Pseudomonas Taiwan ZZRQ2, biological denitrifying agent and application of pseudomonas Taiwan ZZRQ2

Abstract

The invention provides Pseudomonas Taiwan ZZRQ2, a biological denitrification agent and application of the biological denitrification agent, and belongs to the technical field of environmental microorganisms. The invention provides a strain of pseudomonas Taiwan ZZRQ2, the preservation number of which is CGMCC (China General Microbiological Culture Collection Center) No.27545, and the pseudomonas Taiwan ZZRQ2 has efficient heterotrophic nitrification-aerobic denitrification performance and can be used for removing NH4 < + >-N and NO3 <->-N in a natural water body. The invention provides a biological denitrification bacterial agent. The biological denitrification bacterial agent comprises pseudomonas Taiwan ZZRQ2 and biomass slag. The surface of the biomass slag has a large number of orderly arranged pores, the specific surface area is large, and the adsorption capacity is high. Pseudomonas Taiwan ZZRQ2 is combined with biomass slag to treat sewage, the biomass slag serves as a microbial carrier, the adsorption performance of the carrier is combined with the degradation performance of microorganisms, a synergistic interaction effect is achieved in the aspect of sewage denitrification, and the removal rates of NH4 < + >-N and NO3 <->-N reach 98.9% and 93.4% respectively.

Description

Technical Field

[0001] The invention belongs to the technical field of environmental microorganisms, and in particular relates to a strain of Taiwan Pseudomonas ZZRQ2, a biological denitrifier and applications thereof. Background Art

[0002] In recent years, the eutrophication of urban and rural rivers and lakes has become increasingly prominent. Excessive nitrogen in water is one of the main causes of this phenomenon. The control of nitrogen pollution is considered to be the focus and difficulty of current water ecological restoration work. As a microbial treatment technology, the biofilm method has been widely used in the removal of nitrogen, phosphorus and heavy metals from industrial wastewater. Compared with traditional denitrification methods, the biofilm method has the advantages of higher resistance to ammonia nitrogen shock load and less residual sludge.

[0003] At present, the biofilm method has gradually been applied to the restoration of polluted river water. In the process of the biofilm method being actually applied to the restoration of polluted river water, the added biological agents have the problem of difficulty in survival in the early stage, and the agents are easily lost in large quantities with the water flow. Therefore, the selection of appropriate strains and microbial carriers is the key to improving the effect of the biofilm method in the restoration of polluted river water. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a strain of Taiwan Pseudomonas ZZRQ2, a biological denitrifier and its application. The present invention combines Taiwan Pseudomonas ZZRQ2 with biomass slag to treat sewage, which can effectively remove NH4 + -N and NO3 - -N.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a strain of Taiwan Pseudomonas ZZRQ2, and the preservation number of the Taiwan Pseudomonas ZZRQ2 is CGMCC No.27545.

[0007] The present invention also provides a biological denitrification bacterial agent, comprising the above-mentioned Taiwan Pseudomonas ZZRQ2 and biomass slag.

[0008] Preferably, the biomass slag is obtained by drying, crushing and anaerobic carbonization of waste biomass; the temperature of the anaerobic carbonization is 800-1200° C., and the time is 1.5-2.5 hours.

[0009] More preferably, the waste biomass includes wood, straw, and bamboo.

[0010] More preferably, the crushed particle size is 10 to 30 meshes.

[0011] The present invention also provides a method for preparing the above-mentioned biological denitrifier, comprising: activating the above-mentioned Taiwan Pseudomonas ZZRQ2, mixing it with biomass slag, and heating it at 25-30°C and 100-150 r·min -1 The biodenitrifier is obtained by forming biofilm under the following conditions and freeze-drying.

[0012] Preferably, the activation of Taiwan Pseudomonas ZZRQ2 comprises: inoculating the Taiwan Pseudomonas ZZRQ2 in an activation medium, heating at 25-30°C and 100-150 r·min -1 Culture under the same conditions for 20 to 28 hours.

[0013] Preferably, the biofilm formation time is 4 to 6 days.

[0014] The present invention also provides the use of the above-mentioned Pseudomonas Formosanus ZZRQ2 or the above-mentioned biological denitrifier in sewage denitrification treatment.

