CN114703095B

CN114703095B - Pseudomonas adulthood and application thereof in field of sewage and wastewater purification

Info

Publication number: CN114703095B
Application number: CN202210323795.4A
Authority: CN
Inventors: 刘佳琪; 刘圣鹏
Original assignee: Qingdao Weilan Saide Biotechnology Co ltd
Current assignee: Qingdao Weilan Saide Biotechnology Co ltd
Priority date: 2022-03-29
Filing date: 2022-03-29
Publication date: 2023-09-19
Anticipated expiration: 2042-03-29
Also published as: CN114703095A

Abstract

The invention relates to a strain of adzuki pseudomonas and application thereof in the field of sewage and wastewater purification, wherein the strain is preserved in the China general microbiological culture Collection center (China Committee) for culture Collection of microorganisms, and the addresses are: the collection number of the national institute of microbiology, national academy of sciences, no. 3, north Chen West Lu 1, chao yang, beijing, is: CGMCC No.24300, the preservation date is: the invention provides the adzuki pseudomonas for 2022, 1 month and 14 days, which has the following beneficial effects: the strain can normally reproduce and grow when the salinity in the water body is not more than 8%, and the growth condition is optimal under the salinity condition of below 5%.

Description

Pseudomonas adulthood and application thereof in field of sewage and wastewater purification

Technical Field

The invention relates to a strain of adzuki pseudomonas, a microbial agent containing the adzuki pseudomonas and application of the strain in the field of sewage purification, and belongs to the technical field of environmental microorganisms.

Background

In recent years, with the rapid development of industry and the acceleration of life rhythm in China, urban domestic sewage, wastewater from agriculture and industry and the like cause the water environment pollution problem to become serious. The sewage has the characteristics of high COD value, high ammonia nitrogen content, large odor and the like, and causes serious pollution to surface water, underground water and the like. Ammonia nitrogen in a water body is one of main pollution factors, and a great amount of accumulated ammonia nitrogen can cause aggravation of slow flow closed or semi-closed water body eutrophication, cause environmental deterioration, cause aquatic organism poisoning and even death, and seriously affect ecological balance.

In the current sewage treatment method, the ammonia nitrogen removal method mainly comprises a physical method, a chemical method and a biological method. Compared with the physical method and the chemical method, the biological method has the advantages of safety, economy, no secondary pollution and the like, and has become the most widely applied treatment method.

In the biological treatment method of ammonia nitrogen, the adoption of nitrifying bacteria to remove ammonia nitrogen pollution is the most convenient and efficient denitrification way. Nitrifying bacteria are classified into autotrophic nitrifying bacteria and heterotrophic nitrifying bacteria, wherein the autotrophic nitrifying bacteria have low self-propagation speed, low biological density and severe growth conditions, are difficult to form dominant strains, and are difficult to expand production and popularization and use. Since the isolation of heterotrophic nitrifying strains, more and more heterotrophic nitrifying bacteria have been of great interest.

The research results of the prior art are integrated, and most heterotrophic nitrifying bacteria have the problems of overlong time, non-ideal pilot scale amplification experimental effect and the like in the process of treating high ammonia nitrogen wastewater. Therefore, the method for screening the microbial strain capable of efficiently reducing ammonia nitrogen in the water body has great significance for environmental protection.

Disclosure of Invention

Aiming at the defects of the existing heterotrophic nitrifying bacteria in the process of treating high ammonia nitrogen wastewater, the invention provides a strain of Acidovorax faciens, a microbial agent containing the same and application of the strain in the field of sewage and wastewater purification.

Pseudomonas adulthood (Pseudomonas chengduensis) Y25 deposited in China general microbiological culture Collection center with the following addresses: the collection number of the national institute of microbiology, national academy of sciences, no. 3, north Chen West Lu 1, chao yang, beijing, is: CGMCC No.24300, the preservation date is: in 2022, on month 01 and 14, the 16S rDNA sequence is shown as SEQ ID No. 1, and the P.adulthood is referred to as P.adulthood Y25 strain in the invention without special description.

