Nitrosomonas AOB SX-1 and application thereof in ammonia removal

Abstract

The invention discloses nitrosomonas AOB SX-1 and application thereof in ammonia removal, the nitrosomonas AOB SX-1 is preserved in the China Center for Type Culture Collection (CCTCC), the preservation date is November 13, 2023, and the preservation number is CCTCC NO: M 20232220. The ammonia oxidizing bacteria AOB SX-1 can be used for efficiently and deeply treating different types of ammonia nitrogen sewage with different concentrations in a short time, and can be applied to the field of ammonia removal. A dominant strain can be provided for the subsequent development of a novel broad-spectrum autotrophic nitrifying bacterium agent, and the method has a huge research and development prospect.

Description

Technical Field

[0001] This invention relates to a highly efficient broad-spectrum ammonia-oxidizing bacterium, Nitrosomonas sp. AOB SX-1, and its application in ammonia removal in wastewater treatment. Background Technology

[0002] With economic development and social progress, human water consumption for production and daily life is increasing daily. Unsustainable development and uncontrollable pollution are currently "hot issues" in water resource management. Ammonia nitrogen, as the second largest pollutant in wastewater treatment, is discharged in the millions of tons nationwide every year. Excessive ammonia nitrogen discharge into water bodies not only causes foul odors and eutrophication, but also harms aquatic life and human health.

[0003] Currently, ammonia removal in wastewater is mainly achieved through physicochemical and biological technologies. However, physicochemical technologies have limited efficiency, high costs, and are prone to generating secondary pollutants. Therefore, environmentally friendly and economically viable biological technologies are gaining popularity. Common ammonia-removing microorganisms include heterotrophic nitrifying bacteria and autotrophic nitrifying bacteria. Heterotrophic nitrifying bacteria utilize organic matter as a carbon source and remove ammonia nitrogen through assimilation or oxidation. These microorganisms are highly susceptible to the type of carbon source and the C / N ratio, resulting in low ammonia nitrogen removal rates in actual wastewater treatment and competition with existing indigenous bacterial communities, limiting their practical value. Autotrophic nitrifying bacteria include ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). Ammonia-oxidizing bacteria utilize simple inorganic carbon sources (CO2, CO32-). 2- CO3H - It oxidizes ammonia nitrogen into nitrite to achieve ammonia removal. Its ammonia removal capacity is far higher than that of heterotrophic nitrifying bacteria, and it has extremely high industrial development value.

[0004] However, screening and isolating these microorganisms is difficult, and most of the currently reported patented ammonia-oxidizing bacteria have drawbacks such as inefficient and limited ability to remove ammonia nitrogen at depth and the limited range of ammonia-nitrogen-containing wastewater types they can treat. For example, patent CN102268386A discloses that the ammonia-oxidizing bacteria AP-8, after treating 100 mg / L simulated ammonia-nitrogen wastewater, still left 3.67 mg / L of ammonia nitrogen residue after 17 days; patent CN 103074278A reports that the nitrosomonas cetacean CCTCC M2012456, when inoculated at a 10% inoculum into a medium containing 180 mg / L ammonia nitrogen and cultured for 5 days, still left 6.4 mg / L of ammonia nitrogen residue in the culture system. Therefore, providing an ammonia-oxidizing bacterium capable of efficiently and deeply removing different types of ammonia-nitrogen-containing wastewater is of great significance for the development of novel broad-spectrum autotrophic nitrifying bacterial agents and the treatment of various types of ammonia-nitrogen-containing wastewater. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of current microbial ammonia removal technologies and to provide a highly efficient, broad-spectrum ammonia-oxidizing bacterium and its application in ammonia removal in different types of wastewater treatment.

[0006] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0007] In a first aspect, the present invention provides a strain of bacteria for efficient and deep ammonia removal and oxidation in different types of ammonia nitrogen wastewater. The strain is Nitrosomonas sp. AOB SX-1, which is deposited at the China Center for Type Culture Collection (CCTCC) on November 13, 2023, with accession number CCTCC NO: M 20232220, address: Wuhan University, Wuhan, China, postal code: 430072.

[0008] Secondly, the present invention provides the application of the above-mentioned Nitrosomonas sp. AOB SX-1 in ammonia removal, especially in the removal of ammonia from wastewater.

