Klebsiella oxytoca n5 strain and its application in ammonia nitrogen degradation

Abstract

The application belongs to the technical field of agricultural pollutant degradation treatment, and particularly relates to a Klebsiella oxytoca N5 strain and application thereof in ammonia nitrogen degradation. The Klebsiella oxytoca N5 strain has a preservation number of CCTCC NO: M 2024658. Under a low ammonia nitrogen concentration condition, the ammonia nitrogen degradation rate of the strain can reach 96.40% (under an ammonia nitrogen concentration of 106 mg / L), and the strain has a good application prospect, especially under a high concentration ammonia nitrogen condition, and also has a good ammonia nitrogen degradation effect (ammonia nitrogen degradation rates of 72% and 65.43% under ammonia nitrogen concentrations of 742 mg / L and 848 mg / L, respectively). The result shows that the strain has good tolerance and good utilization to ammonia nitrogen. This provides a possibility for effective treatment of high-concentration ammonia nitrogen manure in farms and the like, and has good practical value for related agricultural pollutant treatment.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of agricultural pollutant degradation treatment, and particularly relates to a Klebsiella N5 strain and application thereof in ammonia nitrogen degradation. BACKGROUND

[0002] In large-scale livestock and poultry breeding, the manure sewage at breeding sites is extremely easy to cause water pollution due to its high concentration of ammonia nitrogen, and therefore is considered as an agricultural pollutant. Therefore, the manure sewage must be properly treated before being discharged. In the treatment of such agricultural pollutants, fermentation treatment of the manure sewage by using related microbial strains through proper composting (mainly degrading ammonia nitrogen and the like) is a relatively common treatment method. However, in actual application, due to the diversity of microbial strains in the composting process and the low ammonia nitrogen degradation capacity, the composting treatment often needs a long time and the composting effect is not consistent.

[0003] In existing research, screening of ammonia nitrogen degradation strains with high ammonia nitrogen degradation capacity is a better treatment method for improving the ammonia nitrogen degradation effect in the composting process. For example, Liu Zhishui et al. screened an acid-producing Klebsiella M169-3, which has a good degradation effect on ammonia nitrogen in sewage (Research on Screening and Identification of Klebsiella with High Ammonia Nitrogen Degradation Capacity and Suitable Conditions, Heilongjiang Journal of Animal Husbandry and Veterinary Medicine, 2023); and for another example, a kind of high-efficiency denitrifying microorganism and its application (application number: 201710937789.7) provides an acid-producing Klebsiella (Klebsiella oxytoca) TN-10, which also has a good degradation rate on ammonia nitrogen in wastewater. However, it is found through analysis that the related Klebsiella screened in the prior art only shows a high degradation rate in low-concentration ammonia nitrogen wastewater (below 120 mg / L), and the ammonia nitrogen degradation rate is greatly reduced (the highest ammonia nitrogen degradation rate is only about 58.41%) under the condition of high-concentration ammonia nitrogen (above 250 mg / L). This shows that high-concentration ammonia nitrogen conditions have a great influence on the growth of the strain and the actual degradation effect.

[0004] Since agricultural waste (especially manure sewage at breeding sites) is a high-concentration ammonia nitrogen environment, it is of great technical significance to continue screening ammonia nitrogen degradation strains suitable for specific high-concentration ammonia nitrogen environments for effectively treating agricultural waste. SUMMARY

[0005] Based on the screening of related ammonia nitrogen degradation strains, the application aims to provide a Klebsiella (Klebsiella) N5 strain for ammonia nitrogen degradation, thereby laying a certain technical foundation for ammonia nitrogen degradation in agricultural waste. Klebsiella oxytoca

[0006] The technical solution adopted by the application is mainly as follows. ​

[0007] A Klebsiella N5 strain, the Klebsiella ( Klebsiella oxytoca The N5 strain, with accession number CCTCC NO: M 2024658, was submitted to the China Center for Type Culture Collection (CCTCC) (postcode 430072) at Wuhan University, Wuhan, China for preservation on April 10, 2024, and its viability was confirmed by testing on April 17, 2024.

[0008] The ammonia nitrogen-degrading bacterial agent prepared using the Klebsiella N5 strain.

