CN106350471B - Method for directionally and rapidly screening and enriching broad-spectrum nitrobacteria - Google Patents
Method for directionally and rapidly screening and enriching broad-spectrum nitrobacteria Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 24
- 241000108664 Nitrobacteria Species 0.000 title claims abstract description 20
- 238000012216 screening Methods 0.000 title claims abstract description 12
- 239000010802 sludge Substances 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 18
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims abstract description 17
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims abstract description 10
- 230000005764 inhibitory process Effects 0.000 claims abstract description 5
- 241000894006 Bacteria Species 0.000 claims description 33
- 230000001546 nitrifying effect Effects 0.000 claims description 13
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 235000013619 trace mineral Nutrition 0.000 claims description 12
- 239000011573 trace mineral Substances 0.000 claims description 12
- 239000010865 sewage Substances 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 239000007836 KH2PO4 Substances 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 6
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 6
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 6
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 6
- 230000000694 effects Effects 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- 238000005273 aeration Methods 0.000 claims description 4
- 230000014759 maintenance of location Effects 0.000 claims description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 3
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 claims description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 3
- 229910018890 NaMoO4 Inorganic materials 0.000 claims description 3
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 claims description 3
- 229910052927 chalcanthite Inorganic materials 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 3
- 229910052564 epsomite Inorganic materials 0.000 claims description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 claims description 3
- 230000001133 acceleration Effects 0.000 claims description 2
- 230000008859 change Effects 0.000 claims description 2
- 235000016709 nutrition Nutrition 0.000 claims 1
- 238000006396 nitration reaction Methods 0.000 abstract description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- 241001453382 Nitrosomonadales Species 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000002351 wastewater Substances 0.000 description 4
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000012136 culture method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000012851 eutrophication Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004065 wastewater treatment Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- General Health & Medical Sciences (AREA)
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Abstract
A method for directionally and rapidly screening and enriching broad-spectrum nitrobacteria belongs to the field of water treatment. The activated sludge is adopted as a seed source, so that the structural characteristics of the original AOB and NOB flora are maintained; a high DO maintaining method is adopted, so that the high efficiency of nitration is maintained; under the condition of high DO, controlling FA and FNA to be lower than the inhibition concentration of nitrobacteria by utilizing pH according to the concentrations of ammonia nitrogen and nitrite, keeping the high nitrification efficiency and realizing the rapid culture of the nitrobacteria.
Description
Technical Field
The invention belongs to the field of water treatment, and particularly relates to a technology for directionally and rapidly screening and enriching broad-spectrum nitrobacteria, which aims to rapidly enrich and culture the broad-spectrum nitrobacteria in a short time, wherein the broad-spectrum nitrobacteria comprise Ammonia Oxidizing Bacteria (AOB) and Nitrite Oxidizing Bacteria (NOB), and the culture of the flora has wide application prospects in preparation of nitrifying bioactive fillers and improvement of ammonia nitrogen nitrification performance of sewage plants.
Background
Along with the development of urban modernization, industry and agriculture, the pollution of nitrogen-containing wastewater is more severe, and the over-high ammonia pollution content can cause the eutrophication of water bodies, so that algae can be propagated in large quantities, thereby harming organisms in water. And even destroy the ecological balance. The reduction of the nitrogen concentration in the water body and the pollution caused by the nitrogen in the water body are important subjects in the modern wastewater treatment technology. The traditional method for treating ammonia nitrogen wastewater is biological nitrification and denitrification. In the existing sewage treatment process, the nitrification efficiency of the active sludge system of the existing sewage treatment plant is only 15 mg/(L.h), and a large amount of heterotrophic bacteria, denitrifying bacteria and other strains are associated in the flora, so that the nitrifying bacteria cannot become dominant flora, and the treatment efficiency of the ammonia nitrogen wastewater is low. In the existing experimental research for improving the nitrification efficiency, a single strain of high-efficiency nitrifying bacteria is mostly taken as a core, and a technical route of screening, culturing and enriching the single strain and adding the single strain to a system is adopted. The problems with this technical route are: the single strain has poor adaptability to the environment. The single-strain efficient nitrobacteria hardly form ecological advantages in a sewage treatment system with complex multiple types of bacteria.
