Application of 3-methyl-2-oxazolidinone as a nitration inhibitor and methods for inhibiting nitration.

Abstract

This invention provides the application of 3-methyl-2-oxazolidinone as a nitrification inhibitor and a method for inhibiting nitrification, relating to the technical field of soil remediation. Specifically, this invention relates to the application of 3-methyl-2-oxazolidinone as a nitrification inhibitor in soil treatment and / or wastewater treatment and in inhibiting the nitrification activity of ammonia-oxidizing bacteria. The method for inhibiting nitrification in this invention includes treating soil or water containing ammonium nitrogen and ammonia-oxidizing bacteria with 3-methyl-2-oxazolidinone. The concentration of 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v, based on 100% of the volume of the soil or water. This invention discovers the nitrification-inhibiting activity of 3-methyl-2-oxazolidinone, thereby enriching the selection of nitrification inhibitors.

Description

Technical Field

[0001] This invention relates to the technical field of soil remediation, and more specifically, to the application of 3-methyl-2-oxazolidinone as a nitrification inhibitor and methods for inhibiting nitrification. Background Technology

[0002] Nitrification refers to the process of reacting ammonium nitrogen (NH4) with nitrogen. + -N) is gradually oxidized into nitrate (NO3) through the metabolic activities of microorganisms. - The process of nitrification (NH4+) is a key step in the nitrogen cycle in the natural environment. Nitrification is mainly accomplished by two types of nitrifying microorganisms: ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). Ammonia-oxidizing bacteria convert ammonium nitrogen (NH4+) into nitrogen. + -N) is oxidized to nitrite (NO2) - Nitrification (nitrification) further oxidizes nitrite to nitrate by nitrite-oxidizing bacteria. Nitrification is widespread in ecosystems such as soil, freshwater, and marine environments, and has a significant impact on the ecological environment and agricultural production.

[0003] Excessive nitrification can lead to a series of environmental problems, such as groundwater pollution caused by nitrate leaching and reduced nitrogen fertilizer utilization. Therefore, effectively regulating the nitrification process to reduce adverse effects has become a crucial issue that urgently needs to be addressed in agriculture and environmental management. In recent years, research and development of novel 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 nitration inhibitor to enrich the selection of nitration inhibitors.

[0005] To address the above problems, this invention provides an application of 3-methyl-2-oxazolidinone as a nitration inhibitor and a method for inhibiting nitration.

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

[0007] As a second aspect, the present invention also relates to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrification 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 Nitrifying Monotrophus.

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

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

[0014] As a sixth aspect, the present invention also relates to a nitration inhibitor comprising 3-methyl-2-oxazolidinone.

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

[0016] The advantages of this invention compared to the prior art are:

[0017] This invention, through extensive research and experimentation, has discovered the nitrification inhibitory activity of 3-methyl-2-oxazolidinone. Testing showed that 3-methyl-2-oxazolidinone can inhibit the growth of ammonia-oxidizing bacteria, such as *Nitrosomonas*, at a low dose (0.1‰ w / v), thereby effectively inhibiting nitrification in soil or water, reducing nitrogen loss from farmland and N2O emissions from soil and water. Furthermore, at a concentration of 1‰ w / v, 3-methyl-2-oxazolidinone achieved a nitrification inhibition rate of over 90% against *Nitrosomonas*. In addition, the 3-methyl-2-oxazolidinone of this invention does not exhibit ecotoxicity or potential bioaccumulation, has minimal impact on other heterotrophic microorganisms in the soil, and demonstrates good environmental friendliness. Attached Figure Description

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

[0019] Figure 2 This is a graph showing the changes in nitrite concentration in the control group and each experimental group during the first four days in Example 2 of this invention. Detailed Implementation

[0020] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Although some embodiments of the present invention are shown in the drawings, it should be understood that the present invention can be implemented in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided to provide a more thorough and complete understanding of the present invention. It should be understood that the accompanying drawings and embodiments of the present invention are for illustrative purposes only 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 meaning as commonly understood by one of ordinary skill in the art to which this application pertains; the terminology used in the description of this application is for the purpose of describing particular embodiments only and is not intended to limit this application.

[0022] The term "comprising" and its variations as used herein are open-ended, meaning "including but not limited to"; the term "based on" means "at least partially based on"; the term "one embodiment" means "at least one embodiment"; the term "another embodiment" means "at least one additional embodiment"; the term "some embodiments" means "at least some embodiments"; and the term "optionally" means "optional embodiments". Definitions of other terms will be given in the description below. It should be noted that the concepts of "first," "second," etc., mentioned in this invention are used to distinguish different objects, not to describe a specific order or hierarchy. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined with "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this application, unless otherwise stated, "a plurality of" means two or more.

