CN111268807A - Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora - Google Patents
Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora Download PDFInfo
- Publication number
- CN111268807A CN111268807A CN202010070019.9A CN202010070019A CN111268807A CN 111268807 A CN111268807 A CN 111268807A CN 202010070019 A CN202010070019 A CN 202010070019A CN 111268807 A CN111268807 A CN 111268807A
- Authority
- CN
- China
- Prior art keywords
- indigenous
- microbial flora
- leachate
- denitrification
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P10/00—Technologies related to metal processing
- Y02P10/20—Recycling
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for removing ammonia nitrogen in residual ammonium salt leachate in a rare earth leaching site by utilizing an indigenous denitrification microbial community. Firstly, collecting a soil sample and a leachate sample from a rare earth leaching site, then separating, enriching and screening a single or composite indigenous denitrification microbial flora from the soil sample and the leachate sample, and then inoculating the obtained microbial flora into the residual ammonium salt leachate of the rare earth leaching site to be treated for culture. The indigenous denitrification microbial community used in the invention has strong adaptability to the residual ammonium salt leachate of the rare earth leaching field and high denitrification rate, has the advantages of simple process, low production cost, environmental protection and the like, and is suitable for large-scale denitrification treatment of the residual ammonium salt leachate of the rare earth leaching field.
Description
Technical Field
The invention relates to the technical field of sewage treatment and rare earth mining, in particular to a method for removing ammonia nitrogen in residual ammonium salt leachate of a rare earth leaching site by utilizing an indigenous denitrification microbial community.
Background
The ionic rare earth ore resources in China have great advantages in the world, seven southern provinces are main production areas in China, and the existing mining process of the ionic rare earth ore is developed from pond leaching and heap leaching to in-situ leaching. The in-situ leaching process does not excavate surface soil and damage vegetation, but arranges a liquid injection well on the surface of a mountain, injects an ammonium sulfate solution into an ore body as a leaching agent, collects rare earth ions and ammonium ions into a hydrometallurgy workshop through a liquid collecting project after the rare earth ions and the ammonium ions are subjected to ion reaction, and then carries out precipitation and enrichment by adding ammonium bicarbonate and the like to finally obtain a carbonate rare earth product.
The method has the advantages that although damage to the ecological environment is relieved to a great extent, a great amount of ammonia nitrogen is remained in mountains after in-situ leaching, more residual ammonium salts exist in a rare earth leaching site, the residual ammonium salts cause certain pollution to a mining area and the surrounding environment, the national special provision for the rare earth industry ensures that the direct ammonia nitrogen discharge limit is 15mg/L, compared with ammonia nitrogen wastewater of other industries, the residual ammonium salt leachate of the rare earth leaching site has the characteristics of low ① C/N ratio, large span range of ② leachate ammonia nitrogen concentration value, up to 3000mg/L ammonia nitrogen concentration, as low as 50-100mg/L ammonia nitrogen concentration, ③ has certain salinity, wherein the ammonia nitrogen concentration value contains a great amount of sulfate ions and a small amount of metal ions such as aluminum, iron, silicon, calcium, lead and the like, ④ pH is acidic, and therefore, the problem of how to treat the residual ammonium salt leachate of the rare earth leaching site efficiently and discharge the ammonia nitrogen to reach the standard is still solved with low energy consumption, and the current rare earth.
With the continuous development and progress of the technology, the treatment technology of the ammonia nitrogen which is a common pollutant in the wastewater is continuously upgraded and broken through. The existing treatment method of ammonia-containing wastewater can be roughly divided into a physical-chemical method and a biological method, and the method which is widely applied mainly comprises a biological method, an air stripping method, a membrane separation method, a chemical method, an ion exchange method and the like. The stripping method is suitable for treating high-concentration ammonia nitrogen wastewater, has the defects of high energy consumption, low ammonia nitrogen removal rate and the like, and is generally used for pretreatment of the high-concentration ammonia nitrogen wastewater. The ammonia nitrogen removal efficiency of the ion exchange method is general, and the membrane separation method and the chemical method have higher cost and narrower application range. The biological method has unique advantages compared with a chemical method and a physical method, has excellent treatment effect on the ammonia nitrogen wastewater with medium and low concentration, can reach the national discharge standard after the ammonia nitrogen wastewater is treated by the biological method, and has the advantages of low cost, environmental friendliness and the like. At present, the biological method is widely applied to the treatment of ammonia nitrogen wastewater and has wide development prospect.
