CN112678950A - Method for synchronously strengthening anaerobic ammonium oxidation bacteria culture and enhancing stability by coupling nano zero-valent iron with magnetic field - Google Patents
Method for synchronously strengthening anaerobic ammonium oxidation bacteria culture and enhancing stability by coupling nano zero-valent iron with magnetic field Download PDFInfo
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Abstract
The invention relates to a method for synchronously strengthening the culture and stability of anaerobic ammonia oxidation bacteria by a nano zero-valent iron coupling magnetic field, which utilizes an nZVI coupling magnetic field to strengthen the culture of the anaerobic ammonia oxidation bacteria, the surface of the zero-valent iron can be passivated by the applied magnetic field intensity, the nano zero-valent iron is converted into iron ions under the magnetic field and is used as a trace element for microorganisms, the start of the anaerobic ammonia oxidation is effectively promoted, simultaneously the magnetic field is matched to influence the active sites of proteins in ammonia monooxygenase and hydroxylamine oxidase of the ammonia oxidation bacteria, the activity of the enzyme is improved, the enrichment rate of the ammonia oxidation bacteria is further improved, the start time of a reactor is greatly shortened, and the stability of the anaerobic ammonia oxidation reactor is enhanced. The method has simple process and mild reaction condition, and can be carried out under the condition of normal anaerobic ammoxidation; is safe and harmless to the environment and is convenient for engineering application.
Description
Technical Field
The invention relates to a method for strengthening the culture and stability of anaerobic ammonium oxidation bacteria by a nano zero-valent iron (nZVI) coupling magnetic field, belonging to the technical field of environmental microorganisms and wastewater treatment.
Background
The anaerobic ammonia oxidation process is widely concerned as an environment-friendly autotrophic nitrogen removal technology with low energy consumption. The process mainly depends on anaerobic ammonium oxidation bacteria under the anoxic condition to use NO2 --N is an electron acceptor, NH4 +Conversion of-N to N2. Compared with the traditional biological denitrification process, the shortcut nitrification-anaerobic ammonia oxidation process does not need organic matters as carbon sources, simultaneously reduces the oxygen demand by about 60 percent, reduces the residual sludge yield by about 90 percent, occupies small area and can save the operation cost by about 60 percent. In addition, the anaerobic ammonia oxidation process has higher nitrogen removal rate, is very suitable for treating high ammonia nitrogen wastewater, and reduces the discharge amount of greenhouse gases by about 90 percent compared with the traditional biological denitrification process. However, the anaerobic ammonium oxidation bacteria grow slowly, have long generation period, are sensitive to environmental change and can generate nitrate Nitrogen (NO) in the reaction process3 --N), etc., which are difficult to be widely used in engineering. Therefore, how to shorten the enrichment culture time of the anaerobic ammonia oxidation sludge and accelerate the starting process of the anaerobic ammonia oxidation is of great significance to the practical application of the technology.
The nanometer zero-valent iron (nZVI) has strong oxidation-reduction characteristics, so that the nanometer zero-valent iron is widely applied to environmental management. When nZVI is added to an anaerobic bioreactor, it can undergo the following redox reaction (formula 1) in a neutral anaerobic water body, slowly releasing Fe2+And creates an environment favorable for the survival of anaerobic microorganisms. Zero-valent iron and Fe in environment2+And can also participate in the reduction of nitrate nitrogen under the mediation of microorganisms (formula 2).
Fe0+2H2O→Fe2++H2+2OH-(formula 1)
However, as the oxidation reaction proceeds, goethite (α -FeOOH) and hematite (α -Fe) are formed on the surface of the zero-valent iron2O3) And the like, which causes the problems of zero-valent iron aging, activity reduction and the like.
Chinese patent document CN108557994A discloses a method for treating ammonia nitrogen wastewater, which is improved in that nitrogen-containing wastewater is treated by using montmorillonite-loaded zero-valent iron and anaerobic ammonium oxidation bacteria together. The method adopts the coupling of the montmorillonite-loaded zero-valent iron (MMT-ZVI) and the activated sludge to treat the nitrogen-containing wastewater, and compared with the method of not adding the MMT-ZVI and only adding the zero-valent iron (ZVI), the method has the advantages that the active sites of the zero-valent iron are increased after the montmorillonite is loaded, the stability is enhanced, and the denitrification efficiency is higher. The method cannot solve the problems of aging and activity reduction of zero-valent iron, cannot promote the growth of the anammox bacteria for a long time, and cannot enhance the stability of the anammox process.
