 1 mL and more respectively through the ferrous ammonium sulphate 0.1g of ultraviolet disinfection and L-cysteine hydrochloride 0.5 g; SRB bacterial classification (Desulfotomaculum) 3 ~ 4 ring preserved from solid slope transfering loop picking is inoculated in SRB substratum, cultivates 48 hours, obtain the SRB bacterium liquid of fresh enrichment culture at 65 DEG C.
The JSHD-4 of preservation is used at 75 DEG C NRB liquid nutrient medium (the NRB liquid nutrient medium with in example 1) activation culture 1 d simultaneously, obtain the JSHD-4 bacterium liquid of fresh enrichment culture, then JSHD-4 bacterium liquid and SRB bacterium liquid is inoculated in the substratum of improvement SRB (assay flask is the anaerobism bottle of 250ml) by the inoculative proportion of 2%, and the SODIUMNITRATE adding 10 mmol/L is cultivated at 65 DEG C, simultaneously with only inoculate SRB bacterium liquid and add 10 mmol/L SODIUMNITRATE and do not inoculate nitrate reduction bacterium for control group, relatively nutrient solution change, by detecting H
2the content of S characterizes JSHD-4 in multifunctional bio microbial inoculum to the rejection ability of SRB activity, and detected result is as shown in table 6).After cultivating 7 d, only add the H in the control group of the SODIUMNITRATE of SRB and interpolation 10 mmol/L
2s content is very high, and is adding H in the inhibition system containing JSHD-4
2s concentration all keeps very low at the 3rd d and the 7th d, shows that the activity of SRB in experimental group is obviously suppressed.
Embodiment 3
First denitrifying bacterium JSHD-4 being prepared brood cell's concentration by conventional microbiological aerobic fermentation is 1.0 × 10
6cfu/mL industrial fermentation liquid, this industrial fermentation liquid can use 20L plastic tank to store, and then gets SODIUMNITRATE, Sodium Nitrite and SODIUM PHOSPHATE, MONOBASIC and is commercially available technical grade product, mix in proportion during use, also can add target oil well produced liquid successively.
JSHD-4 multifunctional bio microbial inoculum of the present invention is dropped into Jiangsu oilfield Wang39Jing in following ratio, and before process, the sulfide concentration of this well production fluid is 28mg/L.Dosage is: JSHD-4 industrial fermentation liquid (1.0 × 10
6cfu/mL), dosage is 0.5% of Produced Liquid content; SODIUMNITRATE: 20mg/L; Sodium Nitrite: 50mg/L; SODIUM PHOSPHATE, MONOBASIC: 5mg/L
When adding above-mentioned multifunctional bio microbial inoculum in target oil well, first well casing gas is pressed to casing pressure back to zero, JSHD-4 industrial fermentation liquid directly adopts impact type to add to add in oil well casing, first be dissolved in the water of 25L after SODIUMNITRATE, Sodium Nitrite and SODIUM PHOSPHATE, MONOBASIC weigh in proportion, then adopting impact type to add adds in oil well casing, finally in oil well casing, adds 50L water again.Irregularly detect sulfide content simultaneously, and supplement interpolation multifunctional bio microbial inoculum in good time; To keep inhibition.Inhibition sulfide change curve as shown in Figure 1,60d inhibiting rate reaches 93.14%.
Embodiment 4
Difference from Example 3 is, implementing oil well is Jiangsu oilfield Wei2-53Jing, and sulfide 128.4mg/L, JSHD-4 industrial fermentation liquid spore concentration is 1.3 × 10
10cfu/mL, dosage is 1% of oil well produced liquid hold-up; SODIUMNITRATE: 30mg/L; Sodium Nitrite: 150mg/L; SODIUM PHOSPHATE, MONOBASIC: 10mg/L.Inhibition sulfide change curve as shown in Figure 1,60d inhibiting rate reaches 97.73%.
Embodiment 5
Difference from Example 3 is, implementing oil well is the yellow 88-39 well of Jiangsu oilfield, and sulfide 56mg/L, JSHD-4 industrial fermentation liquid concentration is 1.3 × 10
8cfu/mL, dosage is 0.8% of Produced Liquid content; SODIUMNITRATE: 25mg/L; Sodium Nitrite: 75mg/L; SODIUM PHOSPHATE, MONOBASIC: 8mg/L.Inhibition sulfide change curve as shown in Figure 1,60d inhibiting rate reaches 94.93%.
