Application of 3-methyl-2-oxazolidinone as nitrification inhibitor and method for inhibiting nitrification

Abstract

The invention provides application of 3-methyl-2-oxazolidinone as a nitrification inhibitor and a nitrification inhibition method, and relates to the technical field of soil treatment. Specifically, the invention relates to application of 3-methyl-2-oxazolidinone as a nitrification inhibitor in soil treatment and / or sewage treatment and inhibition of nitrosation activity of ammonia oxidizing bacteria. The method for inhibiting nitrification comprises the step of treating soil or water containing ammonium nitrogen and ammonia oxidizing bacteria by using 3-methyl-2-oxazolidinone. The concentration of the 3-methyl-2-oxazolidinone is not less than 0.01% w / v by taking the volume of the soil or the water body as 100%. The nitrification inhibition activity of the 3-methyl-2-oxazolidinone is found, so that the variety selection of nitrification inhibitors is enriched.

Description

Technical Field

[0001] The invention relates to the technical field of soil treatment, and in particular to application of 3-methyl-2-oxazolidinone as a nitrification inhibitor and a method for inhibiting nitrification. Background Art

[0002] Nitrification refers to the process of ammonium nitrogen (NH4 + -N) is gradually oxidized to nitrate (NO3 - -N) is one of the key links in the nitrogen cycle in the natural environment. Nitrification is mainly completed by two types of nitrifying microorganisms: ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). Ammonia-oxidizing bacteria convert ammonium nitrogen (NH4 + -N) is oxidized to nitrite (NO2 - -N), and nitrite oxidizing bacteria further oxidize nitrite to nitrate. Nitrification is widely present in ecosystems such as soil, freshwater and ocean, and has an important impact on the ecological environment and agricultural production.

[0003] Excessive nitrification may bring about a series of environmental problems, such as groundwater pollution caused by nitrate leaching and reduced nitrogen fertilizer utilization. Therefore, how to effectively regulate the nitrification process to reduce adverse effects has become an important issue that needs to be solved in the fields of agriculture and environmental management. In recent years, research and development of new nitrification inhibitors to selectively inhibit the activity of ammonia oxidizing bacteria has become an important direction for achieving efficient nitrogen utilization and environmental protection. Summary of the invention

[0004] The present invention aims to develop a novel nitrification inhibitor to enrich the selection of nitrification inhibitors.

[0005] In order to solve the above problems, the present invention provides an application of 3-methyl-2-oxazolidinone as a nitrification inhibitor and a method for inhibiting nitrification.

[0006] As a first aspect, the present invention relates to the use of 3-methyl-2-oxazolidinone as a nitrification inhibitor.

[0007] As a second aspect, the present invention also relates to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of ammonia oxidizing bacteria.

[0008] As a third aspect, the present invention also relates to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of Nitrosomonas.

[0009] Optionally, the nitrosomonas includes Nitrosomonas sp. CZ-4 and / or Nitrosomonasnitrosa strain WH-1.

[0010] As a fourth aspect, the present invention also relates to the use of 3-methyl-2-oxazolidinone in soil treatment and / or sewage treatment.

[0011] As a fifth aspect, the present invention also relates to a method for inhibiting nitrification, comprising: treating soil or water containing ammonium nitrogen and ammonia oxidizing bacteria with 3-methyl-2-oxazolidinone.

[0012] Optionally, based on the volume of the soil or water body as 100%, the concentration of the 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v.

[0013] Optionally, based on the volume of the soil or water body being 100%, the concentration of the 3-methyl-2-oxazolidinone is 1‰ w / v.

[0014] As a sixth aspect, the present invention also relates to a nitrification inhibitor, wherein the component of the nitrification inhibitor includes 3-methyl-2-oxazolidinone.

[0015] As a seventh aspect, the present invention also relates to the use of the above-mentioned nitrification inhibitor in soil treatment and / or sewage treatment.

[0016] The beneficial effects of the present invention compared to the prior art are:

[0017] The present invention has found the nitrification inhibition activity of 3-methyl-2-oxazolidinone through a large number of studies and experiments. After testing, 3-methyl-2-oxazolidinone can inhibit the growth of ammonia oxidizing bacteria exemplified by Nitrosomonas at a lower dose (0.1‰w / v), thereby efficiently inhibiting nitrification in soil or water, reducing farmland nitrogen loss and N2O emissions in soil and water. And at a concentration of 1‰w / v, the nitrification inhibition rate of 3-methyl-2-oxazolidinone to Nitrosomonas reaches more than 90%. In addition, the 3-methyl-2-oxazolidinone of the embodiment of the present invention does not have ecotoxicity and potential bioaccumulation, has less influence on other heterotrophic microorganisms in the soil, and has good environmental friendliness. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a graph showing changes in nitrite nitrogen concentrations in the control group and each experimental group on the 4th day in Example 1 of the present invention;

[0019] Figure 2 This is a graph showing changes in nitrite nitrogen concentration in the control group and each experimental group in Example 2 of the present invention during the first 4 days. DETAILED DESCRIPTION

[0020] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. Although certain embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be interpreted as being limited to the embodiments described herein. On the contrary, these embodiments are provided to provide a more thorough and complete understanding of the present invention. It should be understood that the drawings and embodiments of the present invention are only for exemplary purposes and are not intended to limit the scope of protection of the present invention.

[0021] Unless otherwise defined, all technical and scientific terms used in this application have the same meanings as those commonly understood by those skilled in the art to which this application belongs; the terms used in the specification of this application are only for the purpose of describing specific implementation methods and are not intended to limit this application;

[0022] The term "including" and its variations used in this article are open inclusions, that is, "including but not limited to"; the term "based on" is "based at least in part on"; the term "one embodiment" means "at least one embodiment"; the term "another embodiment" means "at least one other embodiment"; the term "some embodiments" means "at least some embodiments"; the term "optionally" means "optional embodiments". The relevant definitions of other terms will be given in the following description. It should be noted that the concepts of "first" and "second" mentioned in the present invention are used to distinguish different objects, rather than to describe a specific order or a primary and secondary relationship. In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of this application, unless otherwise specified, the meaning of "multiple" is two or more.

[0023] In order to enrich the types of nitrification inhibitors, the embodiments of the present invention provide a use of 3-methyl-2-oxazolidinone as a nitrification inhibitor and a method for inhibiting nitrification.

[0024] As a first aspect, the present invention relates to the use of 3-methyl-2-oxazolidinone as a nitrification inhibitor, specifically, to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of ammonia oxidizing bacteria, and more specifically, to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of Nitrosomonas. Wherein, Nitrosomonas may include Nitrosomonas sp.CZ-4 (preservation number is CCTCCNO: M 2018813) and / or Nitrosomonas nitrosastrain WH-1 (preservation number is CCTCCNO: M 2020526). Further, based on the activity of 3-methyl-2-oxazolidinone in inhibiting nitrification, the present invention also relates to the use of 3-methyl-2-oxazolidinone in soil treatment and / or sewage treatment.

[0025] The present invention also specifically relates to a method for inhibiting nitrification, comprising: treating soil or water containing ammonium nitrogen and ammonia oxidizing bacteria with 3-methyl-2-oxazolidinone. Specifically, based on 100% volume of the soil or water, the concentration of 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v, and further, the concentration of 3-methyl-2-oxazolidinone is preferably 1‰ w / v.

[0026] The present invention also relates to a nitrification inhibitor, wherein the component of the nitrification inhibitor includes 3-methyl-2-oxazolidinone. Specifically, 3-methyl-2-oxazolidinone can be used as the active component of the nitrification inhibitor. Based on the nitrification inhibition activity of the nitrification inhibitor, the present invention also relates to the use of the nitrification inhibitor in soil treatment and / or sewage treatment.

[0027] The present invention has found the nitrification inhibition activity of 3-methyl-2-oxazolidinone through a large number of studies and experiments. After testing, 3-methyl-2-oxazolidinone can inhibit the growth of ammonia oxidizing bacteria exemplified by Nitrosomonas at a lower dose (0.1‰w / v), thereby inhibiting the nitrosation activity of ammonia oxidizing bacteria. And at a concentration of 1‰w / v, the nitrification inhibition rate of 3-methyl-2-oxazolidinone to Nitrosomonas reaches more than 90%. In addition, the 3-methyl-2-oxazolidinone of the embodiment of the present invention does not have ecotoxicity and potential bioaccumulation, and has less influence on other heterotrophic microorganisms in the soil, indicating that it has good environmental friendliness.

[0028] The present invention is described in detail below in conjunction with specific embodiments:

[0029] It should be noted that the solutions and preparation methods used in the following examples are as follows:

[0030] Trace element solution: includes the following components in mass concentrations: 0.5 g / L EDTA, 0.075 g / L CuSO4·5H2O, 0.3 g / L ZnSO4·7H2O, 0.375 g / L CoCl2·6H2O, 0.3 g / L MnCl2·2H2O, 0.014 g / L H3BO4 and 0.22 g / L NaMoO4·2H2O.

[0031] Ammonia oxidizing bacteria medium (AOB): FeSO4·7H2O, MgSO4·7H2O, K2HPO4, NaHCO3, CaCl2·2H2O, NaCl, NH4Cl and trace element solution are dissolved in water. The mass concentration of FeSO4·7H2O, MgSO4·7H2O, K2HPO4, NaHCO3, NH4Cl and trace element solution are controlled to be 0.4 g / L, 0.5 g / L, 0.1 g / L, 1.5 g / L, 0.5 g / L, 2.0 g / L, 0.4 g / L and 1 mL / L, respectively. The pH of the ammonia oxidizing bacteria medium is controlled to be 7.5, and then sterilized at high temperature and high pressure (121°C, 20 min) and stored for later use.

[0032] 3-Methyl-2-oxazolidinone mother solution (10000 mg / L): 1 g of 3-methyl-2-oxazolidinone (CAS: 19836-78-3, purchased from Adamas / Titan) was weighed using an analytical balance, and a small amount of water was added to dissolve the mixture, and the mixture was transferred to a 100 mL volumetric flask to obtain a 3-methyl-2-oxazolidinone mother solution. The 3-methyl-2-oxazolidinone mother solution was then filtered and sterilized using a 0.22 μm filter membrane and stored for later use.

[0033] It should be further explained that in the embodiments of the present invention, the ammonia nitrogen concentration detection method adopts the "Water Quality - Determination of Ammonia Nitrogen - Nessler's Reagent Spectrophotometric Method" (HJ 535-2009); the nitrite nitrogen concentration is determined by the "Water Quality - Determination of Nitrite Nitrogen - Spectrophotometric Method" (GB / T 7493-1987).

[0034] It should be further explained that in the embodiment of the present invention, the nitrification inhibition rate is calculated using the following formula:

[0035]

[0036] Example 1

[0037] This example relates to an experiment to evaluate the nitrification inhibition effect of 3-methyl-2-oxazolidinone on the bacterial species Nitrosomonas sp. CZ-4 (hereinafter referred to as CZ-4).

[0038] Take 1 mL of CZ-4 bacterial liquid into fresh AOB medium and culture it in a shaking incubator at 30°C and 140 r / min. Take samples every 24 hours to test the ammonia nitrogen concentration and nitrite nitrogen concentration until the growth rate of nitrite nitrogen concentration reaches 2 mg / L / h.

[0039] The 3-methyl-2-oxazolidinone mother solution was added to the AOB medium containing the CZ-4 bacterial solution at different concentrations, and the CZ-4 culture system was prepared according to the formula in Table 1. Among them, the CZ-4 culture system without the addition of the 3-methyl-2-oxazolidinone mother solution was used as the control group, and the other CZ-4 culture systems were used as the experimental group. Then, the culture was carried out in a shaking table at 30°C and 140r / min.

[0040] Table 1 CZ-4 experimental system formula for the control group and experimental group in Example 1

[0041]

[0042]

[0043] After 4 days, samples were taken to measure the concentrations of ammonia nitrogen and nitrite nitrogen in the control group and each experimental group. The concentrations of nitrite nitrogen in each control group and each experimental group on the 4th day are shown in Table 2 and Figure 1 As shown:

[0044] Table 2 Nitrite nitrogen concentrations of the control group and the experimental group in Example 1 on the 4th day

[0045]

[0046] From Table 2 and Figure 1 It can be seen that the nitrite nitrogen concentration of the control group reached 84.24 mg / L on the 4th day, indicating that the bacteria grew well. After the addition of 3-methyl-2-oxazolidinone, the increase in nitrite nitrogen concentration in the culture system was inhibited to varying degrees, 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 0.001‰w / v, there was no inhibition. The nitrification inhibition rates at 0.01‰w / v and 0.1‰w / v were 8.98% and 7.51% respectively. At a concentration of 1‰, the nitrification inhibition rate reached 95.58%.

[0047] Example 2

[0048] This example relates to an experiment to evaluate the nitrification inhibition effect of 3-methyl-2-oxazolidinone on the bacterial strain Nitrosomonas nitrosa WH-1 (hereinafter referred to as WH-1).

[0049] Take 1 mL of WH-1 bacterial solution and add it to fresh AOB medium. Culture it in a shaking incubator at 30°C and 140 r / min. Take samples every 24 hours to test the ammonia nitrogen concentration and nitrite nitrogen concentration until the growth rate of nitrite nitrogen concentration reaches 2 mg / L / h.

[0050] The mother solution of 3-methyl-2-oxazolidinone was added to the AOB medium containing the WH-1 bacterial solution at different concentrations, and the WH-1 culture system was prepared according to the formula in Table 3. Among them, the WH-1 culture system without the mother solution of 3-methyl-2-oxazolidinone was used as the control group, and the WH-1 culture system at other concentrations was used as the experimental group. Then, the culture was carried out in a shaking table at 30°C and 140r / min, and samples were taken every day to measure the concentrations of ammonia nitrogen and nitrite nitrogen.

[0051] Table 3 WH-1 experimental system formula of the neutralization experimental group in Example 2

[0052]

[0053] The nitrite nitrogen concentrations of the control group and the experimental group on d1, d2, d3 and d4 are shown in Table 4 and Figure 2 As shown:

[0054] Table 4 Nitrite nitrogen concentrations in the control group and each experimental group in Example 2

[0055]

[0056] From Table 4 and Figure 2 It can be seen that the nitrite nitrogen concentration in the control group began to rise sharply on the 3rd day, tended to be flat on the 4th day, and entered a stable period. After adding 3-methyl-2-oxazolidinone, the increase of nitrite nitrogen in the WH-1 culture system was inhibited to varying degrees, indicating that the growth of WH-1 was inhibited to varying degrees, and the inhibition ability decreased with the decrease of concentration.

[0057] From Table 4 and Figure 2 It can be further seen that the WH-1 experimental system was not inhibited at concentrations of 0.001‰ w / v and 0.01‰ w / v of 3-methyl-2-oxazolidinone; the WH-1 experimental system was weakly inhibited at a concentration of 0.1‰ of 3-methyl-2-oxazolidinone, and the nitrification inhibition rate was 4.69%; at a concentration of 1‰ of 3-methyl-2-oxazolidinone, the nitrification inhibition rate of the WH-1 experimental system reached 90.81%, and the inhibition began to show obvious effects on the second day.

[0058] Although the present invention is disclosed as above, the protection scope of the present invention is not limited thereto. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. Application of 3-methyl-2-oxazolidinone as a nitrification inhibitor. Application of 2.3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of ammonia oxidizing bacteria.

3. Application of 3-methyl-2-oxazolidinone in inhibiting the nitrosation activity of Nitrosomonas.

4. The use according to claim 3, characterized in that: The nitrosomonas include Nitrosomonas sp. CZ-4 and / or Nitrosomonas nitrosa strain WH-1.

5. Application of 3-methyl-2-oxazolidinone in soil treatment and / or sewage treatment.

6. A method for inhibiting nitrification, characterized in that: include: Use 3-methyl-2-oxazolidinone to treat soil or water containing ammonium nitrogen and ammonia oxidizing bacteria.

7. The method for inhibiting nitrification according to claim 6, characterized in that: Based on the volume of the soil or water body as 100%, the concentration of the 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v.

8. The method for inhibiting nitrification according to claim 6, characterized in that: Based on the volume of the soil or water body being 100%, the concentration of the 3-methyl-2-oxazolidinone is 1‰ w / v.

9. A nitrification inhibitor, characterized in that: The nitrification inhibitor component includes 3-methyl-2-oxazolidinone.

10. Use of the nitrification inhibitor according to claim 9 in soil treatment and / or sewage treatment.

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