The invention provides the beneficial effects of the adzuki pseudomonas:

1) The ammonia nitrogen degradation capability is strong, the effect is quick, and the ammonia nitrogen degradation rate of 72 hours reaches 94-95% for the initial ammonia nitrogen concentration of 100mg/L at 30 ℃;

2) Can tolerate high-concentration ammonia nitrogen, and the ammonia nitrogen degradation rate of 72 hours reaches more than 92 percent aiming at the initial ammonia nitrogen concentration of 100-400mg/L at the temperature of 30 ℃; when the ammonia nitrogen concentration reaches 600mg/L, the ammonia nitrogen degradation rate in 72 hours is only about 20%, so that the highest ammonia nitrogen tolerance concentration of the strain is 600mg/L;

3) Salt tolerance, the strain can normally reproduce and grow when the salinity in the water body is not more than 8%, and the growth condition is optimal under the salinity condition below 5%;

4) The strain has the advantages of simple culture method, strong environmental adaptability, rapid propagation and high safety.

Microbial agents comprising Pseudomonas adzuki are also claimed.

The invention also claims a fermentation process of pseudomonas ready for use, comprising the steps of:

(1) Primary seed culture: inoculating Pseudomonas into liquid LB culture medium under aseptic condition, culturing at 25-35deg.C and 150-300rpm for 12-24 hr to obtain first seed culture solution;

(2) Secondary seed culture: inoculating the primary seed culture solution into a liquid LB culture medium according to an inoculum size of 1-10vol% under a sterile condition, and culturing for 12-24h at 25-35 ℃ and 150-300rpm to obtain a secondary seed culture solution;

(3) Fermentation: inoculating the secondary seed culture solution into the fermentation culture medium according to the inoculation amount of 1-10vol% after the fermentation culture medium in the fermentation tank is disinfected, controlling the temperature to be 25-35 ℃ and the rotating speed to be 150-300rpm, fermenting under the condition that the ventilation ratio is 1 (1-2), and stopping fermentation when dissolved oxygen starts to rise to obtain fermentation liquor; the fermentation medium comprises the following components: 40-60g/L carbon source, 15-30g/L, PO nitrogen source ₄ ^3- 0.8-1.5g/L、K ⁺ 0.5-1.0g/L、Mg ²⁺ 0.05-0.2g/L、Na ⁺ 0.1-0.3g/L、Mn ²⁺ 0.03-0.1g/L of solventWater, ph=6 to 8.

The aeration ratio in the present invention means the ratio of the volume of air introduced into the fermenter per minute to the total volume of the fermentation liquid.

Wherein the composition of the liquid LB culture medium is as follows: 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride and water as a solvent, wherein the pH=7-7.5.

Preferably, the PO ₄ ^3- And K ⁺ The source of (a) is dipotassium hydrogen phosphate or monopotassium phosphate, and the Mg ²⁺ The source of the Na is one or more of magnesium sulfate, magnesium nitrate or magnesium chloride ⁺ The source of the Mn is one or more of sodium chloride, sodium nitrate or sodium sulfate ²⁺ The source of the catalyst is one or more of manganese sulfate monohydrate, manganese sulfate tetrahydrate, manganese nitrate or manganese chloride.

Further, the carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium succinate.

Further, the nitrogen source is selected from one or more of yeast extract, peptone, urea, ammonium sulfate or potassium nitrate.

In the practical application process, the form of the final product of the adzuki pseudomonas can be determined according to the practical use and storage requirements, when the liquid product is required to be used, the fermentation liquor is diluted to the required concentration to be directly used, when the solid product is required to be used, the fermentation liquor can be centrifuged to obtain bacterial mud, and then the solid bacterial powder is prepared by adopting a spray drying or freeze drying process.

The invention also claims a method for purifying sewage and wastewater by using the activated liquid of the pseudomonas adulthood or the microbial agent containing the pseudomonas adulthood, which comprises the step of inoculating the activated liquid of the pseudomonas adulthood or the microbial agent containing the pseudomonas adulthood into the sewage and wastewater.

Further, the ammonia nitrogen concentration in the sewage and wastewater is 600mg/L or less, more preferably 400mg/L or less, and most preferably 100-200mg/L.

Further, the salinity of the sewage and wastewater is 8% or less, more preferably 6% or less, still more preferably 5% or less, and most preferably 3% or less.

Further, the inoculum size of the activated liquid of P.adulthood or the microbial agent containing P.adulthood is 100ppm or more, preferably 100-2000ppm, most preferably 100-1000ppm.

