CN102520080A - Method for detecting anammox reaction rate in water body sediment and/or soil - Google Patents
Method for detecting anammox reaction rate in water body sediment and/or soil Download PDFInfo
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Abstract
The invention discloses a method for detecting an anammox reaction rate in water body sediment and/or soil. The method comprises the following steps of: (1), culturing a to-be-detected sample under an anaerobic condition to obtain a pre-treated sample, and dividing the pre-treated sample into three parts, respectively the pre-treated sample a, the pre-treated sample b and the pre-treated sample c; (2), reacting by adding (15NH4)2SO4 into the pre-treated sample a, and detecting that no nitrogen is generated through the reaction; (3), reacting by adding (15NH4)2SO4 and Na14NO3 into the pre-treated sample b, and detecting that 29N2 is generated through the reaction; and (4), reacting by adding (14NH4)2SO4 and Na15NO3 into the pre-treated sample c, detecting amounts of 28N2, 29N2 and 30N2 generated through the reaction under a certain time point, thereby obtaining the total amount Atotal of nitrogen generated through an anammox reaction in the to-be-detected sample according to the formula (a) described in the specification, and obtaining the anammox reaction rate in the to-be-detected sample under the certain time point according to the obtained total amount Atotal of the nitrogen. The method provided by the invention is scientific, accurate and effective and can be widely used for researching anammox activities and denitrification contributions in natural water body sediment and soil, thus, development of anammox research is promoted.
Description
Technical field
The present invention relates to a kind of method that detects Anammox reaction rate in water body deposit and/or the soil.
Background technology
The discovery of Anammox reaction is the important breakthrough of nearly ten years environmental sciences, makes people re-recognize the rate-limiting step in this nitrogen cycle of ammoxidation process, has also changed the understanding of people to global nitrogen cycle flux.The traditional understanding that the nitrogen migration transforms is: ammonia nitrogen under the catalytic condition of ammonia monooxygenase (amoA), is oxidized to nitrite by ammonia oxidizing bacteria (AOB), begins the biological nitrogen circulation thus.And current research is found, the oxidation of ammonia is not only carried out under aerobic conditions, can also under the effect of anaerobic ammonia oxidizing bacteria, under the condition of anaerobic/anoxic, be generated nitrogen by nitrite-oxidizing, and this procedure definition is Anammox (anammox).
Anaerobic ammonia oxidizing bacteria extensively exists at nature, verifies at present, and 24%～67% nitrogen is accomplished by the Anammox reaction in the marine ecosystems.But at present the research about the Anammox reaction is limited in the marine ecosystems more, in the research on top layer, continent also seldom.Anammox is reflected at the active of occurring in nature and is question of common concern to the contribution amount of denitrification process, and how accurately measuring occurring in nature Anammox reaction rate and itself and denitrification process are distinguished still is a difficult problem.