[0015] Preferably, the dosage of the Pseudomonas Formosanus ZZRQ2 is 5 to 10 g / L; the dosage of the biological denitrifier is 5 to 10 g / L.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention provides a strain of Taiwan Pseudomonas ZZRQ2, with a preservation number of CGMCC No. 27545. It has high-efficiency heterotrophic nitrification-aerobic denitrification performance and can effectively remove NH4 + -N and NO3 - -N.

[0018] The present invention also provides a biological denitrification agent, including Taiwan Pseudomonas ZZRQ2 and biomass slag. The surface of the biomass slag of the present invention has a large number of orderly arranged pores, a large specific surface area, and a strong adsorption capacity. Taiwan Pseudomonas ZZRQ2 is used in combination with biomass slag to treat sewage. The biomass slag is used as a microbial carrier. The adsorption performance of the carrier is combined with the degradation performance of the microorganism, which has a synergistic effect in the denitrification of sewage. NH4 + -N、NO3 - The removal rates of -N were as high as 98.9% and 93.4% respectively. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The colony morphology characteristics of Pseudomonas formosanus ZZRQ2;

[0020] Figure 2 This is a scanning electron microscope image of the biomass slag in step (1) of Example 2;

[0021] Figure 3This is a scanning electron microscope image of the biological denitrifier in step (3) of Example 2.

[0022] Biological Deposit Description

[0023] The Pseudomonas taiwanensis ZZRQ2 described in the present invention is recommended to be named Pseudomonas taiwanensis in taxonomy. It was deposited in the General Microbiological Center of China Culture Collection Administration on June 5, 2023, with the address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 27545. DETAILED DESCRIPTION

[0024] The present invention provides a strain of Taiwan Pseudomonas ZZRQ2, and the preservation number of the Taiwan Pseudomonas ZZRQ2 is CGMCC No.27545.

[0025] The Taiwan Pseudomonas ZZRQ2 described in the present invention is isolated from the river water environment in the urban area, has high-efficiency heterotrophic nitrification-aerobic denitrification performance, and can effectively remove NH4 in natural water bodies. + -N and NO3 - -N.

[0026] The present invention also provides a biological denitrification bacterial agent, comprising the above-mentioned Taiwan Pseudomonas ZZRQ2 and biomass slag.

[0027] The biomass slag of the present invention is obtained by drying, crushing and anaerobic carbonization of waste biomass; the temperature of the anaerobic carbonization is preferably 800-1200°C, more preferably 1000°C, and the time is preferably 1.5-2.5h, more preferably 2h; the waste biomass preferably includes wood, straw, and bamboo; the particle size of the crushed particle is preferably 10-30 mesh, more preferably 20 mesh.

[0028] After high-temperature modification, the biomass slag prepared by the present invention can increase the specific surface area of ​​the carrier, increase the number of functional groups on the carrier surface, increase the carrier's ability to remove ammonia nitrogen, provide more attachment interfaces for microorganisms, and have a large number of orderly arranged pores on the surface, and have strong adsorption capacity. Therefore, using biomass slag as a microbial carrier, combining the adsorption performance of the carrier with the degradation performance of the microorganism, can better remove ammonia nitrogen in water.

[0029] The present invention also provides a method for preparing the above-mentioned biological denitrifier, comprising: activating the above-mentioned Taiwan Pseudomonas ZZRQ2, mixing it with biomass slag, and heating it at 25-30°C and 100-150 r·min -1 The biodenitrifier is obtained by forming biofilm under the following conditions and freeze-drying.

[0030] The activation of Taiwan Pseudomonas ZZRQ2 of the present invention preferably comprises: inoculating Taiwan Pseudomonas ZZRQ2 into an activation medium, heating at 25-30°C and 100-150 r·min -1 The temperature is preferably 28°C, and the rotation speed is preferably 130 rpm. -1 , the time is more preferably 24h.

[0031] The present invention has no special limitation on the composition of the activation medium, and conventional commercial products in the art can be used. As an implementation method, the activation medium includes the following components at the following concentrations: 10 g / L peptone, 3 g / L beef extract powder, 5 g / L sodium chloride, and a pH value of 7.2.

[0032] The biofilm formation time of the present invention is preferably 4 to 6 days, more preferably 5 days.

