Artificial microbial multicellular high-ammonia nitrogen wastewater treatment flora, promoter, and construction method and application thereof

Abstract

The invention discloses an artificial microbial multicellular high-ammonia nitrogen wastewater treatment bacterial community, a promoter, a construction method and an application thereof. The bacterial community comprises at least two of Bacillus haynesii strain ZJB21169, Bacillus subtilis strain ZJB21170, Stenotrophomonas rhizophila strain ZJB21172, Proteus alimentorum strain ZJB21171 and Paracoccus kondratievae strain ZJB21134. The composite bacterial community is added to a prepared nutrient agent to obtain a microbial nutrient agent, which is used for denitrification in a sewage treatment system. The nutrient enhancer comprises, by weight, 245-450 parts of amino acids, 90-223 parts of metal salts, 0-50 parts of choline and 0-50 parts of B vitamins. The present invention adds a self-developed promoter to assist the GZBS process in strengthening the denitrification process of micro-aerobic activated sludge, and establishes an artificial microbial multicellular biochemical treatment system with synchronous nitrification and denitrification bacteria as the community advantage. The system has high denitrification efficiency and no secondary pollution. It has broad application prospects in biological denitrification treatment, especially for the denitrification treatment of high-ammonia nitrogen landfill leachate.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology, and in particular relates to an artificial microbial multicellular high-ammonia nitrogen wastewater treatment flora, a promoter, and a construction method and application thereof. Background Art

[0002] The treatment of landfill leachate is a recognized challenge both domestically and internationally. As a typical high-ammonia nitrogen wastewater, it has become a major challenge in the water treatment field due to its complex composition, difficult degradation, high organic matter content, and frequently fluctuating water quality indicators. Currently, the only wastewater treatment process that meets these requirements is membrane treatment technology. However, membrane treatment technology also faces two difficult challenges: membrane fouling and concentrate disposal. Traditional nitrification-denitrification denitrification processes are widely used, but the carbon-nitrogen ratio of landfill leachate is very low, requiring the addition of an additional carbon source for denitrification, resulting in high operating costs. Furthermore, high concentrations of ammonia nitrogen inhibit microbial growth and metabolism, resulting in low denitrification and COD removal efficiencies. In recent years, short-cut nitrification has emerged as a new biological denitrification technology for wastewater. Compared to traditional full-cut nitrification, it offers advantages such as a 25% reduction in aeration energy consumption, a 40% reduction in denitrification carbon sources, and reduced sludge production. It can also be coupled with anaerobic ammonium oxidation (Anammox) to achieve full-process autotrophic denitrification. It is currently the most cost-effective wastewater denitrification process with broad application prospects.

[0003] The GZBS (GenotyeZooloea Biochemical System) process flow is based on an "AT-BC system + secondary Fenton + secondary BAF" system, primarily consisting of front-end biochemical treatment and back-end advanced treatment. The front-end biochemical treatment is based on an improved and optimized version of Japan's AT-BC technology; the back-end advanced treatment primarily utilizes a Fenton + biological aerated filter (BAF) process, further removing remaining pollutants through advanced oxidation processes to ultimately meet emission standards. The AT-BC system consists of a rotating mesh microbial contactor, with Bacillus as the dominant bacterial species. By regulating the dissolved oxygen concentration in the system and adding a nutrient solution to the bacteria, the growth of the Bacillus bacteria is enhanced. The relationship between the membrane plate bacterial growth, the sludge thickness on the membrane plate, the rotating disc speed, and the influent water concentration is adjusted, and the recirculation ratio is controlled to ultimately optimize the entire system.

[0004] The GZBS process uses a "micro-aerobic activated sludge / two-stage Fenton-AF-BAF" combined process to treat landfill leachate. Micro-aerobic activated sludge is based on the theory of short-term nitrification-denitrification. It achieves short-term nitrification and denitrification through the control of dissolved oxygen. The anti-impact capacity is enhanced by effluent and sludge reflow, forming a highly efficient system for removing organic pollutants and TN, removing most of the organic matter and total nitrogen in the influent. The effluent of micro-aerobic activated sludge still contains a part of total nitrogen and difficult-to-biodegrade organic matter. The Fenton-AF-BAF deep treatment combined process is adopted. Fenton improves the biodegradability of the wastewater while reducing the color well. Then, by adding carbon source methanol, the total nitrogen and organic matter in the wastewater are removed by nitrification-denitrification reaction in AF-BAF, so that the effluent meets the "Pollution Control Standard for Municipal Waste Landfill GB16887-2008".