Therefore, aiming at the problems, the invention provides a method for directionally and rapidly screening and enriching broad-spectrum nitrobacteria. Because the broad-spectrum nitrobacteria have high ecological stability, the ecological advantage is easily formed in a sewage treatment system. The strain is derived from the return sludge of the existing sewage treatment plant and originally has a nitrification function. The broad-spectrum nitrifying bacteria are classified into the single screened bacteria, and the population distribution characteristics of the original sludge can be maintained. The invention produces bacteria with population characteristics by enriching a large amount of nitrifying bacteria. Based on the premise, the enrichment culture broad-spectrum nitrobacteria can be used for preparing biological nitrification active fillers, can also be added into a reaction tank to directly treat nitrogen-containing wastewater, and can also be used as a source of an enzyme preparation. Different from the traditional single high-efficiency nitrifying bacteria, the technology takes the number as the advantage, and realizes the improvement of the nitrification efficiency in the active sludge system of the existing sewage treatment plant.
Disclosure of Invention
The invention aims to develop a rapid enrichment culture method based on broad-spectrum nitrobacteria, which realizes the large consumption of heterotrophic bacteria and the maintenance of high nitrification rate through high DO; the high oxidation rate of ammonia nitrogen and nitrite is kept through the effective control of Free Ammonia (FA) and Free Nitrous Acid (FNA) inhibiting factors and the effective setting of high dissolved oxygen DO (between 1.0mg/L and 2.0 mg/L), thereby realizing the rapid culture of nitrobacteria.
The invention adopts the activated sludge as the seed source to keep the structural characteristics of the original AOB and NOB flora; a high DO maintaining method is adopted, so that the high efficiency of nitration is maintained; under the condition of high DO, controlling FA and FNA to be lower than the inhibition concentration of nitrobacteria by utilizing pH according to the concentrations of ammonia nitrogen and nitrite, and keeping high nitrification efficiency, thereby realizing the rapid culture of the nitrobacteria.
The enrichment culture method of the broad-spectrum nitrobacteria comprises the following steps:
(1) the method comprises the steps that returned sludge of a sewage plant is used as a strain source, the DO is maintained to be 1.5-2.0mg/L through continuous aeration to realize the large consumption of heterotrophic bacteria, under the condition of no ammonia nitrogen addition, the change of COD and DO of precipitated supernatant of mixed liquor is used as an indication index of a heterotrophic bacteria consumption process, in the process, the COD content of original sludge is low, the COD content of the precipitated supernatant of the mixed liquor is increased along with the death of the heterotrophic bacteria and fluctuates up and down, when the consumption of the heterotrophic bacteria is exhausted, the COD value of the precipitated supernatant of the mixed liquor is kept relatively stable and is maintained to be 40-80 mg/L, and the DO of the precipitated supernatant of the mixed liquor is continuously increased and finally tends to be stable; the sludge concentration is reduced along with the process, and the death of heterotrophic bacteria is reflected from another angle;
(2) performing inorganic culture on the sludge obtained in the step (1), wherein the inlet water of the inorganic culture comprises NH4Cl base solution and inorganic culture solution, wherein NH4The Cl adding amount is along with NH in a sludge culture system4 +Variation in the concentration of N, the inorganic substance in the inorganic culture medium comprising Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2O、CaCl2·2H2O, trace elements; wherein Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2The concentration of O in the sludge culture system is 20mg/L, CaCl2·2H2O10 mg/L, wherein the trace elements are 0.5mL of trace element solution added into each liter of sludge culture system, and the composition and concentration of the trace element solution are as follows: zn2SO4·7H2O is 0.12mg/L, NaMoO4·2H2O is 0.12mg/L, CoCl2·6H2O is 0.15mg/L, Mn2SO4·H2O is 0.12mg/L, NiCl2·6H2O is 0.10mg/L, CuSO4·5H2O is 0.03mg/L, FeCl3·6H2O is 1.5 mg/L.
The inorganic culture solution is added intermittently or continuously to maintain the basic nutrient condition of nitrifying bacteria.