A great number of denitrifying microorganisms exist in nature, such as autotrophic nitrifying bacteria, heterotrophic nitrifying bacteria, denitrifying bacteria, anaerobic ammonium oxidizing bacteria and the like, and the denitrifying microorganisms are the basis for treating ammonia nitrogen wastewater by a biological method. The soil and leachate of the ionic rare earth ore leaching site contain a large amount of ammonia nitrogen, and meanwhile, a large amount of indigenous microorganisms (including denitrifying microorganisms) live in the soil and leachate of the mining area for a long time, so that the indigenous microorganisms have good adaptability and strong tolerance to complex components of residual ammonium salt leachate of the rare earth ore leaching site. Therefore, soil and leachate samples are collected from the ionic rare earth ore leaching field, the indigenous denitrification microbial flora is screened out and used for removing ammonia nitrogen in the residual ammonium salt leachate in the rare earth ore leaching field, and the method is an important research direction for treating ammonia nitrogen wastewater pollution in the rare earth industry.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for removing ammonia nitrogen in residual ammonium salt leachate in a rare earth leaching site by using an indigenous denitrification microbial community. The method has the advantages of good adaptability of the selected microorganisms, strong denitrification capability, high denitrification efficiency, good treatment effect, simple process, low production cost, environmental friendliness and the like. In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the method for removing ammonia nitrogen in the residual ammonium salt leachate of the rare earth leaching site by using the indigenous denitrification microbial flora specifically comprises the following steps: (a) collecting a soil sample and a leachate sample of a rare earth leaching site, and separating and enriching microbial floras in the samples to obtain an indigenous microbial floras enrichment culture solution; (b) denitrifying, screening and culturing the indigenous microbial flora enrichment culture solution obtained in the step (a) to obtain an indigenous denitrifying microbial flora; (c) inoculating the indigenous denitrification microbial community obtained in the step (b) into the residual ammonium salt leachate of the rare earth leaching field, adding a denitrification culture medium for culturing, and discharging the residual ammonium salt until the ammonia nitrogen content reaches the standard.
Further, the sample collection method in the step (a) is specifically as follows: randomly selecting at least 3 points from a rare earth leaching site for sampling, and collecting 100-200g soil samples at each point; randomly selecting at least 3 points from a liquid collecting pool and a liquid collecting channel of the leaching liquid in the rare earth leaching site for sampling, and collecting 100-200mL leaching liquid samples at each point; and storing the collected soil sample and leachate sample in an environment at 0-4 ℃ for later use.
Further, the method for separating and enriching the indigenous microbial flora in the soil sample in the step (a) comprises the following specific steps: uniformly mixing the collected soil sample with sterile water according to the proportion of 200-; then, uniformly mixing the initial bacterial suspension of the soil sample and an enrichment culture medium according to the volume ratio of 1:2-3, and then culturing at constant temperature (the culture temperature is 28-30 ℃, the rotating speed is 150-; uniformly mixing the soil microbial flora enrichment culture solution enriched for the first time with an enrichment culture medium under the same culture condition according to the volume ratio of 1:3-4, and culturing at constant temperature again to obtain a soil microbial flora enrichment culture solution enriched for the second time; uniformly mixing the soil microbial flora enrichment culture solution enriched for the second time with an enrichment culture medium under the same culture condition according to the volume ratio of 1:4-5, and carrying out constant temperature culture for the third time; the culture is repeated for 3-5 times to obtain the soil indigenous microbial flora enrichment culture solution.
Further, the method for separating and enriching the indigenous microbial flora in the leachate sample in the step (a) comprises the following steps: uniformly mixing the collected leachate sample with an enrichment medium according to the volume ratio of 1:2-3, and culturing at constant temperature (the culture temperature is 28-30 ℃, the rotation speed is 150-; uniformly mixing the first-time enriched leachate microbial flora enrichment culture solution with an enrichment culture medium under the same culture condition according to the volume ratio of 1:3-4, and culturing at constant temperature to obtain a second-time enriched leachate microbial flora enrichment culture solution; uniformly mixing the leachate microbial flora enrichment culture solution enriched for the second time with an enrichment culture medium under the same culture condition according to the volume ratio of 1:4-5, and then carrying out constant temperature culture for the third time; repeating the culture for 3-5 times to obtain the culture solution rich in indigenous microbial flora of the leachate.
Furthermore, the indigenous microbial flora enrichment culture solution is selected from one of a soil indigenous microbial flora enrichment culture solution and a leachate indigenous microbial flora enrichment culture solution, or a mixture prepared from the two in any proportion. Experiments show that the latter has better effect.
Further, the enrichment medium comprises the following components in parts by weight: 8-12 parts of peptone, 8-12 parts of NaCl, 3-7 parts of yeast extract, 1000 parts of distilled water and 7-7.5 of pH value.
Further, the denitrification screening culture in the step (b) comprises the following specific processes: uniformly mixing the indigenous microbial flora enrichment culture solution obtained in the step (a) with a denitrification function screening culture medium according to the volume ratio of 1:3-6, and then culturing at constant temperature (the culture temperature is 28-30 ℃, the rotating speed is 150-; mixing the first screened indigenous denitrification microbial flora and a denitrification function screening culture medium under the same culture condition according to the volume ratio of 1:4-8, and culturing at constant temperature to obtain a second screened indigenous denitrification microbial flora; mixing the second screened indigenous denitrification microbial community and the denitrification function screening culture medium under the same culture condition according to the volume ratio of 1:5-10, and carrying out third constant temperature culture; the culture is repeated for 3-5 times to obtain indigenous denitrifying microorganism flora.