The case of Anammox reaction rate is less in the present domestic mensuration physical environment.The domestic researcher of having adds NH in anaerobic ammoxidation activity mud
4 +, NO
2 -Carry out the water bath with thermostatic control shaking table with trace element and cultivate, with beginning experiment behind the nitrogen stripping mud 30min.During sampling, stop shaking table and shake and make sludge settling, feed nitrogen supernatant is discharged.In the experimentation each hour sampling and testing, through measuring each substrate concentration in supernatant constantly, draw the substrate concentration change curve and obtain maximum slope and be maximum Anammox speed.Because NH
4 +And NO
2 -Change in concentration receive the influence of other factors such as denitrification etc., so the Anammox reaction rate that the method is measured is inaccurate, and can not react the contribution of Anammox in denitrification process.Isotope tracer technique can make a distinction Anammox and anti-nitration reaction, records both speed respectively, and this thinking obtains Preliminary Applications in oceanic sediment and the research of water Anammox.This method main thought is following:
Anti-nitration reaction be tradition nitrated-part of denitrification process; Under scarce/anaerobic condition, carry out, 4 pilot process are by the catalysis completion respectively of nitre nitrogen reductase (NAR), nitrite nitrogen reductase (NIR), nitric oxide reduction enzyme (NO) and nitrous oxide reductase (NOS).Course of reaction is suc as formula shown in (1):
Anammox be with ammonia nitrogen and nitrite nitrogen respectively as electron donor and acceptor, under anaerobic directly generate the process of nitrogen by the anaerobic ammonia oxidizing bacteria of autotrophy, azanol and buzane are main intermediate products.Concrete course of reaction is seen formula (2):
Can find out that by the course of reaction formula under the environment of anoxic or anaerobism, anti-nitration reaction and Anammox reaction all produce nitrogen, but mechanism there are differences.Anti-nitration reaction produces nitrogen by nitre nitrogen through a series of intermediate reactions, and two nitrogen-atoms of nitrogen all come from NO
3-The Anammox reaction produces nitrogen as electron donor and acceptor respectively by ammonia nitrogen and nitrite nitrogen, and nitrogen molecule is by NH
4 +And NO
2 -Respectively provide a nitrogen-atoms to form.When carrying out
15During the isotopic spike of N, the difference of anti-nitration reaction and Anammox reaction can be through producing in the nitrogen
28N
2,
29N
2With
30N
2Composition be able to embody.Through calculating the nitrogen amount that denitrification and Anammox produce separately, denitrification and Anammox speed obtain through the recurrence to the time of nitrogen gas generation amount separately.
The mensuration that isotope tracer technique is applied to Anammox reaction rate in the natural ecosystems accurately and reliably; And can Anammox reaction and anti-nitration reaction be distinguished; Be reflected in the biological denitrification process and contribute thereby manifest Anammox, the method for simple possible is provided for the research of this discovery of Anammox reaction.But because oceanic sediment and water body deposit, soil property have than big-difference at aspects such as the content of organic matter, nitrate contents, so this method is inappropriate in the mensuration that is applied directly to water body deposit and soil anaerobic ammoxidation reaction speed.Do the improvement of science to having the method for measuring the Anammox reaction rate now, the mensuration with more being applicable to water body deposit, soil anaerobic ammoxidation reaction speed improves determination efficiency simultaneously.
Summary of the invention
The purpose of this invention is to provide a kind of method that detects Anammox reaction rate in water body deposit and/or the soil, accurate, feasible, high-efficiency method are provided for probing into the contribution that Anammox is reflected in the nature biotechnology denitrification process.
A kind of method that detects Anammox reaction rate in water body deposit and/or the soil provided by the invention comprises the steps:
(1) detected sample is obtained pretreatment sample after cultivating under the oxygen free condition; Said pretreatment sample is divided into three parts, is respectively pretreatment sample a, pretreatment sample b and pretreatment sample c;
(2) in said pretreatment sample a, add (
15NH
4)
2SO
4React, do not have nitrogen to generate through detecting said reaction;
(3) in said pretreatment sample b, add (
15NH
4)
2SO
4And NaNO
3React, generate through detecting said reaction
29N
2
(4) in said pretreatment sample c, add (NH
4)
2SO
4And Na
15NO
3React, detect that under certain time point said reaction generates
28N
2,
29N
2With
30N
2Amount, the nitrogen total amount A that Anammox reaction produces in the then said testing sample
Total(a) promptly gets according to formula; By the nitrogen total amount A that obtains
TotalCan obtain under certain time point Anammox reaction rate in the detected sample;
A
total＝F
N -1×[P
29+2×(1-F
N -1)×P
30] (a)
In the following formula, F
NExpression Na
15NO
3In
15The abundance of N, P
29Reaction described in the expression step (4) generates
29N
2Total amount, P
30Reaction described in the expression step (4) generates
30N
2Total amount.
In the above-mentioned method, oxygen free condition described in the step (1) can be and adds ammonium water through the nitrogen aeration.