The invention also claims the application of the pseudomonas and the microbial agent containing the pseudomonas in the field of sewage and wastewater purification.

Preferably, the Pseudomonas adzuki and the microbial inoculant comprising the Pseudomonas adzuki are used for degrading nitrogen-containing substances in the wastewater, more preferably, substances containing ammoniacal nitrogen in the wastewater.

Drawings

FIG. 1 is a photograph of a colony of Pseudomonas adulthood;

FIG. 2 is a photograph of a colony of P.adulthood magnified by a 1000-fold microscope.

Detailed Description

The principles and features of the present invention are described below in connection with examples, which are set forth only to illustrate the present invention and not to limit the scope of the invention.

Example 1 isolation and purification and identification of Pseudomonas Chenopodii

1. Separation and purification of strains

1. Enrichment of strains

Taking 100ml of water samples from shrimp ponds and fish ponds of a certain aquatic farm in Jiangsu, mixing the water samples of the culture ponds, inoculating 10ml of water samples into a triangular flask filled with 100ml of enrichment medium (ammonium sulfate 0.5g, sodium succinate 5.62g, vickers salt solution 50ml, using sterile water to fix the volume to 1L and pH=7), shaking and culturing for 5 days at 30 ℃ under 200r/min, measuring ammonia nitrogen value every other day, and regulating pH; if the ammonia nitrogen value is reduced, transferring to the next stage of enrichment, namely taking 10ml of culture solution after 5 days of culture, inoculating into 100ml of enrichment medium again, and repeating the culture for 5 times.

2. Screening and isolation of strains

Taking 0.5ml of the culture solution of the last stage, and carrying out gradient dilution to 10 by using sterile water ^-4 ，10 ^-5 ，10 ^-6 ，10 ^-7 And 10 ^-8 And (3) respectively taking diluted culture solutions, coating the diluted culture solutions on LB solid culture medium plates, placing the culture solutions in a 30 ℃ incubator, observing morphological characteristics of the bacterial colonies after the bacterial colonies grow to a proper size, picking single bacterial colonies, carrying out streak purification, carrying out three generations of purification, and then carrying out inclined plane preservation at 4 ℃ to totally separate 8 bacterial strains which are named as Y21-Y28 respectively.

3. Double screen

The 8 strains obtained by the preliminary screening were inoculated into basic media (peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L, water as solvent, pH=7), shake-cultured at 30℃for 24 hours, 10ml of the culture broth was centrifuged at 4000rpm for 10 minutes, the supernatant was discarded, and each of the culture broth was inoculated into 100ml of evaluation media (ammonium sulfate 0.5g, sodium succinate 5.62g, vickers salt solution 50ml, constant volume with sterile water to 1L, pH=7, ammonia nitrogen content of 100 mg/L), shake-cultured at 30℃with three groups of parallel groups of test media were set, and the change in ammonia nitrogen concentration in the evaluation media of each group of test media was periodically detected using sterile water as a blank control, and the results are shown in Table 1.

Table 1 degradation effects of Ammonia on Each strain obtained by preliminary screening

The results in Table 1 show that Y25 strain has a fast effect on ammonia nitrogen degradation compared to other strains, while the removal effect is significantly better than other strains, so that further identification is performed as a preferred strain.

2. Identification of species

1. Morphological observation

Strain Y25 was inoculated on LB solid medium plates and the morphological characteristics of colonies were observed. The colony photograph of the Y25 strain is shown in FIG. 1, the microscopic photograph at 1000 times is shown in FIG. 2, and the colony is punctiform, semitransparent, milky white, smooth in edge and convex.

2. Molecular biological identification

Extracting genome DNA of strain Y25, amplifying 16S rDNA with the extracted genome DNA as template,

primer: 27F 5'-AGAGTTTGATCCTGGCTCAG-3',

1492r:5’-CTACGGCTACCTTGTTACGA-3’；

the PCR products were then extracted and DNA sequenced using a sequencer ABI 3730-XL. And (3) comparing the spliced sequence file with data in an NCBI 16S database by using an NCBI Blast program to obtain species information with the maximum sequence similarity with a species to be detected, and identifying that the Y25 strain belongs to Pseudomonas and is Pseudomonas chenopodii (Pseudomonas chengduensis).