Further, the method can be used for preparing a novel materialThe denitrification function screening culture medium comprises the following components in parts by weight: 5-20 parts of glucose or sodium citrate, (NH)4)2SO40.5-2 parts of MgSO (MgSO)4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5.
Further, the ammonia nitrogen concentration of the residual ammonium salt leachate in the rare earth leaching site in the step (c) is not more than 300mg/L, and the pH value is 4-6. Before treatment, the pH of the solution is adjusted to 7-7.5 by using aqueous solution of sodium hydroxide, sodium carbonate or sodium bicarbonate.
Further, the specific process of step (c) is as follows: inoculating the indigenous denitrification microbial community obtained by screening in the step (b) into a denitrification microbial culture medium for culturing (the culture temperature is 28-30 ℃, the rotating speed is 150-; uniformly mixing the prepared denitrification microorganism culture solution, the denitrification culture medium and the residual ammonium salt leachate of the rare earth leaching field according to the volume ratio of 1:2-3:4-5, and continuously culturing at constant temperature (the culture temperature is 28-30 ℃, the rotating speed is 150-170r/min, and the culture time is 2-3 days).
Further, the denitrification microorganism culture medium comprises the following components in parts by weight: 3-6 parts of sodium citrate, (NH)4)2SO40.3-0.5 part of MgSO (MgSO)4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5.
Further, the denitrification culture medium comprises the following components in parts by weight: 5-20 parts of glucose or sodium citrate and MgSO4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5.
Compared with the prior similar technology, the invention has the beneficial effects that: (1) the high-efficiency indigenous denitrification microbial community used by the invention is from soil and leachate of a rare earth leaching field, local materials are used, and the microbial cultivation cost is reduced; (2) the high-efficiency indigenous denitrification microbial community used by the method can be well adapted to the residual ammonium salt leachate environment of the rare earth leaching field, the tolerance to non-ammonia nitrogen impurities in the leachate is strong, and the denitrification rate reaches more than 90%; (3) the method provided by the invention does not need to allocate the carbon-nitrogen ratio of the wastewater, can keep a good treatment effect aiming at the wastewater with high ammonia nitrogen content, has the advantages of simple process, low production cost, environmental friendliness and the like, and is particularly suitable for large-scale treatment of the residual ammonium salt leachate in the rare earth leaching field.
Detailed Description
In order to make those skilled in the art fully understand the technical solutions and advantages of the present invention, the following embodiments are further described.
The specific formulas of several culture media used for enriching and screening indigenous microbial flora of the invention are as follows:
enrichment culture medium: 10g of peptone, 10g of NaCl, 5g of yeast extract and 1000mL of distilled water, and the pH value is 7-7.5.
And (3) denitrification function screening culture medium: sodium citrate 15g, (NH)4)2SO41.5g，MgSO4·7H2O 0.5g，NaCl 2g，FeSO4·7H2O 0.04g，MnSO4·4H2O 0.01g，K2HPO41g, 1000mL of distilled water, and a pH value of 7-7.5.
Denitrified microbial culture medium: sodium citrate 5g, (NH)4)2SO40.5g，MgSO4·7H2O 0.5g，NaCl2g，FeSO4·7H2O 0.04g，MnSO4·4H2O 0.01g，K2HPO41g, 1000mL of distilled water, and pH 7-7.5.
The denitrification culture medium has the same formula as the denitrification functional screening culture medium, and the difference is that (NH) is removed4)2SO4And (4) components.
The residual ammonium salt leachate of the rare earth leaching field to be treated is from an ionic rare earth leaching field in Gangzhou city, Jiangxi province, Gangzhou province, and multiple multipoint sampling tests show that the initial concentration of ammonia nitrogen is below 350mg/L and the pH value is between 4 and 6. The soil samples and leachate samples in the following examples are from the rare earth ore leaching sites.
Example 1
1. Randomly selecting points from a rare earth leaching field for sampling, collecting soil samples of 5 points, and collecting 150g of soil samples of each point. 5 soil samples are uniformly mixed and put into a plastic freshness protection package, the plastic freshness protection package is put into an ice box at 4 ℃, and the ice box is quickly brought back to a laboratory within 24 hours.
2. Weighing 300g of soil sample, mixing with 1L of sterile water, and placing into a constant-temperature shaking table for shaking culture at the rotation speed of 160r/min and the temperature of 28 ℃ for 30 min. Standing for 10min after oscillation is finished, and taking supernatant to obtain initial bacterial suspension of the soil sample.