[0009] The Klebsiella N5 strain is used in the degradation of ammonia nitrogen. When used, the strain is suitable for ammonia nitrogen-containing waste with a nitrogen content not exceeding 1000 mg / L (specifically, for example, not exceeding 850 mg / L).

[0010] In this application, the inventors screened and obtained a Klebsiella N5 strain. This strain achieved a maximum ammonia nitrogen degradation rate of 96.40% under low ammonia nitrogen concentration conditions (based on N, at an ammonia nitrogen concentration of 106 mg / L, the corresponding (NH4)2SO4 concentration is 0.5 g / L), demonstrating good application prospects. Especially under high ammonia nitrogen concentration conditions, it also showed good ammonia nitrogen degradation effects (72% and 65.43% degradation rates at 742 mg / L and 848 mg / L, respectively). This result indicates that the strain has good tolerance and utilization of ammonia nitrogen. This provides a possibility for the effective treatment of high-concentration ammonia nitrogen manure from livestock farms, and has good practical value and technical significance for the treatment of related agricultural waste. Attached Figure Description

[0011] Figure 1 This is a diagram showing the colony morphology of strain N5 on LB medium.

[0012] Figure 2 Agarose gel electrophoresis image of the PCR amplification product of strain N5's 16S rDNA;

[0013] Figure 3 The results are based on homology alignment of the 16S rDNA sequence.

[0014] Figure 4 This is a growth curve of the strain;

[0015] Figure 5 This is a standard curve for ammonia nitrogen.

[0016] Figure 6 Comparison of ammonia nitrogen degradation rate detection results for different strains;

[0017] Figure 7 Results of ammonia nitrogen degradation rate within 72 hours under different inoculum amounts;

[0018] Figure 8 The change of ammonia nitrogen degradation rate within 7 days for ammonia nitrogen concentration of 106 mg / L;

[0019] Figure 9 The ammonia nitrogen degradation rate results under different ammonia nitrogen concentrations;

[0020] Figure 10 The results of the influence of different culture temperatures on the ammonia nitrogen degradation rate of the strain under high ammonia nitrogen concentration;

[0021] Figure 11 The results of the optimal application pH range of the strain. DETAILED DESCRIPTION

[0022] The present application will be further explained and described below in conjunction with examples. Before introducing the specific examples, the following will be briefly introduced and described in relation to the experimental background of the following examples.

[0023] Main culture medium:

[0024] LB liquid medium: tryptone 10 g, yeast extract 5 g, NaCl 10 g, water 1000 mL, pH 7.0-7.2;

[0025] LB solid medium: tryptone 10 g, yeast extract 5 g, NaCl 10 g, agar powder 16 g, water 1000 mL, pH 7.0-7.2;

[0026] Screening medium (ammonification liquid medium): glucose 10.0 g, (NH4)2SO4 2.0 g, NaCl 1.0 g, MgSO4·7H2O 1.0 g, K2HPO4·2H2O 1.0 g, FeSO4•7H2O 0.4 g, trace element solution 1.0 ml, water 1000 mL, pH 7.2-7.4;

[0027] Ammonification solid medium: glucose 10.0 g, (NH4)2SO4 2.0 g, NaCl 1.0 g, MgSO4·7H2O 1.0 g, K2HPO4·2H2O 1.0 g, FeSO4•7H2O 0.4 g, trace element solution 1.0 ml, agar 20.0 g, water 1000 mL, pH 7.2-7.4;

[0028] The trace element solution, wherein: ZnSO4·7H2O 3.90 g, CaCl2·2H2O 7.00 g, MnSO4·4H2O 5.10 g, (NH4)2MoO4 1.10 g, CuSO4·5H2O 1.60 g, CoCl2·6H2O 1.60 g, after dissolution, constant volume to 1L;

[0029] Ashby culture medium: K2HPO4 0.2 g, MgSO4·7H2O 0.2 g, CaSO4·2H2O 0.1 g, NaCl 0.2 g, CaCO3 5.0 g, mannitol 10.0 g, agar 18.0 g, water 1000 mL, pH 7.2-7.4;

[0030] After the preparation of the above culture medium, sterilize at 121℃ for 20 min.

[0031] Example 1

[0032] This example briefly introduces the screening of Klebsiella oxytoca N5 strain which is claimed in the present application.

[0033] The source of the screening sample: fecal samples were collected from a dairy farm in Anyang, Henan in April 2023;

[0034] Screening process:

[0035] Inoculate 5 g of the sample to be screened into 50 mL of screening medium, and culture at 37℃, 180 r / min on a shaking table for 7 days. During the culture, add 1 ml of 5% ammonium sulfate solution to the culture medium every 24 hours to eliminate microorganisms that cannot utilize NH 4+ , and enrich microorganisms that can utilize NH 4+ (During the screening process, 3-6 groups were set each time to eliminate the influence of operation errors and other factors during the test, and to improve the efficiency of screening bacteria);

[0036] After dilution of the culture liquid after preliminary screening and enrichment, take 1.0×10 -6 , 1.0×10 -7 , and 1.0×10 -8 of the dilution liquid, respectively, and inoculate 100 µL of each into the ammoniation solid culture medium (used for ammonia-tolerant bacteria selection medium) and Ashby culture medium (used for nitrogen-fixing bacteria culture), and culture at 37℃ for 18-24 hours to observe the growth condition of the bacteria (in order to isolate the strain); select the strain with fast growth, large colony, and large number, and further purify it on the ammoniation solid culture medium, and obtain single colonies with uniform size and growth after purification for 6-9 generations for further identification;

[0037] After identification, the colonies were inoculated into LB liquid medium, cultured at 37 ℃ and 180 r / min for 10 h, 1 mL of bacterial liquid was taken and mixed with 1 mL of sterile glycerol, and then stored at -80 ℃ for long-term preservation.

[0038] Strain identification:

[0039] During strain identification, the preliminary identification results of the growth morphology and biochemical identification of the strain and the related gene sequencing comparison results (final identification) were combined for identification.

[0040] As for Klebsiella (Klebsiella) N5 claimed in the present application, the related physicochemical identification results are shown in Table 1, and the related colony growth morphology is shown in Klebsiella oxytoca As can be seen from the colony morphology, the colony morphology is round, white and opaque, the edge is neat, and the upper part is protruding. Combined with these physicochemical identification results and colony growth morphology results, it can be preliminarily determined that it belongs to Klebsiella (Klebsiella) bacteria. Figure 1 Klebsiella oxytoca

[0041] Table 1, partial physicochemical identification results of Klebsiella (Klebsiella) N5 claimed in the present application Klebsiella oxytoca

[0042] .

[0043] During further sequencing identification, the genomic DNA of the strain to be identified (reference to the conventional operation of the prior art) was first extracted, and then the species of the strain to be identified was identified based on the sequencing and comparison results of the PCR amplification product of the specific primer pair (27F, 1492R);

[0044] The primer pair (shown as SEQ ID No. 1, SEQ ID No. 2, respectively) is specifically as follows:

[0045] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3'

[0046] 1492R: 5'-CTACGGCTACCTTGTTACGA-3'.

[0047] As for Klebsiella (Klebsiella) N5 claimed in the present application, the electrophoresis result of the amplification product (16S rDNA) is shown in Klebsiella oxytoca As can be seen from the colony morphology, the colony morphology is round, white and opaque, the edge is neat, and the upper part is protruding. Combined with these physicochemical identification results and colony growth morphology results, it can be preliminarily determined that it belongs to Klebsiella (Klebsiella) bacteria. Figure 2

[0048] ​​​​The PCR products were then sequenced by Pasenuo Biotechnology Co., Ltd. The sequences were assembled based on the sequencing results and compared with data in the existing 16S database of related microorganisms. Finally, species identification was performed based on the comparison results.

[0049] The Klebsiella pneumoniae for which protection is sought in this application ( Klebsiella oxytoca For N5, the alignment results of its amplification products with existing nucleic acid sequences in the GenBank database are as follows: Figure 3 As shown, the 16S rDNA sequence of this strain shares 99% homology with that of *Klebsiella oxytoca*. Combined with the aforementioned preliminary morphological identification results, the isolated strain can be definitively identified as... Klebsiella oxytoca kind.

[0050] Example 2

[0051] Based on the strains obtained from screening in Example 1 (the inventors simultaneously screened Enterococcus faecalis N1, Enterococcus faecalis N2, Bacillus subtilis N3, Bacillus licheniformis N4, and Klebsiella pneumoniae as described in this application), Klebsiella oxytoca The inventors further measured and analyzed the growth characteristics and actual ammonia nitrogen degradation capacity of the ammonia nitrogen (N5). The relevant experimental results are briefly described below.

[0052] (a) Growth status

[0053] Single colonies purified from different strains were inoculated into LB liquid medium and cultured in a constant temperature shaker at 37°C and 150 rpm. Samples were taken every 2 hours, and their optical density (OD) values ​​were measured at 600 nm using a UV-Vis spectrophotometer. The relationship between culture time and OD was plotted. 600 The relationship curve (growth curve) is used to reflect the growth characteristics of the strain.

[0054] The Klebsiella pneumoniae for which protection is sought in this application ( Klebsiella oxytoca For strain N5 (other strains are not discussed further as they are not relevant to this application), its growth curve is as follows: Figure 4 As shown in the figure. Analysis reveals that:

[0055] This strain is in the logarithmic growth phase from 1 to 8 hours, with the fastest growth rate from 4 to 8 hours. After 10 hours, the bacteria almost stop growing, and their ability to metabolize and utilize nutrients decreases to a minimum, gradually entering the stationary phase. Based on these growth characteristics, activated bacterial culture after approximately 8-10 hours was used as seed culture for further expansion and application in subsequent experiments.

[0056] (II) Preliminary determination of ammonia nitrogen degradation rate of different strains

[0057] Using the screening medium (initial ammonia nitrogen concentration of 742 mg / L, ammonia nitrogen concentration refers to N, corresponding to the addition of (NH4)2SO4 in the medium of 3.5 g / L), inoculate different strains (Enterococcus faecalis N1, Enterococcus faecalis N2, Bacillus subtilis N3, Bacillus licheniformis N4, Klebsiella N5), 180r / min shaking bed 37℃ culture 24h, after the end of the culture, the strain OD600 is about 0.6~0.8, using the nessler reagent spectrophotometry, the absorbance value at 420nm is determined and the ammonia nitrogen concentration in the bacterial solution and the ammonia nitrogen degradation rate are calculated according to the standard curve;

[0058] Ammonia nitrogen degradation rate = [(ammonia nitrogen concentration in the medium without inoculating strains - ammonia nitrogen concentration in the medium inoculated with bacteria) / ammonia nitrogen concentration in the medium without inoculating strains] x 100%.

[0059] The standard curve is drawn by using the nessler reagent spectrophotometry (HJ 535-2009) and referring to the following operation:

[0060] First, 0.00 mL, 0.10 mL, 0.20 mL, 0.40 mL, 0.80 mL, 1.20 mL, 1.60 mL and 2.00 mL of ammonia nitrogen standard working solution (10 μg / mL) are added to 8 clean cuvettes respectively, and the corresponding ammonia nitrogen content is 0.0 μg, 1.0 μg, 2.0 μg, 4.0 μg, 8.0 μg, 12.0 μg, 16.0 μg and 20.0 μg respectively, and the volume is made up to 10 mL;

[0061] Then, 200 μL of potassium sodium tartrate solution (500 g / L) is added, shaken well, and then 200 μL of nessler reagent is added, shaken well; after 10 min, 200 μL of each is taken to a 96-well plate, and the wavelength is 420 nm;

[0062] The water is used as a blank control to measure the absorbance;

[0063] The ammonia nitrogen standard curve y=ax+b; wherein: x is the solubility of ammonia nitrogen, y is the absorbance value corresponding to each ammonia nitrogen concentration at a wavelength of 420, several points are linearly fitted to obtain the straight line equation a representing the slope of the straight line and b representing the intercept, and according to the absorbance y, a and b, the solubility x of ammonia nitrogen can be calculated;

[0064] Finally, the ammonia nitrogen standard curve is drawn with the ammonia nitrogen concentration in the solution as the horizontal coordinate and the solution absorbance as the vertical coordinate.

[0065] The standard curve is shown in Figure 5 It can be seen that the curve equation is: y=0.07911x+0.015, the correlation coefficient R2 =0.998; this result shows that the absorbance of the solution (OD420) has a good linear relationship with the corresponding solution ammonia nitrogen concentration, and the actual ammonia nitrogen concentration can be calculated based on the absorbance.

[0066] As shown in the following table, the ammonia nitrogen degradation of different strains under the condition of a specific high ammonia nitrogen concentration (742 mg / L) was determined. Figure 6 It can be seen that Klebsiella N5 has good ammonia nitrogen degradation ability at different times, especially at 36h, the ammonia nitrogen degradation rate can reach 72%, showing good application potential.

[0067] (Three) Ammonia nitrogen degradation rate within 72h under the condition of low ammonia nitrogen concentration and different inoculation amounts

[0068] Based on the above results, the inventors further determined the ammonia nitrogen degradation rate of Klebsiella N5 strain under the condition of low ammonia nitrogen concentration (106 mg / L) and different inoculation amounts, and the specific experimental conditions are briefly introduced as follows.

[0069] According to the growth curve described above, the bacterial solution after 8h of preliminary fermentation was taken as the seed solution, and then inoculated into the ammoniation liquid medium (the initial ammonia nitrogen concentration was 106 mg / L, and the (NH4)2SO4 0.5g / L was added to the medium) at inoculation amounts of 1%, 3% and 5% respectively, and cultured at 37°C and 180r / min on a shaking table for 72h. During the culture, samples were taken every 24h, and the ammonia nitrogen concentration of the bacterial solution was determined and the ammonia nitrogen degradation rate was calculated according to the previous description.

[0070] The determination results are shown in the following table. Figure 7 It can be seen that the ammonia nitrogen degradation rate of Klebsiella N5 at an inoculation amount of 1% increases with the increase of culture time, and the degradation rate is highest at 78.02% after 72h of culture. The ammonia nitrogen degradation rate at an inoculation amount of 3% is highest and changes little with the culture time, and the degradation rate is about 80.01%. The ammonia nitrogen degradation rate at an inoculation amount of 5% first increases and then decreases, and the highest degradation rate is 78.54%.

[0071] (Four) Ammonia nitrogen degradation rate change under the condition of low ammonia nitrogen concentration and optimal inoculation amount

[0072] According to the above "(Three) Ammonia nitrogen degradation rate within 72h under the condition of low ammonia nitrogen concentration and different inoculation amounts", Klebsiella N5 was inoculated into the ammoniation liquid medium (the initial ammonia nitrogen concentration was 106 mg / L) at an inoculation amount of 3%, and the culture was continued for 7d. During the culture, samples were taken every 24h for the first 3d and every 48h for the last 4d, and the ammonia nitrogen concentration of the bacterial solution was determined and the ammonia nitrogen degradation rate was calculated.

[0073] The results are shown in the following table. Figure 8As shown in Table 5, it can be seen that the ammonia nitrogen degradation rate of the strain N5 was stable at 80% in the first 3d, and after 7d of culture, the strain reached the highest ammonia nitrogen degradation efficiency of 96.40%. This result shows that the strain has a good and relatively stable ammonia nitrogen degradation effect in the early stage, and its ammonia nitrogen degradation effect can be further improved with the extension of fermentation time.

[0074] (V) Effect of different ammonia nitrogen concentrations on ammonia nitrogen degradation rate

[0075] Referring to the aforementioned "(III) Ammonia nitrogen degradation rate within 72h under different inoculation amounts at low ammonia nitrogen concentration", Klebsiella N5 was inoculated into different ammonification liquid media (initial ammonia nitrogen concentration was 106 mg / L, 212 mg / L and 848 mg / L, respectively) at an inoculation amount of 3%, and cultured for 72h. The ammonia nitrogen concentration of the bacterial solution was determined every 24h, and the ammonia nitrogen degradation rate was calculated.

[0076] The results are shown in Table 6. Figure 9 As can be seen from Table 6:

[0077] The strain N5 has the highest ammonia nitrogen degradation rate of 80.00% at an initial ammonia nitrogen concentration of 106 mg / L, but the ammonia nitrogen degradation rate does not increase significantly with time within 72h;

[0078] When the initial ammonia nitrogen concentration is 212 mg / L, the ammonia nitrogen degradation rate increases with the increase of culture time (the degradation rate is 74.44% at 24h of culture, and the degradation rate is 74.51% at 48h of culture), and the highest degradation rate is 76.23% at 72h of culture;

[0079] When the initial ammonia nitrogen concentration is 848 mg / L, the ammonia nitrogen degradation rate first increases and then decreases (the degradation rate is 61.55% at 24h of culture), and the highest degradation rate is 65.43% at 48h of culture (the degradation rate is 63.06% at 72h of culture).

[0080] The above results show that the ammonia nitrogen degradation rate of the strain N5 is negatively correlated with the initial ammonia nitrogen concentration, that is, as the initial ammonia nitrogen concentration increases, the ammonia nitrogen degradation rate also decreases to a certain extent, but overall, the strain N5 has a relatively stable degradation effect under different ammonia nitrogen concentrations.

[0081] (VI) Effect of different temperatures on ammonia nitrogen degradation rate under high ammonia nitrogen concentration (848 mg / L)

[0082] According to the foregoing "(three) ammonia nitrogen degradation rate within 72h under low ammonia nitrogen concentration and different inoculation amount", Klebsiella N5 was inoculated into the ammoniation liquid medium (initial ammonia nitrogen concentration was 848 mg / L) at 3% inoculation amount, and was cultured at 25°C, 35°C and 40°C under 180 r / min shaking bed for 72h. During the culture, the sample was taken every 24h, and the ammonia nitrogen concentration of the bacterial solution was determined and the ammonia nitrogen degradation rate was calculated.

[0083] The results are shown in Figure 10 It can be seen that under high ammonia nitrogen concentration (848 mg / L), temperature has a certain effect on ammonia nitrogen degradation efficiency;

[0084] Under 25°C, the ammonia nitrogen degradation rate increased with the increase of culture time, and the highest degradation rate was 55.31%;

[0085] Under 35°C, the ammonia nitrogen degradation rate did not increase significantly with the increase of culture time, and the highest degradation rate was 65.40%;

[0086] Under 40°C, the ammonia nitrogen degradation rate had a decreasing trend with the increase of culture time, and the highest degradation rate was 53.47%.

[0087] This result shows that the growth characteristics of strain N5 are obviously affected by temperature, resulting in a certain fluctuation of ammonia nitrogen degradation rate.

[0088] (7) Low ammonia nitrogen concentration, pH tolerance test

[0089] Considering the pH environment of ammonia nitrogen waste, according to the foregoing "(three) ammonia nitrogen degradation rate within 72h under low ammonia nitrogen concentration and different inoculation amount", Klebsiella N5 was inoculated into the ammoniation liquid medium (initial ammonia nitrogen concentration was 106 mg / L) at 3% inoculation amount, and the pH in the medium was adjusted to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12, respectively. The culture was carried out at 35°C under 180 r / min shaking bed for 24h, and the effect of different pH values on the ammonia nitrogen degradation ability of the strain was determined.

[0090] The results are shown in Figure 11 It can be seen that:

[0091] The ammonia nitrogen degradation rate was zero when the pH value was 1-2, indicating that the strain was not resistant to acidic environment; when the pH value was 3-8, the ammonia nitrogen degradation rate of Klebsiella N5 gradually increased, and when the pH value was 8, the ammonia nitrogen degradation rate reached the highest (about 80%); when the pH value was more than 8, the ammonia nitrogen degradation rate began to decrease; overall, when the pH value was 7-9, the ammonia nitrogen degradation rate maintained at about 65%-80%; this result showed that the ammonia nitrogen degradation rate of the strain was higher in the alkaline environment, and in the higher acidic environment and alkaline environment, the activity of Klebsiella may be affected, thereby affecting the ammonia nitrogen degradation rate. That is, in practical application, the suitable application pH value of the strain is between 7 and 9.

Claims

1. A Klebsiella strain N5 (ATCC PTA- 1 1 1 1 ) Klebsiella oxytoca ) strain, characterized in that, The Klebsiella N5 strain, with the preservation number CCTCC NO: M 2024658, has been preserved in the China Center for Type Culture Collection (CCTCC) located in Wuhan University, Wuhan, China on April 10, 2024, and the detection certificate proves that it is alive on April 17, 2024.

2. An ammonia nitrogen degrading microbial agent prepared by using the Klebsiella N5 strain of claim 1.

3. Application of the Klebsiella N5 strain of claim 1 in ammonia nitrogen degradation.

4. The use of Klebsiella N5 strain in the degradation of ammonia nitrogen according to claim 3, characterized in that, When applied, the strain is suitable for ammonia nitrogen-containing waste with N content not exceeding 1000 mg / L.

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