Stutzerimonas frequens IMPWW-003 strain and application thereof
By developing Stutzerimonas frequency IMPWW-003 strain with good salt tolerance, the problem of nitrogen treatment in high-salt-containing sewage was solved, and efficient sewage nitrogen removal effect was achieved, which was suitable for the treatment of a variety of sewage scenarios.
Patent Information
- Application Number
- CN202510404546.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-01
- Publication Date
- 2025-08-15
AI Technical Summary
The prior art is difficult to effectively treat ammonia, nitrate and nitrosity nitrogen in high-salt wastewater. Conventional biological treatment methods are inhibited by salts, and pretreatment has high cost and poor feasibility to reduce salt concentration.
A Stutzerimonas frequency IMPWW-003 strain was developed, which has good salt tolerance and can efficiently remove ammonia nitrogen, nitrate nitrogen and nitrosity nitrogen in salt-containing sewage. It was prepared by bacterial agents or bacterial balls to apply it to wastewater nitrogen removal treatment.
It has achieved efficient degradation of various inorganic nitrogen in water under high salt content. It is suitable for nitrogen removal treatment of domestic sewage, seawater aquaculture tail water and industrial wastewater. It has broad application prospects and is simple and convenient for industrialization.
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of microorganisms, in particular to a Stutzerimonas frequens IMPWW-003 strain and application thereof. Background Art
[0002] Due to the rapid development of industry, agriculture, and aquaculture, as well as the advent of urbanization, how to treat the generated sewage is an important issue that needs to be addressed urgently. In the process of sewage treatment, the treatment of nitrogen is the key and difficult point. Excessive nitrogen will cause eutrophication of water bodies, make water bodies black and smelly, have toxic effects on organisms, destroy the ecological balance of water bodies, and reduce the biodiversity of water bodies. At present, there are mainly physical, chemical and biological methods for the treatment of nitrogen-containing sewage. The biological method is widely used in sewage treatment in various scenarios because of its wide range of applications, good treatment effects and no secondary pollution. The core of biological denitrification is microorganisms. In the process of water pollution control, the growth of microorganisms can utilize the nitrogen element in sewage and reduce the nitrogen content in the water body, thereby achieving the effect of purifying the water body.
[0003] High-salinity wastewater has a significant inhibitory effect on conventional biological treatments. Industrial production, marine aquaculture, biomedicine, and other industries generate varying concentrations of saline wastewater. The presence of salt inhibits microbial growth and metabolism, ultimately affecting the treatment of pollutants in the water. However, pre-treating high-salinity wastewater to reduce its salt concentration is costly and unfeasible. Therefore, isolating salt-tolerant bacteria for biological denitrification of high-salinity wastewater is of great significance.
[0004] Previous studies have shown that *Stutzerimonas frequens* RL-XB02 exhibits good tolerance to temperature, pH, salinity, and organic solvents, and can efficiently degrade triphenyl phosphate. *Stutzerimonas frequens* strain XQ-5 demonstrates significant CO2 fixation capacity, but its denitrification capabilities in water bodies have not been mentioned. Therefore, it remains necessary to develop new strains with good salt tolerance and efficient removal of various nitrogen forms. Summary of the Invention
[0005] One of the objectives of the present invention is to provide a new Stutzerimonas frequens strain capable of efficiently removing ammonia nitrogen, nitrate nitrogen and nitrite nitrogen from saline wastewater.
[0006] The invention provides a Stutzerimonas frequens IMPWW-003 strain, whose preservation number is CCTCC NO: M 20242433.
[0007] The Stutzerimonas frequens IMPWW-003 of this invention was deposited on November 5, 2024, at the China Center for Type Culture Collection (CCTCC, address: Wuhan University, Wuhan, China, postcode: 430072), classified as Stutzerimonas frequens IMPWW-003, with accession number: CCTCC NO: M 20242433.
[0008] The novel strain Stutzerimonas frequens IMPWW-003 of this invention is an aerobic strain that can efficiently remove ammonia nitrogen, nitrate nitrogen, and nitrite nitrogen from saline wastewater. It can be used for wastewater denitrification treatment in various scenarios such as domestic sewage, marine aquaculture tailwater, and industrial wastewater, and has broad application prospects.
[0009] In the present invention, the salt in "salt content", "salt tolerance" and "salinity" refers to NaCl.