[0005] The GZBS process uses bacteria to replace traditional activated sludge in biochemical treatment, resolving issues such as the difficulty of mud-water separation in landfill leachate treatment, as well as low temperatures, high ammonia and nitrogen concentrations. Advanced treatment utilizes advanced oxidation processes and membrane biological treatment to oxidize, bioflocculate, and filter organic pollutants, fully decomposing nitrogen and carbon pollutants in the wastewater and completing the water purification process. However, the entire process faces challenges: the dominant bacterial community structure in the leachate treatment system is unstable and susceptible to factors such as changes in raw water quality and ambient temperature. This results in poor denitrification efficiency and stability in the treatment system, severely restricting the application and promotion of the process. Summary of the Invention

[0006] In response to the problems existing in the prior art, the purpose of the present invention is to provide a method for constructing a synchronous nitrification-denitrification artificial multicellular microbial community. By exploring the synergy and coupling of the aerobic nitrification and heterotrophic denitrification metabolic pathways in the denitrification process of high-ammonia nitrogen wastewater, different denitrification functional bacteria are coordinated and designed to obtain an artificial multicellular system with high denitrification performance for high-concentration ammonia nitrogen. Furthermore, according to the nutritional requirements of the multicellular system, good nutritional promoters and stabilizers are matched to achieve stable storage of the artificial multicellular community. The constructed artificial microbial multicellular community with synchronous nitrification and high-efficiency denitrification was applied in the engineering process of high-ammonia nitrogen wastewater treatment.

[0007] The specific technical solutions are as follows:

[0008] An artificial microbial multicellular high-ammonia nitrogen wastewater treatment bacterial community comprises at least two of Bacillus haynesii ZJB21169, Bacillus subtilis ZJB21170, Stenotrophomonas riizophila ZJB21172, Proteus alimentorum ZJB21171, and Paracoccus kondratievae ZJB21134. The preservation number of Bacillus haynesii ZJB21169 is CCTCC NO: M20221659, and the preservation number of Bacillus subtilis ZJB21170 is CCTCC NO: M20221659. NO: M20221658, the preservation number of Stenotrophomonas riizophila ZJB21172 is CCTCC NO: M20221657, the preservation number of Proteus alimentorum ZJB21171 is CCTCC NO: M20221656, and the preservation number of Paracoccus kondratievae ZJB21134 is CCTCC NO: M20211514.

[0009] A method for constructing an artificial microbial multicellular high-ammonia nitrogen wastewater treatment bacterial community comprises the following steps:

[0010] 1) Screening of simultaneous nitrification-denitrification bacteria: Take a sample of sewage and pour it into an EP tube containing sterile water. Shake and mix thoroughly. Use the doubling dilution method to make a concentration gradient of 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 The mixture of different concentration gradients was inoculated onto Stephenson medium plates using a spreader rod and cultured at 30°C for several days. The bacteria cultured on the plates with different concentration gradients were distinguished by judging the color, size, and shape of the colonies to separate the different strains by streaking. The plates were streaked three times to obtain pure colonies. The obtained pure colonies were inoculated into LB liquid medium for activation, and then inoculated into Stephenson liquid medium and cultured until turbid. Finally, the bacterial solution was transferred to a glycerol tube and stored at -80°C.

[0011] 2) further conducting denitrification tests on the single bacteria obtained in step 1) to determine efficient denitrification strains, compounding the efficient denitrification single functional bacteria, and screening and optimizing the synergy and stability of the composite bacterial community to construct an artificial multicellular synchronous nitrification-denitrification reaction bacterial community.

[0012] The components of the above-mentioned Stephenson medium include (NH4)2SO4, K2HPO4, NaH2PO4, MnSO4·4H2O, CaCO3, MgSO4·7H2O and H2O.