3. Measuring 20mL of initial bacterial suspension of the soil sample, mixing the initial bacterial suspension with 50mL of enrichment medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 160r/min to obtain a first-time enrichment culture solution of the soil microbial flora. And mixing 10mL of the first-enriched soil microbial flora enrichment culture solution with 40mL of enrichment culture medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture at 28 ℃ and 160r/min for 2 days to obtain a second-enriched soil microbial flora enrichment culture solution. And mixing 10mL of the soil microbial flora enrichment culture solution enriched for the second time with 40mL of enrichment culture medium, placing the mixture in a constant-temperature shaking table, and performing fermentation culture for 2 days at the temperature of 28 ℃ and at the speed of 160r/min to obtain the soil indigenous microbial flora enrichment culture solution.
4. Taking 10mL of soil indigenous microbial flora enrichment culture solution, mixing with 40mL of denitrification function screening culture medium, placing in a constant temperature shaking table, and carrying out shaking culture at 28 ℃ and 160r/min for 2 days to obtain the soil indigenous denitrification microbial flora screened for the first time. 10mL of the soil indigenous denitrification microbial flora screened for the first time is mixed with 40mL of denitrification function screening culture medium and then placed in a constant temperature shaking table for shaking culture for 2 days at 28 ℃ and 160r/min, so as to obtain the soil indigenous denitrification microbial flora screened for the second time. And mixing 10mL of the soil indigenous denitrification microbial flora subjected to the secondary screening with 50mL of the denitrification function screening culture medium, placing the mixture in a constant-temperature shaking table, and performing fermentation culture for 2 days at the temperature of 28 ℃ and at the speed of 160r/min to obtain the soil indigenous denitrification microbial flora.
5. Taking a proper amount of residual ammonium salt leachate of the rare earth leaching site to be treated, and measuring the initial ammonia nitrogen concentration to be 290.5mg/L and the initial pH to be 4.9. The pH was adjusted to 7.1 by the addition of aqueous sodium hydroxide.
6. Inoculating the soil indigenous denitrification microbial flora obtained in the step (4) into a denitrification microbial culture medium, and culturing for 12h at 28 ℃ and 160r/min to reach a logarithmic phase to obtain a soil indigenous denitrification microbial culture solution. Mixing the culture solution of the indigenous denitrification microorganisms in the soil, the denitrification culture medium and the residual ammonium salt leachate of the rare earth leaching field according to the volume ratio of 1:2:5, placing the mixture in a constant temperature shaking table, and carrying out shaking culture for 3 days at the temperature of 28 ℃ and at the speed of 160 r/min.
And sampling and detecting the ammonia nitrogen concentration in the solution, and comparing the ammonia nitrogen concentration with the ammonia nitrogen concentration in the residual ammonium salt leachate of the original rare earth leaching site, wherein the result shows that: the ammonia nitrogen concentration in the solution is reduced to 12.8mg/L from 290.5mg/L before treatment, and the ammonia nitrogen removal rate of the indigenous denitrification microbial community of the soil can reach 95.6 percent by calculation.
Example 2
1. Randomly selecting points from a rare earth leaching field for sampling, collecting soil samples of 8 points, and collecting 200g of soil samples of each point. The 8 soil samples were mixed well, packed into plastic bags, packed into ice boxes at 0 ℃ and brought back to the laboratory quickly.
2. Weighing 400g of soil sample, mixing with 1L of sterile water, and placing into a constant-temperature shaking table for shaking culture at the rotation speed of 165r/min and the temperature of 30 ℃ for 25 min. Standing for 15min after oscillation is finished, and taking supernatant to obtain initial bacterial suspension of the soil sample.
3. Measuring 25mL of initial bacterial suspension of the soil sample, mixing the initial bacterial suspension with 50mL of enrichment medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture for 1 day at the temperature of 30 ℃ and at the speed of 165r/min to obtain a first-time enrichment culture solution of the soil microbial flora. And (3) mixing 20mL of the first-enriched soil microbial flora enrichment culture solution with 60mL of enrichment culture medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture for 1 day at 30 ℃ and 165r/min to obtain a second-enriched soil microbial flora enrichment culture solution. And mixing 10mL of the soil microbial flora enrichment culture solution enriched for the second time with 50mL of enrichment culture medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture for 1 day at 30 ℃ and 165r/min to obtain the soil indigenous microbial flora enrichment culture solution.
4. Taking 10mL of soil indigenous microbial flora enrichment culture solution, mixing with 50mL of denitrification function screening culture medium, placing in a constant temperature shaking table, and carrying out shaking culture at 30 ℃ and 165r/min for 2 days to obtain the soil indigenous denitrification microbial flora screened for the first time. 10mL of the soil indigenous denitrification microbial flora screened for the first time is mixed with 50mL of denitrification function screening culture medium and then placed in a constant temperature shaking table for shaking culture for 2 days at 30 ℃ and 165r/min, so as to obtain the soil indigenous denitrification microbial flora screened for the second time. And mixing 10mL of the soil indigenous denitrification microbial flora subjected to the secondary screening with 100mL of the denitrification function screening culture medium, placing the mixture in a constant-temperature shaking table, and performing fermentation culture for 2 days at 30 ℃ and 165r/min to obtain the soil indigenous denitrification microbial flora.