[0010] The present invention also provides a bacterial agent containing the above-mentioned Stutzerimonas frequens IMPWW-003 strain.
[0011] The bacterial agent of the present invention can be a liquid bacterial agent or a solid bacterial agent, and in addition to the Stutzerimonas frequens IMPWW-003 strain, it can also contain other strains with the same or different functions.
[0012] The present invention also provides a bacterial pellet, which is prepared by fermenting the above-mentioned Stutzerimonas frequens IMPWW-003 strain.
[0013] Specifically, it can be obtained by continuously circulating and culturing sewage / culture medium containing inorganic nitrogen.
[0014] The preparation method of the bacterial balls of the present invention includes: (1) Activate Stutzerimonas frequens IMPWW-003 strain; (2) Aerobic fermentation in DM-1 medium; every 24 hours, replace 1 / 3 of the original medium volume with medium containing inorganic nitrogen; The inorganic nitrogen-containing culture medium is preferably one or more of DM-1, AM or ADM; DM-1 medium includes: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.7218 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; AM medium included: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, (NH4)2SO4 0.4714 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; ADM medium included: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.3609 g / L, (NH4)2SO4 0.2357 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; Trace elements include: EDTA 50.0 g / L, ZnSO4 2.2 g / L, CaCl2 5.5 g / L, MnCl2·4H2O 5.06 g / L, FeSO4·7H2O 5.0 g / L, CuSO4·5H2O 1.57 g / L, CoCl2·6H2O 1.61 g / L; Further preferably, the culture medium used in step (1) is NB culture medium containing 3% NaCl; And / or, the aerobic fermentation temperature in step (2) is 30±1°C and the shaking flask speed is 160-220 rpm.
[0015] Those skilled in the art can select a specific culture time based on the biomass accumulation during pellet preparation.
[0016] The inorganic nitrogen-containing culture medium used to replace 1 / 3 of the original culture medium can be one or more of DM-1, AM or ADM. For example, one of DM-1, AM or ADM can be used in the early stage of culture, and in the middle stage of culture, an inorganic nitrogen-containing culture medium different from that in the early stage can be used instead, and in the late stage of culture, an inorganic nitrogen-containing culture medium different from both the early and middle stages can be used instead.
[0017] The present invention also provides a method for preparing the above-mentioned bacterial pellets, and the preparation method is as described above.
[0018] The present invention also provides use of the above-mentioned Stutzerimonas frequens IMPWW-003 strain, bacterial agent or bacterial pellet in degrading nitrate nitrogen, ammonia nitrogen, and / or nitrite nitrogen.
[0019] The present invention also provides the use of the above-mentioned Stutzerimonas frequens IMPWW-003 strain, bacterial agent or bacterial pellet in treating nitrogen-containing wastewater.
[0020] In the application of the present invention, the nitrogen-containing sewage is domestic sewage or seawater aquaculture tail water.
[0021] In this invention, the main indicators of domestic sewage include: total nitrogen 70.3-86.9 mg / L, ammonia nitrogen 64.3-74 mg / L, nitrate nitrogen 1.4-9.7 mg / L, nitrite nitrogen 0-9.5 mg / L, and COD. cr 110-147.75 mg / L.
[0022] The main indicators of marine aquaculture wastewater include: total nitrogen 22-32.9 mg / L, ammonia nitrogen 11-24.1 mg / L, nitrate nitrogen 0-2.61 mg / L, nitrite nitrogen 0-0.125 mg / L, and COD. cr 343.6-591.8 mg / L.
[0023] The present invention also provides a water treatment agent comprising the above-mentioned Stutzerimonas frequens IMPWW-003 strain, agent, or pellet.
[0024] The present invention also provides a method for degrading inorganic nitrogen in water, wherein the above-mentioned water treatment agent is inoculated into the water to be treated and cultured.
[0025] The preferred culture conditions are: 30°C, 200 rpm.
