Application of 5-methylisoxazole nitrification inhibitor and nitrification inhibition method of 5-methylisoxazole nitrification inhibitor

Abstract

The invention belongs to the technical field of soil treatment and sewage treatment, and particularly relates to application of a 5-methylisoxazole nitrification inhibitor and a nitrification inhibition method of the 5-methylisoxazole nitrification inhibitor. The application comprises the application in inhibiting nitrosation activity of ammonia oxidizing bacteria and the application in inhibiting nitrification activity of nitrite oxidizing bacteria. The nitrification inhibition activity of 5-methylisoxazole is found through a large number of researches and experiments, and full-path regulation and control of the nitrification effect are achieved. Experiments show that the compound can inhibit the ammoxidation activity of typical nitrifying strains at a low dosage (more than or equal to 0.01 per thousand w / v), and can effectively inhibit the activity of nitrite oxidizing bacteria when the concentration is lower than 0.1 per thousand m / v. The pH value of an aqueous solution of the compound is neutral, and the compound has no obvious inhibition effect on non-target heterotrophic microorganisms and has high selectivity and environmental friendliness.

Description

Technical Field

[0001] The invention belongs to the technical field of soil treatment and sewage treatment, and particularly relates to application of a 5-methylisoxazole nitrification inhibitor and a method for inhibiting nitrification. Background Art

[0002] Nitrification is one of the core processes of nitrogen biogeochemical cycle, which refers to the process of ammonium nitrogen (NH4 + -N) is converted into nitrate nitrogen (NO3 - -N) biochemical pathway. This process is catalyzed by two types of chemoautotrophic nitrifying microorganisms in sequence: Ammonia-Oxidizing Bacteria (AOB) first converts NH4 + -N is oxidized to nitrite nitrogen (NO2 - -N), which is then further converted into NO3 by nitrite-oxidizing bacteria (NOB). - As a key link in the transformation of nitrogen forms, nitrification is widely distributed in habitats such as soil, freshwater and marine ecosystems. Its metabolic intensity directly affects the global nitrogen cycle flux, greenhouse gas emissions and nitrogen use efficiency of agricultural production systems.

[0003] Currently, the enhanced nitrification caused by excessive nitrogen input has become a significant environmental threat. Its risk chain reaction is specifically manifested as follows: (1) Nitrate nitrogen leads to nitrate contamination of groundwater through soil leaching; (2) Intensified denitrification process induces the release of strong greenhouse gas N2O; (3) Reduced nitrogen fertilizer utilization efficiency leads to non-point source pollution at the watershed scale. Based on this, how to achieve a dynamic balance between agricultural production and ecological protection through precise regulation of nitrification process has become a core proposition to be solved in the coordinated development of environmental microbiology and sustainable agriculture. How to further reduce the dosage of inhibitors and enrich the types of inhibitors is of great significance to improving agricultural economic benefits. Summary of the Invention

[0004] The object of the present invention is to provide an application of a 5-methylisoxazole nitrification inhibitor capable of inhibiting the growth of ammonia oxidizing bacteria and nitrite oxidizing bacteria and effectively reducing the production of nitrification, and a method for inhibiting nitrification.

[0005] In order to solve the problems existing in the prior art, the present invention adopts the following technical solutions:

[0006] The use of 5-methylisoxazole nitrification inhibitor, including use in any of the following:

[0007] (1) Application in inhibiting the nitrosation activity of ammonia oxidizing bacteria;

[0008] (2) Application in inhibiting the nitrification activity of nitrite oxidizing bacteria.

[0009] In the above-mentioned use of the 5-methylisoxazole nitrification inhibitor, the ammonia oxidizing bacteria include Nitrosomonas sp. CZ-4 or Nitrosomonas nitrosa strain SN-6.

[0010] In the above-mentioned application of the 5-methylisoxazole nitrification inhibitor, the nitrite oxidizing bacteria is the Nitrobacter enrichment culture N-winogradskyiWY-8.

[0011] A method for inhibiting nitrification using a 5-methylisoxazole nitrification inhibitor comprises treating soil or water containing ammonia oxidizing bacteria and / or nitrite oxidizing bacteria with 5-methylisoxazole, wherein the concentration of the 5-methylisoxazole is greater than or equal to 0.01‰ w / v, based on the volume of the soil or water being 100%.

[0012] In the above-mentioned method for inhibiting nitrification using a 5-methylisoxazole nitrification inhibitor, the ammonia oxidizing bacteria include Nitrosomonas sp. CZ-4 and Nitrosomonas nitrosa strain SN-6.

[0013] In the above-mentioned method for inhibiting nitrification using a 5-methylisoxazole nitrification inhibitor, the nitrite oxidizing bacteria is the Nitrobacter enrichment culture N-winogradskyiWY-8.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention has discovered the nitrification inhibitory activity of 5-methylisoxazole through a large number of studies and experiments. After testing, 5-methylisoxazole can inhibit the growth of ammonia oxidizing bacteria such as Nitrosomonas at a relatively low dose (0.01‰w / v), thereby effectively inhibiting nitrification in soil or water, reducing nitrogen loss in farmland and N2O emissions in soil and water. And at a concentration of 0.1‰w / v, the inhibition rate of 5-methylisoxazole on the ammonia oxidation activity of Nitrosomonas CZ-4 reached more than 98%. At a concentration of 0.1‰m / v, it also has a good inhibitory effect on the nitrification activity of Nitrobacter enrichment culture. Its aqueous solution is neutral, has no significant inhibitory effect on other bacteria while not affecting the pH of water / soil, and is highly environmentally friendly. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0017] Figure 1 The effect of different concentrations of 5-methylisoxazole on the growth of Nitrosomonas sp.CZ-4.

[0018] Figure 2 The effects of different concentrations of 5-methylisoxazole on the growth of Nitrosomonas nitrosa SN-6.

[0019] Figure 3 The effect of 5-methylisoxazole on the growth of Nitrosomonas sp. CZ-4 at low concentrations.

[0020] Figure 4 This is the nitrite nitrogen consumption of N-winogradskyiWY-8 when treated with different concentrations of 5-methylisoxazole on the 4th day.

[0021] Figure 5 It is the accumulation of nitrate nitrogen when N-winogradskyiWY-8 is treated with different concentrations of 5-methylisoxazole on the 4th day.

[0022] Figure 6 The effect of 5-methylisoxazole at a concentration of 0.05‰ m / v on the growth of heterotrophic microorganisms in activated sludge.

[0023] Figure 7 The effect of different concentrations of 5-methylisoxazole on ammonia nitrogen concentration in domestic sewage.

[0024] Figure 8 The effect of different concentrations of 5-methylisoxazole on the concentration of nitrite nitrogen in domestic sewage.

[0025] Figure 9 The effect of different concentrations of 5-methylisoxazole on nitrate nitrogen concentration in domestic sewage.

[0026] Figure 10 The effect of 5-methylisoxazole on ammonia nitrogen concentration in soil. DETAILED DESCRIPTION

[0027] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the embodiments of the specification.

[0028] In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention may also be implemented in other ways different from those described herein. Those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0029] Secondly, the term "one embodiment" or "embodiment" herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The phrase "in one embodiment" appearing in various places throughout this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive of other embodiments.

[0030] The culture medium and 5-methylisoxazole mother solution used in the following examples were prepared as follows:

[0031] AOB medium: FeSO4·7H2O 0.4 g / L, MgSO4·7H2O 0.5 g / L, K2HPO4 0.1 g / L, NaHCO3 1.5 g / L, CaCl2·2H2O 1.5 g / L, NaCl 2.0 g / L, trace element solution 1 mL / L, pH 7.5. Trace element solution: EDTA 0.5 g / L, CuSO4·5H2O 0.075 g / L, ZnSO4·7H2O 0.3 g / L, CoCl2·6H2O 0.375 g / L, MnCl2·2H2O 0.3 g / L, H3BO4 0.014 g / L, NaMoO4·2H2O 0.22 g / L. After high temperature and high pressure sterilization (121°C, 20 min), add appropriate amount of filtered sterilized HEPES mother solution and NH4Cl mother solution, and adjust the pH of the culture medium to 7.8 with 1 mol / L NaOH and HCl before use.

[0032] NOB medium: K2HPO4 1.3 g / L, K2HPO4 1 g / L, NaHCO3 0.4 g / L, trace elements 1 mL. Trace element solution: EDTA 1.25 g / L, MgSO4·7H2O 44.4 g / L, CuSO4·5H2O 0.43 g / L, ZnSO4·7H2O 0.55 g / L, FeCl3·6H2O 1.25 g / L, CoCl2·6H2O 0.4 g / L, CaCl2·2H2O 1.375 g / L, NaMoO4·2H2O 0.05 g / L, anhydrous MnCl 2, 0.81 g / L. Add an appropriate amount of NaNO2 mother solution and adjust the pH of the culture medium to 7.8 with 1 mol / L NaOH and HCl before use.

[0033] Beef Peptone Medium (BPM): purchased from Qingdao Haibo Biological, prepared at a concentration of 18 g / L, sterilized at high temperature and high pressure (121°C, 20 min) and stored for future use.

[0034] 5-Methylisoxazole mother liquor: 0.5 g of 5-methylisoxazole mother liquor (CAS: 5765-44-6, purchased from Aladdin) was weighed using an analytical balance. A small amount of water was added to dissolve the mixture, and the mixture was transferred to a 50 mL volumetric flask, fixed to volume, and filtered through a 0.22 μm organic filter membrane. The 5-methylisoxazole mother liquor was sterilized by filtration and stored for later use.

[0035] It should be noted that in the present invention, the method for detecting ammonia nitrogen concentration in water adopts the "Water Quality - Determination of Ammonia Nitrogen - Nessler's Reagent Spectrophotometric Method" (HJ 535-2009); the determination of nitrite nitrogen concentration adopts the "Water Quality - Determination of Nitrite Nitrogen - Spectrophotometric Method" (GB / T 7493-1987). The determination of nitrate nitrogen concentration adopts the "Water Quality - Determination of Nitrite Nitrogen Spectrophotometric Method" (HJ / T 346-2007). The method for detecting ammonia nitrogen concentration in soil is to extract with 1 mol / L KCl solution and then use a continuous flow instrument to measure. It should be further noted that in the embodiments of the present invention, the nitrification inhibition rate is calculated using the following formula:

[0036] (1) Nitrite nitrogen accumulation (mg / L):

[0037] Nitrite nitrogen accumulation = C t-1 -C1

[0038] Where C1 is the initial nitrite nitrogen concentration, C t-1 is the nitrite nitrogen concentration on that day.

[0039] (2) Nitrite nitrogen consumption (mg / L):

[0040] Nitrite nitrogen consumption = C1-C t-1

[0041] Where C1 is the initial nitrite nitrogen concentration, C t-1 is the nitrite nitrogen concentration on that day.

[0042] (3) Nitrate nitrogen accumulation (mg / L):

[0043] Nitrate nitrogen accumulation = C t-1 -C1

[0044] Where C1 is the initial nitrate nitrogen concentration, C t-1 is the nitrate nitrogen concentration on that day.

[0045] (4) Inhibition rate of ammonia oxidation activity (%)

[0046]

[0047] Among them, C k is the nitrite nitrogen accumulation of the control group at a certain moment, C t is the nitrite nitrogen accumulation of the experimental group at the same time.

[0048] Example 1

[0049] This example relates to an experiment to evaluate the inhibitory effect of 5-methylisoxazole nitrification inhibitor on the ammonia oxidizing bacteria species Nitrosomonas sp. CZ-4 (hereinafter referred to as CZ-4).

[0050] Take 1 mL of CZ-4 bacterial liquid and add it to AOB medium. Culture it in a shaking incubator at 30°C and 140 r / min. Take samples every 24 hours to test the concentrations of ammonia nitrogen and nitrite nitrogen until the growth rate of nitrite nitrogen is greater than 30 mg / L / d. Use this as the inoculum.

[0051] Figure 1 The implementation method described in Table 2 was as follows: 5-methylisoxazole mother solution was added at varying concentrations to AOB culture medium with an initial ammonia nitrogen concentration of 100 mg / L, CZ-4 inoculum was added at 1% v / v, and cultured in a shaking incubator at 30°C and 140 rpm. The specific operation method is shown in Table 1. The CZ-4 culture system without 5-methylisoxazole served as the control group, and the CZ-4 culture systems with varying concentrations of 5-methylisoxazole served as the experimental groups.

[0052] Table 1 CZ-4 experimental system of the control group and experimental group in Example 1

[0053]

[0054] Samples were taken every 24 hours to measure the concentrations of ammonia nitrogen and nitrite nitrogen in the control group and each experimental group. Figure 1 Table 2 shows the change trend of nitrite nitrogen in each group and Table 3 shows the nitrite nitrogen concentration and ammonia oxidation inhibition rate in each group at 3 days.

[0055] Table 2 Nitrite nitrogen concentration and ammonia oxidation activity inhibition rate of each group on the 3rd day in Example 1

[0056]

[0057] From Table 2 and Figure 1It can be seen that the nitrite nitrogen concentration of the control group reached 101.26 mg / L on the 3rd day, indicating that the bacteria were in good growth condition. The nitrite nitrogen concentrations of the experimental groups with the addition of 5-methylisoxazole were lower than those of the control group, indicating that the growth of CZ-4 was inhibited to varying degrees. And as the concentration decreased, the inhibition ability decreased. At a concentration of 0.01‰w / v, the inhibition rate of ammonia oxidation activity was 15.00%. At concentrations of 0.1‰ and 1‰w / v, the inhibition rate of ammonia oxidation activity can reach 95.85% and 98.12%, and the ammonia oxidation activity is almost completely inhibited. Example 1 shows that 5-methylisoxazole can effectively inhibit the ammonia oxidation activity of CZ-4 at a concentration greater than 0.1‰w / v (inhibition rate>95%), and can also show a slight inhibitory effect at a concentration of 0.01‰w / v.

[0058] Example 2

[0059] This example relates to an experiment to evaluate the inhibitory effect of 5-methylisoxazole nitrification inhibitor on the ammonia oxidizing bacteria species Nitrosomonas nitrosa SN-6 (hereinafter referred to as SN-6) in water.

[0060] Figure 2 The implementation method of Table 3 was as follows: the strain CZ-4 was replaced by SN-6, and the experimental system and culture method were the same as those in Example 1. Samples were taken every 24 hours to measure the concentrations of ammonia nitrogen and nitrite nitrogen in the control group and each experimental group. Figure 3 Table 3 shows the change trend of nitrite nitrogen in each group and Table 4 shows the nitrite nitrogen concentration and ammonia oxidation inhibition rate in each group on the 3rd day.

[0061] Table 3 Nitrite nitrogen concentration and ammonia oxidation activity inhibition rate of each control group and each experimental group in Example 2 on day 3

[0062]

[0063] From Table 3 and Figure 2 It can be seen that the nitrite nitrogen concentration of the control group reached 112.33 mg / L on the third day, indicating that the bacteria were growing well. The nitrite nitrogen concentrations of the experimental groups with the addition of 5-methylisoxazole were all lower than those of the control group, indicating that the growth of SN-6 was inhibited to varying degrees. As the concentration decreased, the inhibition ability decreased, and no inhibition occurred at a concentration of 0.01‰w / v. At concentrations of 0.1‰ and 1‰w / v, the ammonia oxidation activity inhibition rate could reach 86.65% and 98.74%, respectively, which is a high inhibition rate. Example 2 shows that 5-methylisoxazole can effectively inhibit the ammonia oxidation activity of SN-6 at a concentration greater than 0.1‰w / v, and can also inhibit the growth of SN-6 at a concentration of 0.1‰-0.01‰w / v.

[0064] Example 3

[0065] The purpose of this study was to investigate the inhibitory effect of 5-methylisoxazole on CZ-4 at lower concentrations (0.1‰-0.01‰ w / v).

[0066] 1 mL of CZ-4 bacterial culture was added to AOB medium and cultured in a shaker at 30°C and 140 rpm. Samples were collected every 24 hours to measure ammonia nitrogen and nitrite nitrogen concentrations until the nitrite nitrogen growth rate exceeded 30 mg / L / day. This sample was then used as the inoculum. The purpose of this experiment was to investigate the inhibitory effect of 5-methylisoxazole on CZ-4 at relatively low concentrations (0.1‰-0.01‰ w / v).

[0067] Figure 3 The implementation method shown in Table 4 was as follows: Each group was inoculated with 1% logarithmic-phase CZ-4 cells at a volume ratio of 1% into 100 mL of AOB medium with an initial ammonia nitrogen concentration of 200 mg / L. 5-methylisoxazole stock solution was then added to the experimental group to concentrations of 0.075‰, 0.05‰, 0.025‰, and 0.01‰ (w / v), respectively. No 5-methylisoxazole was added to the control group. Ammonia nitrogen and nitrite nitrogen concentrations were monitored every 24 hours. Figure 3 Table 4 shows the change trend of nitrite nitrogen in each group and the nitrite nitrogen concentration and ammonia oxidation activity inhibition rate in each group at 3 days.

[0068] Table 4 3-day nitrite nitrogen concentration and ammonia oxidation activity inhibition rate of each control group and each experimental group in Example 3

[0069]

[0070] Depend on Figure 3 As shown in Table 4, the nitrite nitrogen concentration in the control group reached 156.04 mg / L on day 3. In the experimental group, inhibition rates at concentrations of 0.075‰, 0.05‰, 0.025‰, and 0.01‰ w / v were 86.75%, 76.72%, 48.08%, and 7.93%, respectively. Example 3 demonstrates that even at a low concentration of 0.01‰ w / v, a weak inhibitory effect was still detectable, and the inhibition rate increased with increasing concentration.

[0071] Example 4

[0072] This example relates to an experiment to evaluate the inhibitory effect of 5-methylisoxazole on an enriched culture of Nitrobacter, N. winogradskyi WY-8 (hereinafter referred to as WY-8).

[0073] Take 1 mL of WY-8 bacterial liquid and place it in NOB medium with an initial nitrite nitrogen concentration of 300 mg / L. Place it in an environment of 30℃ and 140 r / min for cultivation. Take samples every 24 hours to test the nitrite nitrogen concentration and nitrate nitrogen concentration until the nitrite nitrogen concentration increases at a rate greater than 50 mg / L / d. Use this as the inoculum.

[0074] Figure 4 The implementation method described in Table 5 is as follows: 5-methylisoxazole stock solution is added to NOB medium at varying concentrations, supplemented with an appropriate amount of sterile water, and inoculated with WY-8 at a 1% ratio. The WY-8 culture system is prepared according to the recipe in Table 4. A CZ-4 culture system without 5-methylisoxazole serves as a control group, while other CZ-4 culture systems serve as experimental groups. Cultures are then shaken at 30°C and 140 rpm.

[0075] Table 4 WY-8 experimental system of the control group and experimental group in Example 4

[0076]

[0077] Samples were taken on day 0 and day 4 to measure the concentrations of nitrite and nitrate in the control group and each experimental group, respectively. Figure 4 is the nitrite nitrogen consumption of each group at 4 days. Figure 5 Table 5 shows the nitrite nitrogen consumption and nitrate nitrogen accumulation of each group on the 4th day.

[0078] Table 5 Nitrite nitrogen consumption and nitrate nitrogen accumulation in the control group and experimental group on the 4th day in Example 4

[0079]

[0080] From Table 5 and Figure 4 、 Figure 5 The consumption of nitrite and accumulation of nitrate nitrogen indicate that 5-methylisoxazole also has an inhibitory effect on WY-8, and the inhibitory effect is proportional to the concentration. At 5-methylisoxazole concentrations of 1‰ and 0.1‰ w / v, nitrate nitrogen accumulation was 7.3% and 68.8% of the control group, respectively. No inhibitory effect was observed at 0.01‰ w / v. In summary, 5-methylisoxazole also has a certain inhibitory effect on WY-8.

[0081] Example 5

[0082] The purpose of this study is to explore the degree of inhibition of 5-methylisoxazole on heterotrophic microorganisms in water bodies.

[0083] Figure 6The implementation method is as follows: take 20mL beef extract peptone culture medium, inoculate activated sludge (from municipal sewage aerobic tank) at a ratio of 1 / 1000, add 0.05‰ w / v 5-methylisoxazole to the experimental group, and do not add 5-methylisoxazole to the control group. Incubate in a shaking incubator at 30℃ and 140r / min. Samples are taken at 0h and 24h to measure OD 600 .

[0084] Depend on Figure 6 It can be seen that after adding 0.05‰w / v of 5-methylisoxazole, the OD 600 There was no significant difference from the control group, indicating that 0.05‰ of 5-methylisoxazole had no inhibitory effect on other heterotrophic microorganisms in the soil. Example 5 demonstrates that the target compound effectively inhibits nitrifying bacteria while not interfering with the metabolic activity of non-target heterotrophic microorganisms, highlighting the selectivity advantage of its mechanism of action.

[0085] Example 6

[0086] The purpose of this implementation is to explore the specific application effect of 5-methylisoxazole in complex scenarios (domestic sewage).

[0087] Figure 7 、 Figure 8 、 Figure 9 The method was to inoculate 20 mL of domestic sewage into an aerobic tank with activated sludge at a 1% inoculum ratio. 5-methylisoxazole was added to the experimental group at a ratio of 0.5 / 0.2 / 0.1 / 0.05‰ w / v, while no 5-methylisoxazole was added to the control group. The incubation was performed in a shaker at 30°C and 140 rpm. Ammonia nitrogen, nitrite nitrogen, and nitrate nitrogen were measured every 24 hours. Figure 7 The changing trend of ammonia nitrogen, Figure 8 is the changing trend of nitrite nitrogen, Figure 9 The changing trend of nitrate nitrogen.

[0088] Depend on Figure 7 、 Figure 8 、 Figure 9 It can be seen that after the addition of 5-methylisoxazole, nitrification was significantly inhibited. The consumption of ammonia nitrogen began to slow down, and the accumulation rate of nitrate nitrogen was also inhibited. For example, at 48 hours, the experimental group added with 0.05‰ w / v of methylisoxazole had an ammonia nitrogen consumption of 53.7% of the control group, and the accumulation of nitrate nitrogen was only 14.63% of the control group. Moreover, the effect of 5-methylisoxazole at a concentration of 0.5‰-0.05‰ w / v was equivalent. The above results show that in complex scenarios, 5-methylisoxazole at a concentration of less than 0.05‰ w / v can reduce the consumption of ammonia nitrogen and reduce the accumulation of nitrate nitrogen.

[0089] Example 7

[0090] The purpose of this implementation is to explore the specific application effect of 5-methylisoxazole in soil. Figure 10 The method was to take 50g of soil and supplement it with ammonium sulfate to 100mg / kg. The experimental group was treated with 0.1‰ w / v 5-methylisoxazole, while the control group was treated with the same amount of sterile water. Water was added regularly every day to maintain a moist state. Soil ammonia nitrogen levels were measured on the 0th and 14th days.

[0091] Depend on Figure 10 As can be seen, the experimental group treated with 5-methylisoxazole consumed significantly less ammonia nitrogen on the 14th day than the control group. On the 14th day, the control group consumed 27.66 mg / kg of ammonia nitrogen, while the experimental group consumed 45% of that. These results demonstrate that 5-methylisoxazole not only has a significant effect in water but also slows ammonia nitrogen loss in soil.

[0092] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

The application of 1.5-methylisoxazole nitrification inhibitor is characterized in that: Applications included in any of the following: (1) Application in inhibiting the nitrosation activity of ammonia oxidizing bacteria; (2) Application in inhibiting the nitrification activity of nitrite oxidizing bacteria.

2. The use of the 5-methylisoxazole nitrification inhibitor according to claim 1, characterized in that: The ammonia oxidizing bacteria include Nitrosomonas sp. CZ-4 or Nitrosomonas nitrosa strain SN-6.

3. The use of the 5-methylisoxazole nitrification inhibitor according to claim 1, characterized in that: The nitrite oxidizing bacteria is the enriched culture of Nitrobacter N-winogradskyi WY-8.

4. The method for inhibiting nitrification using the 5-methylisoxazole nitrification inhibitor according to any one of claims 1 to 3, characterized in that: include: The soil or water containing ammonia oxidizing bacteria and / or nitrite oxidizing bacteria is treated with 5-methylisoxazole, wherein the concentration of the 5-methylisoxazole is greater than or equal to 0.01‰ w / v, based on the volume of the soil or water being 100%.

5. The method for inhibiting nitrification using a 5-methylisoxazole nitrification inhibitor according to claim 4, wherein: The ammonia oxidizing bacteria include Nitrosomonas sp. CZ-4 and Nitrosomonas nitrosa strain SN-6.

6. The method for inhibiting nitrification using a 5-methylisoxazole nitrification inhibitor according to claim 4, wherein: The nitrite oxidizing bacteria is the enriched culture of Nitrobacter N-winogradskyi WY-8.

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