5. Taking a proper amount of residual ammonium salt leachate of the rare earth leaching site to be treated, and measuring the initial ammonia nitrogen concentration of the leachate to be 300mg/L and the initial pH value of the leachate to be 4.3. Adding sodium carbonate aqueous solution to adjust the pH value to 7.4 for standby.
6. Inoculating the soil indigenous denitrification microbial flora obtained in the step (4) into a denitrification microbial culture medium, and culturing for 12h at 30 ℃ and 165r/min to reach a logarithmic phase to obtain a soil indigenous denitrification microbial culture solution. Mixing the culture solution of the indigenous denitrification microorganisms in the soil, the denitrification culture medium and the residual ammonium salt leachate of the rare earth leaching field according to the volume ratio of 1:2:5, placing the mixture in a constant temperature shaking table, and carrying out shaking culture for 3 days at the temperature of 30 ℃ and at the speed of 165 r/min.
And sampling and detecting the ammonia nitrogen concentration in the solution, and comparing the ammonia nitrogen concentration with the ammonia nitrogen concentration in the residual ammonium salt leachate of the original rare earth leaching site, wherein the result shows that: the ammonia nitrogen concentration in the solution is reduced to 14.9mg/L from 300mg/L before treatment, and the calculation shows that the ammonia nitrogen removal rate of the indigenous denitrification microbial community of the soil reaches 95%.
Example 3
1. Randomly selecting points from a liquid collecting pool of the rare earth leaching field for sampling, collecting leachate samples of 5 points, and collecting 100mL leachate samples of each point. And (3) uniformly mixing the 5 leachate samples, putting the mixture into a plastic preservation bottle, putting the plastic preservation bottle into an ice box at the temperature of 3 ℃, and quickly bringing the plastic preservation bottle back to the laboratory.
2. Weighing 25mL of the leachate sample, mixing with 50mL of enrichment medium, and placing the mixture in a constant temperature shaking table for shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 170r/min to obtain a leachate microbial flora enrichment culture solution enriched for the first time. Taking 10mL of the first-enriched leachate microbial flora enrichment culture solution, mixing with 40mL of enrichment culture medium, placing in a constant-temperature shaking table, and carrying out shaking culture at 28 ℃ and 170r/min for 2 days to obtain a second-enriched leachate microbial flora enrichment culture solution. Taking 10mL of the second-time enriched leachate microbial flora enrichment culture solution, mixing with 50mL of enrichment culture medium, placing in a constant-temperature shaking table, and performing fermentation culture for 2 days at 28 ℃ at 170r/min to obtain the leachate microbial flora enrichment culture solution.
4. Taking 10mL of the leacheate indigenous microbial flora enrichment culture solution, mixing the leacheate indigenous microbial flora enrichment culture solution with 45mL of the denitrification function screening culture medium, placing the mixture in a constant-temperature shaking table, and carrying out shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 170r/min to obtain the first screened leacheate indigenous denitrification microbial flora. 10mL of the first screened indigenous denitrification microbial flora of the leachate is mixed with 50mL of the denitrification function screening culture medium and then placed in a constant temperature shaking table for shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 170r/min, so as to obtain the second screened indigenous denitrification microbial flora of the leachate. And mixing 10mL of the second screened indigenous denitrification microbial flora with 100mL of the denitrification function screening culture medium, placing the mixture in a constant temperature shaking table, and fermenting and culturing for 2 days at 28 ℃ and 170r/min to obtain the indigenous denitrification microbial flora of the leachate.
5. Taking a proper amount of residual ammonium salt leachate of the rare earth leaching site to be treated, and measuring the initial ammonia nitrogen concentration of the leachate to be 185.6mg/L and the initial pH value to be 5.6. Aqueous sodium bicarbonate was added to adjust the pH to 7.3 for use.
6. Inoculating the obtained indigenous denitrification microbial community of the leachate in the denitrification microbial culture medium, and culturing at 28 ℃ and 170r/min for 12h to reach logarithmic phase to obtain an indigenous denitrification microbial culture solution of the leachate. Mixing the leaching liquid indigenous denitrification microorganism culture solution with a denitrification culture medium and the rare earth leaching site residual ammonium salt leaching liquid according to the volume ratio of 1:2:5, placing the mixture in a constant temperature shaking table, and carrying out shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 170 r/min.