[0009] Furthermore, the wastewater refers to textile dyeing wastewater, mixed industrial wastewater, municipal domestic wastewater, aquaculture wastewater, or eutrophic river water.

[0010] Under aerobic conditions, the ammonia-oxidizing bacteria can efficiently and deeply remove ammonia from simulated ammonia nitrogen wastewater, textile dyeing wastewater of different concentrations, mixed industrial wastewater, municipal domestic sewage, aquaculture wastewater, and eutrophic river water.

[0011] Furthermore, the method of application is as follows: the fermentation broth of Nitrosomonas sp. AOB SX-1 is added to the wastewater.

[0012] In one embodiment of the present invention, the OD of the fermentation broth 600 =2.0. Further, the volume ratio of the fermentation liquid to the wastewater is 1:100 to 1000.

[0013] Those skilled in the art will know that the fermentation broth refers to the liquid obtained by fermenting Nitrosomonas sp. AOB SX-1 after inoculating it into a fermentation medium.

[0014] Specifically, the application method is as follows: (1) Nitrosomonas sp. AOBSX-1 is inoculated into a fermentation medium and cultured for the first time at 30°C and 150 rpm for 5 days to obtain seed liquid; during the first shaking culture, the pH of the system is controlled to be 7.5-8.0 and the ammonia nitrogen concentration is 100 mg / L.

[0015] (2) Inoculate the seed culture from step (1) into fresh fermentation medium at a volume inoculation rate of 10%, and incubate with a second shaking incubation at 30°C and 150 rpm (until OD). 600 =2.0), to obtain the fermentation broth; during the second shaking culture, the pH of the system was controlled at 7.5-8.0 and the ammonia nitrogen concentration was 100 mg / L;

[0016] (3) Add the fermentation liquid from step (2) into the wastewater to remove ammonia.

[0017] Furthermore, the volume ratio of the fermentation liquid to the wastewater in step (3) is 1:100 to 1000, and in the embodiment of the present invention it is 1:100.

[0018] In one embodiment of the present invention, the pH of the system in step (1) or (2) is controlled by adding 5% Na2CO3 aqueous solution; the ammonia nitrogen concentration is controlled by adding 5% (NH4)2SO4 aqueous solution.

[0019] In one embodiment of the present invention, the fermentation medium in step (1) or (2) is composed of the following components at final concentrations: 1.5 g / L (NH4)2SO4, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, and 0.03 g / L MgSO4·7H2O; the solvent is water, and the pH is 7.5-8.0.

[0020] Furthermore, the conditions for ammonia removal in step (3) are: 30°C, 150 rpm shaking for 1-48 hours.

[0021] Compared with the prior art, the beneficial effects of the present invention are mainly reflected in:

[0022] Most patented ammonia-oxidizing bacteria and their nitrifying bacteria agents currently suffer from drawbacks such as inefficient deep removal of ammonia nitrogen and limited treatment options for ammonia-nitrogen-rich wastewater. Therefore, in actual wastewater treatment, multiple ammonia-oxidizing bacteria need to be added, and the treatment time is relatively long. However, the ammonia-oxidizing bacteria AOB SX-1 of this invention can efficiently and deeply treat different types and concentrations of ammonia nitrogen wastewater in a short time. It can provide a superior strain for the subsequent development of novel broad-spectrum autotrophic nitrifying bacteria agents and has enormous research and development potential. Attached Figure Description

[0023] Figure 1 The morphology of Nitrosomonas sp. AOB SX-1 strain.

[0024] Figure 2Phylogenetic tree of Nitrosomonas sp. AOB SX-1.

[0025] Figure 3 The effect of AOB SX-1 bacteria on deep ammonia removal from textile dyeing and printing wastewater with low ammonia nitrogen concentration was studied.

[0026] Figure 4 The effect of AOB SX-1 bacteria on deep ammonia removal from textile dyeing and printing wastewater with medium ammonia nitrogen concentration was investigated.

[0027] Figure 5 The effect of AOB SX-1 bacteria on ammonia removal in textile dyeing and printing wastewater with high ammonia nitrogen concentration was investigated.

[0028] Figure 6 The effect of AOB SX-1 bacteria on ammonia removal from mixed industrial wastewater.

[0029] Figure 7 The ammonia removal effect of AOB SX-1 bacteria on municipal sewage.

[0030] Figure 8 The effect of AOB SX-1 bacteria on ammonia removal in aquaculture wastewater.

