CN115058379A - Enrichment culture method of anaerobic ammonium oxidation bacteria - Google Patents
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Abstract
The invention discloses an enrichment culture method of anaerobic ammonium oxidation bacteria, which takes activated sludge containing ammonium oxidation bacteria as an inoculum and high-salinity wastewater as a culture solution to carry out enrichment culture by stages, and specifically comprises the following steps: (1) preparing a culture medium of anaerobic ammonium oxidation bacteria, placing the culture medium in an anaerobic reactor, extracting the anaerobic ammonium oxidation bacteria from sludge with nitrification-denitrification functions, inoculating the anaerobic ammonium oxidation bacteria into the anaerobic reactor, adding 01-0.05% of gibberellin and 0.01-0.03% of erythromycin, adjusting the pH value of the system to 7.0-8.0, and carrying out anaerobic reaction at 30-40 ℃ for 12-24 hours to obtain a bacterial liquid; (2) the method comprises the steps of taking high-salt wastewater as a nutritional agent, inoculating bacterial liquid into an anaerobic reactor for strain acclimation, adding high-salt wastewater with corresponding concentration every 2-4 days at the temperature of 36-38 ℃, the pH value of 8-9 and the hydraulic retention time of the anaerobic reactor for 6-8 hours, observing the bacterial cell structure in sludge by an electron microscope, and identifying the anaerobic ammonium oxidation bacteria as anaerobic ammonium oxidation bacteria by 16S rDNA. The method can obtain the salt-tolerant anaerobic ammonium oxidation bacteria.
Description
Technical Field
The invention belongs to the technical field of microbial culture, and particularly relates to an enrichment culture method of anaerobic ammonium oxidation bacteria.
Background
Bacteria participating in the Anammox process are called Anammox bacteria, which belong to the phylum pumila, and "rhodobacter" is a common name for Anammox bacteria in the industry, and can convert ammonia nitrogen contained in sewage into nitrogen for removal through biochemical reaction. They are of great importance for the global nitrogen cycle and also important bacteria in sewage treatment.
In anaerobic environment, Anammox bacteria (Anammox) is treated with CO 2 As a carbon source, with NO 2 - Is an electron acceptor, NH 4 + As an electron donor, ammonium ion (NH) 4 + ) With Nitrite (NO) 2 - ) Oxidation to nitrogen N 2 The anammox reaction is to oxidize NO 2 - -N and NH 4 + -an efficient way of permanently detaching N from the ecosystem. The anaerobic ammonia oxidation process of anaerobic ammonia oxidizing bacteria (Anammox) greatly shortens the process of oxidizing and reducing ammonia nitrogen into nitrogen, thereby reducing the material and energy consumption of biological denitrification, occupying small area and providing a new development platform for the progress of biological denitrification technology.
The high-salinity wastewater mainly comes from food processing, pharmacy, petroleum and natural gas processing, tanning, preserved szechuan pickle pickling, sewage directly discharged by seawater utilization in coastal areas and the like. The quality of life of people is continuously improved, and the discharge of high-salinity wastewater is increased year by year, and if the wastewater is directly discharged, the mineralization degree of the water quality of rivers is improved, so that the soil, surface water and underground water are seriously polluted, and the ecological environment is further endangered. The high salinity can cause the osmotic pressure of microbial cells to be disordered, damage the cell walls and enzyme systems of the microbes and further cause the activity of the microbes to be reduced or the microbes to die. Therefore, high salinity is always a troublesome problem to be solved urgently in the biological treatment of sewage.
Anaerobic ammonia oxidation is used as a new generation biological denitrification technology, an electron donor is not required to be additionally added, aeration and oxygenation are not required, and compared with other technologies, the operation cost advantage is obvious. Therefore, the anaerobic ammonia oxidation has wide application prospect. However, the anammox technology has some defects, such as weak impact resistance, for example, high salinity (more than 20 g.L) in the waste water -1 ) In the process, the activity of the anammox bacteria is inhibited, so that the treatment efficiency of the whole anammox system is deteriorated, and the system is difficult to recover after being unstable. When the salinity of the wastewater entering the anaerobic ammonia oxidation reactor is high, the activity of anaerobic ammonia oxidation bacteria can be seriously inhibited.