[0026] The beneficial effects of this invention are at least as follows: The present invention provides a new strain Stutzerimonas frequens IMPWW-003 with good salt tolerance and application in the degradation of various inorganic nitrogen species in water. The strain can efficiently degrade ammoniacal nitrogen, nitrate nitrogen, and nitrite nitrogen in water, has the potential for application in sewage treatment, and can provide a reference for the treatment of domestic sewage, industrial sewage, marine aquaculture tail water, etc. The culture medium required for the activation and expansion of the strain of the present invention is simple in components, and the preparation process is relatively easy, facilitating industrial production and subsequent applications. BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 The morphological characteristics of the strain Stutzerimonas frequens IMPWW-003 of the present invention; Figure 2 The phylogenetic tree of the strain Stutzerimonas frequens IMPWW-003 of this invention; Figure 3 The results of the treatment of the strain Stutzerimonas frequens IMPWW-003 of the present invention on a culture medium containing only nitrate nitrogen are shown; Figure 4The results of the treatment of the strain Stutzerimonas frequens IMPWW-003 of the present invention on a culture medium containing only ammonia nitrogen are shown; Figure 5 The results of the treatment of the strain Stutzerimonas frequens IMPWW-003 of the present invention on a culture medium containing only nitrite nitrogen are shown; Figure 6 The results of treating nitrification medium with the strain Stutzerimonas frequens IMPWW-003 of the present invention under different salinity conditions are shown; Figure 7 is a photograph of the sludge particles of the present invention; Figure 8 The monitoring data of the inorganic nitrogen content by the aerobic bacterial ball culture system of the present invention; Figure 9 This is the treatment result of domestic sewage by the aerobic bacteria balls of the present invention; Figure 10 This is the treatment result of the aerobic bacteria pellets of the present invention on marine aquaculture tail water. Detailed Implementation
[0028] The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.
[0029] Unless otherwise specified, the experimental methods used in the following examples are all conventional methods. Unless otherwise specified, the materials, reagents, etc. used in the following examples are all commercially available or prepared according to conventional methods in the art.
[0030] Materials required in the following embodiments: 1. Bacterial source and wastewater samples The experimental materials used for strain screening were sludge samples from a local marine aquaculture tailwater treatment plant. Domestic sewage samples were obtained from a local domestic sewage treatment plant, and marine aquaculture tailwater samples were obtained from a local seafood aquaculture factory.
[0031] 2. Culture medium Nutrient broth (NB): 3 g beef extract powder, 10 g peptone, a certain amount of NaCl, 1000 ml water, pH 7.2-7.4.
[0032] Nitrification medium (AM): KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, (NH4)2SO4 0.4714 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, a certain amount of NaCl, pH 7.0±0.05.
[0033] Denitrification medium 1 (DM-1): KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.7218 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, a certain amount of NaCl, pH 7.0±0.05.
[0034] Denitrification medium 2 (DM-2): KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, NaNO2 0.564 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, a certain amount of NaCl, pH 7.0 ± 0.05.
[0035] Simultaneous nitrification-denitrification medium (ADM): KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.3609 g / L, (NH4)2SO4 0.2357 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, a certain amount of NaCl, pH 7.0±0.05.
[0036] Domestic sewage culture medium (SW): Add 3.0 g / L of sodium acetate and prepare it using domestic sewage samples, pH 6.5±0.05. The main indicators of domestic sewage are: total nitrogen 84.77±1.46 mg / L, ammonia nitrogen 72.64±1.06 mg / L, nitrate nitrogen 4.39±2.75 mg / L, nitrite nitrogen 9.00±0.11 mg / L, COD cr 115.72±2.62 mg / L.
[0037] Mariculture tail water medium (OW): Add sodium acetate 1.025 g / L, prepare using mariculture tail water sample, pH 6.5±0.05, the main indicators of mariculture tail water: total nitrogen 23.17±1.82 mg / L, ammonia nitrogen 11.18±0.22 mg / L, nitrate nitrogen -0.32±0.23 mg / L, nitrite nitrogen 0.125±0 mg / L, COD cr 379.55±17.33 mg / L.
[0038] Trace elements include: EDTA 50.0 g / L, ZnSO4 2.2 g / L, CaCl2 5.5 g / L, MnCl2·4H2O 5.06 g / L, FeSO4·7H2O 5.0 g / L, CuSO4·5H2O 1.57 g / L, and CoCl2·6H2O 1.61 g / L.
[0039] After the above culture medium is prepared, it needs to be sterilized at 121°C for 30 minutes and then used after cooling. For solid culture medium, 2 g of agar should be added per 100 mL.