[0031] Figure 9 The effect of AOB SX-1 bacteria on ammonia removal in eutrophic river water. Detailed Implementation

[0032] The present invention will be further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited thereto:

[0033] The culture medium involved in this invention is as follows:

[0034] 1. Enrichment and acclimatization culture medium: 2 g / L (NH4)2SO4, 1 g / L Na2CO3, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, 0.03 g / L MgSO4·7H2O;

[0035] 2. Screening medium: 1 g / L (NH4)2SO4, 0.5 g / L Na2CO3, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, 0.03 g / L MgSO4·7H2O, 20 g / L agar;

[0036] 3. Fermentation medium: 1.5 g / L (NH4)2SO4, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, 0.03 g / L MgSO4·7H2O, solvent is water;

[0037] 4. Simulated ammonia nitrogen wastewater culture medium: 0.5 g / L (NH4)2SO4, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, 0.03 g / L MgSO4·7H2O, with water as the solvent.

[0038] All the above culture media were water-based solvents. The pH of the system was adjusted to 7.5-8.0 and all media were sterilized at 121℃ for 30 minutes.

[0039] Example 1: Screening and identification of Nitrosomonas AOB SX-1

[0040] (1) Collection of inoculum

[0041] Eight different sludge samples were collected from wastewater treatment plants in Jiangxi, Shaoxing, and Shanghai, and stored at 4°C for microbial screening.

[0042] (2) Sludge acclimation and enrichment

[0043] Take 1g of different sludge samples and add them to 100mL of enrichment and acclimatization medium. Incubate at 30℃ and 150rpm with shaking. Take 1mL of sample every 24h and qualitatively monitor the ammonia nitrogen removal by using wide-range pH test paper and adding Sodium's reagent. If the pH decreases significantly and the solution does not change color or has a very light color after adding Sodium's reagent, it indicates that there are ammonia-oxidizing bacteria in the sludge sample. Save the sample for subsequent strain screening.

[0044] (3) Isolation of ammonia-oxidizing bacteria

[0045] Take the culture medium after acclimatization and enrichment, centrifuge at 8000×g for 5 min, discard the supernatant, collect the bacterial cells, and dilute them with sterile water to a concentration of 10. -1 10 -2 10 -3 and 10 -4 Four gradients of dilution were applied to screening medium plates, which were then incubated at 30°C. The growth of the strains on the plates was observed daily, and colonies of different morphologies and colors were selected and streaked multiple times until the colonies on the plates exhibited a uniform morphology. Ultimately, six initial screening strains were obtained from different sludge samples.

[0046] (4) Comparison of ammonia oxidation efficiency of initially screened strains

[0047] The six strains initially screened were inoculated into 100 mL of simulated ammonia nitrogen wastewater culture medium and cultured at 30℃ with shaking at 150 rpm. 1 mL samples were periodically taken and qualitatively monitored for ammonia nitrogen removal using wide-range pH test paper and by adding Sodium's reagent. Ultimately, it was found that, except for the AOBSX-1 experimental group, all other experimental groups showed a reddish-brown color after adding Sodium's reagent, and the pH of the system did not decrease significantly. Therefore, AOBSX-1 showed the best ammonia nitrogen removal effect.

[0048] (5) Strain identification

[0049] like Figure 1 As shown, strain AOB SX-1 appears pinkish-yellow on the selection medium plate with a dry surface. Strain AOB SX-1 was sent to Hangzhou Qingke Biotechnology Co., Ltd. for 16S rDNA sequencing. The 16S rDNA nucleotide sequence is shown in SEQ ID NO. 1. The full-length sequence of strain AOB SX-1 was found to be 1479 bp. BLAST comparison with the NCBI database showed that this strain had the highest similarity to Nitrosomonas sp. ENI-11, with a homology of 99.93%. Therefore, AOB SX-1 belongs to the genus *Nitrosomonas*. Multiple sequence alignment analysis was performed using MEGA software, and a phylogenetic tree was constructed using the Neighbor-Joining method. The phylogenetic tree of (Nitrosomonas sp.) AOB SX-1 based on 16S rDNA sequence homology is shown below. Figure 2 As shown. The Nitrosomonas sp. AOB SX-1 is deposited at the China Center for Type Culture Collection (CCTCC) on November 13, 2023, with accession number CCTCC NO: M 20232220, address: Wuhan University, Wuhan, China, postcode: 430072.