Disclosure of Invention
Aiming at the defects of the prior art, in particular to the problems that the prior anaerobic ammonia oxidation bacteria have long culture time, the zero-valent iron is easy to age and the activity is reduced, and the practical application of the anaerobic ammonia oxidation process is seriously hindered, the invention provides a method for strengthening the culture and the stability of the anaerobic ammonia oxidation bacteria by coupling a nano zero-valent iron with a magnetic field.
The method can avoid the aging of the nano zero-valent iron, improve the activity of the nano zero-valent iron, promote the quick start of the anaerobic ammonia oxidation, promote the growth of anaerobic ammonia oxidizing bacteria, shorten the time for enriching and culturing the anaerobic sludge into the anaerobic ammonia oxidizing sludge and enhance the stability of the anaerobic ammonia oxidation reactor.
In order to solve the problems, the invention is realized by the following technical scheme:
a method for strengthening the culture and stability of anaerobic ammonium oxidation bacteria by a nano zero-valent iron coupling magnetic field comprises the following steps:
(1) adding nano zero-valent iron particles and anaerobic granular sludge into an upflow anaerobic sludge bed reactor, wherein the adding amount of the nano zero-valent iron particles is 1-30g/L of ammoniacal nitrogen and nitrite nitrogen to-be-treated wastewater;
(2) two permanent magnets are symmetrically arranged on the outer side of the reactor, and form a magnetic field in the horizontal direction in the reactor, and the magnetic field intensity is 5-120 mT;
(3) introducing wastewater to be treated containing ammonia nitrogen and nitrite nitrogen from the bottom of the upflow anaerobic sludge blanket reactor by using a peristaltic pump, stirring the wastewater to be treated at the anaerobic and light-proof condition of 30-34 ℃ to ensure that the anaerobic ammonia oxidation sludge in the reactor and the wastewater to be treated fully react, adjusting the rotating speed of the peristaltic pump according to the volume of the upflow anaerobic sludge blanket reactor to control the inflow velocity, and ensuring that the sewage to be treated hydraulically stays in the reactor for 6-24 hours to realize the quick start of anaerobic ammonia oxidation.
According to the invention, in the step (1), the addition amount of the nano zero-valent iron particles is 1-10g/L of ammoniacal nitrogen and nitrite nitrogen in the wastewater to be treated.
Further preferably, in the step (1), the addition amount of the nano zero-valent iron particles is 1-5g/L of ammoniacal nitrogen and nitrite nitrogen in the wastewater to be treated.
Most preferably, the addition amount of the nano zero-valent iron particles is 3g/L of ammoniacal nitrogen and nitrite nitrogen to-be-treated wastewater.
Preferably, in step (1), the nano zero-valent iron particles have a particle size of 50 to 70 nm.
According to the invention, in the step (1), the addition amount of the anaerobic granular sludge is 5450-5550 mgMLSS/L of ammoniacal nitrogen and nitrite nitrogen in the wastewater to be treated.
The anaerobic granular sludge is the granulated sludge which is generated in the organic wastewater treatment process and is rich in various anaerobic microorganism populations in the prior art, and is taken from the active anaerobic granular sludge of a corn starch sewage treatment plant.
Preferably, in step (2), the permanent magnet has a length of 12 to 15cm, a height of 8 to 12cm and a thickness of 2 to 3 cm.
Preferably, according to the invention, in step (2), the magnetic field strength is in the range of 50 to 100 mT.
Further preferably, in the step (2), the magnetic field strength is 50 to 70 mT.
According to the invention, in the step (2), the molar ratio of ammoniacal nitrogen to nitrous nitrogen in the wastewater to be treated containing ammoniacal nitrogen and nitrous nitrogen is 1: 1.2.
Since the anaerobic ammonium oxidation bacteria contain a plurality of iron storage 'sites', the stored iron element is usually Fe2+In the form ofThe coenzyme factors of Fe-S protein and hemoprotein participate in the electron transfer process of anammox bacteria, including NH4 +-N to N2And can promote metabolism of anammox bacteria. The nanometer zero-valent iron (nZVI) has strong oxidation-reduction characteristics, so that the nanometer zero-valent iron is widely applied to environmental management. When the nZVI is added into the anaerobic bioreactor, the nZVI can generate oxidation-reduction reaction in neutral anaerobic water body and slowly release Fe2+And an environment beneficial to the survival of anaerobic microorganisms is created, however, with the progress of oxidation reaction, the problems of the aging, the activity reduction and the like of the nano zero-valent iron nZVI can occur. The applied magnetic field strength can passivate the surface of zero-valent iron, nano zero-valent iron is converted into iron ions under the magnetic field and is used as a trace element for microorganisms, so that the anaerobic ammonia oxidation starting is effectively promoted, and simultaneously, the magnetic field is matched to influence the active sites of proteins in ammonia monooxygenase and hydroxylamine oxidase of ammonia oxidizing bacteria, so that the activity of the enzyme is improved, the enrichment rate of the ammonia oxidizing bacteria is further improved, and the starting time of the reactor is greatly shortened. The result shows that the nanometer zero-valent iron coupling magnetic field can greatly improve the enrichment rate of ammonia oxidizing bacteria, obviously shorten the starting process of anaerobic ammonia oxidation, ensure that the removal rate of ammoniacal nitrogen and nitrite nitrogen reaches more than 85 percent in a short time, and obviously enhance the stability of the anaerobic ammonia oxidation reactor.
The invention has the following characteristics and effects:
1. according to the invention, the culture of anaerobic ammonium oxidation bacteria is enhanced by using an nZVI coupling magnetic field, the surface of zero-valent iron can be passivated by applying a magnetic field intensity, nano zero-valent iron is converted into iron ions under the magnetic field and is used as a trace element for microorganisms, the anaerobic ammonium oxidation start is effectively promoted, meanwhile, the active sites of proteins in ammonia monooxygenase and hydroxylamine oxidase of the ammonium oxidation bacteria can be influenced by matching with the magnetic field, the activity of the enzyme is improved, the enrichment rate of the ammonium oxidation bacteria is further improved, the start time of a reactor is greatly shortened, and the method has the characteristics of high efficiency and safety.
2. The method utilizes the nZVI coupling magnetic field to strengthen the culture of the anaerobic ammonia oxidation bacteria and the stability of the process, can effectively shorten the starting time and strengthen the stability of the anaerobic ammonia oxidation reactor, is efficient, safe and mild in condition, and can be used under the conventional anaerobic ammonia oxidation condition. The method of the invention has low operation cost, which is mainly reflected in the consumption of nZVI. For example, 3g of nZVI is added into a sewage treatment system with an effective volume of 1L by one time by adopting the method, and a magnetic field with the strength of 65 +/-10 mT is continuously applied, so that the obvious promotion effect can be obtained, the consumption of the nZVI is low, the cost is low, and the nZVI is easy to purchase, safe and harmless to the environment and convenient for engineering application.
Drawings
FIG. 1 is a graph showing the effect of the anaerobic ammonia oxidation start-up process of R1 and R4 reactors in different treatments; a is ammonia nitrogen concentration, and b is nitrite nitrogen concentration; the abscissa is time (unit: d), and the ordinate is nitrogen concentration (unit: mg/L); the nitrogen concentration curves of R1 and R4 are the change curves of the nitrogen concentration in the R1 and R4 reactors along with time respectively.
FIG. 2 is a graph showing the effect of nitrogen removal in the load ramp-up phase of R1 and R4 reactors in different processes; a is ammonia nitrogen concentration, and b is nitrite nitrogen concentration; the abscissa is time (unit: d), and the ordinate is nitrogen concentration (unit: mg/L); the nitrogen concentration curves of R1 and R4 are the change curves of the nitrogen concentration in the R1 and R4 reactors along with time respectively.
FIG. 3 shows the copy number of the amamox bacteria 16S rRNA gene and the functional gene Hzo in different reactors at different stages, a is the copy number of the amamox bacteria 16S rRNA gene, and b is the copy number of the amamox bacteria functional gene Hzo.
Detailed Description
The invention is further illustrated by the following examples, but the scope of the invention as claimed is not limited to the scope described in the detailed description.
The raw materials used in the embodiment of the invention are all commercial products, and the used equipment is conventional equipment.
Examples the reactor used for sewage treatment was an Upflow Anaerobic Sludge Blanket (UASB), which is a commercially available conventional water treatment facility, and practical applications are not limited to the UASB reactor. In the examples, the R1 reactor (UASB) was used as a control (no nZVI and magnetic field added), the R2 reactor was a reactor to which only nano zero-valent iron was added, the R3 reactor was a reactor to which only a magnetic field was applied, and the R4 reactor (UASB) was a reactor to which a nano zero-valent iron-coupled magnetic field was added.
Example the contaminant in the Nitrogen-containing wastewater was NH4 +-N and NO2 --N, NH in the feed water4 +the-N is kept between 50 and 200 mg/L.
Example 1
A method for strengthening the culture and stability of anaerobic ammonium oxidation bacteria by a nano zero-valent iron coupling magnetic field comprises the following steps:
(1) adding nano zero-valent iron particles and anaerobic granular sludge into an upflow anaerobic sludge bed reactor R4, wherein the adding amount of the nano zero-valent iron particles is 3g/L of ammoniacal nitrogen and nitrite nitrogen wastewater to be treated; the grain size of the nano zero-valent iron particles is 60 nm; the addition amount of the anaerobic granular sludge is 5450-5550 mg MLSS/L of wastewater to be treated containing ammoniacal nitrogen and nitrite nitrogen;
(2) two permanent magnets with the length of 14cm, the height of 10cm and the thickness of 2.5cm are symmetrically arranged on the outer side of the reactor, a horizontal magnetic field is formed in the reactor by the two permanent magnets, and the magnetic field intensity is 65 mT;
(3) introducing wastewater to be treated containing ammoniacal nitrogen and nitrite nitrogen from the bottom of an upflow anaerobic sludge blanket reactor by using a peristaltic pump, adding 2L of ammoniacal nitrogen and nitrite nitrogen wastewater to be treated every day into the upflow anaerobic sludge blanket reactor, controlling the temperature to be 32 ℃, stirring under the condition of keeping out of the sun to ensure that anaerobic ammonia oxidation sludge in the reactor and the wastewater to be treated fully react, aerating for 10 minutes every day by using nitrogen, adjusting the rotating speed of the peristaltic pump according to the volume of the upflow anaerobic sludge blanket reactor to control the inflow velocity, ensuring that the hydraulic retention time of the wastewater to be treated in the reactor is 12 hours, and improving the degradation efficiency of pollutants in the wastewater; adding 2L of ammoniacal nitrogen and nitrite nitrogen to-be-treated wastewater every day during operation; through detection and calculation, the total nitrogen removal rate of the R4 reactor reaches more than 85 percent in 50 days.
Application example 1:
the R1 reactor was set up as a control experiment. In the experiment, the nitrogen-containing wastewater with the same volume as that of the R4 is added into the R1, and the rest conditions are the same as those of the R4 reactor in example 1, except that the nano zero-valent iron particles and the magnetic field are not added into the R1 reactor.
1. The effect of anaerobic ammonia oxidation starting process of R1 and R4 reactors with different treatments is shown in figure 1, from 4 days after starting to the end, the total nitrogen removal rate of R4 is always higher than that of R1, the total nitrogen removal rate of the R4 reactor reaches more than 85% in 40 days, and the total nitrogen removal rate of the R1 reactor reaches 85% in 60 days. The comparison shows that the method shortens the time of enrichment culture from anaerobic ammonia oxidation; the start-up time is obviously shortened relative to R1, namely, the addition of 3g/LnZVI to couple 65mT magnetic field can obviously accelerate the start-up process of anaerobic ammonia oxidation.
2. After the R1 and R4 reactors are started, the NH of the inlet water is gradually increased4 +-N and NO2 -The concentration of-N was used to increase the nitrogen load to further observe the stability of the reactor under different process conditions. As shown in fig. 2, R4 was the highest in total nitrogen removal during days 90 to 180; NH in R4 throughout the nitrogen load ramp-up phase4 +-N and NO2 -The removal rate of-N was consistently better than that of the R1 reactor. It can be seen that not only the start-up time of the reactor can be shortened but also the load impact resistance can be increased by the coupling effect of nZVI and the magnetic field. Therefore, the invention can promote the start-up of the anaerobic ammonia oxidation process and enhance the stability of the anaerobic ammonia oxidation process.
Application example 2:
4 reactors were set up: r1, R2, R3 and R4. R1 is blank control group (namely: no nano zero-valent iron particles and magnetic field are added in the reactor), R4 is experimental group (the reactor of example 1), and R2 and R3 are set as controls for further defining the action mechanism of the nano zero-valent iron coupling magnetic field. R2 is consistent with R1 except that no magnetic field is added; except that no nano zero-valent iron is added to R3, the other conditions are consistent with those of R1; the other conditions were identical for all 4 reactors.
In order to research the influence of the nZVI and the magnetic field on the anammox bacteria, the copy number of the 16S rRNA gene of the anammox bacteria in reactors at different stages is determined by adopting a qPCR technology; the copy number of the anammox bacterial functional gene Hzo (encoding hydrazine oxidoreductase) was determined.
Anammox bacteria in different stage reactorThe copy number of 16S rRNA gene is shown in FIG. 3a, and it can be seen from the figure that the copy number of anammox16S rRNA gradually increases with the culture time, and the four reactors show obvious difference. Initially, the copy amounts of the anammox bacteria 16S rRNA in R1, R2, R3 and R4 were all 1.01X 106copy amounts of anammox16S rRNA in R1, R2, R3 and R4 were increased to 5.5X 10 respectively by culturing copies/ng DNA for 180 days6、6.9×106、6.7×106、7.8×106copies/ng DNA. Compared with R1, the copy number of 16S rRNA of anammox bacteria cultured for 180 days in the experimental group (reactor of example 1) of R4 is the highest copy number of 16S rRNA, and the stability in culturing anammox bacteria is strong, as can be seen from FIG. 3.
In order to further explore the influence of the nano-zero-valent iron and the magnetic field on the anammox bacteria, quantitative measurement was carried out on the functional gene Hzo of the anammox bacteria at day 120, and the test results are shown in FIG. 3b, from which it can be seen that the copy numbers of Hzo genes in R1, R2, R3 and R4 are 2.4 × 105、3.5×105、3.3×105、4.0×105copies/ng DNA; compared with R1, the content of the compound is respectively increased by 50.0%, 37.5% and 66.7%. Obviously, R4 is an experimental group (reactor of example 1) nanometer zero-valent iron and a magnetic field can obviously promote the propagation of the anammox bacteria.
Example 2
A method for strengthening the culture and stability of anaerobic ammonium oxidation bacteria by a nano zero-valent iron coupling magnetic field comprises the following steps:
(1) adding nano zero-valent iron particles and anaerobic granular sludge into an upflow anaerobic sludge bed reactor, wherein the adding amount of the nano zero-valent iron particles is 5g/L of ammoniacal nitrogen and nitrite nitrogen wastewater to be treated; the grain size of the nano zero-valent iron particles is 60 nm; the addition amount of the anaerobic granular sludge is 5450-5550 mg MLSS/L of wastewater to be treated containing ammoniacal nitrogen and nitrite nitrogen;
(2) two permanent magnets with the length of 14cm, the height of 10cm and the thickness of 2.5cm are symmetrically arranged on the outer side of the reactor, a horizontal magnetic field is formed in the reactor by the two permanent magnets, and the magnetic field intensity is 75 mT;
(3) introducing wastewater to be treated containing ammoniacal nitrogen and nitrite nitrogen from the bottom of an upflow anaerobic sludge blanket reactor by using a peristaltic pump, adding 2L of ammoniacal nitrogen and nitrite nitrogen wastewater to be treated every day into the upflow anaerobic sludge blanket reactor, controlling the temperature to be 32 ℃, stirring under the condition of keeping out of the sun to ensure that anaerobic ammonia oxidation sludge in the reactor and the wastewater to be treated fully react, aerating for 10 minutes every day by using nitrogen, adjusting the rotating speed of the peristaltic pump according to the volume of the upflow anaerobic sludge blanket reactor to control the inflow velocity, ensuring that the hydraulic retention time of the wastewater to be treated in the reactor is 12 hours, and improving the degradation efficiency of pollutants in the wastewater; 2L of ammoniacal nitrogen and nitrite nitrogen are added to treat wastewater every day during operation.
Claims (10)
1. A method for strengthening the culture and stability of anaerobic ammonium oxidation bacteria by a nano zero-valent iron coupling magnetic field comprises the following steps:
(1) adding nano zero-valent iron particles and anaerobic granular sludge into an upflow anaerobic sludge bed reactor, wherein the adding amount of the nano zero-valent iron particles is 1-30g/L of ammoniacal nitrogen and nitrite nitrogen to-be-treated wastewater;
(2) two permanent magnets are symmetrically arranged on the outer side of the reactor, and form a magnetic field in the horizontal direction in the reactor, and the magnetic field intensity is 5-120 mT;
(3) introducing wastewater to be treated containing ammonia nitrogen and nitrite nitrogen from the bottom of the upflow anaerobic sludge blanket reactor by using a peristaltic pump, stirring the wastewater to be treated at the anaerobic and light-proof condition of 30-34 ℃ to ensure that the anaerobic ammonia oxidation sludge in the reactor and the wastewater to be treated fully react, adjusting the rotating speed of the peristaltic pump according to the volume of the upflow anaerobic sludge blanket reactor to control the inflow velocity, and ensuring that the sewage to be treated hydraulically stays in the reactor for 6-24 hours to realize the quick start of anaerobic ammonia oxidation.
2. The method according to claim 1, wherein the nano zero-valent iron particles are added in the step (1) in an amount of 1-10g/L of ammoniacal nitrogen and nitrous nitrogen wastewater to be treated.
3. The method according to claim 1, wherein the nano zero-valent iron particles are added in the step (1) in an amount of 1-5g/L of ammoniacal nitrogen and nitrous nitrogen wastewater to be treated.
4. The method according to claim 1, wherein the nano zero-valent iron particles are added in an amount of 3g/L of ammoniacal nitrogen and nitrous nitrogen wastewater to be treated.
5. The method according to claim 1, wherein in the step (1), the nano zero-valent iron particles have a particle size of 50 to 70 nm.
6. The method according to claim 1, wherein in the step (1), the amount of the anaerobic granular sludge added is 5450-5550 mg MLSS/L ammoniacal nitrogen and nitrite nitrogen wastewater to be treated.
7. The method according to claim 1, wherein in the step (2), the permanent magnet has a length of 12 to 15cm, a height of 8 to 12cm and a thickness of 2 to 3 cm.
8. The method according to claim 1, wherein in step (2), the magnetic field strength is 50-100 mT.
9. The method of claim 1, wherein in step (2), the magnetic field strength is 50-70 mT.
10. The method according to claim 1, wherein in the step (2), the molar ratio of ammoniacal nitrogen to nitrous nitrogen in the wastewater to be treated containing ammoniacal nitrogen and nitrous nitrogen is 1: 1.2.
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| CN113683186A (en) * | 2021-08-31 | 2021-11-23 | 浙江工业大学 | Iron-carbon fiber coupling filler and application thereof in sewage treatment |
| CN114012104A (en) * | 2021-11-05 | 2022-02-08 | 哈尔滨工业大学 | Preparation method of excess sludge composite nano zero-valent iron |
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| US20190084858A1 (en) * | 2016-03-22 | 2019-03-21 | Chinese Research Academy Of Environmental Sciences | Method for treating and recycling organic wastewater and apparatus |
| CN109970216A (en) * | 2019-04-26 | 2019-07-05 | 西安交通大学 | PVA base gel beads promote the cultural method of anaerobic ammonium oxidation sludge granulating |
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| CN104193002A (en) * | 2014-09-24 | 2014-12-10 | 山东大学 | Method for efficiently treating nitrogen-containing wastewater based on zero-valent iron coupled anaerobic ammonia oxidation |
| CN105236565A (en) * | 2015-07-02 | 2016-01-13 | 中国环境科学研究院 | Magnetic-field-strengthening method for increasing COD (Chemical Oxygen Demand) removal efficiency of anaerobic wastewater treatment by iron powder |
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| CN113683186A (en) * | 2021-08-31 | 2021-11-23 | 浙江工业大学 | Iron-carbon fiber coupling filler and application thereof in sewage treatment |
| CN114012104A (en) * | 2021-11-05 | 2022-02-08 | 哈尔滨工业大学 | Preparation method of excess sludge composite nano zero-valent iron |
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Application publication date: 20210420 |