WO2023035339A1 - 一种电化学驯化厌氧氨氧化细菌的方法及装置 - Google Patents

一种电化学驯化厌氧氨氧化细菌的方法及装置 Download PDF

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WO2023035339A1
WO2023035339A1 PCT/CN2021/120951 CN2021120951W WO2023035339A1 WO 2023035339 A1 WO2023035339 A1 WO 2023035339A1 CN 2021120951 W CN2021120951 W CN 2021120951W WO 2023035339 A1 WO2023035339 A1 WO 2023035339A1
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reactor
carbon fiber
sludge
nitrogen
microbial electrochemical
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French (fr)
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李姗蔚
高琦
吴智仁
周向同
刘志刚
韦静
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Jiangsu University
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    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/005—Combined electrochemical biological processes
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
    • C12M27/02—Stirrer or mobile mixing elements
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
    • 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
    • C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2203/00—Apparatus and plants for the biological treatment of water, waste water or sewage
    • C02F2203/004—Apparatus and plants for the biological treatment of water, waste water or sewage comprising a selector reactor for promoting floc-forming or other bacteria
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/06—Controlling or monitoring parameters in water treatment pH
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/14—NH3-N
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/15—N03-N
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2209/00—Controlling or monitoring parameters in water treatment
    • C02F2209/22—O2

Definitions

  • the invention relates to the technical field of water treatment, in particular to a method and device for electrochemically domesticating anammox bacteria.
  • Sewage treatment methods can be divided into physical chemical methods and biological methods, among which the biological method is widely used because of its good treatment effect; at the same time, the traditional biological method needs to add carbon sources to increase secondary pollution. Therefore, a new biological denitrification technology - anaerobic ammonia oxidation technology is a new direction to solve this problem.
  • Anaerobic ammonium oxidation (ANAMMOX) technology has been widely studied as a new biological denitrification technology. Compared with traditional nitrification and denitrification technology, it has many advantages such as no need for external carbon source, no need for aeration, and low sludge yield. Under anaerobic conditions, anammox bacteria can use nitrite (N0 2 - -N) and ammonia nitrogen (NH 4 + -N) as electron acceptor and electron donor, respectively, to generate nitrogen in one step. However, there are some deficiencies in the practical application of anammox technology.
  • the purpose of the present invention is to provide a method and device for electrochemical domestication of anammox bacteria, to solve the problems of the above-mentioned prior art, to promote the metabolism of anammox bacteria, to accelerate the proliferation and rapid growth of anammox bacteria Enrichment.
  • the present invention provides the following scheme:
  • the invention provides a device for electrochemically domesticating anammox bacteria, comprising a reactor body, a water inlet system connected to the reactor body pipeline, and a sludge return device connected to the reactor body pipeline;
  • the reactor main body includes a microbial electrochemical reactor, and an anode carbon fiber brush, a reference electrode, and a cathode carbon fiber brush are arranged in the microbial electrochemical reactor; connected.
  • the reactor main body also includes a magnetic stirrer, and the rotor of the magnetic stirrer is an octagonal rotor.
  • anode carbon fiber brush and the cathode carbon fiber brush are wound and fixed on the top of the microbial electrochemical reactor through titanium wires, and are connected to the potentiostat through titanium wires.
  • the reference electrode is an Ag/AgCl electrode.
  • the water inlet system includes a nitrogen cylinder, DO/pH monitor I, water inlet pump and water storage tank; an aeration pipe is provided at the bottom of the water storage tank, and the aeration pipe is connected to the nitrogen cylinder through a pipeline ;
  • the upper part of the water storage tank is provided with a water injection port and a one-way outlet valve; the inside of the water storage tank is provided with a heating rod.
  • the sludge return system includes an outlet peristaltic pump, a return sludge peristaltic pump and a secondary settling tank, and the secondary settling tank is provided with an upper water intake of the secondary settling tank.
  • the present invention also provides a method for electrochemically domesticating anammox bacteria, said method adopts the above-mentioned device for electrochemically domesticating anammox bacteria, comprising the following steps:
  • the hydraulic retention time is 23 hours, including 15 minutes for the influent and effluent.
  • the inoculated sludge is anaerobic activated sludge, anoxic activated sludge and anaerobic activated sludge with a volume ratio of 5:4:1; the MLSS of the inoculated sludge The value is 2000-4000mg/L.
  • step (2) after the nitrogen-containing sewage is blown off to remove oxygen, the DO value is lower than 0.5 mg/L, and the pH is adjusted between 7.5-8.2 by adding acid and alkali.
  • step (3) the average ammonia nitrogen content of the influent nitrogen-containing sewage is 50 mg/L, and the average nitrite nitrogen content is 66 mg/L.
  • the carbon fiber brush used in the invention has good electrical conductivity, has the advantages of large biological interception, and provides a place for anaerobic ammonia oxidation bacteria to attach.