[0023] To enrich the selection of nitration inhibitors, this invention provides an application of 3-methyl-2-oxazolidinone as a nitration inhibitor and a method for inhibiting nitration.

[0024] In 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 nitrification activity of ammonia-oxidizing bacteria, and more specifically, to the use of 3-methyl-2-oxazolidinone in inhibiting the nitrification activity of nitrosomonas bacteria. The nitrosomonas bacteria may include Nitrosomonas sp. CZ-4 (accession number CCTCCNO: M 2018813) and / or Nitrosomonas nitrosastrain WH-1 (accession number CCTCCNO: M 2020526). Furthermore, 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 wastewater 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, the concentration of 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v based on 100% of the volume of the soil or water, and further preferably, the concentration of 3-methyl-2-oxazolidinone is 1‰ w / v.

[0026] This invention also relates to a nitrification inhibitor, the component of which includes 3-methyl-2-oxazolidinone. Specifically, 3-methyl-2-oxazolidinone can be used as the active ingredient of this nitrification inhibitor. Based on the nitrification-inhibiting activity of the nitrification inhibitor, this invention also relates to the application of the above-mentioned nitrification inhibitor in soil treatment and / or wastewater treatment.

[0027] This invention, through extensive research and experimentation, has discovered the nitrification inhibitory activity of 3-methyl-2-oxazolidinone. Testing showed that 3-methyl-2-oxazolidinone can inhibit the growth of ammonia-oxidizing bacteria, such as *Nitrosomonas*, at a low dose (0.1‰ w / v), thereby inhibiting the nitrification activity of these bacteria. Furthermore, at a concentration of 1‰ w / v, 3-methyl-2-oxazolidinone achieved a nitrification inhibition rate of over 90% against *Nitrosomonas*. In addition, the 3-methyl-2-oxazolidinone of this invention exhibits no ecotoxicity or potential bioaccumulation, and has minimal impact on other heterotrophic microorganisms in the soil, indicating its good environmental friendliness.

[0028] The present invention will be described in detail below with reference to specific embodiments:

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

[0030] Trace element solution: comprising the following components at the following 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 culture medium (AOB): FeSO4·7H2O, MgSO4·7H2O, K2HPO4, NaHCO3, CaCl2·2H2O, NaCl, NH4Cl, and trace element solution are dissolved in water. The concentrations of FeSO4·7H2O, MgSO4·7H2O, K2HPO4, NaHCO3, CaCl2·2H2O, NaCl, and NH4Cl are controlled at 0.4 g / L, 1.5 g / L, 0.5 g / L, 2.0 g / L, and 0.4 g / L, respectively. The volume concentration of the trace element solution is 1 mL / L. The pH of the ammonia-oxidizing bacteria culture medium is controlled at 7.5. The medium is then sterilized under high temperature and high pressure (121℃, 20 min) and stored for later use.

[0032] 3-Methyl-2-oxazolidinone stock solution (10000 mg / L): Weigh 1 g of 3-methyl-2-oxazolidinone (CAS: 19836-78-3, purchased from Adamas) using an analytical balance, dissolve it in a small amount of water, and transfer it to a 100 mL volumetric flask and dilute to volume to obtain the 3-methyl-2-oxazolidinone stock solution. Then, filter the 3-methyl-2-oxazolidinone stock solution through a 0.22 μm filter membrane for sterilization and store for later use.

[0033] It should be further noted that in the embodiments of the present invention, the ammonia nitrogen concentration detection method adopts the Nessler's reagent spectrophotometric method for the determination of ammonia nitrogen in water quality (HJ 535-2009); the nitrite nitrogen concentration is determined by the spectrophotometric method for the determination of nitrite nitrogen in water quality (GB / T 7493-1987).

[0034] Furthermore, in this embodiment of the invention, the nitrification inhibition rate is calculated using the following formula:

[0035]

[0036] Example 1

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

[0038] Take 1 mL of CZ-4 bacterial culture into fresh AOB medium and culture it in a shaker at 30℃ and 140 r / min. Take samples every 24 h to detect the concentration of ammonia nitrogen and nitrite nitrogen until the nitrite nitrogen concentration increases at a rate of 2 mg / L / h.