In the above-mentioned method; The time of cultivating described in the step (1) can be 20 hours～and 24 hours; Specifically can be 20 hours or 24 hours, the present invention adopts isotope tracer technique to detect Anammox reaction rate in water body deposit and/or the soil, so remaining NO in the sample
3 -Can handle exerting an influence to follow-up tagging, should thoroughly remove; Can make NO in the said incubation step
3 -Be removed through anti-nitration reaction: under anaerobic, NO
3 -With organic matter respectively as electron accepter and electron donor generation anti-nitration reaction, finally generate nitrogen.Because the content of organic matter is starkly lower than water body deposit and soil in the marine bottom sediment; And lower organic concentration can suppress carrying out smoothly of anti-nitration reaction; So when measuring in the marine bottom sediment Anammox reaction rate, it is longer to cultivate pretreatment time, usually more than 30 hours.In addition, nitrate content is higher than water body deposit and soil in the marine bottom sediment, so the time that nitrate consumption in the marine bottom sediment is needed has fully further increased.For water body deposit and soil, less nitrate anti-nitration reaction under the condition that more organic matter exists can carry out smoothly, can reach the remaining NO of removal so shorten the cultivation pretreatment time
3 -Purpose, improved conventional efficient again.
In the above-mentioned method, the temperature of cultivating described in the step (1) can be 22～25 ℃, specifically can be 22 ℃ or 25 ℃.
In the above-mentioned method, the temperature of reaction can be 22～25 ℃ described in step (2), step (3) and the step (4), specifically can be 22 ℃ or 25 ℃.
In the method provided by the invention, the said reaction of step (2) has been removed remaining NO in water body deposit and/or the pedotheque as negative control in order to confirm this reaction system
3 -Interference; The said reaction of step (3) is as positive control, in order to confirm to exist in this reaction system the Anammox reaction and
30N
2Only produce by anti-nitration reaction; Produce Anammox reaction and anti-nitration reaction in the step (4), can produce
28N
2,
29N
2With
30N
2, wherein the nitrogen of Anammox reaction generation is by NH
4 +And NO
3 -Respectively provide a nitrogen-atoms to form; The nitrogen that Anammox reaction in this step produces does
28N
2With
29N
2, the amount of the two is by adding NO
3 -In
15NO
3 -Ratio decision, can get by formula (3) calculating,
A
28＝A
total×(1-F
N)，A
29＝A
total×F
N?(3)
Wherein, A
28And A
29Represent respectively that Anammox reaction produces
28N
2With
29N
2Amount, F
NFor
15NO
3 -In
15The abundance of N.
Two nitrogen-atoms that anti-nitration reaction in this step produces nitrogen all come from NO
3 -Therefore, the nitrogen that produces of the anti-nitration reaction of this step should for
28N
2,
29N
2With
30N
2, their amount can be calculated and can get by formula (4),
D
28＝D
total×(1-F
N)
2，D
29＝D
total×2×(1-F
N)×F
N，D
30＝D
total×F
N 2?(4)
Wherein, D
28, D
29And D
30Represent anti-nitration reaction respectively
28N
2,
29N
2With
30N
2Amount, D
TotalThe nitrogen total amount that the expression anti-nitration reaction produces,
To detect and produce in the step (4)
29N
2With
30N
2Total amount be labeled as P
29And P
30, and D
30=P
30, convolution (3) and (4) can get:
D
total＝F
30×F
N -2 (5)
D
29＝P
30×2×(1-F
N)×F
N -1 (6)
A
29＝P
29-D
29＝P
29-P
30×2×(1-F
N)×F
N -1 (7)
A
28＝A
29×F
N -1×(1-F
N) (8)
Superposing type (7) and (8) promptly get formula (a).