[0033] Before forming biofilm, the biomass slag is preferably filtered through 20 meshes and placed in sterile physiological saline for thorough shaking and washing to remove free bacteria attached to the surface.

[0034] The present invention also provides the use of the above-mentioned Pseudomonas Formosanus ZZRQ2 or the above-mentioned biological denitrifier in sewage denitrification treatment.

[0035] The sewage of the present invention preferably includes nitrogen-containing sewage, and more preferably includes one or more of domestic sewage, industrial wastewater, farmland drainage and aquaculture sewage.

[0036] The dosage of the Pseudomonas Formosanus ZZRQ2 of the present invention is preferably 5-10 g / L, more preferably 8 g / L; the dosage of the biological denitrifier is 5-10 g / L, more preferably 8 g / L.

[0037] The temperature of the sewage treated in the present invention is preferably 20-30°C, more preferably 25°C, and the pH value is preferably 7.0-7.3, more preferably 7.2.

[0038] The Taiwan Pseudomonas ZZRQ2 provided by the present invention can be used alone or in combination with biomass slag. The Taiwan Pseudomonas ZZRQ2 of the present invention and biomass slag are used in combination to have a synergistic effect in wastewater denitrification. + -N and NO3 - The removal rates of -N were as high as 98.9% and 93.4% respectively.

[0039] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0041] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0042] Example 1

[0043] Isolation, Screening and Identification of Pseudomonas Taiwanense ZZRQ2

[0044] 1. Enrichment and domestication

[0045] After the sludge and riverbed mud are taken back to the laboratory, they are promptly placed in the refrigerator. Shake them properly before use. Use a pipette to absorb 10mL of the mud-water mixture and add it to 90mL of normal saline. Weigh 10g of the bacterial agent and put it into 90mL of normal saline. Put it into a shaker and shake for 8h before taking it out. Let it stand for 10min first, then take 2mL of the supernatant and inoculate it into a 250mL conical flask containing 98mL of sterile LB culture medium. Seal the bottle mouth with a sealing film and place it in a constant temperature shaker at 30℃ and 150r / min for cultivation until obvious turbidity appears. Then take 2mL of LB bacterial solution and inoculate it into 98mL of heterotrophic nitrification culture medium. After obvious turbidity appears, transfer it to fresh sterile heterotrophic nitrification culture medium. Repeat this process. While transferring, increase the concentration of ammonium sulfate in the culture medium in turn until the content reaches 3.0g / L, and then perform plate separation.

[0046] 2. Separation

[0047] Take the culture solution that has been domesticated under the condition of ammonium sulfate content of 3.0g / L, and dilute it with sterile water in a gradient manner (10 -1 ~10 -8 ), then take 100 μl of the gradient diluted bacterial solution and evenly spread it on a solid heterotrophic nitrification medium plate with an ammonia nitrogen content of 200 mg / L, mark them with numbers, and culture them in a 30°C biochemical incubator for 2 to 3 days. After the colonies grow, pick colonies with different morphologies and separate them by streaking on the plate. In order to ensure the purity of the strain, repeat streaking two to three times. After the purification work is completed, a single colony is inoculated on an LB slant and stored at 4°C.

[0048] 3. Screening of dominant strains and preliminary determination of ammonia nitrogen and nitrate nitrogen degradation capabilities

[0049] Six purified colonies were picked from the plates, inoculated into 100 mL of sterilized LB liquid medium, and cultured in a shaker at 30 °C and 150 r / min for 10 h. Then, 2 mL of the bacterial solution was inoculated into 98 mL of heterotrophic nitrification medium (Na2HPO4·7H2O 7.9 g, KH2PO4 1.5 g, MgSO4·7H2O 0.1 g, C4H4Na2O4·6H2O 10.806 g, (NH4)2SO4 0.943 g, trace element solution 2 mL / L) and aerobic denitrification medium (Na2HPO4·7H2O 7.9 g, KH2PO4 1.5 g, MgSO4·7H2O 0.1 g, C6H4Na2O4·6H2O 10.806 g, (NH4)2SO4 0.943 g, trace element solution 2 mL / L). 12 O6·H2O 8.257g, KNO31.444g, trace element solution 2mL / L), placed in a shaker at 30℃ and 150r / min for 24h, sampled and tested the concentrations of ammonia nitrogen and nitrate nitrogen, and compared the degradation ability of each purified strain. All 6 strains have a certain ammonia nitrogen and nitrate nitrogen degradation effect, among which ZZRQ2 strain has the best degradation effect, with 24h ammonia nitrogen and nitrate nitrogen degradation rates of 58.54% and 64.33% respectively. Therefore, ZZRQ2 strain was taken as the object of further study.