And sampling and detecting the ammonia nitrogen concentration in the solution, and comparing the ammonia nitrogen concentration with the ammonia nitrogen concentration in the residual ammonium salt leachate of the original rare earth leaching site, wherein the result shows that: the ammonia nitrogen concentration in the solution is reduced to 13.56mg/L from 185.6mg/L before treatment, and the calculation shows that the ammonia nitrogen removal rate of the indigenous denitrification microbial community of the soil reaches 92.7%.
Example 4
1. And (3) preparing the residual ammonium salt leachate of the rare earth leaching site to be treated in the same batch in advance, and measuring that the initial ammonia nitrogen concentration is 212.5mg/L and the initial pH is 5.2. The pH of the leachate was adjusted to 7.3 by adding aqueous sodium hydroxide solution and then divided into 7 groups of at least 3 samples per group.
2. The indigenous denitrification microbial flora and the leachate indigenous denitrification microbial flora prepared in examples 1 and 3 were mixed uniformly in a ratio of 1:1, 1:2, 2:1, 1:5, 5:1, 1:10 and 10:1, respectively, to obtain 7 complex indigenous denitrification microbial flora.
3. Respectively inoculating the 7 composite indigenous denitrification microbial floras obtained in the step (2) into a denitrification microbial culture medium, and culturing for 12h at 28 ℃ and 165r/min to reach a logarithmic phase, thereby respectively obtaining 7 composite indigenous denitrification microbial floras culture solutions. Mixing the prepared culture solution of the composite indigenous denitrification microbial community with a denitrification culture medium and a residual ammonium salt leachate of a rare earth leaching field according to the volume ratio of 1:2:5, placing the mixture in a constant temperature shaking table, and carrying out shaking culture for 2 days at the temperature of 28 ℃ and at the speed of 165 r/min.
And sampling and detecting the ammonia nitrogen concentration in the solution, and comparing the ammonia nitrogen concentration with the ammonia nitrogen concentration in the residual ammonium salt leachate of the original rare earth leaching site, wherein the result shows that: the ammonia nitrogen concentration in the solution is respectively reduced to 10.54mg/L, 9.43mg/L, 11.35mg/L, 9.72mg/L, 14.67mg/L, 8.97mg/L and 12.08mg/L from 212.5mg/L before treatment. The calculation shows that the ammonia nitrogen removal rate of the composite indigenous denitrification microbial community is 95%, 95.6%, 94.7%, 95.4%, 93.1%, 95.8% and 94.3% in sequence, and all the ammonia nitrogen removal rate reaches more than 90%.
Claims (10)
1. The method for removing ammonia nitrogen in the residual ammonium salt leachate of the rare earth leaching site by using the indigenous denitrification microbial flora is characterized by comprising the following steps of: (a) collecting a soil sample and a leachate sample of a rare earth leaching site, and separating and enriching microbial floras in the samples to obtain an indigenous microbial floras enrichment culture solution; (b) denitrifying, screening and culturing the indigenous microbial flora enrichment culture solution obtained in the step (a) to obtain an indigenous denitrifying microbial flora; (c) inoculating the indigenous denitrification microbial community obtained in the step (b) into the residual ammonium salt leachate of the rare earth leaching field, and adding a denitrification culture medium for culture.
2. The method of claim 1, wherein: the sample collection method in the step (a) is specifically as follows: randomly selecting at least 3 points from a rare earth leaching site for sampling, and collecting 100-200g soil samples at each point; randomly selecting at least 3 points from a liquid collecting pool and a liquid collecting channel of the leaching liquid in the rare earth leaching site for sampling, and collecting 100-200mL leaching liquid samples at each point; and storing the collected soil sample and leachate sample in an environment at 0-4 ℃ for later use.
3. The method of claim 1, wherein: the method for separating and enriching the indigenous microbial flora in the soil sample in the step (a) comprises the following steps: uniformly mixing the collected soil sample with sterile water according to the proportion of 200-500g:1L, carrying out constant-temperature shaking culture, and standing to obtain a supernatant to obtain an initial bacterial suspension of the soil sample; then uniformly mixing the initial bacterial suspension of the soil sample with an enrichment culture medium according to the volume ratio of 1:2-3, and then culturing at constant temperature to obtain a first-time enrichment culture solution of the soil microbial flora; uniformly mixing the soil microbial flora enrichment culture solution enriched for the first time with an enrichment culture medium under the same culture condition according to the volume ratio of 1:3-4, and culturing at constant temperature again to obtain a soil microbial flora enrichment culture solution enriched for the second time; uniformly mixing the soil microbial flora enrichment culture solution enriched for the second time with an enrichment culture medium under the same culture condition according to the volume ratio of 1:4-5, and carrying out constant temperature culture for the third time; the culture is repeated for 3-5 times to obtain the soil indigenous microbial flora enrichment culture solution.