[0050] Nitrosomonas sp. AOB SX-1 16S rDNA sequence

[0051]

[0052] Example 2: Culture of Nitrosomonas sp. AOB SX-1

[0053] A single colony of *Nitrosomonas SX-1* was picked up with a long toothpick and inoculated into 100 mL of fermentation medium. The culture was incubated at 30°C with shaking at 150 rpm. During the incubation period, pH and ammonia nitrogen concentrations were measured every 6 hours during the day and every 12 hours at night. The pH (7.5-8.0) and ammonia nitrogen concentration (approximately 100 mg / L) of the culture system were maintained by adding appropriate amounts of 5% Na₂CO₃ solution and 5% (NH₄)₂SO₄. After 5 days of incubation, a bacterial suspension was obtained. Then, 100 mL of the *Nitrosomonas SX-1* bacterial suspension was inoculated into 1 L of fermentation medium, and the same incubation method with shaking was performed for 5 days to obtain the fermentation broth of this strain.

[0054] Example 3: Treatment of simulated ammonia nitrogen wastewater by Nitrifying Monotropha AOB SX-1 and nitrifying bacteria agents from different environmental protection companies.

[0055] The efficiency of AOB SX-1 strain, Shanghai GandeW-NI solid powder ammonia nitrogen removal agent, and Wuhan Shuizhiguo Environmental Protection Co. liquid autotrophic nitrifying bacteria (BP100) in removing ammonia nitrogen was compared using simulated ammonia nitrogen wastewater. The specific experimental design is shown in Table 1 below.

[0056] Table 1: Experimental schemes for the removal of ammonia nitrogen from simulated wastewater by AOB SX-1 strain and different nitrifying agents

[0057]

[0058] The simulated ammonia nitrogen wastewater consisted of 0.5 g / L (NH4)2SO4, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, and 0.03 g / L MgSO4·7H2O. Both the control and experimental groups were cultured at 30℃ and 150 rpm. The pH of each culture system was monitored in real time during the treatment, and the pH was maintained between 7.5 and 8.0 by adding an appropriate amount of 5% Na2CO3 solution.

[0059] The experimental results are shown in Table 2 below. The results indicate that although the initial concentration of AOB SX-1 strain was low, after approximately 5 days of aerobic treatment (30℃, 150 rpm) on simulated ammonia nitrogen wastewater, the ammonia nitrogen concentration in the system was only 0.266 mg / L, with a removal rate exceeding 99%. Extending the treatment time (6 days), ammonia nitrogen was almost completely removed. In contrast, an ammonia nitrogen removal agent from a Shanghai company achieved a removal rate of only 26.59% after 7 days. At Wuhan Shuizhiguo, after 5 days of aerobic treatment with autotrophic nitrifying bacteria, the ammonia nitrogen concentration in the system was still 34.12 mg / L. After 7 days of aerobic treatment, the ammonia nitrogen concentration was 0.512 mg / L, with a removal rate of 99.53%. Therefore, the Nitrifying Monotroph AOB SX-1 strain has a more rapid effect on removing ammonia nitrogen from water bodies.

[0060] Table 2: Changes in ammonia nitrogen concentration (mg / L) under different treatment regimens

[0061]

[0062] Table 2 (continued): Changes in ammonia nitrogen concentration (mg / L) under different treatment regimens

[0063]

[0064] Example 4: Application of Nitrifying Monotropha AOB SX-1 in the Advanced Treatment of Textile Dyeing Wastewater with Low Ammonia Nitrogen Concentration

[0065] The ammonia nitrogen concentration in the biochemical effluent of the wastewater treatment plant of Shaoxing Yongfeng Textile Printing and Dyeing Company has long remained at 3-10 mg / L. To verify the deep ammonia nitrogen removal capacity of *Nitrosomonas* AOB SX-1, 100 mL of the company's biochemical effluent was taken and 1 mL of *Nitrosomonas* AOB SX-1 fermentation broth (OD) was added. 600 =2.0), cultured at 30℃ and 150rpm with shaking, and samples were taken periodically to detect ammonia nitrogen content. The control group was set as the dyeing biochemical effluent without bacteria.