[0039] The 3-methyl-2-oxazolidinone stock solution was added to AOB medium containing CZ-4 bacterial culture at different concentrations, specifically according to the formulations in Table 1. The CZ-4 culture system without the addition of the 3-methyl-2-oxazolidinone stock solution served as the control group, while the other CZ-4 culture systems served as the experimental groups. The cultures were then incubated in a shaker at 30℃ and 140 rpm.

[0040] Table 1. Formulas of CZ-4 experimental systems for the control and experimental groups in Example 1.

[0041]

[0042]

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

[0044] Table 2. Nitrite concentrations in the control and experimental groups on day 4 of Example 1.

[0045]

[0046] From Table 2 and Figure 1 It was found that the nitrite concentration in the control group reached 84.24 mg / L on day 4, indicating good bacterial growth. After the addition of 3-methyl-2-oxazolidinone, the increase in nitrite concentration in the culture system was inhibited to varying degrees, indicating that the growth of CZ-4 was inhibited to varying degrees. Furthermore, the inhibitory effect decreased with decreasing concentration; at a concentration of 0.001‰ w / v, there was no inhibitory effect; the nitrification inhibition rates at 0.01‰ w / v and 0.1‰ w / v were 8.98% and 7.51%, respectively; and at a concentration of 1‰, the nitrification inhibition rate reached 95.58%.

[0047] Example 2

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

[0049] Take 1 mL of WH-1 bacterial culture into fresh AOB medium and culture it in a shaker at 30℃ and 140 r / min. Take samples every 24 h to detect the concentration of ammonia nitrogen and nitrite nitrogen until the nitrite nitrogen concentration increases at a rate of 2 mg / L / h.

[0050] The 3-methyl-2-oxazolidinone stock solution was added to AOB medium containing WH-1 bacterial culture at different concentrations, specifically according to the formulations in Table 3. The WH-1 culture system without the addition of the 3-methyl-2-oxazolidinone stock solution served as the control group, while the WH-1 culture systems at other concentrations served as the experimental groups. The cultures were then incubated in a shaker at 30℃ and 140 rpm, with daily samples taken to measure ammonia nitrogen and nitrite nitrogen concentrations.

[0051] Table 3. Formulation of the WH-1 experimental system in Example 2.

[0052]

[0053] The nitrite concentrations of each control group and each experimental group on days 1, 2, 3, and 4 are shown in Table 4. Figure 2 As shown:

[0054] Table 4. Nitrite 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 concentration in the control group began to rise sharply on the 3rd day, and then leveled off on the 4th day, entering a stable period. After the addition of 3-methyl-2-oxazolidinone, the increase of nitrite 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 inhibitory effect decreased with decreasing concentration.

[0057] From Table 4 and Figure 2 Further analysis revealed that the WH-1 experimental system was not inhibited at concentrations of 0.001‰ w / v and 0.01‰ w / v for 3-methyl-2-oxazolidinone; the WH-1 experimental system was slightly inhibited at a concentration of 0.1‰ for 3-methyl-2-oxazolidinone, with a nitration inhibition rate of 4.69%; and the nitration inhibition rate of the WH-1 experimental system reached 90.81% at a concentration of 1‰ for 3-methyl-2-oxazolidinone, with the inhibitory effect becoming apparent from the second day onwards.

[0058] While the present invention has been disclosed above, its scope of protection is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention, and all such changes and modifications will fall within the scope of protection of the present invention.

Claims

1. Use of 3-methyl-2-oxazolidinone for inhibiting the nitrification activity of Nitrosomonas.

2. Use according to claim 1, characterized in that, The Nitrosomonas includes Nitrosomonas sp. CZ-4 and / or Nitrosomonas nitrosa strain WH-1.

3. Use of 3-methyl-2-oxazolidinone for inhibiting the nitrification activity of Nitrosomonas in soil treatment and / or sewage treatment.

4. A method of inhibiting nitration, characterized by, Comprising: treating soil or water containing ammonium nitrogen and Nitrosomonas with 3-methyl-2-oxazolidinone.

5. The method of inhibiting nitration of claim 4, wherein, The concentration of the 3-methyl-2-oxazolidinone is not less than 0.1‰ w / v, based on 100% of the volume of the soil or water.

6. The method of inhibiting nitration of claim 4, wherein, The concentration of the 3-methyl-2-oxazolidinone is 1‰ w / v, based on 100% of the volume of the soil or water.

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