  • Electrochemical technology is used for the domestication of anammox bacteria with high efficiency and operational flexibility. It can change the morphology and structure of anammox bacteria, catalyze some proteases in anammox bacteria cells, and accelerate the growth of anammox bacteria cells.
  • the transfer between the internal electrons and the electrodes, through the three-electrode microbial electrochemical device promotes the proliferation of anammox bacteria, shortens the start-up time of the anammox process, and the ultimate goal is to improve the denitrification ability of anammox bacteria.
  • the sludge return system avoids the loss of biomass without setting up a three-phase separator.
  • the invention uses carbon fiber with good conductivity, good biological affinity, economical and environmental protection as the biofilm carrier, uses the principle of microbial electrochemistry, strengthens the activity of anammox bacteria, shortens the start-up time of the anammox process and realizes long-term stable operation , to provide a theoretical basis for the wide application of new biological denitrification technology.
  • the microbial electrochemical system can improve the biological activity of functional bacteria.
  • the present invention uses microbial electrochemical technology to domesticate anammox bacteria, and develops a new method for electrochemical domestication of anammox bacteria, which can solve the problem of existing anaerobic bacteria.
  • the limitations of ammonia oxidation technology have practical implications.
  • Fig. 1 is the structural representation of the device of domesticating anammox bacterium by electrochemical method
  • nitrogen cylinder 1 heating rod 2
  • aeration tube 3 DO/pH monitor I 4
  • water inlet pump 5 water storage tank 6
  • water injection hole 7 one-way outlet valve 8
  • anode carbon fiber brush 9 indwelling port 10
  • cathode Carbon fiber brush 11 reference electrode 12
  • octagonal rotor 13 air outlet check valve 14
  • DO/pH monitor II heating rod 16
  • water outlet peristaltic pump 17 return sludge peristaltic pump 18
  • secondary sedimentation tank 19 secondary sedimentation Water intake 20 at the upper part of the pool, potentiostat 21, magnetic stirrer 22, microbial electrochemical reactor 23;
  • Fig. 2 is the change map of ammonia nitrogen (NH 4 + -N) in the domestication process
  • Fig. 3 is the change diagram of nitrite nitrogen (NO 2 - -N) during the domestication process
  • Fig. 4 is the diagram of the change of nitrate nitrogen (NO 3 - -N) during the acclimation process.
  • An embodiment of the present invention provides a device for electrochemically acclimating anammox bacteria, including a reactor body, a water inlet system connected to the reactor body pipeline, and a sludge return device connected to the reactor body pipeline; wherein, The main body of the reactor includes a microbial electrochemical reactor 23 , and the microbial electrochemical reactor 23 is provided with an anode carbon fiber brush 9 , a reference electrode 12 and a cathode carbon fiber brush 11 .
  • the microbial electrochemical reactor 23 is an airtight cylinder made of acrylic plate, and the outside adopts a shading cloth for shading.
  • the wall thickness of the container is 2cm, the effective height is 10-15cm, preferably 15cm, and its inner diameter is 15-20cm , preferably 15cm, and an effective volume of 1.5L;
  • a circular indwelling port 10 with a diameter of 3cm is provided on the top of the microbial electrochemical reactor 23, and a rubber stopper is used to block the inside of the reactor for observing changes in the operation and adjusting the operation status , and adjust the DO or pH value of the sewage;
  • a DO/pH monitor II 15 is installed on the upper part of the microbial electrochemical reactor 23, which is used to detect the DO value and pH value of the sewage during the reaction process, so as to adjust the water condition of the sewage at any time.
  • the anode carbon fiber brush 9 and the cathode carbon fiber brush 11 are cylindrical, with a diameter of 3cm, and a bristle length of 13-15cm, preferably 13cm, which is fixed on the microbial electrochemical reactor 23 top by winding titanium wire, titanium The wire extends out of the microbial electrochemical reactor 235cm so that the titanium wire is connected with the alligator clip of the potentiostat 21; the volume of the two carbon fiber brushes accounts for 40% of the microbial electrochemical reactor 23.
  • the reference electrode 12 is an Ag/AgCl electrode, which has a stable electrode potential and can be used for a long time in the microbial electrochemical reactor 23, and has the advantages of long service life and good reproducibility;
  • the working electrode and The counter electrodes are two carbon fiber brushes.
  • the potentiostat 21 is a CHI1030C multi-channel potentiostat, which provides electrode potentials to carbon fiber brushes, and can be used for electrochemical tests under the simultaneous operation of multiple reaction devices and multiple working electrodes to improve the overall operation. efficiency.
  • the carbon fiber content of its single carbon brush is 10g, and the carbon fiber lower end is pasted on the bottom of the microbial electrochemical reactor 23 by a glue stick, and the carbon fiber has high temperature resistance, Various properties such as electrical conductivity and thermal conductivity provide a convenient place for the attachment, habitat and metabolism of anammox bacteria.
  • the reactor main body also includes a magnetic stirrer 22, the rotor of the magnetic stirrer 22 is an octagonal rotor 13, and the octagonal rotor is used to reduce the pressure on the microbial electrochemical reactor 23 on the one hand and protect the Reactor; on the other hand, the octagonal rotor 13 rotates more stably than ordinary rotors, which is conducive to the full and uniform mixing of anammox sludge and sewage, accelerating the efficiency of nitrogen treatment and improving the operating efficiency of the reactor.
  • the water inlet system includes nitrogen cylinder 1, DO/pH monitor I4, water inlet pump 5 and water storage tank 6; the bottom of water storage tank 6 is provided with aeration pipe 3, and aeration pipe 3 passes through pipeline and nitrogen
  • the bottle 1 is connected to blow off and deoxygenate nitrogen-containing sewage; the upper part of the water storage tank 6 is provided with a water injection port 7 and a one-way air outlet valve 8; the inside of the water storage tank 6 is provided with a heating rod 2 for water temperature adjustment of nitrogen-containing sewage.
  • the aeration pipe 3 is placed in a circle at the bottom of the water storage tank 6, which is conducive to uniform aeration, increases the aeration efficiency, and shortens the aeration time.
  • the sludge return system includes an outlet peristaltic pump 17, a return sludge peristaltic pump 18, and a secondary settling tank 19.
  • the secondary settling tank 19 is provided with a water intake 20 on the upper part of the secondary settling tank for collecting incoming and outgoing water samples. , to monitor the growth of anammox bacteria.
  • the sludge return system is connected with the microbial electrochemical reactor 23 through two channels, which is used for the return of sewage sludge and prevents part of the sludge from being discharged into the microbial electrochemical reactor 23 with the effluent.
  • This setting effectively ensures the sludge concentration in the reactor. It provides a favorable guarantee for the long-term stable operation of anaerobic ammonium oxidation.
  • the embodiment of the present invention also provides a method for electrochemically domesticating anammox bacteria, the method adopts the above-mentioned device for electrochemically domesticating anammox bacteria, and includes the following steps:
  • Nitrogenous sewage is injected into the microbial electrochemical reactor 23 through the water inlet pump 5, the total volume of the inoculated sludge and nitrogenous sewage is 1.5L, the anode carbon fiber brush 9 and the cathode carbon fiber brush 11 are submerged, and the reference electrode 12 is placed 3cm below the water surface;
  • the MLSS value of the inoculated sludge is 2000-4000 mg/L, preferably 3500 mg/L.
  • the average ammonia nitrogen content of the influent nitrogen-containing sewage is 50 mg/L, and the average nitrite nitrogen content is 66 mg/L.
  • R1, R2, R3, and R4 Four identical microbial electrochemical reactors were constructed, named R1, R2, R3, and R4, respectively, and R1 was not applied with a potential (as a control group), that is, the open circuit (OC) mode.
  • R2, R3, R4 applied electrode potentials of 0.003V, -0.197V, -0.397V (vs. Ag/AgCl) respectively (three experimental groups).
  • Shanghai Chenhua's CHI1030C multi-channel potentiostat applies corresponding electrode potentials to each reactor.
  • Each reactor was inoculated with anoxic pond sludge, anaerobic pond sludge and anammox sludge, anoxic pond sludge, anaerobic pond sludge and anaerobic ammonia
  • the inoculum volumes of oxidized sludge were 50ml, 40ml and 10ml, respectively.
  • the water storage tank is injected with simulated sewage.
  • composition of simulated sewage is as follows: 2mg/L KH 2 PO 4 , 110mg/L KHCO 3 , 20mg/L MgSO 4 7H 2 O, 20mg/L CaCl 2 2H 2 O, 1.5mL/L trace element I and 1.5mL /L trace element II.
  • Trace element I is 5g/L EDTA ⁇ 2Na and 5g/L FeSO 4 ⁇ 7H 2 O; trace element II is 15g/L EDTA, 0.25g/L CuSO 4 ⁇ 5H 2 O, 0.99g/L MnCl 2 ⁇ 4H 2 O, 0.43 g/L ZnSO 4 ⁇ 7H 2 O, 0.24 g/L CoCl 2 ⁇ 6H 2 O, 0.19 g/L NiCl 2 ⁇ 6H 2 O, and 0.22 g/L NaMoO 4 ⁇ 2H 2 O.
  • NH 4 + -N is 50mg/L, NO 2 - -N is 66mg/L.
  • the substrates NH 4 + -N and NO 2 ⁇ -N are provided by NH 4 Cl and NaNO 2 .
  • the average influent NH 4 + -N and average influent NO 2 - -N are 50mg/L and 66mg/L.
  • the sludge that had just been inoculated into the microbial electrochemical reactor failed to adapt to the new environment, resulting in bacterial lysis and autolysis, and the concentration of NH 4 + -N in the effluent was higher than that of the influent. No NO 2 - -N was detected in the effluent at this stage. The reason was that the denitrifying bacteria used the NO 2 - -N in the influent to undergo a strong denitrification reaction.
  • the anammox reaction produces nitrate nitrogen
  • the activity of anammox can be calculated by monitoring the content of nitrate nitrogen in the effluent. During the hydrolysis period of the bacteria, the content of nitrate nitrogen was not detected, and the denitrification reaction of nitrate nitrogen as the substrate may have occurred or the anaerobic ammonium oxidation activity did not appear.
  • the NH 4 + -N removal rate of the three reactors with potential applied is higher than that of the reactor without potential applied.
  • the removal rate of NH 4 + -N is as high as 96.73%.
  • the removal rate was 25.3% higher than that of the R1 reactor with no potential applied.
  • the NO 2 - -N removal rates in reactors R1, R2, R3, and R4 gradually decreased from 98%, 94%, 99%, and 92% at the beginning of this stage to 17% at the 22d, 16% of 19d, 28% of 19d, 39% of 19d.
  • the NO 2 - -N removal rate of R1 at 22d, R2, R3, R4 at 19d is the lowest. After the 19th day for R1, and after the 22nd day for R2, R3, and R4, the removal rate of NO 2 - -N increased.
  • the reason for the sudden drop of NO 2 - -N removal rate is that the reduction of denitrifying bacteria and the disappearance of autolysis phenomenon promote the residual accumulation of NO 2 - -N; Increased, the abundance and activity of anammox bacteria increased, and the order of recovery speed was R1 ⁇ R2 ⁇ R3 ⁇ R4, which indicated that the applied electrode potential could accelerate NO 2 - -N removal.
  • the substrate NO 2 - -N is consumed by the anammox bacteria as electron acceptor, and the turning point also represents the improvement of the activity of the anammox bacteria.
  • the applied potential has a significant promotion effect on the denitrification ability of anammox.

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Abstract

一种电化学驯化厌氧氨氧化细菌的方法及装置，属于水处理技术领域，该装置包括反应器主体、与反应器主体管路连接的进水系统和与反应器主体管路连接的污泥回流装置；其中，反应器主体包括微生物电化学反应器（23），微生物电化学反应器（23）内设置有阳极碳纤维刷（9）、参比电极（12）和阴极碳纤维刷（11）；通过使用导电性好、生物亲和性好、经济环保的碳纤维作为生物膜载体，运用微生物电化学原理，强化了厌氧氨氧化细菌的活性，缩短了厌氧氨氧化工艺启动时间和实现了长期平稳运行。

Description

一种电化学驯化厌氧氨氧化细菌的方法及装置 技术领域
本发明涉及水处理技术领域，特别是涉及一种电化学驯化厌氧氨氧化细菌的方法及装置。
背景技术
污水处理方法可以分为物理化学法和生物法，其中生物法因其处理效果好而被广泛应用；同时，传统的生物法需要投加碳源，增加二次污染。因此，新型的生物脱氮技术--厌氧氨氧化技术是解决此问题的新方向。
厌氧氨氧化(ANAMMOX)技术作为一种新型生物脱氮技术被广泛地研究，与传统的硝化反硝化技术相比，具有无需外加碳源、无需曝气、污泥产率低等诸多优点。在厌氧条件下，厌氧氨氧化细菌能分别利用亚硝酸盐(N0 2 --N)和氨氮(NH 4 +-N)作为电子受体和电子供体，一步转化生成氮气。然而，厌氧氨氧化技术在实际应用中存在一些不足，由于厌氧氨氧化细菌生长缓慢、细胞倍增时间较长，导致厌氧氨氧化工艺启动时间较长；而且厌氧氨氧化细菌对环境条件极其敏感，如污废水中的有机物、溶解氧、盐度和重金属离子等。因此，厌氧氨氧化细菌的快速富集一直是该类工艺首要突破的难题，研究新的富集方法和装置，对推动厌氧氨氧化工艺大规模的应用具有重要的意义。
发明内容
本发明的目的是提供一种电化学驯化厌氧氨氧化细菌的方法及装置，以解决上述现有技术存在的问题，促进厌氧氨氧化细菌的新陈代谢，加速厌氧氨氧化细菌的增殖和快速富集。
为实现上述目的，本发明提供了如下方案：
本发明提供一种电化学驯化厌氧氨氧化细菌的装置，包括反应器主体、与所述反应器主体管路连接的进水系统和与所述反应器主体管路连接的污泥回流装置；其中，所述反应器主体包括微生物电化学反应器，所述微生物电化学反应器内设置有阳极碳纤维刷、参比电极和阴极碳纤维刷；所述阳极碳纤维刷和阴极碳纤维刷分别与恒电位仪相连接。
进一步地，所述反应器主体还包括磁力搅拌器，所述磁力搅拌器的转子为八角型转子。
进一步地，所述阳极碳纤维刷和阴极碳纤维刷通过钛丝缠绕固定于所述微生物电化学反应器顶部，并通过钛丝与所述恒电位仪相连接。
进一步地，所述参比电极为Ag/AgCl电极。
进一步地，所述进水系统包括氮气瓶、DO/pH监测仪Ⅰ、进水泵和贮水箱；所述贮水箱底部设有曝气管，所述曝气管通过管路与所述氮气瓶连接；所述贮水箱上部设有注水口和单向出气阀；所述贮水箱内部设有加热棒。
进一步地，所述污泥回流系统包括出水蠕动泵、回流污泥蠕动泵和二沉池，所述二沉池上设有二沉池上部取水口。
本发明还提供一种电化学驯化厌氧氨氧化细菌的方法，所述方法采用上述的电化学驯化厌氧氨氧化细菌的装置，包括以下步骤：
(1)将接种污泥接种到微生物电化学反应器中；
(2)通过加热棒控制贮水箱中含氮污水的水温在35℃；开启氮气瓶，通过曝气管吹脱除氧20-30min；
(3)将含氮污水通过进水泵注入微生物电化学反应器中；
(4)开启磁力搅拌器，转速100-300r/min；
(5)监测微生物电化学反应器内的pH，使pH保持在7.5-8.2之间；
(6)水力停留时间为23h，其中包括进水和出水各静置15min。
进一步地，在步骤(1)中，所述接种污泥为体积比5:4:1的厌氧活性污泥、缺氧活性污泥和厌氧氨氧化污泥；所述接种污泥的MLSS值为2000-4000mg/L。
进一步地，在步骤(2)中，所述含氮污水吹脱除氧后，使DO值小于0.5mg/L，通过外加酸碱调节pH在7.5-8.2之间。
进一步地，在步骤(3)中，进水含氮污水的平均氨氮含量为50mg/L，平均亚硝态氮含量为66mg/L。
本发明公开了以下技术效果：
本发明所用的碳纤维刷具有良好的导电性，具有生物截留量大的优点，为厌氧氨氧化细菌提供了附着的场所。电化学技术用于厌氧氨氧化细菌的驯化具有高效性和操作灵活性，可以改变厌氧氨氧化细菌的形态结构、催化厌氧氨氧化细菌细胞内的一些蛋白酶、加速厌氧氨氧化细菌细胞内电子和电极之间的传递，通过三电极微生物电化学装置，促进厌氧氨氧化细菌的增殖，缩短厌氧氨氧化工艺启动时间，最终目的是提高厌氧氨氧化细菌的脱氮能力。此外，在不设置三相分离器的前提下，污泥回流系统避免了生物量的流失。
本发明使用导电性好、生物亲和性好、经济环保的碳纤维作为生物膜载体，运用微生物电化学原理，强化厌氧氨氧化细菌的活性，缩短厌氧氨 氧化工艺启动时间和实现长期平稳运行，为新型生物脱氮技术的广泛运用提供理论基础。
微生物电化学系统能提高功能性细菌的生物活性,本发明利用微生物电化学技术来驯化厌氧氨氧化细菌，开发了一种电化学驯化厌氧氨氧化细菌的新方法，对解决现有厌氧氨氧化技术的局限性具有现实意义。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为电化学方法驯化厌氧氨氧化细菌的装置的结构示意图；
其中，氮气瓶1、加热棒2、曝气管3、DO/pH监测仪Ⅰ4、进水泵5、贮水箱6、注水孔7、单向出气阀8、阳极碳纤维刷9、留置口10、阴极碳纤维刷11、参比电极12、八角型转子13、出气单向阀14、DO/pH监测仪Ⅱ15、加热棒16、出水蠕动泵17、回流污泥蠕动泵18、二沉池19、二沉池上部取水口20、恒电位仪21、磁力搅拌器22、微生物电化学反应器23；
图2为驯化过程的氨氮(NH 4 +-N)变化图；
图3为驯化过程的亚硝酸盐氮(NO 2 --N)变化图；
图4为驯化过程的硝酸盐氮(NO 3 --N)变化图。
具体实施方式
现详细说明本发明的多种示例性实施方式，该详细说明不应认为是对本发明的限制，而应理解为是对本发明的某些方面、特性和实施方案的更 详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式，并非用于限制本发明。另外，对于本发明中的数值范围，应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明，否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料，但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入，用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时，以本说明书的内容为准。
在不背离本发明的范围或精神的情况下，可对本发明说明书的具体实施方式做多种改进和变化，这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见的。本发明说明书和实施例仅是示例性的。
关于本文中所使用的“包含”、“包括”、“具有”、“含有”等等，均为开放性的用语，即意指包含但不限于。
本发明实施例提供一种电化学驯化厌氧氨氧化细菌的装置，包括反应器主体、与反应器主体管路连接的进水系统和与反应器主体管路连接的污泥回流装置；其中，反应器主体包括微生物电化学反应器23，微生物电化 学反应器23内设置有阳极碳纤维刷9、参比电极12和阴极碳纤维刷11。
其中，微生物电化学反应器23为由亚克力板制成的密闭圆柱体，外部采用遮光布遮光，容器的壁厚为2cm，有效高度为10-15cm，优选为15cm，其内直径为15-20cm，优选为15cm，有效容积为1.5L；在微生物电化学反应器23上部设有直径为3cm的圆形留置口10，使用橡胶塞堵塞，用于观察运行时反应器内部的变化和调节运行状况，以及调节污水DO或pH值；同时，微生物电化学反应器23上部设有DO/pH监测仪Ⅱ15，用于检测反应过程中污水的DO值和pH值，以便随时对污水水况进行调整。
作为本发明的优选方案，阳极碳纤维刷9和阴极碳纤维刷11为圆柱形，直径为3cm，刷毛长度为13-15cm，优选为13cm，通过钛丝缠绕固定于微生物电化学反应器23顶部，钛丝伸出微生物电化学反应器235cm，以便钛丝与恒电位仪21的鳄鱼夹连接；两个碳纤维刷的体积占微生物电化学反应器23的40％。
作为本发明的优选方案，参比电极12为Ag/AgCl电极，其电极电势稳定，可在微生物电化学反应器23中长时间使用，具有使用寿命长、重现性好的优点；工作电极和对电极则为两个碳纤维刷。
作为本发明进一步的优选方案，恒电位仪21为CHI1030C多通道恒电位仪，对碳纤维刷提供电极电势，并可用于多反应装置、多工作电极同时运行条件下的电化学测试，提高整体的运行效率。
作为本发明进一步的优选方案，对于阳极碳纤维刷9和阴极碳纤维刷11，其单个碳刷的碳纤维含量为10g，碳纤维下端通过胶棒粘贴在微生物电化学反应器23的底部，碳纤维具有耐高温、导电和导热等多种特性，为 厌氧氨氧化细菌的附着、栖息和新陈代谢提供便利的场所。
作为本发明的优选方案，反应器主体还包括磁力搅拌器22，磁力搅拌器22的转子为八角型转子13，采用八角型的转子，一方面减小对微生物电化学反应器23的压强，保护反应器；另一方面，八角型转子13比普通转子旋转更稳定，有利于厌氧氨氧污泥和污水充分均匀混合，加速氮素处理效率和提高反应器的运行效率。
作为本发明的优选方案，进水系统包括氮气瓶1、DO/pH监测仪Ⅰ4、进水泵5和贮水箱6；贮水箱6底部设有曝气管3，曝气管3通过管路与氮气瓶1连接，用于含氮污水的吹脱除氧；贮水箱6上部设有注水口7和单向出气阀8；贮水箱6内部设有加热棒2，用于含氮污水的水温调整。曝气管3围成圆形置于贮水箱6底部，有利于曝气均匀，增加曝气效率，缩短曝气时间。
作为本发明的优选方案，污泥回流系统包括出水蠕动泵17、回流污泥蠕动泵18和二沉池19，二沉池19上设有二沉池上部取水口20，用于采集进出水样，监测厌氧氨氧化细菌的生长状况。
污泥回流系统与微生物电化学反应器23双通路连接，用于污水污泥的回流，避免部分污泥随出水排出微生物电化学反应器23，该设置有效保证了反应器中的污泥浓度，为厌氧氨氧化长期平稳运行提供了有利保证。
本发明实施例还提供电化学驯化厌氧氨氧化细菌的方法，方法采用上述的电化学驯化厌氧氨氧化细菌的装置，包括以下步骤：
(1)将江苏省镇江市城市生活污水厂的厌氧活性污泥、缺氧活性污泥和厌氧氨氧化污泥按体积比5:4:1接种到微生物电化学反应器23内；
(2)将含氨氮(NH 4 +-N)和亚硝酸盐氮(NO 2 --N)的模拟含氮污水注入贮水箱6，通过加热棒2维持温度为35℃；打开氮气瓶1，使用纯度99.5％的氮气通过曝气管3吹脱除氧20-30min，优选25min，使得贮水箱6中含氮污水的DO值维持在0.5mg/L以下，并使用1M碳酸氢钠溶液或1M稀盐酸调节含氮污水pH在7.5-8.2之间；
(3)将含氮污水通过进水泵5注入微生物电化学反应器23中，接种污泥和含氮污水的总体积为1.5L，浸没阳极碳纤维刷9和阴极碳纤维刷11，参比电极12放置在水面以下3cm处；
(4)开启磁力搅拌器22，转速100-300r/min，优选为120r/min；
(5)监测微生物电化学反应器23内的pH，并通过留置口10进行调节(用1M碳酸氢钠溶液或1M稀盐酸进行调节)，使pH保持在7.5-8.2之间；
(6)设置水力停留时间为23h，其中静置30min，包括进水和出水各15min。
作为本发明进一步的优选方案，接种污泥的MLSS值为2000-4000mg/L，优选为3500mg/L。
作为本发明进一步的优选方案，在步骤(3)中，进水含氮污水的平均氨氮含量为50mg/L，平均亚硝态氮含量为66mg/L。
实施例1
(1)构建4个相同的微生物电化学反应器，分别命名为R1、R2、R3、R4，其中R1未施加电势(作为对照组)，即开路(OC)模式。R2、R3、R4分别施加0.003V、-0.197V、-0.397V(vs.Ag/AgCl)的电极电势(三个实 验组)。上海辰华的CHI1030C多通道恒电位仪为各反应器施加相应的电极电势。
(2)各反应器分别接种江苏省镇江市城市污水厂的缺氧池污泥、厌氧池污泥和厌氧氨氧化污泥，缺氧池污泥、厌氧池污泥和厌氧氨氧化污泥的接种体积分别为50ml、40ml和10ml。
(3)贮水箱注入模拟污水。
模拟污水的组成如下：2mg/L KH 2PO 4、110mg/L KHCO 3、20mg/L MgSO 4·7H 2O、20mg/L CaCl 2·2H 2O、1.5mL/L微量元素Ⅰ和1.5mL/L微量元素Ⅱ。
微量元素I为5g/L EDTA·2Na和5g/L FeSO 4·7H 2O；微量元素II为15g/L EDTA、0.25g/L CuSO 4·5H 2O、0.99g/L MnCl 2·4H 2O、0.43g/L ZnSO 4·7H 2O、0.24g/L CoCl 2·6H 2O、0.19g/L NiCl 2·6H 2O和0.22g/L NaMoO 4·2H 2O。
NH 4 +-N为50mg/L，NO 2 --N为66mg/L。
底物NH 4 +-N和NO 2 --N由NH 4Cl和NaNO 2提供。
(4)脱氮效果评价：在整个驯化过程中，每日监测NH 4 +-N、NO 2 --N、NO 3 --N的浓度变化情况。
实验结果
1)菌体水解期：R2和R3为第1-3天，R1和R4为第1-6天，
平均进水NH 4 +-N和平均进水NO 2 --N为50mg/L和66mg/L。刚接种到微生物电化学反应器的污泥，未能适应新的环境，导致细菌裂解自溶，出水的NH 4 +-N浓度高于进水。此阶段未检测到出水中有NO 2 --N，其原因在于， 反硝化细菌利用进水中的NO 2 --N发生强烈的反硝化反应。此外，厌氧氨氧化反应是产硝酸盐氮的，通过监测出水硝酸盐氮的含量，可以推算出厌氧氨氧化的活性。在菌体水解期，未检测到硝酸盐氮的含量，可能发生了硝酸盐氮作为基质的反硝化反应或者厌氧氨氧化活性未显现。
NO 2 -+3H(电子供给体-有机物)→0.5N 2+H 2O+OH -
NO 3 -+5H(电子供给体-有机物)→0.5N 2+2H 2O+OH -
2)活性提高期：R2、R3、R4第7-28天，R1第11-28天。
随着驯化天数的增加，出水NH 4 +-N浓度逐渐降低，由于R1反应器未施加电势，进入厌氧氨氧化活性提高期的时间慢于施加三种不同电极电势的R2、R3、R4反应器。从图2中可以看出，NH 4 +-N去除率不断递增，NH 4 +-N去除率顺序是R1<R2<R3<R4。在第20d-27d，R1、R2、R3、R4反应器的平均NH 4 +-N去除率分别是71.43％、93.36％、94.76％和96.73％。由此可见，施加电势的三个反应器NH 4 +-N去除率高于未施加电势的反应器。其中在最佳电势-0.397V下，NH 4 +-N去除率高达96.73％。比未施加电势的R1反应器去除率高25.3％。
如图3所示，R1、R2、R3、R4反应器中NO 2 --N去除率逐渐降低由本阶段起始的98％、94％、99％、92％下降至第22d的17％、第19d的16％、第19d的28％、第19d的39％。在本实验中R1第22d，R2、R3、R4第19d的NO 2 --N去除率最低。R1在第19d后、R2、R3、R4在第22d后，NO 2 --N去除率反升。NO 2 --N去除率骤降的原因在于，反硝化细菌的减少和自溶现象消失，促使NO 2 --N出现了剩余积累；之后又出现去除率回升的原因在于，随着驯化时间的增加，厌氧氨氧化细菌的丰度和活性提高，回升的速度顺序为 R1<R2<R3<R4，说明施加的电极电势可以加速NO 2 --N去除。促使底物NO 2 --N被厌氧氨氧化细菌当做电子受体而被消耗，转折点也代表了厌氧氨氧化细菌活性的提高。有研究表明，NO 2 --N去除率从100％逐渐降低，说明反硝化作用减弱，所以NO 2 --N的去除率先升高后降低、再由低升高，表明厌氧氨氧化反应活性增强。
如图4所示，R1、R2、R3、R4中NO 3 --N的产生量是逐渐增加的，并且上升速度的顺序是R4>R3>R2>R1。通过厌氧氨氧化反应式推断，当施加-0.397V的电极电势时，厌氧氨氧化细菌活性最高。第24d-28d，R1、R2、R3、R4平均出水NO 3 --N浓度分别为6mg/L、9mg/L、10mg/L和15mg/L，可以看出R4硝酸盐氮的出水浓度最高。再次证实了当施加-0.397V电势时，厌氧氨氧化细菌活性最高。
由此可见施加电势对厌氧氨氧化脱氮能力有明显的促进作用。这说明微生物电化学系统对生物新陈代谢起到促进作用，加速厌氧氨氧化细菌的增殖，说明了本发明技术方案的可行性。
以上所述的实施例仅是对本发明的优选方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种变形和改进，均应落入本发明权利要求书确定的保护范围内。

Claims (10)

  1. 一种电化学驯化厌氧氨氧化细菌的装置，其特征在于，包括反应器主体、与所述反应器主体管路连接的进水系统和与所述反应器主体管路连接的污泥回流装置；其中，所述反应器主体包括微生物电化学反应器(23)，所述微生物电化学反应器(23)内设置有阳极碳纤维刷(9)、参比电极(12)和阴极碳纤维刷(11)；所述阳极碳纤维刷(9)和阴极碳纤维刷(11)分别与恒电位仪(21)相连接。
  2. 根据权利要求1所述的装置，其特征在于，所述反应器主体还包括磁力搅拌器(22)，所述磁力搅拌器(22)的转子为八角型转子(13)。
  3. 根据权利要求1所述的装置，其特征在于，所述阳极碳纤维刷(9)和阴极碳纤维刷(11)通过钛丝缠绕固定于所述微生物电化学反应器(23)顶部，并通过钛丝与所述恒电位仪(21)相连接。
  4. 根据权利要求1所述的装置，其特征在于，所述参比电极(12)为Ag/AgCl电极。
  5. 根据权利要求1所述的装置，其特征在于，所述进水系统包括氮气瓶(1)、DO/pH监测仪Ⅰ(4)、进水泵(5)和贮水箱(6)；所述贮水箱(6)底部设有曝气管(3)，所述曝气管(3)通过管路与所述氮气瓶(1)连接；所述贮水箱(6)上部设有注水口(7)和单向出气阀(8)；所述贮水箱(6)内部设有加热棒(2)。
  6. 根据权利要求1所述的装置，其特征在于，所述污泥回流系统包括出水蠕动泵(17)、回流污泥蠕动泵(18)和二沉池(19)，所述二沉池(19)上设有二沉池上部取水口(20)。
  7. 一种电化学驯化厌氧氨氧化细菌的方法，其特征在于，所述方法采 用权利要求1-6任一项所述的电化学驯化厌氧氨氧化细菌的装置，包括以下步骤：
    1)将接种污泥接种到微生物电化学反应器(23)中；
    2)通过加热棒(2)控制贮水箱(6)中含氮污水的水温在35℃；开启氮气瓶(1)，通过曝气管(3)吹脱除氧20-30min；
    3)将含氮污水通过进水泵(5)注入微生物电化学反应器(23)中；
    4)开启磁力搅拌器(22)，转速100-300r/min；
    5)监测微生物电化学反应器(23)内的pH，使pH保持在7.5-8.2之间；
    6)水力停留时间为23h，其中包括进水和出水各静置15min。
  8. 根据权利要求7所述的方法，其特征在于，在步骤1)中，所述接种污泥为体积比5:4:1的厌氧活性污泥、缺氧活性污泥和厌氧氨氧化污泥；所述接种污泥的MLSS值为2000-4000mg/L。
  9. 根据权利要求7所述的方法，其特征在于，在步骤2)中，所述含氮污水吹脱除氧后，使DO值小于0.5mg/L，通过外加酸碱调节pH在7.5-8.2之间。
  10. 根据权利要求7所述的方法，其特征在于，在步骤3)中，进水含氮污水的平均氨氮含量为50mg/L，平均亚硝态氮含量为66mg/L。
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