[0066] The results are as follows Figure 3 As shown, after 2 hours of treatment with *A. nitrosomonas* AOB SX-1, the ammonia nitrogen concentration in the biochemical effluent from a dyeing and printing enterprise decreased from 8.9 mg / L to 0.31 mg / L, achieving a removal rate of 96.5%. This result indicates that *A. nitrosomonas* AOB SX-1 can effectively treat dyeing and printing wastewater for deep ammonia removal.

[0067] Example 5: Application of Nitrosomonas AOB SX-1 in the treatment of textile dyeing and printing wastewater with medium ammonia nitrogen concentration

[0068] Take 100mL of biochemical effluent from a dyeing and printing enterprise in Xindu, Zhejiang, and add 1mL of *Nitrosomonas* AOB SX-1 fermentation broth (OD). 600=2.0), cultured at 30℃ and 150rpm with shaking, and samples were taken periodically to detect ammonia nitrogen content. The control group was set as the dyeing biochemical effluent without bacteria.

[0069] The results are as follows Figure 4 As shown, after 16 hours of treatment with Nitrosomonas AOB SX-1, the ammonia nitrogen concentration in textile dyeing wastewater with medium ammonia nitrogen levels decreased from 31.5 mg / L to 0.412 mg / L, achieving a removal rate as high as 98.7%. Therefore, Nitrosomonas AOB SX-1 can be used for deep ammonia removal treatment of textile dyeing wastewater with medium ammonia nitrogen levels.

[0070] Example 6: Application of Nitrosomonas AOB SX-1 in the treatment of textile dyeing and printing wastewater with high ammonia nitrogen concentration

[0071] The ammonia nitrogen concentration in the biochemical effluent of Zhejiang Yinan Printing and Dyeing Co., Ltd. has consistently remained at 100-300 mg / L. To verify the ability of *Nitrosomonas* AOB SX-1 to remove wastewater with high ammonia nitrogen concentrations, 100 mL of the company's biochemical effluent was taken, and 1 mL of *Nitrosomonas* AOB SX-1 fermentation broth (OD) was added. 600 =2.0), cultured at 30℃ and 150rpm with shaking, and samples were taken periodically to detect ammonia nitrogen content. The control group was set as the dyeing biochemical effluent without bacteria.

[0072] The results are as follows Figure 5 As shown, after 48 hours of treatment with Nitrosomonas AOB SX-1, the ammonia nitrogen concentration in textile dyeing wastewater with high ammonia nitrogen levels decreased from 270 mg / L to 1.3 mg / L, achieving a removal rate as high as 99%. Therefore, Nitrosomonas AOB SX-1 has great potential for treating textile dyeing wastewater with high ammonia nitrogen concentrations.

[0073] Example 7: Application of Nitrosomonas AOB SX-1 in the treatment of mixed industrial wastewater

[0074] Take 100 mL of influent from the oxidation ditch of the second phase industrial line of Shaoxing Water Treatment Wastewater Plant and add 1 mL of Nitromonas AOBSX-1 fermentation broth (OD). 600 =2.0), cultured at 30℃ and 150rpm with shaking, and samples were taken periodically to detect ammonia nitrogen content. The control group was set as the influent of the industrial oxidation ditch without bacteria.

[0075] The results are as follows Figure 6 As shown, after 4 hours of treatment with Nitrosomonas AOB SX-1, the ammonia nitrogen level in the mixed industrial wastewater decreased to 0.84 mg / L, achieving a removal rate of 96%. This result indicates that Nitrosomonas AOB SX-1 can effectively perform deep ammonia removal treatment on mixed industrial wastewater.

[0076] Example 8: Application of Nitrosomonas AOB SX-1 in Municipal Wastewater Treatment

[0077] Take the wastewater from the Shaoxing Water Treatment Plant's domestic sewage line A 2 100 mL of O process influent was supplemented with 1 mL of nitrosomonas AOB SX-1 fermentation broth (OD). 600 =2.0), cultured at 30℃ with shaking at 150 rpm and samples were taken periodically to detect ammonia nitrogen content. The control group was set as A without bacteria. 2 O process water inlet.

[0078] The results are as follows Figure 7 As shown, after 4 hours of treatment with Nitrosomonas AOB SX-1, the ammonia nitrogen level in municipal wastewater decreased to 0.11 mg / L, achieving a removal rate of 99%. This result indicates that Nitrosomonas AOB SX-1 can effectively perform deep ammonia removal treatment on municipal wastewater.