[0013] An artificial microbial multicellular high-ammonia nitrogen wastewater treatment bacterial community is used in denitrification, comprising the following steps: 1) preparing a nutrient enhancer, wherein the nutrient enhancer comprises, by weight, 245-450 parts of amino acids, 90-223 parts of metal salts, 0-50 parts of choline, and 0-50 parts of B vitamins, and sterilizing the mixture;

[0014] 2) mixing the fermented simultaneous nitrification-denitrification reaction bacterial community with the nutrient enhancer prepared in step 1), and adding 5% by mass concentration of sodium benzoate and 1% by mass concentration of magnesium chloride, stirring and mixing until uniformly mixed to obtain a microbial nutrient enhancer for denitrification;

[0015] 3) The microbial nutrient enhancer in step 2) is inoculated into the sewage for denitrification treatment.

[0016] Furthermore, the amino acid in step 1) includes at least one of alanine, glutamic acid, valine, proline and leucine; the metal salt includes calcium salt, magnesium salt, ferrous salt and copper salt; and the B vitamins include at least one of pantothenic acid and pyridoxine.

[0017] Furthermore, the amino acids include, by mass, 200-500 parts of alanine, 0-40 parts of glutamic acid, 0-40 parts of valine, 0-40 parts of proline, and 0-40 parts of leucine.

[0018] Furthermore, the metal salts include, by mass, 20-50 parts of calcium salt, 20-30 parts of magnesium salt, 10-25 parts of ferrous salt, and 10-20 parts of copper salt, wherein the calcium salt is CaCl2, the magnesium salt is MgSO4, the ferrous salt is FeSO4, and the copper salt is CuSO4.

[0019] Furthermore, the B vitamins, calculated by weight, include: 0-30 parts of pantothenic acid, 0-20 parts of pyridoxine, and the weight percentage of at least one substance is not 0.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] 1) No need to supplement carbon source: Bacillus is an aerobic and facultative anaerobic bacterium. It uses molecular oxygen as a carrier. When the oxygen supply is insufficient, it can use nitrate as the final electron carrier to produce NO2-N and N2, and remove the nitrate from the system. Operation data shows that the system can achieve stable operation when the carbon-nitrogen ratio is greater than 2.

[0022] 2) Complete decomposition of pollution: Both the biochemical system and the deep treatment system use sewage treatment technology to decompose pollutants, and homemade nutrient solution can assist the system in eliminating pollution.

[0023] 3) No odor and secondary pollution: Bacillus uses its rich enzymes to strongly decompose nitrogen and sulfur pollutants, purifying the water while eliminating the odor generated during sewage treatment.

[0024] 4) Less sludge generated: The project operation data show that the nutrient enhancer-assisted GZBS process technology of the present invention generates less sludge during the treatment of landfill leachate. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Schematic diagram of the 16S rDNA phylogenetic tree of the strains of the present invention;

[0026] Figure 2 Schematic diagram of ammonia nitrogen degradation rate of simultaneous nitrification and denitrification bacteria of the present invention;

[0027] Figure 3 This is a schematic diagram of a two-by-two combination of simultaneous nitrification and denitrification bacteria according to the present invention;

[0028] Figure 4 This is a schematic diagram of a three-by-three combination of simultaneous nitrifying and denitrifying bacteria according to the present invention;

[0029] Figure 5 Schematic diagram of the four-four and five-five combinations of the synchronous nitrifying and denitrifying bacteria of the present invention;

[0030] Figure 6 This is a schematic diagram of the process for preparing the microbial nutrient solution of the present invention;

[0031] Figure 7 Schematic diagram of the degradation of high-concentration ammonia nitrogen by the simultaneous nitrification and denitrification bacteria of the present invention;

[0032] Figure 8 This is a flow chart of the GZBS process in the present invention;

[0033] Figure 9 Schematic diagram of the changes in ammonia nitrogen concentration in the homemade microbial nutrient solution of the present invention and the nutrient solution of the control group in the GZBS process;

[0034] Figure 10Schematic diagram of COD changes in the homemade microbial nutrient solution of the present invention and the nutrient solution-assisted GZBS process of the control group;