Therefore, how to effectively enrich and culture the salt-tolerant anammox bacteria becomes a difficult point and a key point of the current research. Based on practical experience and professional knowledge of designing and manufacturing the product for many years, the inventor of the invention actively carries out research and innovation by matching with the application of theory so as to create an enrichment culture method of anammox bacteria, and the cultured anammox bacteria have good salt resistance and can improve the denitrification performance of an anammox reactor.
Disclosure of Invention
The invention mainly aims to overcome the defects in the prior art and provide an enrichment culture method of anammox bacteria, which can culture the anammox bacteria with good salt resistance and improve the denitrification performance of an anammox reactor.
The purpose of the invention and the technical problem to be solved are realized by adopting the following technical scheme.
The invention provides an enrichment culture method of anaerobic ammonium oxidation bacteria, which takes activated sludge containing ammonia oxidation bacteria as an inoculum and high-salinity wastewater as a culture solution to carry out enrichment culture in stages, and specifically comprises the following steps:
(1) preparing a culture medium of anaerobic ammonium oxidation bacteria, placing the culture medium in an anaerobic reactor, extracting the anaerobic ammonium oxidation bacteria from sludge with nitrification-denitrification functions, inoculating the anaerobic ammonium oxidation bacteria into the anaerobic reactor, adding 0.01-0.05% of gibberellin and 0.01-0.03% of erythromycin, adjusting the pH value of the system to 7.0-8.0, and carrying out anaerobic reaction at 30-40 ℃ for 12-24 hours to obtain a bacterial liquid;
(2) preparing high-salt wastewater with different concentrations as a nutritional agent, inoculating the obtained bacterial liquid into an anaerobic reactor for strain acclimation, wherein the conditions are that the reaction temperature is 36-38 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 6-8 h, high-salt wastewater with corresponding concentration is added every 2-4 days during acclimation, the bacterial cell structure in sludge is observed through an electron microscope after acclimation, the cell wall surface of the high-salt wastewater is found to have a piloid structure, and the high-salt wastewater is identified as anaerobic ammonium oxidation bacteria through 16S rDNA.
The method for enrichment culture of anammox bacteria, wherein the medium in the step (1) comprises the following components: KHCO 3 ：1.75～2.55g/L；KCl：0.01～0.015g/L；KH 2 PO 4 ：0.004～0.006g/L；VC：0.01~0.02g/L；CaCl 2 ：0.6～0.9g/L；MgSO 4 ·7H 2 O：0.3～0.5g/L；FeSO 4 ·7H 2 O: 0.015-0.025 g/L; NaCl: 0.015-0.018 g/L and 2mL of trace elements.
The enrichment culture method of anammox bacteria, wherein the trace elements comprise: MnCl 2 ·4H 2 O：400~600mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：200~400mg/L；CuCl 2 ：125~225mg/L；CoCl 2 ·6H 2 O：120~225mg/L；NaMoO 4 ·2H 2 O：100~200mg/L；Na 2 SeO 4 ：100~110mg/L；NiCl 2 ·2H 2 O：125~155mg/L；H 3 BO 4 ：20~25mg/L。
In the above enrichment culture method of anammox bacteria, the sludge having nitrification-denitrification function in step (1) is aerobic activated sludge obtained from a municipal sewage treatment plant.
The enrichment culture method of the anaerobic ammonium oxidation bacteria comprises the following steps of (1), wherein the gibberellin is GA3, and the concentration of the gibberellin is 2-4 mg/L; the concentration of the erythromycin is 1-3 mg/L.
The method for enrichment culture of anammox bacteria, wherein the high-salinity wastewater in the step (2) is obtained from wastewater discharged from a factory and diluted by 1 time, 2 times, 4 times, 10 times, 20 times, or 30 times.
The enrichment culture method of anammox bacteria, wherein the nutrient in step (2) further comprises an accelerant, and the accelerant comprises the following substances in percentage by weight: 40-60% of polyamine substance, 5-10% of inorganic hydroxylamine acid and Na 2 SO 3 30~50%。
The method for enrichment culture of anammox bacteria, wherein the polyamine-based substance is selected from spermine, spermidine or a mixture thereof.
In the method for enrichment culture of anammox bacteria, the inorganic hydroxylamine acid is one or a mixture of hydroxylamine hydrochloride, hydroxylamine sulfate and hydroxylamine phosphate.
By the technical scheme, the invention at least has the following advantages: the invention provides an enrichment culture method of salt-tolerant anammox bacteria, which is characterized in that high-concentration salt-containing wastewater is used as a nutrient for strain acclimation, and the screened anammox bacteria have good milk salt performance and can improve the denitrification performance of an anammox reactor. The method has the advantages of low difficulty, easy acquisition, low cost and the like.
In conclusion, the special enrichment culture method for the salt-tolerant anaerobic ammonium oxidation bacteria has the advantages of low difficulty, easiness in obtaining, low cost and the like. The method has the advantages and practical value, does not have similar design publication or use in the similar products and methods, is innovative, has great improvement on the method or the function, has great technical progress, produces good and practical effects, has multiple enhanced efficacies compared with the prior products, is more suitable for practical use, has industrial wide utilization value, and is a novel, improved and practical new design.
The foregoing is a summary of the present invention, and in order to provide a clear understanding of the technical means of the present invention and to be implemented in accordance with the present specification, the following is a detailed description of the preferred embodiments of the present invention.
Detailed Description
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
According to the enrichment culture method of the anaerobic ammonium oxidation bacteria, the activated sludge containing the ammonia oxidation bacteria is used as an inoculum, high-salinity wastewater is used as a culture solution, and enrichment culture is performed in stages, and the enrichment culture method specifically comprises the following steps:
(1) preparing a culture medium of anaerobic ammonium oxidation bacteria, placing the culture medium in an anaerobic reactor, extracting the anaerobic ammonium oxidation bacteria from sludge with nitrification-denitrification functions, inoculating the anaerobic ammonium oxidation bacteria into the anaerobic reactor, adding 0.01-0.05% of gibberellin and 0.01-0.03% of erythromycin, adjusting the pH value of the system to 7.0-8.0, and carrying out anaerobic reaction at 30-40 ℃ for 12-24 hours to obtain a bacterial liquid;
(2) preparing high-salt wastewater with different concentrations as a nutritional agent, inoculating the obtained bacterial liquid into an anaerobic reactor for strain acclimatization, wherein the conditions are that the reaction temperature is 36-38 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 6-8 h, the high-salt wastewater with corresponding concentration is added every 2-4 days during acclimatization, the bacterial cell structure in the sludge is observed through an electron microscope after acclimatization, the cell wall surface of the sludge is found to have a piloid structure, and the sludge is identified as anaerobic ammonium oxidation bacteria through 16S rDNA.
According to the method for enrichment culture of anammox bacteria of the present invention, preferably, the step(1) The medium comprises the following components: KHCO 3 ：1.75～2.55g/L；KCl：0.01～0.015g/L；KH 2 PO 4 ：0.004～0.006g/L；VC：0.01~0.02g/L；CaCl 2 ：0.6～0.9g/L；MgSO 4 ·7H 2 O：0.3～0.5g/L；FeSO 4 ·7H 2 O: 0.015-0.025 g/L; NaCl: 0.015-0.018 g/L and 2mL of trace elements.
According to the enrichment culture method of anammox bacteria, the trace elements preferably comprise: MnCl 2 ·4H 2 O：400~600mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：200~400mg/L；CuCl 2 ：125~225mg/L；CoCl 2 ·6H 2 O：120~225mg/L；NaMoO 4 ·2H 2 O：100~200mg/L；Na 2 SeO 4 ：100~110mg/L；NiCl 2 ·2H 2 O：125~155mg/L；H 3 BO 4 ：20~25mg/L。
According to the enrichment culture method of anammox bacteria, preferably, the sludge with nitrification-denitrification function in the step (1) is aerobic activated sludge taken from a municipal sewage treatment plant.