[0040] 3. Analytical methods Nitrate nitrogen: Nitrate nitrogen content was determined by ultraviolet spectrophotometry according to HJ / T 346-2007; Ammoniacal nitrogen: Determine the ammoniacal nitrogen content using Nessler's reagent method according to HJ 535-2009; Nitrite nitrogen: Nitrite nitrogen content was determined by the naphthylethylenediamine spectrophotometric method according to GB 17378-2007; 30-minute sludge settling ratio (SV30): measured by volume method according to CJ / T221-2023; Mixed liquor sludge concentration (MLSS): determined by weight method according to CJ / T221-2023.
[0041] Total inorganic nitrogen is the sum of nitrate nitrogen, ammonia nitrogen and nitrite nitrogen content. Inorganic nitrogen determination results less than 0 mg / L are considered as 0 mg / L in the calculation of total inorganic nitrogen. The formula for calculating the total inorganic nitrogen removal rate is as follows: Total inorganic nitrogen removal rate = Total inorganic nitrogen residue / Initial total inorganic nitrogen content × 100%.
[0042] The determination process of the relevant indicators in the present invention is completed in a similar time using the same batch of reagents.
[0043] Example 1 Screening, identification and preservation of bacteria 1. Strain enrichment, domestication, isolation and purification After shaking the sludge sample well, take 2 mL and inoculate it into DM-1 and AM without NaCl for enrichment culture. After a period of culture, inoculate it at 2% into DM-1 and AM with higher salt content (calculated as NaCl, the same below) to gradually increase the salinity of the culture medium in order to enrich strains that have both salt tolerance and heterotrophic nitrification-aerobic denitrification (HN-AD) functions.
[0044] The enriched bacterial sample was isolated and purified using 3% salinity (3% NaCl) NB to obtain no less than 50 strains that can grow well at 3% salinity. After the strains were activated using 3% salinity NB, they were inoculated into 3% salinity DM-1 and cultured for 24 hours. The OD was measured. 600 By evaluating the growth of the strain and the nitrate nitrogen removal rate, a strain with salt tolerance and HN-AD function was screened and named IMPWW-003.
[0045] 2. Strain Identification and Preservation (1) Morphological characteristics The strain IMPWW-003 was streaked on 3% salinity (3% NaCl) solid NB, as shown in FIG. Figure 1 As shown, the colonies are irregular, yellow, with wrinkled and raised surfaces, dry, opaque, and serrated edges.
[0046] (2) 16S rDNA sequence analysis The 16S rDNA sequence of the above-mentioned strain IMPWW-003 was analyzed. The 16S rDNA sequence of bacteria is highly conserved among different species, and the sequence is different among different species. Therefore, after amplifying and sequencing this sequence through PCR and comparing it with known sequences in GenBank, the bacterial species can be determined and classified into genus or species. After preparing strain IMPWW-003 according to requirements, it was sent to a sequencing company for 16S rDNA sequencing (universal primers 27F: 5'-AGAGTTTGATCCTGGCTCAG-3', SEQ ID No. 1 and 1492R: 5'-GGTTACCTTGTTACGACTT-3', SEQ ID No. 2). The sequencing results were compared with the NCBI database, and the results showed that IMPWW-003 had 100% sequence identity with Stutzerimonas frequens (CP045416.1), indicating that the strain is most closely related to Stutzerimonas frequens. The sequence was submitted to the GenBank database, and a phylogenetic tree was constructed using the Neighbor-Joining method with MEGA 11 software as shown below. Figure 2 shown.
[0047] Based on morphological characteristics and 16S rDNA sequence analysis, the strain IMPWW-003 was identified as Stutzerimonas frequens, the preservation number of the strain is: CCTCC NO: M 20242433, and the preservation method is -70°C, low-temperature freezing; its 16S rDNA gene sequence is shown in SEQ ID NO.3.
[0048] Example 2 Evaluation of the denitrification effect on inorganic nitrogen The above-mentioned strain Stutzerimonas frequens IMPWW-003 was inoculated into 3% salinity NB and activated in a constant temperature shaker at 30°C and 200 rpm. 2% of the culture medium was inoculated into DM-1, DM-2, and AM for cultivation. 100 mL of the culture medium was placed in a 250 mL shake flask and cultured at 30°C and 200 rpm. Samples were taken at regular intervals and filtered through a 0.22 μm filter membrane. The nitrate nitrogen, ammonia nitrogen, and nitrite nitrogen in the samples were determined.
[0049] The results of the treatment with the only nitrate nitrogen medium (DM-1) are as follows: Figure 3 As shown in the figure, it can be seen that within 12 h, nitrate nitrogen can be basically removed, and there is almost no accumulation of inorganic nitrogen, and the removal rate of total inorganic nitrogen reaches 99.54%.
[0050] The results of the treatment with the only ammonia nitrogen medium (AM) are as follows Figure 4 As shown in the figure, it can be seen that within 16 h, ammonia nitrogen can be basically removed, and there is almost no accumulation of inorganic nitrogen, and the removal rate of total inorganic nitrogen reaches 97.47%.
[0051] The results of the treatment with the only nitrite-nitrogen medium (DM-2) are as follows: Figure 5 As shown in the figure, it can be seen that within 12 h, nitrite nitrogen can be basically removed, and there is almost no accumulation of inorganic nitrogen, and the removal rate of total inorganic nitrogen reaches 98.79%.
[0052] Example 3 Ammonia nitrogen removal at different salinities The strain Stutzerimonas frequens IMPWW-003 was inoculated into NB with salinity ranging from 0% to 7.5% and activated in a constant temperature shaker at 30°C and 200 rpm. A 2% inoculation was then performed into AM with corresponding salinity levels (0% to 7.5%). AM with different salinities was prepared using the same bottle of salt-free AM. 100 mL of the medium was placed in a 250 mL shake flask and incubated at 30°C and 200 rpm. After 24 hours of incubation, samples were collected and filtered through a 0.22 μm filter. Nitrate, ammonia, and nitrite nitrogen content was determined. An initial salt-free AM without the strain was used as a control.
[0053] The results of the test are as follows Figure 6When the salinity is less than 6%, ammonia nitrogen can be basically removed after 24 h of treatment; when the salinity is less than 3%, the removal rate of total inorganic nitrogen exceeds 90%; after 24 h of treatment with strain IMPWW-003, only the treatment group with a salinity of 7.5% has a total inorganic nitrogen removal rate of less than 80%, and after culturing it for another 36 h, its total inorganic nitrogen removal rate increases to 92.84%.
[0054] Experiment Example 4: Cultivation of Aerobic Bacterial Balls After activating the above-mentioned strain *Stutzerimonas frequens* IMPWW-003 in NB medium with a salinity of 3%, it was inoculated at 2% in sterilized DM-1 medium with a salinity of 3%. 200 mL of this medium was placed in a 500 mL shake flask and incubated at 30°C and 200 rpm for 24 h. After allowing the cells to settle completely, one-third of the original culture medium volume was replaced with DM-1 medium with a salinity of 3%. This process was repeated for 7 days. The culture was then continued in the same manner, but this time one-third of the original culture medium volume was replaced with AM medium with a salinity of 3%. This process was repeated for 11 days. After shaking and sampling, the cells were allowed to settle completely, and a portion of the supernatant was decanted to obtain the desired result. Figure 7 The sludge particles shown had a 30-minute settling ratio (SV30) of (25.78±0.24)% and a suspended solids concentration (MLSS) of (4147.2±112.61) mg / L. After shaking and sampling, the bacterial cells were cultured in the same manner, except that 1 / 3 of the original culture medium volume was replaced with 3% salinity ADM. This process was repeated for 13 days.
[0055] Before each replacement of the culture medium, the supernatant was sampled and the inorganic nitrogen content was measured to monitor the operation of the system. The indicator measurement results were as follows: Figure 8 As shown, during system operation, regardless of whether the nitrogen source added to the simulated wastewater culture medium is nitrate nitrogen, ammonia nitrogen, or a mixture of both, as biomass accumulates significantly over time, inorganic nitrogen removal remains good, consistently at a low level. Starting from day 2, the total inorganic nitrogen residue is highest on day 13, at only 4.62 mg / L. After repeating the process for more than 7 days, the biomass in the culture medium significantly increases compared to the initial level, and the mutually adhering bacterial cells form aerobic bacterial spheres with good settling properties.
[0056] Example 5 Application of aerobic bacteria pellets in the removal of inorganic nitrogen from domestic sewage Using the aerobic bacterial balls obtained in Example 3, 1% was inoculated into SW. 100 mL of the solution was placed in a 250 mL shake flask and incubated at 30°C and 200 rpm. Samples were taken at regular intervals for analysis. After filtration through a 0.22 μm filter membrane, the nitrate nitrogen, ammonia nitrogen, and nitrite nitrogen content of the samples were measured. The treatment of SW using aerobic bacterial balls is as follows... Figure 9As shown in the figure, the removal rate of total inorganic nitrogen reached 96.32% in 12 h and 97.49% in 24 h.
[0057] Example 6 Application of aerobic bacteria pellets in the removal of inorganic nitrogen from marine aquaculture tail water The aerobic pellets obtained in Example 3 were inoculated into OW at 1%, and 100 mL was placed in a 250 mL shake flask and cultured at 30°C and 200 rpm. Samples were taken at regular intervals for determination. After filtering with a 0.22 μm filter membrane, the nitrate nitrogen, ammonia nitrogen, and nitrite nitrogen in the samples were determined. Figure 10 As shown in the figure, the removal rate of total inorganic nitrogen reached 85.86% in 12 h and 95.13% in 24 h.
[0058] Although the present invention has been described in detail above using general descriptions and specific embodiments, it will be apparent to those skilled in the art that modifications and improvements may be made based on the present invention. Therefore, such modifications and improvements, which do not depart from the spirit of the present invention, are intended to be within the scope of protection claimed herein.
Claims
1. A strain of Stutzerimonas frequens IMPWW-003, characterized in that The deposit number is CCTCC NO: M 20242433.
2. A bacterial agent, characterized in that Contains the Stutzerimonas frequens IMPWW-003 strain according to claim 1.
3. A bacterial ball, characterized in that The method is prepared by fermenting the Stutzerimonas frequens IMPWW-003 strain according to claim 1.
4. The bacterial ball according to claim 3, characterized in that The preparation method comprises: (1) Activate Stutzerimonas frequens IMPWW-003 strain; (2) Aerobic fermentation in DM-1 medium; every 24 hours, replace 1 / 3 of the original medium volume with medium containing inorganic nitrogen; The inorganic nitrogen-containing culture medium is preferably one or more of DM-1, AM or ADM; DM-1 medium includes: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.7218 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; AM medium included: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, (NH4)2SO4 0.4714 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; ADM medium included: KH2PO4 1.0 g / L, MgSO4·7H2O 0.2 g / L, KNO3 0.3609 g / L, (NH4)2SO4 0.2357 g / L, sodium acetate 3.418 g / L, trace elements 10 mL, NaCl 30 g / L, pH 7.0 ± 0.05; Trace elements include: EDTA 50.0 g / L, ZnSO4 2.2 g / L, CaCl2 5.5 g / L, MnCl2·4H2O 5.06 g / L, FeSO4·7H2O 5.0 g / L, CuSO4·5H2O 1.57 g / L, CoCl2·6H2O 1.61 g / L; Further preferably, the culture medium used in step (1) is NB culture medium containing 3% NaCl; And / or, the aerobic fermentation temperature in step (2) is 30±1°C and the shaking flask speed is 160-220 rpm.
5. A method for preparing a bacterial ball according to claim 3 or 4, characterized in that: The preparation method is as described in claim 4.
6. Use of the Stutzerimonas frequens IMPWW-003 strain according to claim 1, the bacterial agent according to claim 2, or the bacterial pellet according to claim 3 or 4 in degrading nitrate nitrogen, ammonia nitrogen, and / or nitrite nitrogen.
7. Use of the Stutzerimonas frequens IMPWW-003 strain according to claim 1, the bacterial agent according to claim 2, or the bacterial pellets according to claim 3 or 4 in treating nitrogen-containing wastewater.
8. The use according to claim 7, characterized in that The nitrogen-containing sewage is domestic sewage or seawater aquaculture tail water.
9. A water treatment agent, characterized in that It includes the Stutzerimonas frequens IMPWW-003 strain according to claim 1, the bacterial agent according to claim 2, or the bacterial pellet according to claim 3 or 4.
10. A method for degrading inorganic nitrogen in water, characterized in that: The water treatment agent according to claim 9 is inoculated into the water body to be treated for cultivation.