[0035] Figure 11 Schematic diagram of the changes in total nitrogen concentration in the homemade microbial nutrient solution of the present invention and the nutrient solution of the control group assisted GZBS process. DETAILED DESCRIPTION

[0036] It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0037] The present invention is further explained by the following examples, but is not intended to limit the present invention. It should be understood that these examples are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0038] Example 1 Screening of simultaneous nitrification and denitrification bacteria

[0039] Our laboratory took 100 μL of sample from a sewage treatment plant in Hangzhou, poured it into an eppendorf tube containing 900 μL of sterile water, and mixed it by oscillation. The concentration gradient of 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 100 μL of the mixture at different concentration gradients was inoculated onto a Stephenson medium plate using a spreader rod and incubated at 30°C for several days. The bacteria cultured on the plates with different concentration gradients were distinguished by colony color, size, and shape, and then streaked three times to isolate pure colonies. The pure colonies were inoculated into LB liquid medium for activation, then inoculated into Stephenson liquid medium and incubated until turbid. Finally, the culture solution was transferred to a glycerol tube and stored at -80°C.

[0040] The above-mentioned Stephenson medium: (NH4)2SO42g, K2HPO40.75g, NaH2PO40.25g, MnSO4·4H2O0.01g, CaCO35g, MgSO4·7H2O0.03g, H2O1000mL.

[0041] The purified single colony was inoculated into SND medium (sterilized) for screening of synchronous nitrifying and denitrifying bacteria. Each single bacterium was inoculated into LB medium and cultured in a shaker (30°C, 160r / min) for 24h. Then, it was inoculated again as seed liquid (10% v / v) into a conical flask (250mL) containing 100mL liquid culture medium and cultured under the same conditions. After 0h, 24h, 48h, and 72h, samples were taken to test the bacterial density (OD 600 ), nitrate (NO3 - -N), ammonia nitrogen (NH3-N), nitrite nitrogen (NO2 - -N) and total nitrogen (TN) and other indicators, three groups of parallel samples (n = 3) were measured each time.

[0042] LB medium: 10 g tryptone, 5 g yeast powder, 10 g sodium chloride, distilled water to 1000 mL, natural pH.

[0043] The efficient denitrification strain was identified and PCR amplified using the universal bacterial primers 27F: 5′-AGAGTTTGATCMTGGCTCAG-3′, 1492R: 5′-TACGGYTACCTTGTTACGACTT-3′, and verified by agarose gel electrophoresis (1%); electrophoresis detection, gel excision, purification and sequencing were performed by Qingke Biotechnology.

[0044] The PCR reaction system consisted of 50 μl of 25 μl of Phusion High-Fidelity PCR MasterMix with HF Buffer, 3 μl of each 10 μM forward and backward F / R primers, 10 μl of DNA template, and 6 μl of ddH2O. PCR amplification was performed using the following reaction conditions: initial denaturation at 98°C for 30 seconds, followed by 25 cycles of denaturation at 98°C for 15 seconds, annealing at 58°C for 15 seconds, and extension at 72°C for 15 seconds. A final extension at 72°C for 1 minute was performed. PCR products were purified using AMPure XP Beads (Beckman Coulter, Indianapolis, IN) and quantified using the PicoGreen dsDNA Assay Kit (Invitrogen, Carlsbad, CA, USA).

[0045] The 16S rDNA sequence of strains ZJB21169, ZJB21170, ZJB21171, ZJB21172, and ZJB21134 is 1360 bp in length, and their gene sequences are shown in SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, and SEQ ID NO.5.

[0046] The strain sequence results were uploaded to the NCBI database and compared with the existing bacterial 16S rDNA gene sequences in the database. The phylogenetic tree was constructed by the neighbor-joining method of Mega.7.0 software to analyze the genetic characteristics of the strain and determine its species and evolutionary status in genetics. The results are shown in Figure 1 .

[0047] The strain ZJB21169 was named Bacillus haynesii, strain number ZJB21169, deposited in the China Center for Type Culture Collection (CCTCC), with the deposit number CCTCC NO: M20221659, and the deposit date was October 25, 2022.

[0048] The strain ZJB21170 was named Bacillus subtilis, strain number ZJB21170, deposited in the China Center for Type Culture Collection (CCTCC), with the deposit number CCTCC NO: M20221658, and the deposit date was October 25, 2022.

[0049] The strain ZJB21172 was named Stenotrophomonas rhizophila, strain number ZJB21172, deposited in the China Center for Type Culture Collection (CCTCC), with the deposit number CCTCC NO: M20221657, and the deposit date was October 25, 2022.

[0050] The strain ZJB21171 was named Proteus alimentorum, strain number ZJB21171, deposited in the China Center for Type Culture Collection (CCTCC), with the deposit number CCTCC NO: M20221656, and the deposit date was October 25, 2022.

[0051] The strain ZJB21134 was named Paracoccus kondratievae, strain number ZJB21134, deposited in the China Center for Type Culture Collection (CCTCC), with the preservation number CCTCC NO: M20211514, and the preservation date was November 30, 2021.

[0052] Example 2 Construction of simultaneous nitrification and denitrification bacterial community

[0053] The efficient denitrification strains obtained from the preliminary screening were compounded, and the optimal denitrification combination was used as the strain combination for constructing the simultaneous nitrification and denitrification bacterial community. The strain compound numbering method is shown in Table 1, and the combination arrangement method is shown in Table 2.

[0054] Table 1 Correspondence table of strain compound numbers

[0055]

[0056]

[0057] Table 2 Compound permutation and combination table

[0058]

[0059] like Figure 2 Figure 2 shows the ammonia degradation rates of five single bacterial strains. After multiple rounds of enrichment, isolation, and purification, 30 strains with simultaneous nitrification and denitrification capabilities were identified. These strains were inoculated at a 10% inoculum into simultaneous nitrification and denitrification culture medium and incubated at 30°C, 160 rpm, and shaker for 24 hours. Based on water quality testing, strains with ammonia degradation rates exceeding 80% during simultaneous nitrification and denitrification were retained for subsequent experiments. Five highly efficient simultaneous nitrification and denitrification strains were identified, and their ammonia degradation efficiencies are shown in the figure. The ammonia degradation rates of S1, L2, X3, N4, and B5 were 88.98%, 88.11%, 99.10%, 97.76%, and 94.35%, respectively.

[0060] Figure 3 、 4 ,5. Schematic diagram of ammonia nitrogen and nitrate nitrogen degradation with different compound results. Five strains were combined to find the optimal denitrification combination, such as Figure 3 As shown, among the two-by-two combinations, the highest ammonia degradation rate was achieved in combination #1+3, reaching 97.55%. Nitrate degradation rates were also higher in combinations #1+4, #3+4, and #4+5, with degradation rates of 45.49%, 43.31%, and 46.34%, respectively. This suggests that the #1+3 combination may achieve better ammonia oxidation in the early stages of ammonia oxidation, while Paracoccus plays a dominant role in the later stages of denitrification. Furthermore, growth measurements revealed a strong correlation between growth and ammonia degradation rates—the better the bacterial growth, the higher the ammonia degradation rate. However, for nitrate degradation, growth did not seem to be particularly correlated. This may be because ammonia degradation is primarily a process of cellular assimilation. Among the three-by-three combinations, all achieved ammonia degradation rates exceeding 90%. The highest ammonia degradation rate was achieved in combination #3+4+5, while the highest nitrate degradation rate was achieved in combination #1+2+5, with a degradation rate of 69.46%. This corresponds to an ammonia degradation rate of 98.04%. In the 4:4 combination and 5:5 combination, the ammonia nitrogen degradation rates are slightly higher than those in the 2:2 combination and 3:3 combination, and the ammonia nitrogen degradation rates both reach more than 98%. The combination with the highest ammonia nitrogen and nitrate nitrogen degradation rates is #1+2+3+4+5, with values ​​of 98.95% and 29.54% respectively.