According to the enrichment culture method of the anaerobic ammonium oxidation bacteria, preferably, the gibberellin in the step (1) is GA3, and the concentration of the gibberellin is 2-4 mg/L; the concentration of the erythromycin is 1-3 mg/L.
According to the method for enrichment culture of anammox bacteria of the present invention, it is preferable that the high salinity wastewater in the step (2) is taken from a wastewater discharged from a factory and diluted by 1 time, 2 times, 4 times, 10 times, 20 times, or 30 times.
According to the enrichment culture method of the anaerobic ammonia oxidizing bacteria, preferably, the nutrient in the step (2) further comprises an accelerant, and the accelerant comprises the following substances in percentage by weight: 40-60% of polyamine substance, 5-10% of inorganic hydroxylamine acid and Na 2 SO 3 30~50%。
According to the method for enrichment culture of anammox bacteria, the polyamine substances are preferably selected from spermine, spermidine or a mixture thereof.
According to the enrichment culture method of anaerobic ammonium oxidation bacteria, the inorganic hydroxylamine acid is preferably one or a mixture of hydroxylamine hydrochloride, hydroxylamine sulfate and hydroxylamine phosphate.
Example 1
Preparing a culture medium of anaerobic ammonium oxidation bacteria: KHCO 3 ：2g/L；KCl：0.015g/L；KH 2 PO 4 ：0.005g/L；VC：0.01g/L；CaCl 2 ：0.7g/L；MgSO 4 ·7H 2 O：0.4g/L；FeSO 4 ·7H 2 O: 0.015 g/L; NaCl: 0.015 g/L; and 2mL of MnCl 2 ·4H 2 O：500mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：300mg/L；CuCl 2 ：225mg/L；CoCl 2 ·6H 2 O：150mg/L；NaMoO 4 ·2H 2 O：100mg/L；Na 2 SeO 4 ：100mg/L；NiCl 2 ·2H 2 O：155mg/L；H 3 BO 4 ：23mg/L。
Placing the culture medium in an anaerobic reactor, carrying out precipitation treatment for 30 minutes on aerobic sludge with nitrification-denitrification functions obtained from urban sewage treatment plants, discarding supernatant, and taking the lower concentrated high-concentration sludge mixed liquor as inoculum for acclimatization. The suspended solid concentration (MLSS) of the sludge mixed liquor as the inoculum is 40-60 g/L, the volatile suspended solid concentration (MLVSS) of the sludge mixed liquor is 20-42 g/L, and the MLVSS/MLSS is 0.5-0.7, the sludge mixed liquor is inoculated into an anaerobic reactor, 0.033 mg/L of gibberellin GA3 and 0.022 mg/L of erythromycin are added, the pH value of the system is adjusted to 7.0-8.0, and anaerobic reaction is carried out for 24 hours at 30 ℃ to obtain bacterial liquid.
Diluting high-salt wastewater from pharmaceutical factory by 1, 2, 4, 10, 20, and 30 times to obtain nutritional agent, which can be added with promoter comprising spermidine 50 parts, hydroxylamine hydrochloride 10 parts, and Na 100 parts 2 SO 3 40 portions, inoculating the obtained bacterial liquid into an anaerobic reactor for strain acclimatization, and performing strain acclimatization on each strainWhen the sludge is domesticated under the concentration, the conditions are that the reaction temperature is 38 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 7 hours, high-salinity wastewater with corresponding concentration is added every 3 days, after domestication, the bacterial cell structure in the sludge is observed through an electron microscope, the cell wall surface of the sludge is found to have a piloid structure, and the sludge is identified as the anaerobic ammonia oxidizing bacteria through 16S rDNA.
Example 2
Preparing a culture medium of anaerobic ammonium oxidation bacteria: KHCO 3 ：1.75g/L；KCl：0.01g/L；KH 2 PO 4 ：0.006g/L；VC：0.01g/L；CaCl 2 ：0.7g/L；MgSO 4 ·7H 2 O：0.4g/L；FeSO 4 ·7H 2 O: 0.025 g/L; NaCl: 0.018 g/L; and 2mL of MnCl 2 ·4H 2 O：600mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：200mg/L；CuCl 2 ：125mg/L；CoCl 2 ·6H 2 O：120mg/L；NaMoO 4 ·2H 2 O：150mg/L；Na 2 SeO 4 ：110mg/L；NiCl 2 ·2H 2 O：125mg/L；H 3 BO 4 ：20mg/L。
Placing the culture medium in an anaerobic reactor, carrying out precipitation treatment on aerobic sludge with nitrification-denitrification functions obtained from an urban sewage treatment plant for 30 minutes, discarding supernatant, and taking a high-concentration sludge mixed solution concentrated at the lower part as an inoculum for acclimatization. The suspended solid concentration (MLSS) of the sludge mixed liquor used as the inoculum is 40-60 g/L, the volatile suspended solid concentration (MLVSS) of the sludge mixed liquor is 20-42 g/L, and MLVSS/MLSS is 0.5-0.7, the sludge mixed liquor is inoculated into an anaerobic reactor, 0.053 mg/L gibberellin GA3 and 0.023 mg/L erythromycin are added, the pH value of the system is adjusted to 7.0-8.0, and anaerobic reaction is carried out for 18h at 35 ℃ to obtain bacterial liquid.
Diluting high-salt wastewater of an oil and gas processing plant by 1 time, 2 times, 4 times, 10 times, 20 times and 30 times to obtain a nutritional agent, wherein an accelerant can be added into the nutritional agent, and the accelerant comprises 40 parts of spermidine, 10 parts of hydroxylamine hydrochloride and Na by 100 parts of total weight 2 SO 3 50 parts of and mixingInoculating the obtained bacterial liquid into an anaerobic reactor for strain domestication, wherein when domestication is carried out under each concentration, the conditions are that the reaction temperature is 36 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 8h, high-salinity wastewater with corresponding concentration is added every 2 days, after domestication, the bacterial cell structure in sludge is observed through an electron microscope, the surface of the cell wall of the bacterial cell has a piloid structure, and the bacterial cell is identified as anaerobic ammonium oxidation bacteria through 16S rDNA.
Example 3
Preparing a culture medium of anaerobic ammonium oxidation bacteria: KHCO 3 ：2.55g/L；KCl：0.01g/L；KH 2 PO 4 ：0.004g/L；VC：0.01g/L；CaCl 2 ：0.9g/L；MgSO 4 ·7H 2 O：0.5g/L；FeSO 4 ·7H 2 O: 0.02 g/L; NaCl: 0.016 g/L; and 2mL of MnCl 2 ·4H 2 O：400mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：400mg/L；CuCl 2 ：175mg/L；CoCl 2 ·6H 2 O：175mg/L；NaMoO 4 ·2H 2 O：200mg/L；Na 2 SeO 4 ：105mg/L；NiCl 2 ·2H 2 O：130mg/L；H 3 BO 4 ：25mg/L。
Placing the culture medium in an anaerobic reactor, carrying out precipitation treatment on aerobic sludge with nitrification-denitrification functions obtained from an urban sewage treatment plant for 30 minutes, discarding supernatant, and taking a high-concentration sludge mixed solution concentrated at the lower part as an inoculum for acclimatization. The suspended solid concentration (MLSS) of the sludge mixed liquor as the inoculum is 40-60 g/L, the volatile suspended solid concentration (MLVSS) of the sludge mixed liquor is 20-42 g/L, and MLVSS/MLSS is 0.5-0.7, the sludge mixed liquor is inoculated into an anaerobic reactor, 0.014 mg/L gibberellin GA3 and 0.032 mg/L erythromycin are added, the pH value of the system is adjusted to 7.0-8.0, and anaerobic reaction is carried out for 18h at 35 ℃ to obtain bacterial liquor.
Diluting high-salinity wastewater of petroleum and natural gas processing plants by 1 time, 2 times, 4 times, 10 times, 20 times and 30 times to obtain nutritional agent, wherein the nutritional agent can be added with promoter comprising sub-components in 100 parts of total weight60 parts of spermine, 5 parts of hydroxylamine hydrochloride and Na 2 SO 3 35 parts of the bacterial liquid is inoculated into an anaerobic reactor for strain acclimation, when acclimation is carried out at each concentration, the conditions are that the reaction temperature is 37 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 6 hours, high-salinity wastewater with corresponding concentration is added every 3 days, after acclimation, the bacterial cell structure in sludge is observed through an electron microscope, the surface of the cell wall of the bacterial cell structure is found to have a piloid structure, and the bacterial cell structure is identified to be anaerobic ammonia oxidizing bacteria through 16S rDNA.
Although the present invention has been described with reference to a preferred embodiment, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (9)
1. An enrichment culture method of anaerobic ammonium oxidation bacteria, which takes activated sludge containing the ammonium oxidation bacteria as an inoculum and high-salinity wastewater as a culture solution to carry out enrichment culture by stages, and specifically comprises the following steps:
(1) preparing a culture medium of anaerobic ammonium oxidation bacteria, placing the culture medium in an anaerobic reactor, extracting the anaerobic ammonium oxidation bacteria from sludge with nitrification-denitrification functions, inoculating the anaerobic ammonium oxidation bacteria into the anaerobic reactor, adding 0.01-0.05% of gibberellin and 0.01-0.03% of erythromycin, adjusting the pH value of the system to 7.0-8.0, and carrying out anaerobic reaction at 30-40 ℃ for 12-24 hours to obtain a bacterial liquid;
(2) preparing high-salt wastewater with different concentrations as a nutritional agent, inoculating the obtained bacterial liquid into an anaerobic reactor for strain acclimatization, wherein the conditions are that the reaction temperature is 36-38 ℃, the pH value is 8-9, the hydraulic retention time of the anaerobic reactor is 6-8 h, the high-salt wastewater with corresponding concentration is added every 2-4 days during acclimatization, the bacterial cell structure in the sludge is observed through an electron microscope after acclimatization, the cell wall surface of the sludge is found to have a piloid structure, and the sludge is identified as anaerobic ammonium oxidation bacteria through 16S rDNA.
2. The method for enriching culture of anammox bacteria according to claim 1, wherein the medium composition in the step (1) is: KHCO 3 ：1.75～2.55g/L；KCl：0.01～0.015g/L；KH 2 PO 4 ：0.004～0.006g/L；VC：0.01~0.02g/L；CaCl 2 ：0.6～0.9g/L；MgSO 4 ·7H 2 O：0.3～0.5g/L；FeSO 4 ·7H 2 O: 0.015-0.025 g/L; NaCl: 0.015-0.018 g/L and 2mL of trace elements.
3. The method for enrichment culture of anammox bacteria according to claim 2, wherein the trace elements comprise: MnCl 2 ·4H 2 O：400~600mg/L；EDTA：1500mg/L；ZnSO 4 ·7H 2 O：200~400mg/L；CuCl 2 ：125~225mg/L；CoCl 2 ·6H 2 O：120~225mg/L；NaMoO 4 ·2H 2 O：100~200mg/L；Na 2 SeO 4 ：100~110mg/L；NiCl 2 ·2H 2 O：125~155mg/L；H 3 BO 4 ：20~25mg/L。
4. The enrichment culture method of anaerobic ammonium oxidation bacteria according to claim 1, wherein the sludge with nitrification-denitrification function in the step (1) is aerobic activated sludge taken from a municipal sewage treatment plant.
5. The enrichment culture method of anaerobic ammonia oxidizing bacteria according to claim 1, wherein the gibberellin in the step (1) is GA3, and the concentration of the gibberellin is 2-4 mg/L; the concentration of the erythromycin is 1-3 mg/L.
6. The method for culturing anammox bacteria in an enriched state according to claim 1, wherein the high-salinity wastewater obtained in the step (2) is diluted by 1-fold, 2-fold, 4-fold, 10-fold, 20-fold, or 30-fold.
7. The method for enriching and culturing the anammox bacteria according to claim 1, wherein the nutrient in the step (2) further comprises an accelerant, and the accelerant comprises the following substances in percentage by weight: 40-60% of polyamine substance, 5-10% of inorganic hydroxylamine acid and Na 2 SO 3 30~50%。
8. The method for the enrichment culture of anammox bacteria according to claim 7, wherein the polyamine-based substance is selected from spermine, spermidine, or a mixture thereof.
9. The method for enrichment culture of anaerobic ammonium oxidation bacteria according to claim 7, wherein the inorganic acid hydroxylamine is one or a mixture of hydroxylamine hydrochloride, hydroxylamine sulfate and hydroxylamine phosphate.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927125A (en) * | 2023-02-15 | 2023-04-07 | 知和环保科技有限公司 | Production process of high-load short-cut nitrifying bacteria agent and application method of bacteria agent |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102952764A (en) * | 2011-08-31 | 2013-03-06 | 中国石油化工股份有限公司 | Culture method of salt-tolerant anaerobic ammonium oxidation bacteria |
| CN102976489A (en) * | 2012-12-21 | 2013-03-20 | 重庆大学 | Method for starting anaerobic ammonium oxidation reactor to treat high-salt nitrogenous wastewater |
| CN104450558A (en) * | 2014-10-30 | 2015-03-25 | 大连海洋大学 | Enrichment culturing method of seawater candidatus scalindua |
| CN106754618A (en) * | 2015-11-19 | 2017-05-31 | 中国石油化工股份有限公司 | A kind of Enrichment culture method of Anammox flora |
| CN108676767A (en) * | 2018-05-25 | 2018-10-19 | 中国人民大学 | A kind of method and its application promoting anaerobic ammonia oxidizing bacteria proliferation |
| WO2019082071A1 (en) * | 2017-10-23 | 2019-05-02 | Tubitak | A method for enrichment of anaerobic ammonium oxidation bacteria |
-
2022
- 2022-07-07 CN CN202210795818.1A patent/CN115058379A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102952764A (en) * | 2011-08-31 | 2013-03-06 | 中国石油化工股份有限公司 | Culture method of salt-tolerant anaerobic ammonium oxidation bacteria |
| CN102976489A (en) * | 2012-12-21 | 2013-03-20 | 重庆大学 | Method for starting anaerobic ammonium oxidation reactor to treat high-salt nitrogenous wastewater |
| CN104450558A (en) * | 2014-10-30 | 2015-03-25 | 大连海洋大学 | Enrichment culturing method of seawater candidatus scalindua |
| CN106754618A (en) * | 2015-11-19 | 2017-05-31 | 中国石油化工股份有限公司 | A kind of Enrichment culture method of Anammox flora |
| WO2019082071A1 (en) * | 2017-10-23 | 2019-05-02 | Tubitak | A method for enrichment of anaerobic ammonium oxidation bacteria |
| CN108676767A (en) * | 2018-05-25 | 2018-10-19 | 中国人民大学 | A kind of method and its application promoting anaerobic ammonia oxidizing bacteria proliferation |
Non-Patent Citations (1)
| Title |
|---|
| ZHENG-YANG LU等: "Response of anammox granules to the simultaneous exposure to macrolide and aminoglycoside antibiotics: Linking performance to mechanism", JOURNAL OF ENVIRONMENTAL MANAGEMENT, vol. 286, pages 1 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115927125A (en) * | 2023-02-15 | 2023-04-07 | 知和环保科技有限公司 | Production process of high-load short-cut nitrifying bacteria agent and application method of bacteria agent |
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