4. The method of claim 1, wherein: the method for separating and enriching the indigenous microbial flora in the leachate sample in the step (a) comprises the following steps: uniformly mixing the collected leachate sample with an enrichment medium according to the volume ratio of 1:2-3, and then culturing at constant temperature to obtain a leachate microbial community enrichment culture solution enriched for the first time; uniformly mixing the first-time enriched leachate microbial flora enrichment culture solution with an enrichment culture medium under the same culture condition according to the volume ratio of 1:3-4, and culturing at constant temperature to obtain a second-time enriched leachate microbial flora enrichment culture solution; uniformly mixing the secondary enriched leachate microbial flora enrichment culture solution with an enrichment culture medium under the same culture condition according to the volume ratio of 1:4-5, and then carrying out constant-temperature culture; repeating the culture for 3-5 times to obtain the culture solution rich in indigenous microbial flora of the leachate.
5. The method of claim 3 or 4, wherein: the enrichment medium comprises the following components in parts by weight: 8-12 parts of peptone, 8-12 parts of NaCl, 3-7 parts of yeast extract, 1000 parts of distilled water and 7-7.5 of pH value.
6. The method of claim 1, wherein: the indigenous microbial flora enrichment culture solution is selected from one of a soil indigenous microbial flora enrichment culture solution and a leachate indigenous microbial flora enrichment culture solution, or a mixture prepared from the two in any proportion.
7. The method of claim 1, wherein: the denitrification screening culture in the step (b) comprises the following specific processes: uniformly mixing the indigenous microbial flora enrichment culture solution obtained in the step (a) with a denitrification function screening culture medium according to the volume ratio of 1:3-6, and then culturing at constant temperature to obtain a first screened indigenous denitrification microbial flora; mixing the first screened indigenous denitrification microbial flora and a denitrification function screening culture medium under the same culture condition according to the volume ratio of 1:4-8, and culturing at constant temperature to obtain a second screened indigenous denitrification microbial flora; mixing the second screened indigenous denitrification microbial community and the denitrification function screening culture medium under the same culture condition according to the volume ratio of 1:5-10, and culturing at constant temperature; repeatedly culturing for 3-5 times to obtain indigenous denitrified microbial flora; the denitrification function screening culture medium comprises the following components in parts by weight: 5-20 parts of glucose or sodium citrate, (NH)4)2SO40.5-2 parts of MgSO (MgSO)4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl1, and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5.
8. The method of claim 1, wherein: the ammonia nitrogen concentration of the residual ammonium salt leachate in the rare earth leaching site in the step (c) is not more than 300mg/L, and the pH value is 4-6; before treatment, the pH of the solution is adjusted to 7-7.5 by using aqueous sodium hydroxide solution, aqueous sodium carbonate solution or aqueous sodium bicarbonate solution.
9. The method according to claim 1, wherein the step (c) is embodied by inoculating the indigenous denitrifying microorganism flora screened in the step (b) to a denitrifying microorganism culture medium for culture, and obtaining a denitrifying microorganism culture solution when the flora grows to a logarithmic phase; uniformly mixing the prepared denitrification microorganism culture solution, the denitrification culture medium and the residual ammonium salt leachate of the rare earth leaching field according to the volume ratio of 1:2-3:4-5, and continuously culturing at constant temperature; the denitrification microorganism culture medium comprises the following components in parts by weight: citric acid3-6 parts of sodium, (NH)4)2SO40.3-0.5 part of MgSO (MgSO)4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5; the denitrification culture medium comprises the following components in parts by weight: 5-20 parts of glucose or sodium citrate and MgSO4·7H20.3-0.5 part of O, 1.5-3 parts of NaCl and FeSO4·7H20.01 to 0.05 portion of O and MnSO4·4H20.01-0.04 part of O, K2HPO40.5-1.5 parts of distilled water, and the pH value is 7-7.5.