[0050] Example 2

[0051] Preparation of biological denitrifier

[0052] (1) Preparation of biomass slag

[0053] The waste biomass wood is dried, crushed through a 20-mesh sieve, and then placed in a crucible with a cover, subjected to oxygen-deficient carbonization in a muffle furnace, and calcined at 1000° C. for 2 h until the waste biomass is in the form of residue, thereby obtaining biomass slag;

[0054] (2) Activation of ZZRQ2 in Pseudomonas formosanus

[0055] Pseudomonas formosanus ZZRQ2 was inoculated into the activation medium and incubated at 28°C and 130 r·min -1 The culture was carried out in a constant temperature shaking incubator for 24 h under the conditions to obtain an activated bacterial solution (the effective number of viable bacteria is preferably ≥ 6 × 10 10 CFU / mL);

[0056] Activation medium components: peptone 10g / L, beef extract powder 3g / L, sodium chloride 5g / L, pH 7.2;

[0057] (3) Film formation

[0058] The biomass slag prepared in step (1) is passed through a 20-mesh filter and placed in sterile physiological saline for thorough shaking and washing to remove free bacteria attached to the surface;

[0059] The activated bacterial solution prepared in step (2) was uniformly mixed with the cleaned biomass slag with a particle size of 20 mesh and heated at 28°C and 130 r / min. -1 The biomass slag was dried with a freeze dryer for 3 days to obtain a biological denitrifier, which was then stored in a frozen state.

[0060] Example 3

[0061] Preparation of biological denitrifier

[0062] (1) Preparation of biomass slag

[0063] The waste biomass straw is dried, crushed through a 10-mesh sieve, and then placed in a crucible with a lid, subjected to oxygen-deficient carbonization in a muffle furnace, and calcined at 800° C. for 2.5 h until the waste biomass is in the form of residue, thereby obtaining biomass slag;

[0064] (2) Activation of ZZRQ2 in Pseudomonas formosanus

[0065] Pseudomonas formosanus ZZRQ2 was inoculated into the activation medium and cultured at 25°C and 150 r·min -1 The culture was carried out in a constant temperature shaking incubator for 28 h to obtain an activated bacterial solution (effective viable bacteria count ≥ 6 × 10 10 CFU / mL);

[0066] Activation medium components: peptone 10g / L, beef extract powder 3g / L, sodium chloride 5g / L, pH 7.2;

[0067] (3) Film formation

[0068] The biomass slag prepared in step (1) is passed through a 20-mesh filter and placed in sterile physiological saline for thorough shaking and washing to remove free bacteria attached to the surface;

[0069] The activated bacterial solution prepared in step (2) was evenly mixed with the cleaned biomass slag with a particle size of 20 mesh, and heated at 25°C and 150 r·min. -1 The biomass slag was dried with a freeze dryer for 3 days to obtain a biological denitrifier, which was then stored in a frozen state.

[0070] Example 4

[0071] Preparation of biological denitrifier

[0072] (1) Preparation of biomass slag

[0073] The waste biomass bamboo is dried, crushed through a 30-mesh sieve, and then placed in a crucible with a cover, subjected to oxygen-deficient carbonization in a muffle furnace, and calcined at 1200° C. for 1.5 h until the waste biomass is a residue, thereby obtaining a biomass slag;

[0074] (2) Activation of ZZRQ2 in Pseudomonas formosanus

[0075] Pseudomonas formosanus ZZRQ2 was inoculated into the activation medium and heated at 30°C and 100 r·min -1 The culture was carried out in a constant temperature shaking incubator for 20 h under the conditions to obtain an activated bacterial solution (effective viable bacteria count ≥ 6 × 10 10 CFU / mL);

[0076] Activation medium components: peptone 10g / L, beef extract powder 3g / L, sodium chloride 5g / L, pH 7.2;

[0077] (3) Film formation

[0078] The biomass slag prepared in step (1) is passed through a 20-mesh filter and placed in sterile physiological saline for thorough shaking and washing to remove free bacteria attached to the surface;

[0079] The activated bacterial solution prepared in step (2) was evenly mixed with the cleaned biomass slag with a particle size of 20 mesh, and heated at 30°C and 100 r / min. -1 The biomass slag was dried with a freeze dryer for 3 days to obtain a biological denitrifier, which was then stored in a frozen state.