Sequence table
<110> Sinopec Group; Sinopec Co., Ltd. Jiangsu Oil Field Branch
<120> prevents and treats the bacteria agent of SRB in high temperature water body and suppresses the method for SRB
<160>1
<210>1
<211>1012
<212>DNA
<213> artificial sequence
<400>1
ATCATGCAAG TCGAGCGGAC CGAACGGAAG CTTGCTTCTG TTCGGTTAGC GGCGGACGGG 60
TGAGTAACAC GTGGGTAACC TGCCCGTAAG ACCGGGATAA CTCCGGGAAA CCGGGGCTAA 120
TACCGGATAA CACCGAAGAC CGCATGGTCT TTGGTTGAAA GGTGGCTTTT GCTACCACTT 180
ACGGATGGGC CCGCGGCGCA TTAGCTAGTT GGTGAGGTAA CGGCTCACCA AGGCGACGAT 240
GCGTAGCCGG CCTGAGAGGG TGACCGGCCA CACTGGGACT GAGACACGGC CCAGACTCCT 300
ACGGGAGGCA GCAGTAGGGA ATCTTCCGCA ATGGACGAAA GTCTGACGGA GCGACGCCGC 360
GTGAGCGAAG AAGGTCTTCG GATCGTAAAG CTCTGTTGTT AGGGAAGAAG AAGTACCGTT 420
CGAATAGGGC GGTACGGTGA CGGTACCTAA CGAGAAAGCC CCGGCTAACT ACGTGCCAGC 480
AGCCGCGGTA ATACGTAGGG GGCGAGCGTT GTCCGGAATT ATTGGGCGTA AAGCGCGCGC 540
AGGCGGTCC CTTAAGTCTGA TGTGAAAGCC CACGGCTCAA CCGTGGAGGG TCATTGGAAA 600
CTGGGGGACT TGAGTGCAGA AGAGGAGAGC GGAATTCCAC GTGTAGCGGT GAAATGCGTA 660
GAGATGTGGA GGAACACCAG TGGCGAAGGC GGCTCTCTGG TCTGTAACTG ACGCTGAGGC 720
GCGAAAGCGT GGGGGAGCAA ACAGGATTAG ATACCCTGGT AGTCCACGCC GTAAACGATG 780
AGTGCTAAGT GTTAGAGGGG TTTTCCCTTT AGTGCTGTAG CTAACGCGTT AAGCACTCCG 840
CCTGGGGAGT ACGGCGCATG CTGAACTCAA GAATTGACGG GGCCCGCACA AGCGTGAGCA 900
TGTGGTTTAA TTCGAGCACG CGAGACCTTA CAGTCTGACA TCCCTGACAC CTGGAAATGC 960
GTTCCCCCTC GGGGGACAGG TGACAGGGGG GCATGGTGTC GTCACACTCG TG 1012
<210>2
<211>19
<212>DNA
<213> artificial sequence
<400>2
GGTTACCTTGTTACGACTT
<210>3
<211>60
<212>DNA
<213> artificial sequence
<400>3
CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCACGGGGGGAGAGTTTGATCTTGGCTCAG
Claims (5)
1. prevent and treat the bacteria agent of SRB in high temperature water body for one kind, it is characterized in that, described bacteria agent is JSHD-4 ground bacillus, there is the sequence shown in sequence table SEQ ID NO:1, be preserved in Chinese microorganism strain preservation management trust person and understand common micro-organisms preservation center, Latin name is Geobacillus
toebii, deposit number is CGMCC NO 7273 preservation date is on March 5th, 2013.
2. the bacteria agent of SRB in the high temperature water body environment of control oil field according to claim 1, it is characterized in that, it is 1.0 × 10 that described JSHD-4 ground bacillus obtains bacterial concentration by microorganism aerobic fermentation
6~ 1.3 × 10
10the industrial fermentation liquid of cfu/mL, tolerable temperature is 50-95 DEG C, and optimum growth temp is 60-70 DEG C.
3. a bacteria agent according to claim 1 suppresses the method for SRB in the high temperature water body environment of oil field, it is characterized in that, add the multifunctional bio microbial inoculum of following ratio in the amount of oil field high temperature water body: described JSHD-4 ground bacillus is oil field high temperature water body volume 0.5 ~ 1% by the ratio that adds of the industrial fermentation liquid of aerobic fermentation; SODIUMNITRATE 20 ~ 30 mg/L; Sodium Nitrite 50 ~ 150 mg/L; SODIUM PHOSPHATE, MONOBASIC 5 ~ 10 mg/L.
4. bacteria agent according to claim 3 suppresses the method for SRB in the high temperature water body environment of oil field, it is characterized in that, add the multifunctional bio microbial inoculum of following ratio in the amount of oil field high temperature water body: described JSHD-4 ground bacillus is 1.0 ~ 1.3 × 10 by the concentration of the industrial fermentation liquid of aerobic fermentation
8cfu/mL, the ratio of adding is oil field high temperature water body volume 0.8%; SODIUMNITRATE 25 mg/L; Sodium Nitrite 75 mg/L; SODIUM PHOSPHATE, MONOBASIC 8 mg/L.
5. the bacteria agent according to claim 3 or 4 suppresses the method for SRB in the high temperature water body environment of oil field, it is characterized in that, when adding described multifunctional bio microbial inoculum in target oil well, first well casing gas is pressed to casing pressure back to zero, JSHD-4 ground bacillus bacterium liquid directly adopts impact type to add to add in oil well casing, first be dissolved in the water of 25-30L after SODIUMNITRATE, Sodium Nitrite and SODIUM PHOSPHATE, MONOBASIC weigh in proportion, then adopting impact type to add adds in oil well casing, finally in oil well casing, adds 50L water again.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182211A (en) * | 2018-10-16 | 2019-01-11 | 中国石油化工股份有限公司 | The composite bacteria agent and application method of SRB are prevented and treated under wide temperature environment |
| CN111011398A (en) * | 2019-11-12 | 2020-04-17 | 广州航海学院 | Biological bactericide for crude oil transportation pipeline and application thereof |
| CN111997582A (en) * | 2019-05-27 | 2020-11-27 | 惠博普(武汉)生物环保科技有限公司 | Biological treatment technology for hydraulic control sulfur extracted from oil field |
| CN112342167A (en) * | 2020-11-19 | 2021-02-09 | 中海石油(中国)有限公司 | Method for inhibiting sulfate reducing bacteria in high-temperature oil field water body |
| CN113956860A (en) * | 2021-09-16 | 2022-01-21 | 华东理工大学 | Construction method of microbial corrosion control system of oil field system |
| CN115161001A (en) * | 2022-06-23 | 2022-10-11 | 华东理工大学 | Method for inhibiting oil field biological cause hydrogen sulfide |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101235359A (en) * | 2008-03-04 | 2008-08-06 | 武汉科技学院 | Sulfate reducing bacteria resisting microbial preparation and preparation method thereof |
| CN101987770A (en) * | 2010-08-07 | 2011-03-23 | 中国石油化工股份有限公司 | Method for removing hydrogen sulfide from oil field injection water |
| CN103113866A (en) * | 2013-02-06 | 2013-05-22 | 中国石油化工股份有限公司 | Novel biological inhibitor for inhibiting activity of sulfate reducing bacteria in oil well and use method thereof |
-
2014
- 2014-10-23 CN CN201410569095.9A patent/CN104357035B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101235359A (en) * | 2008-03-04 | 2008-08-06 | 武汉科技学院 | Sulfate reducing bacteria resisting microbial preparation and preparation method thereof |
| CN101987770A (en) * | 2010-08-07 | 2011-03-23 | 中国石油化工股份有限公司 | Method for removing hydrogen sulfide from oil field injection water |
| CN103113866A (en) * | 2013-02-06 | 2013-05-22 | 中国石油化工股份有限公司 | Novel biological inhibitor for inhibiting activity of sulfate reducing bacteria in oil well and use method thereof |
Non-Patent Citations (2)
| Title |
|---|
| SIRI MYHR ET AL.: "Denitrovibrio acetiphilus, a novel genus and species of dissimilatory nitrate-reducing bacterium isolated from an oil reservoir model column", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 * |
| 庄文 等: "油田采出液中硝酸盐还原菌的分离培养及对硫酸盐还原菌的抑制研究", 《科学技术与工程》 * |
Cited By (10)
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|---|---|---|---|---|
| CN109182211A (en) * | 2018-10-16 | 2019-01-11 | 中国石油化工股份有限公司 | The composite bacteria agent and application method of SRB are prevented and treated under wide temperature environment |
| CN109182211B (en) * | 2018-10-16 | 2021-09-07 | 中国石油化工股份有限公司 | Composite biological agent for preventing and treating SRB in wide-temperature environment and use method |
| CN111997582A (en) * | 2019-05-27 | 2020-11-27 | 惠博普(武汉)生物环保科技有限公司 | Biological treatment technology for hydraulic control sulfur extracted from oil field |
| CN111011398A (en) * | 2019-11-12 | 2020-04-17 | 广州航海学院 | Biological bactericide for crude oil transportation pipeline and application thereof |
| CN111011398B (en) * | 2019-11-12 | 2021-07-23 | 广州航海学院 | Biological bactericide for crude oil transportation pipeline and application thereof |
| CN112342167A (en) * | 2020-11-19 | 2021-02-09 | 中海石油(中国)有限公司 | Method for inhibiting sulfate reducing bacteria in high-temperature oil field water body |
| CN112342167B (en) * | 2020-11-19 | 2022-03-08 | 中海石油(中国)有限公司 | Method for inhibiting sulfate reducing bacteria in high-temperature oil field water body |
| CN113956860A (en) * | 2021-09-16 | 2022-01-21 | 华东理工大学 | Construction method of microbial corrosion control system of oil field system |
| CN115161001A (en) * | 2022-06-23 | 2022-10-11 | 华东理工大学 | Method for inhibiting oil field biological cause hydrogen sulfide |
| CN115161001B (en) * | 2022-06-23 | 2023-11-03 | 华东理工大学 | Inhibition method of oilfield biological cause hydrogen sulfide |
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