[0061] Example 3 Preparation of a biological nutrient enhancer based on simultaneous nitrification and denitrification bacteria

[0062] 1) Bacillus heinensis, Bacillus subtilis, Stenotrophomonas rhizophila, Proteus, and Paracoccus condrasii were inoculated into LB liquid medium, cultured in a constant shaking incubator at 30°C and 160 rpm for 18-24 hours, and centrifuged at 8000 rpm for 10 minutes;

[0063] 2) Prepare the nutritional supplement according to Table 3, sterilize at 120°C for 20 min, and sterilize the B vitamins by filtration using a 0.22 μm filter membrane. Store at 4°C.

[0064] 3) The fermented mixed strains were mixed with the nutrient components, and 6 g of wet cell mud was added to each 1 L of the solution made from the nutrient components. Then, sodium benzoate with a preservative concentration of 5% and magnesium chloride with a protective agent concentration of 1% were added to the bacterial agent. The mixture was stirred at 100 rpm for 10 minutes and mixed well to obtain the finished product. The number of viable bacteria in the finished product was 10 7 ~10 8 CFU / mL, and can be stored at 4 ° C for more than 50 days. The parameters of the nutritional enhancer are shown in Table 4. The preparation process of the nutritional enhancer is as follows: Figure 6 shown.

[0065] Table 3 Nutritional Enhancer Components (by mass)

[0066] Components parts by mass Alanine 250 glutamate 40 Valine 40 Proline 40 Leucine 4 calcium chloride 30 magnesium sulfate 25 Ferrous sulfate 20 copper sulfate 15 choline 25 Pantothenic acid 25 Pyridoxine 15

[0067] Table 4 Nutrient solution compound addition scheme

[0068]

[0069] Example 4 Degradation of high concentration ammonia nitrogen by microbial nutrient enhancers.

[0070] The formula of high concentration ammonia nitrogen culture medium is: ammonium sulfate ((NH4)2SO4) 2.36g, sodium succinate (C4 H4Na2O4), 16.88 g of potassium dihydrogen phosphate (KH2PO4), 1.5 g of potassium dihydrogen phosphate trihydrate (K2HPO4·3H2O), 6.5 g of trace element solution, 20 mL of distilled water to 1000 mL, pH 7.0-7.5; the trace element solution includes: zinc sulfate (ZnSO4), 4.4 mg of manganese chloride tetrahydrate (MnCl2·4H2O), 10.2 mg of calcium chloride (CaCl2), 11 mg of ferrous sulfate heptahydrate (FeSO4·7H2O), 3.2 mg of copper sulfate pentahydrate (CuSO4·5H2O), 2.2 mg of ammonium molybdate tetrahydrate ((NH4)6Mo7O2·4H2O), 3.2 mg of cobalt chloride hexahydrate (CoCl2·6H2O), 144 mg of EDTA-Na, 1000 mL of distilled water, natural pH.

[0071] The treatment method is: inoculate 2% seed liquid (seed liquid is a bacterial suspension cultured in LB medium at 30°C and 160 rpm for 12 hours), culture at 30°C and 160 rpm. Regular sampling is performed to determine the ammonia nitrogen, nitrite nitrogen, nitrate nitrogen, total nitrogen and OD in the supernatant. 600 Value, see the result Figure 7 .

[0072] Conclusion: As the strain grew, ammonia nitrogen was essentially degraded within 24 hours, reaching a degradation rate of 87.89%. Nitrate and nitrite nitrogen accumulation was not observed, indicating that any NO₂-N or NO₃-N produced by the system was immediately reduced to N₂ and did not accumulate. Furthermore, total nitrogen concentrations fluctuated throughout the process, ultimately decreasing slightly.

[0073] Example 5 Application of microbial nutrient enhancers in biological denitrification treatment of landfill leachate and comparison of their effects

[0074] The nutrient enhancer is applied to the biological treatment section of landfill leachate. The operation mode of the section is as follows: Figure 8 The initial water quality parameters are shown in Table 5. 500 mL of the mother solution of the microbial nutrient enhancer was taken and added to the aeration tank section of the sewage treatment plant at a dilution ratio of 1:3000. The ammonia nitrogen, COD and total nitrogen contents in the sewage in different sections were measured and compared with the denitrification effect of the imported nutrient enhancer. The results are shown in the figure. Figure 9 、 Figure 10 and Figure 11 shown.

[0075] Table 5 Initial water quality parameters of sewage treatment plants

[0076] Chemical oxygen demand Total nitrogen Ammonia nitrogen C / N 10000-15000 2000-3000 2000-2500 5-7

[0077] Conclusion: The data of commercial nutrient solution assisted GZBS process were used as the control group. Figure 9 、 Figure 10 and Figure 11 As shown in the results, within 34 days of operation, the ammonia nitrogen concentration of the homemade nutrient solution group decreased from approximately 2000 mg / L to approximately 50 mg / L; COD decreased from approximately 15,000 in the influent to approximately 500, and the total nitrogen concentration decreased from approximately 2500 mg / L to approximately 100 mg / L. In the control group without the bioaugmentant, the ammonia nitrogen concentration, COD, and total nitrogen concentrations were all slightly higher than those of the homemade nutrient solution. This demonstrates that the microbial nutrient enhancer of the present invention is superior to commercially available nutrient solutions in assisting the denitrification process in the GZBS process.

[0078] The artificial microbial multicellular high-ammonia nitrogen wastewater treatment flora based on simultaneous nitrification and denitrification flora, promoter, and construction method thereof provided by the present invention have been proven to have a good promoting effect on the assisted GZBS process to enhance the micro-aerobic activated sludge denitrification treatment, indicating that this technology has broad application prospects in biological denitrification treatment, especially in landfill leachate denitrification treatment technology.

Claims

1. Artificial microbial multicellular high ammonia nitrogen wastewater treatment flora, characterized by The bacterial group is a composite combination of at least four of Bacillus heinensis ZJB21169, Bacillus subtilis ZJB21170, Stenotrophomonas rhizophila ZJB21172, Proteus amanita ZJB21171 and Paracoccus condrasii ZJB21134, wherein Bacillus heinensis ( Bacillus haynesii ) The deposit number of ZJB21169 is CCTCC NO: M20221659, Bacillus subtilis ( Bacillus subtilis ) The deposit number of ZJB21170 is CCTCC NO: M20221658, and the rhizotrophic bacteria ( Stenotrophomonas riizophila ) The deposit number of ZJB21172 is CCTCC NO: M20221657, Proteus amanita ( Proteus alimentorum ) The deposit number of ZJB21171 is CCTCC NO: M20221656, Paracoccus condellatus ( Paracoccus kondratievae ) The deposit number of ZJB21134 is CCTCC NO: M20211514.

2. An application of the artificial microbial multicellular high-ammonia nitrogen wastewater treatment bacterial community as claimed in claim 1 in denitrification, characterized in that The steps include: 1) preparing a nutritional enhancer comprising, by weight, 245-450 parts of amino acids, 90-223 parts of metal salts, 0-50 parts of choline, and 0-50 parts of B vitamins, and sterilizing the mixture; 2) mixing the bacterial community of claim 1 with the nutrient enhancer prepared in step 1), and adding 5% by mass concentration of sodium benzoate and 1% by mass concentration of magnesium chloride, stirring and mixing until uniformly mixed to obtain a microbial nutrient enhancer for denitrification; 3) The microbial nutrient enhancer in step 2) is inoculated into the sewage for denitrification treatment.

3. The use according to claim 2, characterized in that The amino acid in step 1) includes at least one of alanine, glutamic acid, valine, proline and leucine; the metal salt includes calcium salt, magnesium salt, ferrous salt and copper salt; and the B vitamin includes at least one of pantothenic acid and pyridoxine.

4. The use according to claim 3, characterized in that The amino acids include, by mass, 250 parts of alanine, 40 parts of glutamic acid, 40 parts of valine, 40 parts of proline, and 4 parts of leucine.

5. The use according to claim 3, characterized in that The metal salts are calculated by mass and include: 30 parts of calcium salt, 25 parts of magnesium salt, 20 parts of ferrous salt, and 15 parts of copper salt. The calcium salt is CaCl2, the magnesium salt is MgSO4, the ferrous salt is FeSO4, and the copper salt is CuSO4.

6. The use according to claim 2, characterized in that The B vitamins, calculated by mass, include: 25 parts of pantothenic acid and 15 parts of pyridoxine.

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