Method for improving anaerobic ammonia oxidation denitrification effect of high-ammonia-nitrogen wastewater

Abstract

The invention relates to the technical field of wastewater treatment, and particularly provides a method for improving the anaerobic ammonia oxidation denitrification effect of high-ammonia-nitrogen wastewater. The method is realized by adding one or more enhancer combinations into a reaction system, wherein the enhancer is selected from vitamin B6, siderophore catechin (CAT) and siderophore hydroxyethyl ethylenediamine triacetic acid (HEDTA). By adding the enhancer, the inhibition effect of a high-ammonia-nitrogen environment on anaerobic ammonium oxidation functional flora can be effectively relieved, and the metabolic activity and the proliferation rate of the anaerobic ammonium oxidation flora are remarkably enhanced. The method is simple and convenient to operate and low in cost, can greatly improve the denitrification efficiency and operation stability of the system under high ammonia nitrogen load on the premise of not changing the structure of the existing reaction system, and is suitable for treating high ammonia nitrogen wastewater such as mine leaching tail water, landfill leachate and the like.

Description

Technical Field

[0001] This invention belongs to the field of wastewater treatment technology and relates to a method for improving the denitrification effect of anaerobic ammonia oxidation in high ammonia nitrogen wastewater. In particular, it relates to a method for regulating the denitrification effect by adding one or more of the following in combination as enhancers: vitamin B6, siderophore catechin (CAT), and siderophore hydroxyethyl ethylenediamine triacetic acid (HEDTA) to high ammonia nitrogen wastewater. By alleviating the inhibitory effect of the high ammonia nitrogen environment on the anaerobic ammonia oxidation functional bacteria, the metabolic activity and proliferation rate of the anaerobic ammonia oxidation bacteria are significantly enhanced. Background Technology

[0002] Anaerobic ammonia oxidation occurs under anaerobic conditions, where anaerobic ammonia-oxidizing bacteria utilize NH4+. + -N and NO2 - -N is used as the substrate, which is converted into nitrogen gas, accompanied by a small amount of NO3. - The process of nitrogen (N-) generation. This process breaks through the dependence of traditional nitrification-denitrification on organic carbon sources, achieving autotrophic denitrification without the need for external carbon sources. This characteristic directly brings multiple core advantages: significantly reduced aeration energy consumption, significantly reduced excess sludge production, and a multiplied increase in reactor efficiency through high-load granular sludge technology, thereby saving land and infrastructure investment. While significantly reducing operating costs, this process also possesses excellent carbon emission reduction potential due to its low energy consumption characteristics, making it a key technological path to achieving "carbon neutrality" in wastewater treatment.

[0003] However, when applied to treat wastewater with high ammonia nitrogen concentrations, the denitrification efficiency of anaerobic ammonia oxidation systems shows a significant decline. This is because the high ammonia nitrogen environment severely interferes with the normal physiological activities of the bacterial community: on the one hand, it causes drastic fluctuations in intracellular pH and disrupts proton balance, directly inhibiting the activity of key metabolic enzymes; on the other hand, continuous osmotic stress forces cells to consume large amounts of energy to maintain homeostasis, ultimately severely restricting the metabolic activity and proliferation capacity of the entire bacterial community. Furthermore, since anaerobic ammonia oxidizing bacteria are chemoautotrophic, their slow growth rate and long doubling time further limit their denitrification performance under high ammonia nitrogen loads. Therefore, a method is urgently needed to alleviate the inhibitory effect of high ammonia nitrogen on anaerobic ammonia oxidizing bacteria, improve their activity under high ammonia nitrogen conditions, and enhance their denitrification performance.

[0004] Existing technologies for enhancing anaerobic ammonia-oxidizing bacteria mainly include: adding iron-based materials such as magnetite and hematite to improve the aggregation morphology and physical structure of the bacteria; or supplementing with nutrients such as specific amino acids to enhance the bacteria's ability to resist external stresses such as heavy metals. However, these methods focus on optimizing the physical properties of the bacteria or enhancing specific resistance, failing to directly and efficiently address the problem of metabolic activity inhibition of anaerobic ammonia-oxidizing bacteria by high-ammonia nitrogen wastewater. For example, CN116161787A discloses a method for enhancing the performance of anaerobic ammonia oxidation using magnetite as a reinforcing agent. This method involves adding micron-sized magnetite to an upflow anaerobic sludge blanket (UASB) reactor to promote the secretion of extracellular polymers by bacterial communities. The extracellular polymers and iron nuclei increase the particle size of the bacteria, thereby enhancing the activity of anaerobic ammonia oxidizing bacteria. However, this method has limited applicability, is not highly targeted at high ammonia nitrogen wastewater, and the introduced inorganic solids may cause long-term side effects.

[0005] For example, CN119349765A discloses a method for efficiently treating nitrogen-containing wastewater using an iron promoter. After inoculating the reactor with anaerobic ammonia oxidation active seed sludge, nano-sized hematite is added in a single step, along with trace elements, to increase the sludge particle density of the anaerobic ammonia oxidizing bacteria, thereby improving their growth rate and activity. However, this method has a limited treatment concentration range and insufficient capacity for treating high-ammonia nitrogen wastewater. Its mechanism of action mainly relies on physical conduction, offering limited improvement in the intrinsic metabolic tolerance of the bacterial community. Without the addition of an external promoter, the system's denitrification performance rapidly declines.

[0006] For example, CN119912060A discloses a method for enhancing the resistance of anammox granular sludge to heavy metal stress by adding nutrients, wherein the nutrients include at least one of proline, arginine, glutamic acid, lysine, and nicotinic acid. The nutrient solution is periodically added to an anammox granular sludge system that has been or may be subsequently subjected to heavy metal stress, thereby improving the sludge's ability to resist heavy metal stress. However, this method mainly targets specific stresses and has no significant effect on substrate inhibition caused by ammonia nitrogen itself. Furthermore, the effectiveness of this method is highly dependent on the survival and dominance of a single bacterial species (i.e., anammox bacteria) in the complex and variable actual wastewater, and faces limitations in stability and reliability during long-term operation.

[0007] In summary, while existing enhancement methods can improve some performance aspects of anaerobic ammonia oxidation systems under specific conditions, they do not fundamentally enhance the metabolic activity and tolerance of bacterial communities in high ammonia nitrogen environments. Therefore, how to directly and efficiently alleviate the inhibitory effect of high ammonia nitrogen on the activity of anaerobic ammonia oxidation functional bacteria remains a key issue that urgently needs to be addressed in the practical application of this technology in high ammonia nitrogen wastewater treatment. Summary of the Invention

[0008] To address the shortcomings of existing technologies and practical needs, this invention provides a method for enhancing the denitrification performance of anaerobic ammonia oxidation by adding one or more of the following enhancers in high ammonia nitrogen wastewater: vitamin B6, siderophore CAT, and siderophore HEDTA. This method alleviates the inhibitory effect of the high ammonia nitrogen environment on the functional bacteria of anaerobic ammonia oxidation, thereby significantly enhancing the metabolic activity and proliferation rate of the anaerobic ammonia oxidation bacteria and effectively improving the denitrification performance of the anaerobic ammonia oxidation system.

[0009] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a method for improving the denitrification effect of anaerobic ammonia oxidation in high-ammonia nitrogen wastewater, the method comprising: adding a strengthening agent to high-ammonia nitrogen wastewater containing anaerobic ammonia oxidation synergistic bacteria; The fortifying agent includes vitamin B6 and / or ferrocarrier; The siderophore includes one of catechin (CAT) and hydroxyethyl ethylenediamine triacetic acid (HEDTA); The application methods include: Vitamin B6 may be added alone, or siderophores may be added alone, or a combination of vitamin B6 and one of the siderophores may be added. The stimulant can be added all at the beginning of the anaerobic ammonia oxidation synergistic bacterial culture, or it can be added in batches and evenly throughout the entire culture cycle.

[0010] This invention proposes a biochemical enhancement strategy to address the inhibition of high ammonia nitrogen. By applying specific enhancers and specific dosing patterns, the metabolic activity of anaerobic ammonia-oxidizing bacteria in high ammonia nitrogen wastewater is enhanced, promoting the proliferation of anaerobic ammonia-oxidizing bacteria, reducing the inhibitory effect of high ammonia nitrogen on anaerobic ammonia-oxidizing bacteria, and improving the denitrification performance of anaerobic ammonia-oxidizing bacteria and their operational stability in high ammonia nitrogen environments. This strategy is applicable to actual anaerobic ammonia oxidation wastewater treatment processes.

[0011] Optionally, the anaerobic ammonia oxidation synergistic microbial community includes a core genus. Candidate Brocadia (Brocardia spp.).

[0012] Optionally, the anaerobic ammonia oxidation synergistic microbial community also includes Pseudomonas (Pseudomonas) Macellibacteriodes (Carnivorous Bacteria) Proteocatella (Proteophiles) Comamonadaceae (Trichomonaceae family) or Acetoanaerobic At least one of the genus (acetic acid-producing anaerobic bacteria).

[0013] Optionally, the high ammonia nitrogen wastewater contains NH4 + and NO2 - The concentrations of each are independently 500~1500 mg / L, for example, 600, 700, 800, 900, 1000, 1100, 1200, 1300 or 1400 mg / L, etc.

[0014] Preferably, the fortifying agent is added in combination with vitamin B6 and one of the siderophores, namely, vitamin B6 combined with CAT or vitamin B6 combined with HEDTA. Preferably, the concentrations of vitamin B6, siderophore CAT, and siderophore HEDTA in the combination are the optimal fortifying agent addition amounts at the corresponding concentrations in the single-index experiment.

[0015] Optionally, the concentration of vitamin B6 added is 0.1~5.0 mg / L, for example, it can be 0.5, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0 or 5.0 mg / L, etc. Preferably, the concentration of vitamin B6 added is 1.0 mg / L~3.0 mg / L. More preferably, the concentration of vitamin B6 can be selected according to the ammonia nitrogen concentration. When the ammonia nitrogen concentration is 500 mg / L, the vitamin B6 used is preferably 1.0 mg / L; when the ammonia nitrogen concentration is 1000 mg / L, the vitamin B6 used is preferably 3.0 mg / L; and when the ammonia nitrogen concentration is 1500 mg / L, the vitamin B6 used is preferably 1.0 mg / L.

[0016] Optionally, the concentration of catechin (CAT) added is 12~48 μmol / L, for example, it can be 12, 15, 20, 24, 30, 36, 45 or 48 μmol / L, etc., preferably, the concentration added is 12 μmol / L~36 μmol / L. More preferably, when the ammonia nitrogen concentration is 500 mg / L, the CAT used is preferably 36 μmol / L, and when the ammonia nitrogen concentration is 1000 and 1500 mg / L, the CAT used is preferably 12 μmol / L.

[0017] Optionally, the concentration of hydroxyethyl ethylenediamine triacetic acid (HEDTA) added is 12~48 μmol / L, for example, it can be 12, 15, 20, 24, 30, 36, 45 or 48 μmol / L, etc., preferably, the concentration is 24 μmol / L~36 μmol / L. More preferably, the HEDTA used is preferably 24 μmol / L when the ammonia nitrogen concentration is 500, 1000, or 1500 mg / L.

[0018] Optionally, the concentration range of the vitamin B6 and siderophore combination is 0.1~5.0 mg / L and 12~48 μmol / L, respectively, and the addition ratio is 1 mg / 50 μmol / L~5 mg / 12 μmol / L, for example, it can be 1 mg / 12 μmol / L, 2 mg / 24 μmol / L, 3 mg / 24 μmol / L, 3 mg / 36 μmol / L, 4 mg / 36 μmol / L or 5 mg / 48 μmol / L, etc. The preferred ratio of vitamin B6 to siderophore CAT is 3 mg / 48 μmol~1 mg / 12 μmol, and the preferred ratio of vitamin B6 to siderophore HEDTA is 1 mg / 24 μmol~3 mg / 36 μmol.

[0019] In a second aspect, the present invention provides a method for anaerobic ammonia oxidation denitrification of wastewater, the method comprising: Anaerobic ammonia-oxidizing bacteria were inoculated into the wastewater for cultivation, and the enhancer described in the first aspect was added to the cultivation system.

[0020] This invention, by adding the aforementioned enhancer, effectively strengthens the bacterial activity and system stability of the anaerobic ammonia-oxidizing functional bacteria, significantly improves denitrification efficiency, and broadens the adaptability and treatment capacity of this technology for high-concentration ammonia nitrogen wastewater (such as leaching tailings and landfill leachate). This invention further optimizes the treatment process, significantly reducing energy consumption and the use of other reagents, and improving the utilization efficiency and capacity of the treatment facility.

[0021] Compared with the prior art, the present invention has at least the following beneficial effects: (1) This invention proposes a biochemical enhancement strategy for high ammonia nitrogen inhibition. By adding specific enhancers and adopting corresponding addition modes, the metabolic activity of anaerobic ammonia oxidizing bacteria in high ammonia nitrogen wastewater is improved, the inhibition of anaerobic ammonia oxidizing bacteria by high ammonia nitrogen is alleviated, and their denitrification performance is enhanced. After adding enhancers to the anaerobic ammonia oxidation system, the ammonia nitrogen removal efficiency can be significantly improved, and the microbial community structure can be optimized to promote the function of bacteria. Candidate Brocadia Enrichment.

[0022] (2) This invention optimizes the operation of the anaerobic ammonia oxidation process, significantly reduces energy consumption and the use of other reagents, and improves the utilization efficiency and capacity of the treatment facilities. It has the advantages of flexible operation, significant effect and easy promotion.

[0023] (3) This invention effectively breaks through the tolerance limit of traditional biological methods to high ammonia nitrogen concentration, and provides a new biodegradation option that can replace some physical and chemical treatment technologies. When treating specific high ammonia nitrogen wastewater, it is expected to realize the transformation from high-energy physical and chemical treatment to low-energy biological treatment, and broaden the application scope of biological denitrification process. Attached Figure Description

[0024] Figure 1 Figure 1 shows the results of sludge population analysis in each group after adding vitamin B6 to a medium with an initial ammonia nitrogen concentration of 1000 mg / L. Detailed Implementation

[0025] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. However, the following examples are merely simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention is determined by the claims.

[0026] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased from legitimate channels.

[0027] This invention aims to develop a method to improve the anaerobic ammonia oxidation denitrification effect of high ammonia nitrogen wastewater, so as to effectively alleviate the inhibition of bacterial communities by the high ammonia nitrogen environment, enhance their metabolic activity and proliferation rate, and thus efficiently treat wastewater containing high concentrations of ammonia nitrogen, such as ion-adsorption rare earth mineral leaching wastewater and landfill leachate.

[0028] Specifically, this invention provides a method for improving the denitrification effect of anaerobic ammonia oxidation in high-ammonia nitrogen wastewater, enhancing its denitrification performance. The enhancing agent includes vitamin B6 and / or siderophores (such as CAT and HEDTA). In this invention, specific vitamins and siderophores are designed and used as enhancing agents to promote the proliferation and activity of anaerobic ammonia oxidizing bacteria, thereby helping them recover and maintain denitrification performance in a high-ammonia nitrogen environment. Further, based on the enhancing agent, a method for improving the denitrification effect of anaerobic ammonia oxidation in high-ammonia nitrogen wastewater is developed. The method includes: inoculating the wastewater with anaerobic ammonia oxidizing bacteria for cultivation, and adding the enhancing agent to the cultivation system. This can be done by adding all the agent at the beginning of cultivation or by adding it in stages throughout the cultivation cycle.

[0029] Optionally, the anaerobic ammonia-oxidizing bacterial community includes the genus *Anaerobic ammonia-oxidizing core bacteria*. Candidate Brocadia In addition, it may include Pseudomonas , Macellibacteriodes , Proteocatella , Comamonadaceae , Acetoanaerobic One or more of them.

[0030] Optionally, NH4 in the culture system + The specific concentration can be 500~1500 mg / L, NO2 - The specific concentration can be 500~1500 mg / L.

[0031] Optionally, the concentration of vitamin B6 added is 0.1~5.0 mg / L, which can be adapted according to the initial ammonia nitrogen concentration in the anaerobic ammonia oxidation system. The anaerobic ammonia oxidation system contains NH4... + and NO2 - When the concentration is 500 mg / L, the preferred dosage concentration of vitamin B6 is 0.1~2.0 mg / L. NH4 + and NO2 - When the concentration is 1000 mg / L, the preferred dosage concentration of vitamin B6 is 2.0~5.0 mg / L. NH4 + and NO2 - When the concentration is 1500 mg / L, the preferred concentration of vitamin B6 is 0.1~2.0 mg / L.

[0032] Optionally, the concentration of the enhancer CAT is 12~48 μmol / L, which can be adapted according to the initial ammonia nitrogen concentration in the anaerobic ammonia oxidation system. NH4 + and NO2 - When the concentration is 500 mg / L, the preferred concentration is 12~36 μmol / L, NH4 + and NO2 - When the concentration is 1000 mg / L or 1500 mg / L, the preferred concentration is 12–24 μmol / L. The HEDTA dosage concentration is 12–48 μmol / L, which can be adjusted according to the initial ammonia nitrogen concentration in the anaerobic ammonia oxidation system. NH4 + and NO2 - When the concentrations are 500, 1000 or 1500 mg / L, the preferred HEDTA concentration is 24~36 μmol / L.

[0033] Optionally, the fortifier can be added in the form of a combination of vitamin B6 and the siderophore CAT, in an anaerobic ammonia oxidation system with NH4 + and NO2 - When the concentrations were 500, 1000, or 1500 mg / L, the combination of 1.0 mg / L vitamin B6 and 12 μmol / L CAT as a fortifier showed the most significant effect among all combinations. The fortifier can be added as a combination of vitamin B6 and the siderophore HEDTA, using an anaerobic ammonia oxidation system with NH4+. + and NO2 - When the concentrations were 500, 1000, or 1500 mg / L, the combination of 1.0 mg / L vitamin B6 and 24.0 μmol / L HEDTA as an enhancer showed the most significant effect among all combinations.

[0034] Optionally, the fortifier can be added as a combination of vitamin B6 and the siderophore CAT, and vitamin B6 and CAT can be added in batches (e.g., twice) during the culture period, with the anaerobic ammonia oxidation system NH4 + and NO2 - When the concentrations were 500, 1000, or 1500 mg / L, the ammonia nitrogen removal rate of the batch addition method was better than that of the case where the same amount of enhancer was added at the beginning of the culture.

[0035] In a specific embodiment of the present invention, the experimental procedure for culturing anaerobic ammonia-oxidizing bacteria includes the following steps: Containing anaerobic ammonia-oxidizing bacteria (such as core bacteria) Candidate Brocadia Anaerobic ammonia oxidation granular sludge (taken from the anaerobic tank of an ammonia nitrogen wastewater treatment process in a wastewater treatment plant) was activated and cultured for 7-10 days. Before activation, the particle diameter was between 0.5 and 3 mm. Inoculation was performed in a glove box, and cultivation was carried out on a constant temperature shaker at 200 r / min. During cultivation, the initial culture medium pH was 7.0, a strictly anaerobic environment was maintained, and the cultivation temperature was maintained at 29-31℃. After activation with the nutrient medium, the sludge gradually turned red, exhibiting granular shapes of varying sizes, with the particles growing larger to a diameter between 0.5 and 4 mm, indicating that the anaerobic ammonia oxidation granular sludge possessed denitrification activity, paving the way for further experiments. The nutrient medium formula was as follows (1 L): 3 g tryptone, 0.5 g (NH4)2SO4, 0.25 g NaNO2, 0.84 g NaHCO3, and 5 g NaCl. Add the above-mentioned ingredients to approximately 800 mL of distilled water and stir thoroughly with a magnetic stirrer until all components are dissolved. Measure the pH of the solution in real time using a pH meter, and adjust the pH to 7.0 using NaOH (5 M) or HCl (5 M). Make up the volume to 1 L. Dispense the culture medium into vials and autoclave (121°C, 20 minutes) to ensure thorough sterilization. After sterilization, inoculate in a glove box with an inoculation volume of approximately 10-20% of the culture medium volume. The MLSS concentration of the inoculated sludge should be 10.0-11.0 g / L. Incubate after inoculation, maintaining a strictly anaerobic environment at a temperature of 29-31°C.

[0036] The activated and cultured granular sludge was inoculated with initial NH4. + and NO2 -Subsequent experiments were conducted using anaerobic ammonia oxidation media with concentrations of 500, 1000, and 1500 mg / L. The formulation of the anaerobic ammonia oxidation media with an initial ammonia nitrogen concentration of 500 mg / L was as follows (1L): (NH4)2SO4 1.83g, NaNO2 0.75g, NaHCO3 0.84g, low-magnesium solution 10 mL, Se / W / Mo solution 1 mL, and vitamin stock solution 1 mL; while NH4 + and NO2 - Culture media with concentrations of 1000 and 1500 mg / L were adjusted proportionally to correspond to the masses of (NH4)2SO4 and NaNO2. After preparing the anaerobic ammonia oxidation medium according to the described formula in a 2 L three-necked flask, the medium was boiled using a heating device and purged with nitrogen for approximately 30 min to remove dissolved oxygen. 45 mL of the medium was then transferred to a vial using a pipette, sealed with a rubber stopper and aluminum cap, and autoclaved (121°C, 20 min). Following sterilization, inoculation was performed in a glove box. The magnesium-deficient solution (1 L) contains: NaCl 100 g, MgCl2·6H2O 10 g, KH2PO4 20 g, KCl 30 g, CaCl2·2H2O 1.5 g; the Se / W / Mo solution (1 L) contains: NaSeO3 0.004 g, NaWO4·2H2O 0.008 g, Na2MoO4·2H2O 0.036 g, NaOH 0.5 g; the vitamin stock solution (1 L) contains: D(+) biotin 20 mg, folic acid 20 mg, pyridoxine dihydrochloride monohydrate 252 mg, riboflavin 50 mg, thiamine hydrochloride 50 mg, niacin 50 mg, calcium-D(+)-pantothenic acid 50 mg, para-aminobenzoic acid 50 mg, DL-α-lipoic acid 50 mg, and vitamin B12 50 mg.

[0037] Under an initial ammonia nitrogen concentration of 500 mg / L, the concentrations of ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen in the culture medium were measured every 2 days. Ammonia nitrogen was detected using Nessler's reagent colorimetric method, nitrite nitrogen using N-(1-naphthyl)-ethylenediamine spectrophotometry, and nitrate nitrogen using direct ultraviolet spectrophotometry. Under initial ammonia nitrogen concentrations of 1000 and 1500 mg / L, the detection interval was 3 days. The experiment ended on day 6, and the ammonia nitrogen removal rate was calculated. The calculation formula is: ; in, To provide real-time ammonia nitrogen concentration during the cultivation process, This represents the initial ammonia nitrogen concentration in the culture medium.

[0038] The settings for each embodiment and comparative example are shown in Table 1.

[0039] Examples 1-15 In the glove box, the activated granular sludge was inoculated into vials containing anaerobic ammonia oxidation medium. In the vitamin B6 experiment, the experimental group was divided into 6 groups, each inoculated with the same wet weight sludge, and different concentrations of vitamin B6 were added to the 6 groups: the control group (comparative example) without vitamin B6, and the group with different concentrations of vitamin B6.

[0040] Examples 16-27 In the experiment on CAT (carrier iron), the experimental group was divided into 5 groups, each inoculated with the same wet heavy sludge. Different concentrations of CAT were added to each of the 5 groups: a control group (comparative example) without CAT, and experimental groups with different concentrations of CAT. Furthermore, to ensure that all experimental groups had the same and sufficient iron source, Fe was added to each group at a concentration equal to the molar concentration of the CAT. 3+ (Provided in the form of Fe2(SO4)3), the concentrations of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the wastewater were measured every 3 days under all conditions. The experiment ended on the 6th day and the ammonia nitrogen removal rate was calculated.

[0041] Examples 28-39 In the experiment using HEDTA as a siderophore, the experimental group was divided into 5 groups, each inoculated with the same wet heavy sludge. Different concentrations of HEDTA were added to each of the 5 groups: a control group (comparative example) without HEDTA, and experimental groups with different concentrations of HEDTA. Each group also received Fe2+ at a molar concentration equal to that of the siderophore. 3+ (Provided in the form of Fe2(SO4)3), the concentrations of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the wastewater were measured every 3 days under all conditions. The experiment ended on the 6th day and the ammonia nitrogen removal rate was calculated.

[0042] Examples 40-45 In the experiment of combining vitamin B6 with the siderophore CAT / HEDTA as a fortifier, the concentration of vitamin B6 and siderophore CAT was matched according to the principle of low concentration of vitamin B6 with low concentration of CAT and high concentration of vitamin B6 with high concentration of CAT. The experimental group was divided into 4 groups, each inoculated with the same wet heavy sludge. The 4 groups corresponded to different concentration combinations: 0 mg / L vitamin B6 + 0 μmol / L CAT (comparative example), 1 mg / L vitamin B6 + 12 μmol / L CAT (500 mg / L initial ammonia nitrogen), 3 mg / L vitamin B6 + 48 μmol / L CAT (1000 mg / L initial ammonia nitrogen), and 1 mg / L vitamin B6 + 12 μmol / L CAT (1500 mg / L initial ammonia nitrogen). In the combined experiment of vitamin B6 and the siderophore HEDTA, the experimental group was divided into four groups, each inoculated with the same wet heavy sludge. The four groups corresponded to different concentration combinations: 0 mg / L vitamin B6 + 0 μmol / L HEDTA (comparative example), 1 mg / L vitamin B6 + 24 μmol / L HEDTA (500 mg / L initial ammonia nitrogen), 3 mg / L vitamin B6 + 36 μmol / L HEDTA (1000 mg / L initial ammonia nitrogen), and 1 mg / L vitamin B6 + 24 μmol / L HEDTA (1500 mg / L initial ammonia nitrogen). Furthermore, to ensure that all experimental groups had the same and sufficient iron source, Fe was added to each group at a concentration equal to the molar concentration of the siderophore. 3+ (Provided in the form of Fe2(SO4)3), the concentrations of ammonia nitrogen, nitrite nitrogen and nitrate nitrogen in the wastewater were measured every 3 days under all conditions. The experiment ended on the 6th day and the ammonia nitrogen removal rate was calculated.

[0043] Examples 46-48 An experiment designed for the batch addition of vitamin B6 and siderophore CAT combined fortifier was conducted. Vitamin B6 and CAT were added twice during a 6-day culture period (i.e., half of vitamin B6 and CAT were added in the first batch, and the remaining vitamin B6 and CAT were added in the second batch).

[0044] Comparative Examples 1-3 This is equivalent to the control group without the added reinforcing agent in the aforementioned examples.

[0045] Table 1 The ammonia nitrogen removal rates of the comparative examples were calculated, and the results are shown in Table 1.

[0046] For vitamin B6, in the initial NH4 + and NO2 -At a concentration of 500 mg / L, appropriate addition of vitamin B6 can enhance the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia-oxidizing bacteria. The preferred concentration range is 0.1–2.0 mg / L, with an optimal concentration of 1.0 mg / L. (Initially, NH4...) + and NO2 - When the concentration is 1000 mg / L, the preferred concentration range is 2.0~5.0 mg / L, and the optimal concentration is 3.0 mg / L; in the initial NH4 + and NO2 - When the concentration is 1500 mg / L, the preferred concentration range is between 0.1 and 2.0 mg / L, with an optimal concentration of 1.0 mg / L. Excessive addition (e.g., ≥2.0 mg / L) may actually reduce the promoting effect or even inhibit bacterial activity. Population analysis was performed on the anaerobic ammonia oxidation granular sludge cultured under an initial ammonia nitrogen condition of 1000 mg / L (Comparative Example 2 and Examples 6-10). The results are as follows... Figure 1 As shown, in the experimental group treated with 3.0 mg / L vitamin B6, the anaerobic ammonia-oxidizing bacteria... Candidate Brocadia The highest relative abundance was found in *Broccardia*, reaching 10.95%, followed by other genera: Pseudomonas (Pseudomonas): 35.99%, Macellibacteriodes (Carnivorous Bacteria): 9.57%, Proteocatella (Proteophiles): 10.34%, Comamonadaceae (Trichomonasceae): 7.46%, Acetoanaerobic (Acetogenic anaerobic bacteria): 1.76%. Pseudomonas As key symbiotic bacteria, they primarily support the anaerobic ammonium oxidation (Anammox) process in two ways: firstly, through denitrification, they convert nitrates (NO3-) into nitrogenous compounds. - ) is reduced to nitrite (NO2) - Firstly, it provides key reaction substrates for the Anammox process; secondly, it secretes quorum sensing signaling molecules to regulate and promote the enrichment and metabolic activity of Anammox bacteria. Furthermore, its metabolic processes consume trace amounts of oxygen and degrade some COD, contributing to a more suitable anaerobic environment for Anammox bacteria. Comamonadaceae It has the ability to remove excess NO3 from the system through complete denitrification. - The role of (Anammox byproducts) in optimizing total nitrogen removal. Macellibacteriodes、Proteocatella and Acetoanaerobic Providing carbon sources and related substrates for anaerobic ammonia-oxidizing bacteria helps maintain a stable anaerobic environment. Adding different concentrations of vitamin B6 affects the function of anaerobic ammonia-oxidizing bacteria. Candidate Brocadia The relative abundance of [the bacteria] has an impact and can significantly improve the functional bacteria. Candidate Brocadia The proportion of nitrogen in the community improves the overall nitrogen removal performance of the system.

[0047] For ferrocarrier CAT in initial NH4 + and NO2 - At concentrations of 500, 1000, and 1500 mg / L, appropriate addition of the siderophore CAT improved the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia-oxidizing bacteria. The preferred concentration ranges were 12–36 μmol / L (at 500 mg / L ammonia nitrogen) and 12–24 μmol / L (at 1000 and 1500 mg / L ammonia nitrogen), respectively, with optimal concentrations of 36 μmol / L (at 500 mg / L ammonia nitrogen) and 12 μmol / L (at 1000 and 1500 mg / L ammonia nitrogen). Excessive addition may lead to a decrease in the promoting effect or even a lower removal rate than the control group. For the siderophore HEDTA, at the initial NH4... + and NO2 - When the concentrations are 500, 1000 and 1500 mg / L, the appropriate addition of iron carrier HEDTA can improve the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia oxidizing bacteria. The preferred concentration range is 24~36 μmol / L, and the optimal concentration is 24 μmol / L.

[0048] For the combination of vitamin B6 and the siderophore CAT, in the initial NH4 + and NO2 - At concentrations of 500, 1000, and 1500 mg / L, the addition of appropriate concentrations of vitamin B6 and the siderophore CAT combination all improved the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia oxidizing bacteria. Among all concentration combinations, the combination of 1 mg / L vitamin B6 + 12 μmol / L CAT showed the best overall denitrification effect. For the combination of vitamin B6 and the siderophore HEDTA, at initial culture concentrations of 500, 1000, and 1500 mg / L, the addition of appropriate concentrations of vitamin B6 and the siderophore HEDTA combination all improved the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia oxidizing bacteria. Among all concentration combinations, the combination of 1 mg / L vitamin B6 + 24 μmol / L HEDTA showed the best overall denitrification effect.

[0049] For the phased addition of the combination of vitamin B6 and the siderophore CAT, in the initial NH4 + and NO2 - At concentrations of 500, 1000, and 1500 mg / L, the combined effect of the enhancer was the same as when all the enhancers were added at the beginning of the culture, all of which could improve the removal of ammonia nitrogen and nitrite nitrogen by anaerobic ammonia oxidizing bacteria. However, when the enhancers were added in batches, the enhancers at the same concentration showed a better denitrification effect.

[0050] In summary, this invention utilizes a specific enhancer to improve the activity of anaerobic ammonia-oxidizing bacteria under high ammonia nitrogen conditions, thereby enhancing their nitrogen removal performance. When added to an anaerobic ammonia oxidation system, it significantly improves ammonia nitrogen removal efficiency, optimizes the microbial community structure, and promotes the growth of functional bacteria. Candidate Brocadia Enrichment of vitamin B6 at low, medium, and high initial NH4 levels. + and NO2 - Optimal dosage ranges exist for each concentration; excessive dosage may inhibit bacterial activity. Siderophores CAT and HEDTA also exhibit clear concentration effects under different initial ammonia nitrogen conditions, effectively enhancing the system's denitrification performance. In particular, when vitamin B6 is used in combination with siderophores CAT or HEDTA, or added in batches, the stability and treatment efficiency of the anaerobic ammonia oxidation process can be further enhanced under appropriate concentration combinations, achieving superior ammonia nitrogen removal. The enhancers and their combination strategies provided by this invention are suitable for wastewater treatment with different initial pollution loads, offering advantages such as flexible operation, significant effects, and ease of promotion. It represents a highly efficient and controllable microbial enhanced denitrification technology.

[0051] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. A method for improving the nitrogen removal efficiency of anaerobic ammonia oxidation in high-ammonia nitrogen wastewater, characterized in that, The method includes: adding a strengthening agent to high ammonia nitrogen wastewater containing anaerobic ammonia oxidation synergistic bacteria; The fortifying agent includes vitamin B6 and / or ferrocarrier; The siderophore includes one of catechin (CAT) and hydroxyethyl ethylenediamine triacetic acid (HEDTA); The application methods include: Vitamin B6 may be added alone, or siderophores may be added alone, or a combination of vitamin B6 and one of the siderophores may be added. The stimulant can be added all at the beginning of the anaerobic ammonia oxidation synergistic bacterial culture, or it can be added in batches and evenly throughout the entire culture cycle.

2. The method for improving the anaerobic ammonia oxidation denitrification effect in high-ammonia nitrogen wastewater according to claim 1, characterized in that, The anaerobic ammonia oxidation synergistic bacterial community includes core bacteria. Candidatus Brocadia ; Optionally, the anaerobic ammonia oxidation synergistic microbial community also includes Pseudomonas , Macellibacteriodes , Proteocatella , Comamonadaceae or Acetoanaerobium At least one of them.

3. The method for improving the anaerobic ammonia oxidation denitrification effect in high-ammonia nitrogen wastewater according to claim 1, characterized in that, NH4 in high ammonia nitrogen wastewater + and NO2 - The concentrations of each are independently 500~1500 mg / L.

4. The method for improving the anaerobic ammonia oxidation denitrification effect in high-ammonia nitrogen wastewater according to claim 1, characterized in that, The concentration of vitamin B6 added is 0.1~5.0 mg / L, preferably 1.0 mg / L~3.0 mg / L.

5. The method for improving the anaerobic ammonia oxidation denitrification effect in high-ammonia nitrogen wastewater according to claim 1, characterized in that, The concentration of catechin added is 12~48 μmol / L, preferably 12 μmol / L~36 μmol / L.

6. The method for improving the anaerobic ammonia oxidation denitrification effect in high-ammonia nitrogen wastewater according to claim 1, characterized in that, The concentration of the hydroxyethyl ethylenediamine triacetic acid is 12~48 μmol / L, preferably 24 μmol / L~36 μmol / L.

7. The method for improving the denitrification effect of anaerobic ammonia oxidation in high-ammonia nitrogen wastewater according to claim 1, characterized in that, When the combined form of vitamin B6 and one of the siderophores is used as a strengthening agent, and the initial ammonia nitrogen concentration is 500~1500 mg / L, the addition concentration range of vitamin B6 and siderophore is 0.1~5.0 mg / L and 12~48 μmol / L, respectively. When added in combination, the addition ratio is 1 mg / 50 μmol~5 mg / 12 μmol. The preferred ratio of vitamin B6 to siderophore CAT is 3 mg / 48 μmol~1 mg / 12 μmol, and the preferred ratio of vitamin B6 to siderophore HEDTA is 1 mg / 24 μmol~3 mg / 36 μmol.

Images