[0079] Example 9: Application of Nitrifying Monotropha AOB SX-1 in Aquaculture Wastewater Treatment

[0080] Take 1L of wastewater from a turtle farm in Huzhou and add 1mL of Nitromonas AOB SX-1 fermentation broth (OD). 600 =2.0), cultured at 30℃ and 150rpm with shaking, and samples were taken regularly to detect ammonia nitrogen content. The control group was set as wastewater from a turtle farm without bacteria.

[0081] The results are as follows Figure 8 As shown, after 12 hours of treatment with *Nitrosomonas* AOB SX-1, the ammonia nitrogen concentration in aquaculture wastewater decreased to 0.6 mg / L, and the ammonia nitrogen removal rate increased to 0.24 mg / L / h compared to the control group. This result indicates that *Nitrosomonas* AOB SX-1 can effectively reduce ammonia nitrogen concentrations in aquaculture wastewater.

[0082] Example 10: Application of Nitrifying Monotrophs AOB SX-1 in Eutrophic Water Bodies

[0083] Take 1L of eutrophic river water from a river in Shaoxing, Zhejiang Province, and add 1mL of Nitromonas AOBSX-1 fermentation broth (OD). 600 =2.0), cultured at 30℃ with shaking at 150rpm and samples were taken periodically to detect ammonia nitrogen content. The control group was set as eutrophic river water without bacteria.

[0084] The results are as follows Figure 9 As shown, after 12 hours of treatment with *Nitrosomonas* AOB SX-1, the ammonia nitrogen level in eutrophic river water decreased to 0.321 mg / L, achieving a removal rate of 94.1%. Compared to the control group, the ammonia nitrogen removal rate increased to 0.43 mg / L / h. This result indicates that *Nitrosomonas* AOB SX-1 can effectively reduce ammonia levels in eutrophic river water.

Claims

1. Nitrosomonas sp. AOB SX-1, deposited at the China Center for Type Culture Collection (CCTCC) on November 13, 2023, accession number: CCTCC NO: M 20232220, address: Wuhan University, Wuhan, China, postcode: 430072.

2. The application of Nitrifying Monotroph AOB SX-1 as described in claim 1 in ammonia removal.

3. The application as described in claim 2, characterized in that: The application is for removing ammonia from wastewater.

4. The application as described in claim 3, characterized in that: The wastewater refers to textile dyeing wastewater, mixed industrial wastewater, municipal domestic wastewater, aquaculture wastewater, or eutrophic river water.

5. The application as described in claim 3, characterized in that: The method of application is as follows: the fermentation liquid of the Nitrifying Monotropha AOBSX-1 is added to the wastewater.

6. The application as described in claim 5, characterized in that: The OD of the fermentation broth 600 =2.

0.

7. The application as described in claim 5, characterized in that: The volume ratio of the fermentation liquid to the wastewater is 1:100 to 1000.

8. The application as described in any one of claims 5-7, characterized in that: The method of application is as follows: (1) The Nitrifying Monotroph AOB SX-1 was inoculated into a fermentation medium and cultured for the first time at 30℃ and 150 rpm for 5 days to obtain a seed culture; during the first shaking culture, the pH of the system was controlled at 7.5-8.0 and the ammonia nitrogen concentration was controlled at 100 mg / L. (2) The seed culture obtained in step (1) is inoculated into fresh fermentation medium at a volume inoculation rate of 10%, and cultured for the second time at 30°C and 150 rpm to obtain fermentation culture; during the second culture, the pH of the system is controlled at 7.5-8.0 and the ammonia nitrogen concentration is controlled at 100 mg / L. (3) Add the fermentation liquid from step (2) into the wastewater to remove ammonia.

9. The application as described in claim 8, characterized in that: In step (1) or (2), the pH of the system is controlled by adding 5% Na2CO3 aqueous solution; the ammonia nitrogen concentration is controlled by adding 5% (NH4)2SO4 aqueous solution. The fermentation medium described in step (1) or (2) consists of the following components at the following final concentrations: 1.5 g / L (NH4)2SO4, 0.3 g / L NaCl, 0.24 g / L NaH2PO4, 2.64 g / L Na2HPO4, 0.03 g / L FeSO4·7H2O, and 0.03 g / L MgSO4·7H2O; the solvent is water, and the pH is 7.5-8.

0.

10. The application as described in claim 8, characterized in that: The conditions for ammonia removal in step (3) are: 30℃, 150rpm shaking for 1-48h.

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