[0080] Example 5

[0081] Scanning electron microscope

[0082] The biomass slag obtained in step (1) of Example 2 and the biological denitrification agent obtained in step (3) were scanned by electron microscope. The electron microscope images of the scans are as follows: Figure 2-Figure 3 shown.

[0083] according to Figure 2-Figure 3 It can be seen that since biomass slag has a larger micropore area and rough pore walls, it is easier for microorganisms to be loaded on it, and the amount of biofilm is greatly increased.

[0084] Example 6

[0085] NH4 in simulated wastewater + -N and NO3 - -N removal test

[0086] 8g of the biomass slag from step (1) of Example 2, 8g of the biological denitrifier from Example 2, and 8mL of the activated Taiwan Pseudomonas ZZRQ2 bacterial solution from step (2) of Example 2 were respectively added to two 1L simulated wastewaters (treatment 1 was NH4 + -N wastewater, treatment 2 to NO3 - -N wastewater), treated at 25°C and pH 7.0-7.3, and no substance was added to the control. The simulated wastewater NH4 was sampled and tested every 24 hours.+ -N and NO3 - -N content, and calculate NH4 + -N、NO3 - -N removal effect. Three parallel experiments were performed for each treatment, and the results were averaged. The specific results are shown in Tables 1 to 3.

[0087] The removal rate formula is: Removal rate (%) = (control treatment concentration - test treatment concentration) / control treatment concentration × 100.

[0088] Simulated wastewater components:

[0089] Treatment 1: Heterotrophic nitrification medium (g / L): Na2HPO4·7H2O 7.9, KH2PO4 1.5, MgSO4·7H2O 0.1, C4H4Na2O4·6H2O 10.806, (NH4)2SO4 0.943 (NH4 + -N concentration is 200 mg / L), trace element solution 2 mL / L, pH value 7.0-7.5, sterilization at 121°C.

[0090] Treatment 2: Aerobic denitrification medium (g / L): Na2HPO4·7H2O 7.9, KH2PO4 1.5, MgSO4·7H2O 0.1, C6H 12 O6·H2O 8.257, KNO31.444(NO3 - -N concentration is 200 mg / L), trace element solution 2 mL / L, pH value 7-7.5, sterilization at 121°C.

[0091] Table 1 Effect of different incubation time of biomass slag on NH4 + -N、NO3 - -N removal effect

[0092]

[0093] Table 2 Effect of different culture time of activated bacterial solution of Pseudomonas formosanus ZZRQ2 on NH4 + -N、NO3 - -N removal effect

[0094]

[0095] Table 3 Effect of different cultivation time of biological denitrifier on NH4 + -N、NO3 - -N removal effect

[0096]

[0097] According to Table 1, in the initial stage of the experiment, the biomass slag had a significant effect on the NH4 + -N, NO3 - -N removal rate was only 8.7% and 12.9%. With the increase of adsorption time, the removal rate of nitrogen by biomass slag first increased and then decreased. + -N, NO3 - The -N removal rates reached 78.0% and 52.6%, and the adsorption of nitrogen had reached saturation. After more than 96 hours, the adsorption rate began to decline.

[0098] According to Table 2, strain ZZRQ2 has a strong affinity for NH4 + -N, NO3 - -N has a significant removal effect. During the experiment, as the strain grew, it could significantly remove inorganic nitrogen components in the sewage, including NH4 + -N, NO3 - -N had the highest removal rate, which was 92.9% and 88.8% respectively. Then, as the experimental time increased, some microorganisms died and the removal rate began to decline.