EXAMPLE 2 salinity tolerance test of Pseudomonas Chengonensis

Under aseptic condition, inoculating the Pseudomonas chenopodii into a 250ml conical flask containing 100ml of liquid LB culture medium, and performing shake culture in a shaker at 30 ℃ and 200rpm for 24 hours to perform strain activation to obtain an activated liquid of the Pseudomonas chenopodii.

Taking liquid LB culture medium as basic culture medium (salinity is 1%), adding NaCl 1g,2g,3g,4g,5g,6g,7g,8g and 9g into triangular flask containing 100ml liquid LB culture medium, respectively, further providing culture medium with salinity gradient of 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% and 10%, respectively taking 5ml of each activating solution of Pseudomonas, respectively, inoculating into each culture medium, shake culturing at 30deg.C, and periodically measuring OD ₆₀₀ The values and the number of viable bacteria are shown in Table 2.

OD ₆₀₀ The value represents the absorbance of the solution at 600nm, and the concentration of the bacterial culture is measured by the value, which is commonly used to refer to the cell density of the cells, and the growth of the bacteria can be measured by OD ₆₀₀ Is monitored to determine the extent of growth of the strain.

TABLE 2 OD of Pseudomonas adzuki culture in different salinity media ₆₀₀ Value and viable count

According to the detection results in Table 2, the adult Pseudomonas can grow at a salinity of 1-8%, and the strain growth condition is significantly better than that at a salinity of less than 5% than at a salinity of more than 5%, so that a salinity of less than 5% is suitable for growth, and an optimal growth salinity is less than 3%.

Example 3 determination of Ammonia-nitrogen degradation efficiency of Pseudomonas Chengdu

Under aseptic condition, inoculating the adzuki pseudomonas into a 250ml conical flask containing 100ml of liquid LB culture medium, and carrying out shaking culture for 24 hours at 30 ℃ and 200rpm in a shaking table to obtain an active liquid of the adzuki pseudomonas, wherein the number of active bacteria of the active liquid is 65 hundred million cfu/ml.

10. Mu.l, 100. Mu.l and 1000. Mu.l of each of the activated liquid of Pseudomonas dumi were inoculated into 100ml of sterilized evaluation medium (ammonium sulfate 0.5g, sodium succinate 5.62g, vickers salt solution 50ml, volume-fixed to 1L with sterile water, pH=7, ammonia nitrogen content 100 mg/L) and the respective inoculum amounts of the activated liquid of Pseudomonas dumi were 100ppm, 1000ppm and 10000ppm, respectively, and then each of the evaluation media was subjected to shaking culture in a shaker at 30℃and 200rpm, 3 replicates each were set, and the ammonia nitrogen content in the evaluation medium was measured every 24 hours using sterile water as a blank control, and the results are shown in Table 3.

The ammonia nitrogen detection method is implemented according to HJ535-2009 ammonia nitrogen determination-Nashi reagent spectrophotometry.

TABLE 3 Ammonia nitrogen degradation Effect of Pseudomonas adzuki

As shown in Table 3, the results show that the degradation rate and the degradation capacity of the ammonia nitrogen of the P.adulthood are high, the ammonia nitrogen degradation efficiency of 72 hours can reach more than 94% when the inoculation amount of the P.adulthood activated liquid is more than 100ppm, and the effect of the continuous increase of the inoculation amount on the ammonia nitrogen degradation effect is weak after the inoculation amount reaches 100ppm, so that the preferred inoculation amount of the P.adulthood activated liquid is 100-1000ppm based on the economic consideration.

Example 4 test of resistance of P.adulthood to various ammonia nitrogen concentrations

Under aseptic condition, the adzuki pseudomonas is inoculated into a 250ml conical flask containing 100ml of liquid LB culture medium, and the strain is activated by shaking culture for 24 hours in a shaking table at 30 ℃ and 200rpm, so that an activation solution of the adzuki pseudomonas is obtained, and the number of viable bacteria of the activation solution is 65 hundred million cfu/ml.

Evaluation media (ammonium sulfate content 0.5g, 1g,2g,3g,4g and 5g, sodium succinate 5.62g, vickers salt solution 50ml, volume to 1L with sterile water, pH=7) were prepared with ammonia nitrogen concentrations of 100mg/L, 200mg/L, 400mg/L, 600mg/L, 800mg/L and 1000mg/L, respectively. 100 μl of the activated liquid of Pseudomonas adzuki is inoculated into 100ml of sterilized evaluation culture medium, each of the evaluation culture medium is placed in shaking culture at 30deg.C in a shaker at 200rpm, 3 parallel groups are placed, sterile water is added into the evaluation culture medium as blank control of each experimental group, and ammonia nitrogen content in the evaluation culture medium is detected every 24h, and the experiment is arranged as follows:

experiment group 1: ammonia nitrogen concentration 100mg/L; control group 1: ammonia nitrogen concentration 100mg/L;

experiment group 2: ammonia nitrogen concentration 200mg/L; control group 2: ammonia nitrogen concentration 200mg/L;

experiment group 3: ammonia nitrogen concentration 400mg/L; control group 3: ammonia nitrogen concentration 400mg/L;

experiment group 4: ammonia nitrogen concentration 600mg/L; control group 4: ammonia nitrogen concentration 600mg/L;

experimental group 5: ammonia nitrogen concentration 800mg/L; control group 5: ammonia nitrogen concentration 800mg/L;

experiment group 6: ammonia nitrogen concentration 1000mg/L; control group 6: ammonia nitrogen concentration 1000mg/L;

the ammonia nitrogen detection method is carried out according to HJ535-2009 ammonia nitrogen determination-Nashi reagent spectrophotometry, and the results are shown in Table 4.

TABLE 4 degradation results of P.adulthood on different ammonia nitrogen concentrations

As can be seen from the data in Table 4, the Pseudomonas chenopodii shows better ammonia nitrogen degradation effect at ammonia nitrogen concentrations of 100mg/L, 200mg/L and 400mg/L, the ammonia nitrogen degradation rate of 72h reaches over 92%, and the ammonia nitrogen degradation rate of 72h is only about 20% at the ammonia nitrogen concentration of 600mg/L, so that it is considered that the Pseudomonas chenopodii has an ammonia nitrogen tolerance concentration of 600mg/L, and the strain hardly shows the ammonia nitrogen degradation effect at the ammonia nitrogen concentration of more than 600 mg/L.

EXAMPLE 5 fermentation Process of Pseudomonas Chengdu

The fermentation method of the adzuki pseudomonas comprises the following steps:

(1) Primary seed culture: inoculating Pseudomonas all of which is taken under aseptic condition into a liquid LB culture medium, and shake culturing for 24 hours at 30 ℃ and 200rpm to obtain a first-stage seed culture solution;

(2) Secondary seed culture: inoculating the primary seed culture solution into a liquid LB culture medium according to 10vol% of inoculum size under the aseptic condition, and culturing for 18 hours at 30 ℃ and 200rpm to obtain a secondary seed culture solution;

(3) Fermentation: after the fermentation medium in the fermentation tank is disinfected, inoculating the secondary seed culture solution into the fermentation medium according to the inoculation amount of 1-10vol%, wherein the fermentation medium comprises the following components: 50g/L of glucose, 10g/L of yeast extract powder, 10g/L of peptone, 1g/L of monopotassium phosphate, 1g/L of dipotassium phosphate, 0.5g/L of magnesium sulfate, 0.5g/L of sodium chloride and 0.2g/L of manganese sulfate, fermenting under the conditions that the temperature is controlled to be 25-35 ℃ and the rotating speed is 150-300rpm and the ventilation ratio is 1 (1-2), stopping fermenting when dissolved oxygen starts to rise, and obtaining fermentation liquor of the pseudomonas dui, wherein the viable count of the fermentation liquor is 300 hundred million cfu/ml through test.

Example 6 application of Pseudomonas Chengdu in refuse osmosis liquid treatment engineering

The waste water treatment process includes the steps of leading raw water to an anoxic tank after passing through an adjusting tank and UASB, leading the raw water to a nitrifying tank, and finally, discharging the raw water after membrane treatment. Before the microbial inoculum is added, the aeration is performed intermittently for one week, but the ammonia nitrogen content is not reduced. Then, the fermentation broth of the Geoporia amabilis prepared in the embodiment 5 of the invention is sprinkled into a nitrification tank with the addition volume ratio of 0.01%, the water temperature in the daytime is about 25-30 ℃ in the nitrification process, and the index change of ammonia nitrogen in the nitrification process is monitored, and the result is shown in Table 5.

TABLE 5 Ammonia nitrogen content in landfill leachate with time

Time/h	Ammonia nitrogen content/(mg/L)
		0	392
24	189
		48	97
72	59
		96	33
120	25
		144	27

As can be seen from the data in Table 5, after the addition of the fermentation broth of all Pseudomonas, the ammonia nitrogen in the effluent of the biochemical system is reduced to below 30mg/L within five days, and the ammonia nitrogen degradation rate reaches 94%, which indicates that the all Pseudomonas has high-efficiency ammonia nitrogen removal capability.

The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.

Sequence listing

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aggccaggta aggttcttcg cgttgcttcg aattaaacca catgctccac cgcttgtgcg 540

ggcccccgtc aattcatttg agttttaacc ttgcggccgt actccccagg cggtcaactt 600

aatgcgttag ctgcgccact aagttctcaa ggaacccaac ggctagttga catcgtttac 660

ggcgtggact accagggtat ctaatcctgt ttgctcccca cgctttcgca cctcagtgtc 720

agtatcagtc caggtggtcg ccttcgccac tggtgttcct tcctatatct acgcatttca 780

ccgctacaca ggaaattcca ccaccctcta ccgtactcta gctcgccagt tttggatgca 840

gttcccaggt tgagcccggg gctttcacat ccaacttaac gaaccaccta cgcgcgcttt 900

acgcccagta attccgatta acgcttgcac ccttcgtatt accgcggctg ctggcacgaa 960

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tatcccccac taccaggcag attcctaggc attactcacc cgtccgccgc taaatcaggg 1380

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Claims

1. Pseudomonas of adult strainPseudomonas chengduensis) The method is characterized in that the strain is preserved in China general microbiological culture Collection center (China Committee) with the preservation number: cgmccno.24300.

2. A microbial agent, characterized in that the active ingredient comprises Pseudomonas adzuki of claim 1.

3. The fermentation process of Pseudomonas adzuki of claim 1, comprising the steps of:

(3) Fermentation: after the fermentation medium in the fermentation tank is disinfected, inoculating the secondary seed culture solution into the fermentation medium according to the inoculation amount of 1-10vol%, controlling the temperature to be 25-35 ℃ and the rotating speed to be 150-300rpm, fermenting under the condition that the ventilation ratio is 1 (1-2), stopping fermentation when dissolved oxygen starts to rise, and obtaining the fermentation solution, wherein the fermentation medium comprises the following components: 40-60g/L carbon source, 15-30g/L, PO nitrogen source ₄ ^3- 0.8-1.5g/L、K ⁺ 0.5-1.0g/L、Mg ²⁺ 0.05-0.2g/L、Na ⁺ 0.1-0.3g/L、Mn ²⁺ 0.03-0.1g/L, water as solvent, pH=6-8.

4. A fermentation process according to claim 3, wherein the carbon source is selected from one or more of glucose, sucrose, starch, sodium acetate or sodium succinate;

the nitrogen source is selected from one or more of yeast extract powder, peptone, urea, ammonium sulfate or potassium nitrate.

5. A method for purifying waste water, comprising the step of inoculating the waste water with the activated liquid of pseudomonas adzuki of claim 1 or the microbial agent of claim 2.

6. The method of claim 5, wherein the concentration of ammonia nitrogen in the wastewater is 600mg/L or less.

7. The method of claim 6, wherein the concentration of ammonia nitrogen in the wastewater is 400mg/L or less.

8. The method of claim 7, wherein the concentration of ammonia nitrogen in the wastewater is 100-200mg/L.

9. The method of any one of claims 5-8, wherein the waste water has a salinity of 8% or less.

10. The method of claim 9, wherein the waste water has a salinity of 6% or less.

11. The method of claim 10, wherein the waste water has a salinity of 5% or less.

12. The method of claim 11, wherein the waste water has a salinity of 3% or less.

13. Use of the adzuki pseudomonas as claimed in claim 1 and the microbial agent as claimed in claim 2 in the field of waste water purification.

14. The use according to claim 13, wherein the pseudomonas adulthood and microbial agent are used for degrading nitrogen-containing substances in waste water.

15. The use according to claim 14, wherein the pseudomonas and microbial agents are used to degrade ammonia nitrogen containing materials in waste water.