In the culture process, the reaction temperature is controlled at 25 ℃, DO in a sludge culture system is in the range of 1.0-1.2 mg/L, and the pH is 7.5-8; controlling the inhibition effect of FA and FNA on NOB, namely controlling FA to be between 3mg/L and 12mg/L and FNA to be below 0.01mg/L, but utilizing the acceleration effect of nitrite on NOB, namely the nitrite concentration is above 150mg/L to accelerate NOB; under the premise that the hydraulic retention time is kept unchanged, the concentration of the ammonia nitrogen in the inlet water is gradually increased, so that the nitrification efficiency is continuously increased to 150 mg/(L.h) or even above, and in the process, the concentration of the sludge is continuously increased along with the increase of the culture time, and finally, the screening and high-concentration enrichment of nitrifying bacteria (AOB and NOB) are realized.
Preferably: and (2) under the premise that the hydraulic retention time is kept unchanged, the ammonia nitrogen concentration of the inlet water is gradually increased, so that the nitrification efficiency is continuously increased to 150 mg/(L.h) within 8-12 days of a short time, even more.
Preferably: and (3) the initial concentration of the ammonia nitrogen concentration of the inlet water in the step (2) is 100 mg/L.
The invention has the following beneficial effects:
(1) through continuous aeration at the initial stage of culture and ammonia nitrogen-free addition, a large amount of heterotrophic bacteria die in an endogenous respiration mode, so that the types of bacteria are purified.
(2) By effectively matching pH and ammonia nitrogen adding amount, FA and FNA are effectively controlled to realize rapid screening and enrichment of broad-spectrum nitrobacteria.
(3) In the culture process, after the heterotrophic bacteria are consumed, the growth of NOB is promoted by high-concentration accumulation of nitrite.
(4) The nitrified activated sludge of the sewage treatment plant is taken as a seed source to be screened and enriched, and the composition characteristics of the original AOB and NOB floras are kept.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1
The culture mode has obvious effect through small tests, and the specific operation is as follows:
(1) the returned sludge is cultured by using a barrel type SBR reactor, continuous aeration is carried out for 4 days without adding ammonia nitrogen, the original sludge concentration (MLSS) is 6976mg/L, DO is 1.5-2.0mg/L, COD is 16.084mg/L, MLSS is changed into 5324mg/L after 4 days, DO content is increased to 6.34mg/L, COD fluctuates up and down after straight rising, and finally is stabilized at about 60 mg/L. Apparent conditions show that a large amount of heterotrophic bacteria are completely consumed, and then the enrichment of broad-spectrum nitrobacteria is carried out.
(2) On the basis of the step (1), an intermittent water feeding mode is adopted, and by controlling water feeding components, the temperature in the system is 25 ℃, DO is 1.0-1.2 mg/L, and pH is 7.5-8, so that the optimal conditions are provided for growth of nitrifying bacteria. The feed water comprises NH4Cl base solution and inorganic substance solution, NH4The Cl adding amount is along with NH in a sludge culture system4 +Variation in the concentration of N, inorganic substances including Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2O、CaCl2·2H2O, trace elements; wherein Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2The concentration of O in the sludge culture system is 20mg/L, CaCl2·2H2O10 mg/L, wherein the trace elements are 0.5mL of trace element solution added into each liter of sludge culture system, and the composition and concentration of the trace element solution are as follows: zn2SO4·7H2O is 0.12mg/L, NaMoO4·2H2O is 0.12mg/L, CoCl2·6H2O is 0.15mg/L, Mn2SO4·H2O is 0.12mg/L, NiCl2·6H2O is 0.10mg/L, CuSO4·5H2O is 0.03mg/L, FeCl3·6H2O is 1.5 mg/L.
And determining that the ammonia nitrogen concentration of inlet water is gradually increased to 300mg/L from 100mg/L, controlling the FA concentration not to exceed the inhibition concentration of NOB, controlling the FNA not to exceed 0.01mg/L when the nitrite concentration is less than 150mg/L, and culturing for 10 days to stably increase the sludge concentration by 200mg/L and improve the nitrification efficiency to 150 mg/(L.h) under the condition of continuously discharging sludge.