By formula (5) and (a) obtain D
TotalAnd A
TotalBe the nitrogen gas generation amount of anti-nitration reaction and Anammox reaction in each sample, obtain to obtain anti-nitration reaction and Anammox reaction rate through the recurrence of nitrogen gas generation amount separately to the time.
In the method provided by the invention,, therefore do not consider the influence of carbon source loss because anaerobic ammonia oxidizing bacteria is an autotrophic bacteria; In addition; Also having ignored the isotope fractionation phenomenon of the different nucleic of nitrogen element in reaction in isotope analysis and the computation process (refers between the different nucleic of isotope because mass discrepancy; A kind of nucleic is easier to the phenomenon that transformed through physics, chemistry and biological chemistry action with respect to another kind of nucleic), can this difference be included into systematic error.
The invention provides a kind of method that adopts isotope tracer technique to detect Anammox reaction rate in water body deposit and/or the soil; This methodological science, accurate, efficient; Can be widely used in the research of anaerobic ammoxidation activity and denitrogenation contribution in nature water body deposit, the soil, thereby promote the development of Anammox research.
Description of drawings
Fig. 1 is nitrogen amount and the time relation figure that produces in the step among the embodiment 1 (2).
Fig. 2 is nitrogen amount and the time relation figure that produces in step among the embodiment 1 (3) and (4).
Fig. 3 is nitrogen amount and the time relation figure that produces in the step among the embodiment 2 (2).
Fig. 4 is nitrogen amount and the time relation figure that produces in step among the embodiment 2 (3) and (4).
Embodiment
Employed experimental technique is conventional method like no specified otherwise among the following embodiment.
Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.
The speed of Anammox reaction in embodiment 1, the mensuration Baiyang Lake sediment
(1) the Baiyang Lake sediment about 2.0ml is put into 12.6mL column vial (Labco; UK) in; Add simultaneously when two beaded glasses are beneficial to react and shake up; Fill with then through the abundant aeration of nitrogen add ammonium tap water (ammonia nitrogen concentration is 5mg/L), make it form oxygen-free environment, under 22 ℃ of room temperatures on rotating generator rotating and culturing 20 hours to remove NO wherein
3-, obtain pretreated sediment, should be divided into three parts by pretreated sediment, pre-service sediment a, pre-service sediment b and pre-service sediment c.
(2) in pre-service sediment a, add (
15NH
4)
2SO
4, wherein,
15The abundance F of N
NBe 98.2%, it is reacted under 22 ℃ and take a sample, in each time point sample, inject 200 μ L 7M ZnCl at each time point
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is as shown in Figure 1; Can know by Fig. 1, not see any
29N
2With
30N
2Gas is explained through the processing in step (1), remaining NO in the sediment
3 -Exhaust fully, can not exert an influence subsequent treatment.
(3) in pre-service sediment b, add (
15NH
4)
2SO
4And NaNO
3, wherein,
15The abundance of N is 98.2%, it is reacted under 22 ℃ and takes a sample at each time point, in each time point sample, injects 200 μ L 7MZnCl
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is shown in Fig. 2 (a); Can know by Fig. 2 (a),
29N
2Amount prolong in time and increase, explain that Anammox is reflected at and exist really in this processing form and have certain activity, but
30N
2In this processing form, do not detect explanation
30N
2Only produce by anti-nitration reaction.
(4) in pre-service sediment c, add (NH
4)
2SO
4And Na
15NO
3, wherein,
15The abundance F of N
NBe 99.29%, it is reacted under 22 ℃ and take a sample, in each time point sample, inject 200 μ L7M ZnCl at each time point
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is shown in Fig. 2 (b); Can know by Fig. 2 (b),
29N
2With
30N
2Amount all prolong in time and increase, explained anti-nitration reaction and Anammox the reaction be present in simultaneously in this processing form.
Respectively can be by formula (5) with (a) calculate in this step total nitrogen generation that anti-nitration reaction and Anammox react under each time point.Total nitrogen generation of two processes was returned respectively the time, draw the speed of denitrification process and anaerobic ammonium oxidation process, be respectively 18.96nmol N g
-1h
-1With 1.25nmol N g
-1h
-1Hence one can see that, and the activity of being tried Anammox in the sediment sample is the nitrogen that every gram sediment per hour generates 0.6 nanomole, and anaerobic ammonium oxidation process then is 6.2% to the contribution of total nitrogen generation.
The speed of Anammox reaction in embodiment 2, mensuration and rice field, Jiaxing wetland soil
(1) with putting into 12.6mL column vial (Labco with rice field, Jiaxing wetland soil about 2.0ml; UK) in; Add simultaneously when two beaded glasses are beneficial to react and shake up; Fill with then through the abundant aeration of nitrogen add ammonium tap water (ammonia nitrogen concentration is 5mg/L), make it form oxygen-free environment, under 25 ℃ on rotating generator rotating and culturing 24 hours to remove NO wherein
3-, obtain pretreated soil, should be divided into three parts by pretreated soil, locate soil a, pre-service soil b and pre-service soil c in advance.
(2) in pre-service soil a, add (
15NH
4)
2SO
4, wherein,
15The abundance F of N
NBe 98.2%, it is reacted under 25 ℃ and take a sample, in each time point sample, inject 200 μ L 7M ZnCl at each time point
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is as shown in Figure 3; Can know by Fig. 3, not see any
29N
2With
30N
2Gas is explained through the processing in step (1), remaining NO in the soil
3 -Exhaust fully, can not exert an influence subsequent treatment.
(3) in pre-service soil b, add (
15NH
4)
2SO
4And NaNO
3, wherein,
15The abundance of N is 98.2%, it is reacted under 25 ℃ and takes a sample at each time point, in each time point sample, injects 200 μ L 7MZnCl
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is shown in Fig. 4 (a); Can know by Fig. 4 (a),
29N
2Amount prolong in time and increase, explain that Anammox is reflected at and exist really in this processing form and have certain activity, but
30N
2In this processing form, do not detect explanation
30N
2Only produce by anti-nitration reaction.
(4) in pre-service soil c, add (NH
4)
2SO
4And Na
15NO
3, wherein,
15The abundance F of N
NBe 99.29%, it is reacted under 25 ℃ and take a sample, in each time point sample, inject 200 μ L 7MZnCl at each time point
2Solution is with cessation reaction; Be anaerobic environment in the whole process vial; In the vial of accomplishing reaction, fill helium and extract the 2mL supernatant out, importing is full of in the Labco vial of helium.Acutely rock the vial that supernatant and helium are housed, make the nitrogen in the supernatant between the gas-liquid two-phase of vial, reach balance; Through gas chromatography-isotope-ratio mass spectrometer (Gasbench-MAT253), measure in the gas phase
29N
2With
30N
2Concentration, this reaction in gained
29N
2With
30N
2Gas generated time dependent situation is shown in Fig. 4 (b); Can know by Fig. 4 (b),
29N
2With
30N
2Amount all prolong in time and increase, explained anti-nitration reaction and Anammox the reaction be present in simultaneously in this processing form.
Respectively can be by formula (5) with (a) calculate in this step total nitrogen generation that anti-nitration reaction and Anammox react under each time point.Total nitrogen generation of two processes was returned respectively the time, draw the speed of denitrification process and anaerobic ammonium oxidation process, be respectively 7.16nmol N g
-1h
-1With 1.45nmol N g
-1h
-1Hence one can see that, and the activity of being tried Anammox in the sediment sample is the nitrogen that every gram sediment per hour generates 0.7 nanomole, and anaerobic ammonium oxidation process then is 17.2% to the contribution of total nitrogen generation.
Claims (5)
1. a method that detects Anammox reaction rate in water body deposit and/or the soil comprises the steps:
(1) detected sample is cultivated under oxygen free condition obtained pretreatment sample; Said pretreatment sample is divided into three parts, is respectively pretreatment sample a, pretreatment sample b and pretreatment sample c;
(2) in said pretreatment sample a, add (
15NH
4)
2SO
4React, do not have nitrogen to generate through detecting said reaction;
(3) in said pretreatment sample b, add (
15NH
4)
2SO
4And NaNO
3React, generate through detecting said reaction
29N
2
(4) in said pretreatment sample c, add (NH
4)
2SO
4And Na
15NO
3React, detect that under certain time point said reaction generates
28N
2,
29N
2With
30N
2Amount, the nitrogen total amount A that Anammox reaction produces in the then said testing sample
Total(a) promptly gets according to formula; By the nitrogen total amount A that obtains
TotalCan obtain under certain time point Anammox reaction rate in the detected sample;
A
total＝F
N -1×[P
29+2×(1-F
N -1)×P
30](a)
In the following formula, F
NExpression Na
15NO
3In
15The abundance of N, P
29Reaction described in the expression step (4) generates
29N
2Total amount, P
30Reaction described in the expression step (4) generates
30N
2Total amount.
2. method according to claim 1 is characterized in that: oxygen free condition described in the step (1) is for adding ammonium water through the nitrogen aeration.
3. method according to claim 1 and 2 is characterized in that: the time of cultivating described in the step (1) is 20 hours～24 hours.
4. according to arbitrary described method among the claim 1-3, it is characterized in that: the temperature of cultivating described in the step (1) is 22 ℃～25 ℃.
5. according to arbitrary described method among the claim 1-4, it is characterized in that: the temperature of reaction is 22 ℃～25 ℃ described in step (2), step (3) and the step (4).
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| CN109596796A (en) * | 2019-01-31 | 2019-04-09 | 郑州轻工业学院 | A kind of measuring method of river bottom mud anaerobic ammoxidation activity |
| CN111487365A (en) * | 2020-01-07 | 2020-08-04 | 三峡大学 | Method for in-situ determination of denitrification and anaerobic ammonia oxidation rates of deep-water reservoir sediments |
| CN113156016A (en) * | 2021-04-26 | 2021-07-23 | 中国科学院生态环境研究中心 | Evaluation method for heavy metal pollution degree of sediment and application thereof |
| CN114994237A (en) * | 2022-04-28 | 2022-09-02 | 红菌生物(广东)有限公司 | Method for measuring anammox activity |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104749278A (en) * | 2015-03-26 | 2015-07-01 | 中国科学院生态环境研究中心 | Method for determining N-DAMO (nitrite-dependent anaerobic methane oxidation) rate |
| CN106018737A (en) * | 2016-05-12 | 2016-10-12 | 辽宁石油化工大学 | Method for measuring total nitration contribution rate of edaphon to soil |
| CN106053780A (en) * | 2016-05-12 | 2016-10-26 | 辽宁石油化工大学 | Method for measuring soil denitrification contribution rate of soil microbes |
| CN108985005A (en) * | 2018-07-03 | 2018-12-11 | 中国科学院南海海洋研究所 | A method of ammonia oxidation speed rate and nitrite-oxidizing rate are measured based on mathematical model simultaneously |
| CN109596796A (en) * | 2019-01-31 | 2019-04-09 | 郑州轻工业学院 | A kind of measuring method of river bottom mud anaerobic ammoxidation activity |
| CN111487365A (en) * | 2020-01-07 | 2020-08-04 | 三峡大学 | Method for in-situ determination of denitrification and anaerobic ammonia oxidation rates of deep-water reservoir sediments |
| CN113156016A (en) * | 2021-04-26 | 2021-07-23 | 中国科学院生态环境研究中心 | Evaluation method for heavy metal pollution degree of sediment and application thereof |
| CN114994237A (en) * | 2022-04-28 | 2022-09-02 | 红菌生物(广东)有限公司 | Method for measuring anammox activity |
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