[0099] According to Table 3, the biomass slag with bacteria had a significant effect on the NH4 + -N, NO3 - -N removal rates were only 21.8% and 28.6%, which were lower than those of strain ZZRQ2 for NH4 + -N, NO3 - -N removal rate, because the dead bacteria residues attached to the surface of the carrier increase the NH4 + -N, NO3 - -N concentration. NH4 at 72h + -N, NO3 - -N had the highest removal rates, which were 98.9% and 93.4%, respectively. Then, as the experimental time increased, NH4 + -N and NO3 - The -N removal rate tends to be stable.

[0100] The removal rates of each treatment in Tables 1-3 are summarized, and the results are shown in Tables 4 and 5.

[0101] Table 4 Effect of biomass slag, activated bacterial solution and biological denitrification agent on NH4 + -N removal effect (%)

[0102] deal with 24h 48h 72h 96h 120h Biomass slag 8.7 13.2 65.2 78.0 73.4 Activated bacteria solution 59.2 70.9 92.9 90.5 88.4 Biological denitrifier 21.8 52.2 98.9 97.4 97.2

[0103] Table 5 Effect of biomass slag, activated bacterial solution and biological denitrification agent on NO3 - -N removal effect (%)

[0104] deal with 24h 48h 72h 96h 120h Biomass slag 12.9 22.5 44.2 52.6 51.9 Activated bacteria solution 61.0 85.6 88.8 84.1 83.5 Biological denitrifier 28.6 92.6 93.4 95.3 95.6

[0105] From Tables 4 and 5, we can see that the surface of biomass slag has a large number of orderly arranged micropores, which can effectively absorb NH4 + -N removal rate is 8.7% to 78.0%, and the removal rate of NO3 - The removal rate of -N was 12.9% to 52.6%, and the adsorption rate reached saturation at 96h. + -N removal rate is 59.2% to 92.9%, and the removal rate of NO3 - The removal rate of -N was 61.0% to 88.8%, among which the free bacteria content was the highest at 72h, and the nutrient-deficient strains in the environment began to die after 72h. The biological denitrifier can better remove ammonia nitrogen and nitrate nitrogen in water due to the combination of the adsorption properties of biomass slag itself and the degradation properties of microorganisms. The removal rate of NH4 + -N removal rate is 21.8% to 98.9%, and the removal rate of NO3 - The removal rate of -N is 28.6% to 95.6%, and the removal rate of ammonia nitrogen and nitrate nitrogen in water is kept stable after 72 hours.

[0106] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A strain of Pseudomonas formosanus ZZRQ2, It is characterized in that The deposit number of the Taiwan Pseudomonas ZZRQ2 is CGMCC No.27545.

2. A biological denitrification agent, It is characterized in that It comprises the Taiwan Pseudomonas ZZRQ2 described in claim 1 and biomass slag.

3. The biological denitrifier according to claim 2, It is characterized in that The biomass slag is obtained by drying, crushing and anoxic carbonization of waste biomass; the temperature of the anoxic carbonization is 800-1200° C. and the time is 1.5-2.5 hours.

4. The biological denitrifier according to claim 3, It is characterized in that The waste biomass includes wood, straw and bamboo.

5. The biological denitrifier according to claim 3, It is characterized in that The particle size of the crushed material is 10 to 30 meshes.

6. A method for preparing a biological denitrifier according to any one of claims 2 to 5, It is characterized in that include: The Taiwan Pseudomonas ZZRQ2 of claim 1 is activated, mixed with biomass slag, and heated at 25 to 30°C and 100 to 150 r·min -1 The biodenitrifier is obtained by forming biofilm under the following conditions and freeze-drying.

7. The preparation method according to claim 6, It is characterized in that The activation of Taiwan Pseudomonas ZZRQ2 comprises: inoculating Taiwan Pseudomonas ZZRQ2 in an activation medium, heating at 25-30°C and 100-150 r·min -1 Culture under the same conditions for 20 to 28 hours.

8. The preparation method according to claim 6, It is characterized in that The biofilm formation time is 4 to 6 days.

9. Use of the Pseudomonas Formosanus ZZRQ2 according to claim 1 or the biological denitrifier according to any one of claims 2 to 5 in wastewater denitrification treatment.

10. The use according to claim 9, It is characterized in that The dosage of the Taiwan Pseudomonas ZZRQ2 is 5-10 g / L; the dosage of the biological denitrifier is 5-10 g / L.

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