Claims (3)
1. The method for directionally and rapidly screening and enriching broad-spectrum nitrobacteria is characterized by comprising the following steps:
(1) the method comprises the steps that returned sludge of a sewage plant is used as a strain source, the DO is maintained to be 1.5-2.0mg/L through continuous aeration to realize the large consumption of heterotrophic bacteria, under the condition of no ammonia nitrogen addition, the change of COD and DO of precipitated supernatant of mixed liquor is used as an indication index of a heterotrophic bacteria consumption process, in the process, the COD content of original sludge is low, the COD content of the precipitated supernatant of the mixed liquor is increased along with the death of the heterotrophic bacteria and fluctuates up and down, when the consumption of the heterotrophic bacteria is exhausted, the COD value of the precipitated supernatant of the mixed liquor is kept relatively stable and is maintained to be 40-80 mg/L, and the DO of the precipitated supernatant of the mixed liquor is continuously increased and finally tends to be stable; the sludge concentration is reduced along with the process, and the death of heterotrophic bacteria is reflected from another angle;
(2) performing inorganic culture on the sludge obtained in the step (1), wherein the inlet water of the inorganic culture comprises NH4Cl base solution and inorganic substance culture solution, wherein NH4The Cl adding amount is along with NH in a sludge culture system4 +Variation in the concentration of N, inorganic substances including Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2O、CaCl2·2H2O, trace elements; wherein Na2CO3、K2HPO4、KH2PO4、MgSO4·7H2The concentration of O in the sludge culture system is 20mg/L, CaCl2·2H2O10 mg/L, wherein the trace elements are 0.5mL of trace element solution added into each liter of sludge culture system, and the composition and concentration of the trace element solution are as follows: zn2SO4·7H2O is 0.12mg/L, NaMoO4·2H2O is 0.12mg/L, CoCl2·6H2O is 0.15mg/L, Mn2SO4·H2O is 0.12mg/L, NiCl2·6H2O is 0.10mg/L, CuSO4·5H2O is 0.03mg/L, FeCl3·6H2O is 1.5 mg/L; culturingIn the process, the reaction temperature is controlled at 25 ℃, DO in a sludge culture system is in the range of 1.0-1.2 mg/L, and the pH is 7.5-8; controlling the inhibition effect of FA and FNA on NOB, namely controlling FA to be between 3mg/L and 12mg/L and FNA to be below 0.01mg/L, but utilizing the acceleration effect of nitrite on NOB, namely controlling the nitrite concentration to be above 150 mg/L; under the premise that the hydraulic retention time is kept unchanged, the ammonia nitrogen concentration of the inlet water is gradually increased, so that the nitrification efficiency is continuously increased to be more than or equal to 150 mg/(L.h), and in the process, the sludge concentration is continuously increased along with the increase of the culture time, and finally, the screening and high-concentration enrichment of nitrifying bacteria are realized;
the initial concentration of the ammonia nitrogen of the inlet water in the step (2) is 100 mg/L.
2. The method for directionally and rapidly screening enriched broad-spectrum nitrifying bacteria according to claim 1, wherein an inorganic culture solution is added intermittently or continuously to maintain the basic nutritional conditions of the nitrifying bacteria.
3. The method for directionally and rapidly screening the enriched broad-spectrum nitrifying bacteria according to claim 1, wherein the step (2) is characterized in that the concentration of the ammonia nitrogen in the inlet water is gradually increased under the premise that the hydraulic retention time is kept unchanged, so that the nitrification efficiency is continuously increased to be more than or equal to 150 mg/(L.h) within 8-12 days of short time.
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| CN108726677A (en) * | 2017-04-25 | 2018-11-02 | 湖南大学 | A kind of new process promoting low-carbon-source wastewater treatment |
| CN112746052B (en) * | 2019-10-30 | 2022-08-12 | 中国石油化工股份有限公司 | Culture method for improving resistance of nitrobacteria |
| CN114735811B (en) * | 2022-03-23 | 2023-01-20 | 北京工业大学 | Culture method of short-cut nitrification activated sludge with high accumulation rate |
| CN115261261B (en) * | 2022-06-23 | 2023-12-12 | 浙江巨能环境工程有限公司 | Autotrophic ammonia oxidizing bacteria enrichment method |
| CN116354506B (en) * | 2023-03-21 | 2024-01-19 | 北京工业大学 | Method for realizing heterotrophic nitrification-aerobic denitrification high-efficiency denitrification through stress of high-concentration quorum sensing inhibitor |
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| CN104726376A (en) * | 2015-03-27 | 2015-06-24 | 中蓝连海设计研究院 | Continuous large-scale culture method for high-concentration salt-resistant nitrifying bacteria |
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