10. The method according to claim 1, wherein the culture conditions in steps (a) to (c) are as follows: the culture temperature is 28-30 ℃, the rotation speed is 150-.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010070019.9A CN111268807B (en) | 2020-01-21 | 2020-01-21 | Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010070019.9A CN111268807B (en) | 2020-01-21 | 2020-01-21 | Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111268807A true CN111268807A (en) | 2020-06-12 |
| CN111268807B CN111268807B (en) | 2023-04-07 |
Family
ID=70994957
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010070019.9A Active CN111268807B (en) | 2020-01-21 | 2020-01-21 | Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111268807B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112266074A (en) * | 2020-09-07 | 2021-01-26 | 武汉工程大学 | Method for enhancing denitrification of heterotrophic nitrification-aerobic denitrification strain by magnesium salt modified biomass charcoal |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911332A (en) * | 2014-04-04 | 2014-07-09 | 范敏 | Method for culturing nitrogen and phosphorus removal composite bacteria of black and odorous water body |
| CN104862257A (en) * | 2015-06-02 | 2015-08-26 | 湖北工业大学 | Enrichment culture method of indigenous nitrogen transforming microorganism and application of same to treatment of water ammonia-nitrogen pollution |
| CN106676022A (en) * | 2015-11-06 | 2017-05-17 | 丹阳市尚德生物科技有限公司 | Method for preparing microbe bacteria liquid for treating black-odor riverway |
| CN109055293A (en) * | 2018-08-06 | 2018-12-21 | 浙江大学 | A kind of screening of river regulation bacterial strain and acclimation method |
| CN110624949A (en) * | 2019-09-30 | 2019-12-31 | 武汉工程大学 | Method for repairing excessive phosphorus pollution of phosphorite waste land by combining indigenous microorganisms and plants |
-
2020
- 2020-01-21 CN CN202010070019.9A patent/CN111268807B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911332A (en) * | 2014-04-04 | 2014-07-09 | 范敏 | Method for culturing nitrogen and phosphorus removal composite bacteria of black and odorous water body |
| CN104862257A (en) * | 2015-06-02 | 2015-08-26 | 湖北工业大学 | Enrichment culture method of indigenous nitrogen transforming microorganism and application of same to treatment of water ammonia-nitrogen pollution |
| CN106676022A (en) * | 2015-11-06 | 2017-05-17 | 丹阳市尚德生物科技有限公司 | Method for preparing microbe bacteria liquid for treating black-odor riverway |
| CN109055293A (en) * | 2018-08-06 | 2018-12-21 | 浙江大学 | A kind of screening of river regulation bacterial strain and acclimation method |
| CN110624949A (en) * | 2019-09-30 | 2019-12-31 | 武汉工程大学 | Method for repairing excessive phosphorus pollution of phosphorite waste land by combining indigenous microorganisms and plants |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112266074A (en) * | 2020-09-07 | 2021-01-26 | 武汉工程大学 | Method for enhancing denitrification of heterotrophic nitrification-aerobic denitrification strain by magnesium salt modified biomass charcoal |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111268807B (en) | 2023-04-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110791444B (en) | Pseudomonas stutzeri, composite microbial inoculum prepared from pseudomonas stutzeri and application of composite microbial inoculum | |
| CN111268857B (en) | Method for removing high-concentration ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by chemical-biological method | |
| CN103805529A (en) | Halomonas campisalis with heterotrophic nitrification aerobic denitrification function and application thereof | |
| CN109810923A (en) | One plant of aerobic denitrifying bacteria SLY2-21 and its application for sewage water denitrification | |
| CN103373764B (en) | Method for quickly starting short-cut nitrification and denitrification of ammonia-containing wastewater | |
| CN109182192A (en) | One plant of aerobic denitrifying bacteria HY3-2 and its application in sewage water denitrification | |
| CN111718864A (en) | Rare earth leaching site soil indigenous high-efficiency denitrification strain pseudomonas flaviviridis K3 and pseudomonas morganii K17 and application thereof | |
| CN111268807B (en) | Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by using indigenous denitrification microbial flora | |
| CN102690765A (en) | Low-temperature aerobic denitrifying strain Pseudomonas psychrophila Den-03 and screening method and application thereof | |
| CN112723558B (en) | Application of paracoccus denitrificans in preparation of microbial agent for degrading ammoniacal nitrogen in landfill leachate | |
| CN109762748B (en) | Preparation method and application of microbial inoculum for removing ammonia nitrogen | |
| CN103184152B (en) | Method for screening natural gas biological desulfurization flora | |
| CN110951647B (en) | Bacillus psychrophilus GBW-HB1901 and application thereof | |
| CN112553095B (en) | Compound microbial inoculum for treating high-concentration kitchen wastewater and preparation method thereof | |
| CN112522152B (en) | Salt-tolerant paracoccus denitrificans GBW-HB1904 and application thereof | |
| CN111849840B (en) | Separated Raoultella and application thereof in preparing flocculant | |
| CN111004749B (en) | Salt-tolerant bacillus lentus GBW-HB1902 and application thereof | |
| CN111268808B (en) | Method for removing ammonia nitrogen in residual ammonium salt leachate of rare earth leaching site by combining indigenous sulfate reducing bacteria and indigenous denitrifying bacteria | |
| CN113060918A (en) | Method for zero-valent iron reinforcement of synergistic anaerobic digestion of excess sludge and landfill leachate | |
| CN110734878A (en) | bacterial strain separation method for high ammonia nitrogen resistant HN-AD | |
| CN111647520B (en) | Indigenous efficient denitrification strains of acinetobacter bailii A9 and Acidobacterium citri A13 of leachate of rare earth leaching field and application thereof | |
| CN109055225B (en) | Enrichment culture method of methane anaerobic oxidizing bacteria flora | |
| CN114854641B (en) | Low-temperature-resistant composite denitrifying bacterium and application thereof in sewage treatment | |
| CN117887593B (en) | Mixed nutrition type denitrifying bacterium Penicillium sp.N8 and application thereof | |
| Wu et al. | Study on characteristics of the sediment release in Chaohu Lake |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |