Method for rapid enrichment of denitrifying anammox bacteria

Abstract

The application discloses a method for rapidly enriching denitrification of anaerobic ammonia oxidation bacteria, and is applied to a short-range nitrification-anaerobic ammonia oxidation process, and active sludge in a secondary sedimentation tank of an urban sewage treatment plant is inoculated in an anaerobic ammonia oxidation reaction tank. 2+ The method comprises an enrichment stage and a sewage treatment stage; Fe 2+ Anthraquinone compounds are added in the water inlet of the enrichment stage, and the ultrasonic broken bacteria liquid of Shewanella oneidensis is added in the anaerobic ammonia oxidation reaction tank; in the sewage treatment stage, the water inlet is the sewage to be treated added with the anthraquinone compounds, and the ultrasonic broken bacteria liquid of Shewanella oneidensis is added in the anaerobic ammonia oxidation reaction tank. 2+ The culture solution is used as the water inlet to domesticate and enrich the anaerobic ammonia oxidation bacteria, the ultrasonic broken bacteria liquid of Shewanella oneidensis and the anthraquinone compounds are simultaneously added, the electron transfer rate and the growth rate of the anaerobic ammonia oxidation bacteria are cooperatively improved, the rapid enrichment culture of the anaerobic ammonia oxidation bacteria is realized, and the denitrification efficiency is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of energy-saving and environment-friendly denitrification technology, and in particular to a method for rapidly enriching anaerobic ammonia-oxidizing bacteria for denitrification, thereby improving denitrification efficiency. BACKGROUND

[0002] The information disclosed in this Background section is only for the purpose of increasing an understanding of the general background of the application and does not necessarily constitute an admission by the patent applicant or the assignee that this information constitutes prior art.

[0003] In recent years, the eutrophication trend of the nearshore sea area has been aggravated. According to the survey, the river into the sea is one of the main sources of total nitrogen pollution in the nearshore sea area. Strengthening the regulation of heavily polluted rivers into the sea in coastal cities, and strengthening the control of total nitrogen emissions according to local conditions and implementing total nitrogen reduction projects into the sea are the current development trend. Total nitrogen control is a key research direction in water pollution control in recent years.

[0004] Anaerobic ammonia oxidation process is a highly efficient biological denitrification process, which has important significance in high ammonia nitrogen wastewater treatment and nitrogen cycle. Anaerobic ammonia oxidation bacteria can oxidize ammonia nitrogen to nitrogen gas under anaerobic conditions with nitrite as the electron acceptor. This process does not require organic matter as an electron donor. Compared with the traditional nitrification-denitrification denitrification process, the anaerobic ammonia oxidation process has the advantages of low energy consumption, low sludge yield, no need for external carbon source, and energy-saving and environment-friendly in denitrification. In the treatment of landfill leachate, aquaculture wastewater, upgrading and reconstruction of municipal sewage treatment plants, and short-cut nitrification-denitrification biological denitrification process, it shows outstanding denitrification capacity.

[0005] Although the anaerobic ammonia oxidation technology has significant denitrification advantages, the anaerobic ammonia oxidation bacteria have the disadvantages of long generation time, generally 10-15 days, low growth rate and cell yield coefficient, long start-up time for engineering application, and sensitivity to environment, which limit its large-scale promotion and application. Future research needs to make breakthroughs in strain enrichment, rapid start-up, process optimization, and improvement of anti-interference ability.

[0006] Numerous studies have shown that increasing the electron transport rate can increase the growth rate and cell yield of anaerobic ammonia oxidation bacteria and improve denitrification efficiency. A large number of studies have added Fe(II) in the culture system, mainly because Fe 2+ can act as an electron donor to reduce NO3 - to N2, which can significantly improve the denitrification efficiency of anaerobic ammonia oxidation bacteria when the iron ion concentration is within a certain range. However, there are still problems such as long start-up time, low bacteria concentration, and sensitivity to environmental factors in the anaerobic ammonia oxidation process. Therefore, it is particularly urgent to develop a method for rapidly enriching ammonia-oxidizing bacteria for treating high ammonia-nitrogen wastewater.

[0007] Cytochrome C is an important electron transfer protein in the process of anaerobic ammonia oxidation, which is involved in the electron transfer process from nitrite reductase to hydrazine synthetase. In the process of anaerobic ammonia oxidation, the increase of cytochrome C concentration can accelerate the transmission rate of electrons and energy, thereby enhancing the activities of nitrite reductase, hydrazine synthetase and hydrazine dehydrogenase, accelerating the anaerobic ammonia oxidation reaction and improving the overall denitrification efficiency. Liu Zhaohua, Dang Yan, et al. in the paper “Enhanced anaerobic ammonia oxidation process by external potential for treatment of short-cut nitrification effluent of landfill leachate” showed that when the concentration of cytochrome C increased from a low level to 52.4 nmol·L -1 , the activities of nitrite reductase, hydrazine synthetase and hydrazine dehydrogenase increased by 47.8%, 17.5% and 48.1% respectively, and the overall denitrification efficiency was significantly improved. Peng Yongzhen team found in the research “Ultrastructure and function of anaerobic ammonium oxidation bacteria cells” that the increase of iron ion concentration can promote the synthesis of cytochrome C, thereby accelerating the electron transfer and promoting the growth of anaerobic ammonia oxidation bacteria. When the concentration of iron ion was 0.075 mmol / L, the maximum removal rates of NO2 - -N and NH4 + -N reached 1.6 and 1.8 times of the original respectively. Cytochrome C plays a key role in electron transfer, energy metabolism and functional enzyme activity promotion in the growth and metabolism of anaerobic ammonia oxidation bacteria, and is an important component indispensable to the process of anaerobic ammonia oxidation. Exogenous addition of cytochrome C can also improve the growth of anaerobic ammonia oxidation bacteria.

[0008] In the process of biochemical reaction, electrons need to be transferred from one reactant to another. However, there may be a large spatial distance between the reactants, and the direct electron transfer efficiency is low, which is more unfavorable for the cultivation of slow-growing anaerobic ammonia oxidation bacteria. Redox mediators can act as electron shuttles to accelerate the electron transfer efficiency between microorganisms and extracellular electron acceptors. In the process of anaerobic ammonia oxidation bacteria cultivation, efficient electron transfer is required for the oxidation-reduction reaction of ammonia nitrogen and nitrite, in order to improve the denitrification efficiency. Quinones have reversible oxidation-reduction characteristics and can convert between oxidized and reduced states, which are a good redox mediator. In the process of anaerobic ammonia oxidation, quinones can act as electron shuttles to transfer the intracellular metabolized electrons to extracellular electron acceptors (such as nitrite), which can provide a more efficient electron transfer pathway for microorganisms, thereby promoting the growth and reproduction of microorganisms.

[0009] Shewanella can produce cytochrome C, and Shewanella oneidensis has a complete cytochrome C expression and maturation system, and can efficiently synthesize and secrete cytochrome C. Cytochrome C produced by Shewanella plays a key role in the electron transfer process, enabling it to utilize a variety of electron acceptors. SUMMARY

[0010] In order to solve the slow growth of anaerobic ammonia oxidation bacteria, the start-up of denitrification in high ammonia nitrogen wastewater such as landfill leachate and aquaculture wastewater is slow, and it is difficult to promote and apply in engineering on a large scale. The present application provides a method for rapid enrichment of anaerobic ammonia oxidation bacteria for denitrification, which is applied to short-cut nitrification-anaerobic ammonia oxidation process and activated sludge in the secondary sedimentation tank of a municipal wastewater treatment plant. When enriching anaerobic ammonia oxidation bacteria, Fe 2+ and anthraquinone compounds, and adding ultrasonic broken bacteria liquid of Shewanella oneidensis in the anaerobic ammonia oxidation reaction tank, the three promote synergistically, and the anaerobic ammonia oxidation bacteria are added Fe 2+ It appears 100 days in advance, and the total nitrogen removal rate increases by 21.3%, realizing rapid enrichment and culture of anaerobic ammonia oxidation bacteria and improvement of total nitrogen removal rate.

[0011] The specific technical solutions are as follows:

[0012] A method for rapid enrichment of anaerobic ammonia oxidation bacteria for denitrification, which is applied to short-cut nitrification-anaerobic ammonia oxidation process, and inoculates activated sludge in the secondary sedimentation tank of a municipal wastewater treatment plant in the anaerobic ammonia oxidation reaction tank;

[0013] The method comprises an enrichment phase and a wastewater treatment phase;

[0014] Fe 2+ (Fe 2+ , preferably from ferrous sulfate) and anthraquinone compounds are added in the influent in the enrichment phase, and ultrasonic broken bacteria liquid of Shewanella oneidensis is added in the anaerobic ammonia oxidation reaction tank;

[0015] In the wastewater treatment phase, the influent is wastewater to be treated added with anthraquinone compounds, and ultrasonic broken bacteria liquid of Shewanella oneidensis is added in the anaerobic ammonia oxidation reaction tank.

[0016] The present application adds Fe 2+ The culture solution is used as the influent to domesticate and enrich anaerobic ammonia oxidation bacteria, and ultrasonic broken bacteria liquid of Shewanella oneidensis and anthraquinone compounds are added at the same time, so as to synergistically improve the electron transfer rate and growth rate of anaerobic ammonia oxidation bacteria, realize rapid enrichment and culture of anaerobic ammonia oxidation bacteria, and improve the denitrification efficiency.

[0017] The activated sludge of the present application contains Fe 2+The culture solution is domesticated and enriched, the ultrasonic broken bacterial solution of the Shewanella oneidensis producing cytochrome C is added, the cytochrome C produced in the cell and outside the cell is fully utilized, the electron transfer rate and the growth rate of the bacteria are improved, the redox mediator anthraquinone compound is added in the activated sludge, the contact distance between the space barrier effect and the reactants is shortened, the electron transfer rate is further improved, and the growth of the anammox bacteria is accelerated. Through the triple synergistic progressive effect, the electron transfer is accelerated, the growth and metabolism of the anammox bacteria are improved, the rapid enrichment culture of the anammox bacteria is realized, and technical support is provided for popularization and application of the anammox bacteria high-efficiency denitrification engineering application.

[0018] In the application, the anthraquinone compound preferably includes one or more of anthraquinone-2,6-disulfonic acid disodium, anthraquinone-2-sulfonic acid sodium and 9,10-anthraquinone, and further preferably includes 9,10-anthraquinone.

[0019] In the application, the anthraquinone compound is preferably added in the form of a solution, for example, the anthraquinone compound is dissolved in an alkaline (for example, pH=8.5) aqueous solution to obtain an anthraquinone compound solution for addition.

[0020] In the application, the Shewanella oneidensis can be obtained through a commercial channel, for example, it can be an ATCC strain purchased, and specifically, it can be a Shewanella oneidensis strain with the number ATCC700550.

[0021] The hydraulic retention time (HRT) of the short-cut nitrification reaction tank is preferably 4-6 hours, and further preferably 5 hours.

[0022] The dissolved oxygen (DO) concentration in the short-cut nitrification reaction tank is preferably 1.5-2.0 mg / L.

[0023] The hydraulic retention time of the anammox reaction tank is preferably 15-35 hours, and further preferably 24 hours.

[0024] The anammox reaction tank is filled with nitrogen to ensure an anaerobic state.

[0025] The short-cut nitrification reaction tank is preferably inoculated with activated sludge from an aerobic tank of a municipal wastewater treatment plant.

[0026] The sludge SV (30-minute sludge settling ratio) inoculated in the short-cut nitrification reaction tank is preferably 45%-55%, and further preferably 50%. 30

[0027] The volume of the sludge inoculated in the short-cut nitrification reaction tank is preferably 30%-35% of the effective volume of the short-cut nitrification reaction tank, and further preferably 1 / 3 of the effective volume of the short-cut nitrification reaction tank, which can be expressed as the inoculation amount of the sludge being 1 / 3 (V / V), and so on. ​

[0028] The sludge inoculated in the anammox reaction tank has an SV of preferably 70% to 80%, further preferably 75%. 30 Preferably, the sludge inoculated in the anammox reaction tank has an SV of 70% to 80%, further preferably 75%.

[0029] Preferably, the volume of the sludge inoculated in the anammox reaction tank is 65% to 70% of the effective volume of the anammox reaction tank, further preferably 2 / 3 of the effective volume of the anammox reaction tank, which can be expressed as the inoculation amount of the sludge being 2 / 3 (V / V), and so on.

[0030] Preferably, the influent in the enrichment stage comprises a nutrient solution and anthraquinones.

[0031] Preferably, the nutrient solution comprises ammonium chloride 267.5 to 1605 mg / L (e.g., 270 mg / L, etc.), sodium nitrite 345 to 2070 mg / L (e.g., 500 mg / L, etc.), calcium chloride 150 mg / L, magnesium sulfate 300 mg / L, potassium dihydrogen phosphate 30 mg / L, potassium bicarbonate 1250 mg / L, trace element mother liquor I 1.25 mL / L, and trace element mother liquor II 1.25 mL / L.

[0032] Preferably, the trace element mother liquor I comprises ethylenediaminetetraacetic acid (EDTA) 15 g / L, zinc sulfate 0.4 g / L, cobalt chloride 0.25 g / L, manganese chloride 1 g / L, copper sulfate 0.23 g / L, sodium selenate 0.23 g / L, sodium molybdate 0.25 g / L, nickel chloride 0.17 g / L, boric acid 0.014 g / L, and sodium tungstate 0.05 g / L.

[0033] Preferably, the trace element mother liquor II comprises ethylenediaminetetraacetic acid 6.25 g / L and ferrous sulfate 6.25 g / L.

[0034] Preferably, the concentration of the anthraquinones in the influent in the enrichment stage is 1 to 3 mg / L, such as 1.5 mg / L, 2 mg / L, 2.5 mg / L, etc., further preferably 2.5 mg / L.

[0035] Preferably, the method for preparing the ultrasonic broken bacterial solution of the Candidatus Accumulibacter phosphatis comprises the following steps:

[0036] 1) Dissolve tryptone, soybean peptone, and sodium chloride in water and adjust the pH to 7.3±0.2, sterilize to obtain a liquid medium;

[0037] 2) Inoculate the Candidatus Accumulibacter phosphatis in the liquid medium and culture on a 35℃ shaking table to obtain a bacterial agent culture solution, with a bacterial concentration of 10 7 ~10 8 CFU / mL;

[0038] 3) The bacterial agent culture solution is placed in an ice bath for ultrasonic disruption treatment, and the ultrasonic disruption bacterial solution of the Shewanella oneidensis is obtained.

[0039] In step 1), the preferred ratio of the amounts of the tryptone, the soybean peptone, the sodium chloride and the water is 10 g:2.5 g:2.5 g:1000 mL.

[0040] In step 1), the preferred sterilization temperature is 121℃, and the preferred sterilization time is 20 min.

[0041] In step 2), the preferred rotation speed of the shaker is 180 rpm, and the preferred shaker culture time is 24-36 hours.

[0042] In step 3), intermittent ultrasonic disruption treatment is preferred, and further preferred, the ultrasonic disruption is performed for 10-30 s (e.g., 15 s, etc.) with an interval of 20-30 s, and the total ultrasonic disruption time is 10-20 min (e.g., 15 min, etc.), and more further preferred, the ultrasonic disruption is performed for 15 s with an interval of 30 s, and the total ultrasonic disruption time is 15 min.

[0043] In the enrichment stage, the preferred ratio of the volume of the single addition of the ultrasonic disruption bacterial solution of the Shewanella oneidensis in the anaerobic ammonia oxidation reaction tank to the effective volume of the anaerobic ammonia oxidation reaction tank is 1‰-5‰, such as 2‰, 3‰, 4‰, etc., and further preferred, 3‰, which can be expressed as the addition amount of the ultrasonic disruption bacterial solution of the Shewanella oneidensis is 3‰ (V / V), and the like.

[0044] In the enrichment stage, the addition frequency of the ultrasonic disruption bacterial solution of the Shewanella oneidensis in the initial anaerobic ammonia oxidation reaction tank is once a day, and when the removal rate of the total nitrogen in the effluent fluctuates within 4%, the addition frequency of the ultrasonic disruption bacterial solution of the Shewanella oneidensis in the anaerobic ammonia oxidation reaction tank is 5-7 times a week.

[0045] The addition conditions of the anthraquinone compound and the ultrasonic disruption bacterial solution of the Shewanella oneidensis in the sewage treatment stage can refer to those in the enrichment stage.

[0046] Preferably, the addition conditions of the anthraquinone compound in the sewage treatment stage are consistent with those in the enrichment stage.

[0047] Preferably, the addition frequency of the ultrasonic disruption bacterial solution of the Shewanella oneidensis in the sewage treatment stage is 5-7 times a week, and the remaining addition conditions are consistent with those in the enrichment stage.

[0048] The temperature in the enrichment stage and the sewage treatment stage of the method for rapidly enriching anaerobic ammonia oxidation bacteria for denitrification is 25-30℃. The method can rapidly enrich anaerobic ammonia oxidation bacteria and significantly improve the denitrification effect at 25-30℃.

[0049] Compared with the prior art, the present application has the following beneficial effects:

[0050] 1. The present application utilizes the characteristics of cytochrome C participating in bacterial electron transfer, accelerating the growth and metabolic rate of anaerobic bacteria, and adds the ultrasonic broken bacteria liquid of Shewanella oneidensis producing cytochrome C in the anaerobic ammonia oxidation bacteria reaction tank, compared with the previous addition of Fe 2+ accelerating electron transfer, further improving the electron transport rate of anaerobic ammonia oxidation bacteria, promoting the rapid growth of anaerobic ammonia oxidation bacteria, and realizing high-efficiency denitrification.

[0051] 2. The present application utilizes the characteristics of redox mediators reducing steric hindrance of electron transfer and accelerating electron transfer, and adds the redox mediator anthraquinone compound in the domesticated culture solution, improves electron transfer and rapid enrichment of anaerobic ammonia oxidation bacteria, and realizes high-efficiency denitrification.

[0052] 3. The present application adds Fe 2+ , the ultrasonic broken bacteria liquid of Shewanella oneidensis producing cytochrome C, and the anthraquinone compound in the domesticated culture solution, and the three are synergistic and progressive, which is helpful for rapid enrichment and improvement of the concentration of anaerobic ammonia oxidation bacteria, and promotes the development and engineering application of high-efficiency, energy-saving and environmentally-friendly denitrification technology. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 It is the enrichment of anaerobic ammonia oxidation bacteria and the total nitrogen removal rate result graph of Example 1.

[0054] Figure 2 It is the enrichment of anaerobic ammonia oxidation bacteria and the total nitrogen removal rate result graph of Example 2.

[0055] Figure 3 It is the enrichment of anaerobic ammonia oxidation bacteria and the total nitrogen removal rate result graph of Example 3.

[0056] Figure 4 It is the enrichment of anaerobic ammonia oxidation bacteria and the total nitrogen removal rate result graph of Example 4.

[0057] Figure 5 It is the total nitrogen removal rate result graph of landfill leachate treatment of Example 5. DETAILED DESCRIPTION

[0058] The present application will be further described below in combination with the drawings and specific examples. It should be understood that these examples are only used to illustrate the present application and are not used to limit the scope of the present application. The operation methods not specified in the following examples are usually carried out according to the conventional conditions or the conditions recommended by the manufacturers.

[0059] Unless otherwise specified, the volume mentioned in the following examples refers to the effective volume.

[0060] Anaerobic ammonia oxidation bacteria are red, and the application reflects the enrichment degree of anaerobic ammonia oxidation bacteria in sludge by observing and calculating the volume proportion of red sludge.

[0061] Example 1

[0062] The nutrient solution contains: 270 mg / L of ammonium chloride, 500 mg / L of sodium nitrite, 150 mg / L of calcium chloride, 300 mg / L of magnesium sulfate, 30 mg / L of potassium dihydrogen phosphate, 1250 mg / L of potassium bicarbonate, 1.25 mL / L of trace element mother liquor I, and 1.25 mL / L of trace element mother liquor II. The composition of trace element mother liquor I is: 15 g / L of EDTA, 0.4 g / L of zinc sulfate, 0.25 g / L of cobalt chloride, 1 g / L of manganese chloride, 0.23 g / L of copper sulfate, 0.23 g / L of sodium selenate, 0.25 g / L of sodium molybdate, 0.17 g / L of nickel chloride, 0.014 g / L of boric acid, and 0.05 g / L of sodium tungstate. The composition of trace element mother liquor II is: 6.25 g / L of EDTA and 6.25 g / L of ferrous sulfate.

[0063] In the short-cut nitrification-anaerobic ammonia oxidation denitrification process, the volume of the short-cut nitrification reaction tank is 10 L, and the volume of the anaerobic ammonia oxidation reaction tank is 20 L. The short-cut nitrification reaction tank is inoculated with activated sludge from the aerobic tank of a municipal wastewater treatment plant, with an SV 30 of 50%, an inoculation amount of 1 / 3 (V / V), an HRT of 4 h, and a DO of 2.0 mg / L. The anaerobic ammonia oxidation reaction tank is inoculated with activated sludge from the secondary sedimentation tank of a municipal wastewater treatment plant, with an SV 30 of 75%, an inoculation amount of 2 / 3 (V / V), an HRT of 24 h, and a nitrogen gas flow rate of 0.5 mL / min to ensure anaerobic conditions.

[0064] The room temperature is maintained at 25℃, and the above-mentioned nutrient solution is used as the influent for acclimation for 300 days, as shown in the table, about 5% of the activated sludge appears sporadic red at 140 days of acclimation, the total nitrogen removal rate is 62.3%, at 300 days, the proportion of red activated sludge is about 23%, and the total nitrogen removal rate reaches 77.3%, and the total nitrogen removal rate fluctuates greatly. Figure 1

[0065] Example 2

[0066] ​Preparation of ultrasonic broken bacteria solution of Shewanella oneidensis: weigh 10 g of tryptone, 2.5 g of soybean peptone, 2.5 g of sodium chloride, and 1000 mL of water, mix well, and adjust pH to 7.3±0.2. 500 mL of the liquid medium is divided into 200 mL in a 500 mL triangular flask and sterilized at 121°C for 20 min. After sterilization, the cooled liquid medium is inoculated with a single colony of Shewanella oneidensis ATCC700550 from the plate culture medium on a sterile operation table. The liquid medium is cultured at 180 rpm and 35°C for 24-36 h to obtain a bacterial agent culture solution with a bacterial concentration of 10 7 ~10 8 CFU / mL. 250 mL of the obtained bacterial agent culture solution is placed in a 500 mL beaker and subjected to ultrasonic crushing in an ice bath for 15 s, with an interval of 30 s, and a total of 15 min.

[0067] Example 2 is carried out synchronously with Example 1. The nutrient solution of Example 1 is used as the influent to acclimate for 300 days, as shown in Figure 2 , 1‰ (V / V) of the ultrasonic broken bacteria solution of Shewanella oneidensis is added to the ANAMMOX reaction tank every day at the beginning, and the addition amount is increased by 1‰ (V / V) every two months, and sequentially increased to 5‰ (V / V). The ultrasonic broken bacteria solution of Shewanella oneidensis is added once every 7 days after 150 days. The remaining operations are the same as those of Example 1.

[0068] As shown in Figure 2 , when acclimated to 70 days, about 5% of the activated sludge in the ANAMMOX reaction tank appears sporadic red, and the total nitrogen removal rate is 56.2%. When acclimated to 300 days, the proportion of red activated sludge is about 60%, and the total nitrogen removal rate reaches 97.3%. After 150 days, the total nitrogen tends to be stable, and the removal rate is 93.4%-96.7%. At this time, the addition amount of the ultrasonic broken bacteria solution of Shewanella oneidensis is 3‰. When the addition amount of the ultrasonic broken bacteria solution of Shewanella oneidensis is 4‰, the proportion of red activated sludge is more than about 55% and tends to be stable. Therefore, when the addition amount of the ultrasonic broken bacteria solution of Shewanella oneidensis is 3‰, the total nitrogen removal rate is 93.4%-96.7% and tends to be stable, which is the optimal condition.

[0069] Example 3:

[0070] Preparation of anthraquinone mother liquor: 30 g of 9,10-anthraquinone is dissolved in tap water with pH of 8.5 to prepare an anthraquinone mother liquor with a concentration of 30 g / L.

[0071] Example 3 is carried out synchronously with Example 1 and Example 2. Anthraquinone is added to the nutrient solution in the form of the above-mentioned mother liquor to form the influent together with the nutrient solution. The final concentration of anthraquinone added to the initial influent is 1 mg / L, as shown in Figure 3, 0.5 mg / L every two months, and then 3 mg / L. The rest is the same as Example 1. The color change of the activated sludge in the ANAMMOX reactor and the total nitrogen removal rate were observed.

[0072] As shown in Figure 3 , when domesticated to 70 days, about 5% of the activated sludge in the ANAMMOX reactor appeared sporadic red, and the total nitrogen removal rate was 62.4%. When domesticated to 200 days, the proportion of red activated sludge was about 21%, and the total nitrogen removal rate reached 73.6%. After 230 days, the total nitrogen tended to be stable, with a removal rate of 79.6%~80.3%, and the proportion of red activated sludge was about 25%~27%, and the anthraquinone addition amount was 2.5 mg / L. Therefore, when the anthraquinone addition amount is 2.5 mg / L, the total nitrogen removal rate is 79.6%~80.3%, which tends to be stable, and is the optimal condition.

[0073] Example 4:

[0074] Referring to Example 1, the difference is that the anthraquinone mother liquor of Example 3 is used, and the anthraquinone is added in the nutrient solution in the form of the above-mentioned mother liquor to form the influent with the nutrient solution, and the final concentration of anthraquinone in the influent is 2.5 mg / L. The ultrasonic broken bacteria solution of the Lake O'Neill Shewanella of Example 2 is used, and 3‰ (V / V) of the ultrasonic broken bacteria solution of the Lake O'Neill Shewanella is added to the ANAMMOX reactor every day at the beginning, and the ultrasonic broken bacteria solution of the Lake O'Neill Shewanella is added once every 7 days after 170 days. The change of the total nitrogen removal rate and the activated sludge was investigated by continuous influent.

[0075] As shown in Figure 4 , when domesticated to 40 days, about 5% of the activated sludge in the ANAMMOX reactor appeared sporadic red, and the total nitrogen removal rate was 52.9%. From 40 days to 230 days, the proportion of red activated sludge was always on the rise, and after 230 days, the proportion reached about 80% and tended to be stable. After 170 days, the total nitrogen tended to be stable, with a removal rate of 96.3%~98.6%.

[0076] Example 5:

[0077] The short-cut nitrification-ANAMMOX process system domesticated and cultured in Example 4 was used, the influent was fresh landfill leachate pretreated and fed into the biochemical tank, the total nitrogen was 453~500 mg / L, the anthraquinone mother liquor of Example 3 was used, and the anthraquinone was mixed into the landfill leachate in the form of the above-mentioned mother liquor to a final concentration of 2.5 mg / L, 3‰ (V / V) of the ultrasonic broken bacteria solution of the Lake O'Neill Shewanella of Example 2 was added to the ANAMMOX reactor, and the addition frequency was once every 7 days. The rest of the operating conditions were the same as those in Example 4, and the total nitrogen removal rate was investigated by continuous influent.

[0078] As shown in Figure 5As shown, the total nitrogen removal rate was 24.4% to 84.3% before 40 days of the experimental operation, and increased rapidly with large fluctuations. During the period of 40 to 80 days, the total nitrogen removal rate was 86.5 to 95.1%, and increased slowly and steadily.

[0079] It is to be understood that even though numerous characteristics and embodiments of the application have been set forth in the foregoing disclosure, the details can be varied without departing from the application, which is defined by the following claims.

Claims

1. A method for rapid enrichment of denitrifying anammox bacteria, characterized in that, The method is applied to a short-cut nitrification-anammox process, and the anammox reaction tank is inoculated with activated sludge from a secondary sedimentation tank of a municipal wastewater treatment plant; The method comprises an enrichment stage and a wastewater treatment stage; Fe was added in the influent of the enrichment stage 2+ and anthraquinones, ultrasonic broken bacteria solution of Shewanella oneidensis in the anammox reactor The wastewater treatment stage is fed with wastewater to be treated added with anthraquinone compounds, and the anammox reaction tank is added with ultrasonic broken bacterial solution of Shewanella oneidensis MR-1.

2. The method of claim 1, wherein, The hydraulic retention time of the short-cut nitrification reaction tank is 4-6 hours; The dissolved oxygen concentration in the short-cut nitrification reaction tank is 1.5-2.0 mg / L; The hydraulic retention time of the anammox reaction tank is 15-35 hours; The anammox reaction tank is filled with nitrogen to ensure an anaerobic state.

3. The method of claim 1, wherein, The activated sludge in the aerobic tank of a municipal wastewater treatment plant was inoculated into the short-cut nitrification reaction tank. The SV of the inoculated sludge was 45% to 55%, and the volume of the inoculated sludge was 30% to 35% of the effective volume of the short-cut nitrification reaction tank. 30 The activated sludge in the aerobic tank of a municipal wastewater treatment plant was inoculated into the short-cut nitrification reaction tank. The SV of the inoculated sludge was 45% to 55%, and the volume of the inoculated sludge was 30% to 4. The method of claim 1, wherein, The sludge SV inoculated in the anammox reaction tank 30 was 70% to 80%, and the volume of the inoculated sludge was 65% to 70% of the effective volume of the anammox reaction tank.

5. The method of claim 1, wherein, The anthraquinone compounds include one or two or more of anthraquinone-2,6-disulfonic acid disodium salt, anthraquinone-2-sulfonic acid sodium salt and 9,10-anthraquinone; The feed of the enrichment stage comprises a nutrient solution and anthraquinone compounds; The nutrient solution contains: ammonium chloride 267.5-1605 mg / L, sodium nitrite 345-2070 mg / L, calcium chloride 150 mg / L, magnesium sulfate 300 mg / L, potassium dihydrogen phosphate 30 mg / L, potassium bicarbonate 1250 mg / L, trace element stock solution I 1.25 mL / L and trace element stock solution II 1.25 mL / L; The trace element stock solution I contains: ethylenediaminetetraacetic acid 15 g / L, zinc sulfate 0.4 g / L, cobalt chloride 0.25 g / L, manganese chloride 1 g / L, copper sulfate 0.23 g / L, sodium selenate 0.23 g / L, sodium molybdate 0.25 g / L, nickel chloride 0.17 g / L, boric acid 0.014 g / L and sodium tungstate 0.05 g / L; The trace element stock solution II contains: ethylenediaminetetraacetic acid 6.25 g / L and ferrous sulfate 6.25 g / L; The concentration of the anthraquinone compounds in the feed of the enrichment stage is 1-3 mg / L.

6. The method of claim 1, wherein, The preparation method of the ultrasonic broken bacterial solution of Shewanella oneidensis MR-1 comprises the following steps: 1) Dissolve tryptone, soybean peptone and sodium chloride in water and adjust the pH to 7.3±0.2, sterilize to obtain a liquid medium; 2) Shewanella oneidensis was inoculated into the liquid medium, and cultured at 35°C on a shaker to obtain a bacterial agent culture solution with a bacterial concentration of 10 7 ~10 8 CFU / mL; 3) Place the bacterial agent culture solution in an ice bath for ultrasonic breaking treatment to obtain the ultrasonic broken bacterial solution of Shewanella oneidensis MR-1.

7. The method of claim 6, wherein, In step 1), the amount ratio of tryptone, soybean peptone, sodium chloride and water is 10 g:2.5 g:2.5 g:1000 mL, the sterilization temperature is 121℃ and the sterilization time is 20 min; In step 2), the shaking bed rotation speed is 180 rpm and the shaking bed culture time is 24-36 hours; In step 3), intermittent ultrasonic breaking treatment is adopted, 10-30 s of ultrasonic breaking is followed by 20-30 s of interval, and the total ultrasonic breaking time is 10-20 min.

8. The method of claim 1, 6 or 7, wherein, In the enrichment stage, the ratio of the single addition volume of the ultrasonic broken bacterial solution of Shewanella oneidensis MR-1 in the anammox reaction tank to the effective volume of the anammox reaction tank is 1‰-5‰. In the enrichment stage, the frequency of adding the ultrasonic broken Shewanella oneidensis bacterial solution in the initial anaerobic ammonia oxidation reaction tank is once a day, and when the removal rate of total nitrogen in the effluent fluctuates within 4%, the frequency of adding the ultrasonic broken Shewanella oneidensis bacterial solution in the anaerobic ammonia oxidation reaction tank is once every 5-7 days.

9. The method of claim 1, wherein, The adding condition of anthraquinone compounds in the sewage treatment stage is consistent with that in the enrichment stage. The frequency of adding the ultrasonic broken Shewanella oneidensis bacterial solution in the sewage treatment stage is once every 5-7 days, and the rest of the adding conditions are consistent with those in the enrichment stage.

10. The method of claim 1, wherein, The temperature in the enrichment stage and the sewage treatment stage is 25-30℃.

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