CN114606131B - Chlorella strain and application thereof in treatment of rare earth ammonia nitrogen wastewater - Google Patents
Chlorella strain and application thereof in treatment of rare earth ammonia nitrogen wastewater Download PDFInfo
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- CN114606131B CN114606131B CN202210333021.XA CN202210333021A CN114606131B CN 114606131 B CN114606131 B CN 114606131B CN 202210333021 A CN202210333021 A CN 202210333021A CN 114606131 B CN114606131 B CN 114606131B
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- 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
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
- C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
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- 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
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- 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
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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Abstract
The invention discloses a green algae strain and application thereof in rare earth ammonia nitrogen wastewater treatment, which is classified and named as Chlamydomonas YC, and the academic name isChlamydomonasYC was preserved in China center for type culture collection (CCTCC NO) in Wuhan at 2021, 5 and 13 days: m2021529. The chlorella strain can be used for treating high-concentration ammonia nitrogen wastewater.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a green alga strain and application thereof in rare earth ammonia nitrogen wastewater treatment.
Background
The existing rare earth mining process mainly adopts an in-situ leaching process, a liquid injection well is arranged on the surface of a mountain, an ammonium sulfate solution prepared into a certain concentration is used as a leaching agent to be injected into a ore body to replace rare earth in rare earth ore, leaching liquid is collected, and carbonate rare earth products are enriched through ammonium bicarbonate precipitation. In-situ leaching does not need to strip surface soil and excavate mountain bodies, so that damage to mountain vegetation is small, and a large amount of tailings are not generated. Because a large amount of ammonium sulfate is added into the leaching, soil and tail water form a huge amount of high-concentration ammonia nitrogen pollution in mountain after ore closing. The rare earth industry is reported to produce more than 20 hundred million tons per year, with ammonia nitrogen concentrations mostly in the range of 300-5000 mg/L, which cannot be adequately handled. The water is collected into streams through surface leakage, rain water leaching and the like, so that serious ecological and social problems are caused around mining areas. It is necessary to treat and utilize such sewage. The pollutant emission standard in the rare earth industry requires that the direct emission concentration is less than or equal to 15 mg/L (GB 26451-2011), and serious environmental pollution can be caused if the treatment is improper.
So far, although the ammonia nitrogen wastewater treatment process method is more and the process is more mature, the method is mainly applicable to domestic sewage and general industrial wastewater. Because of the specificity of the rare earth ammonia nitrogen sewage, such as high ammonia nitrogen concentration, low COD, extremely unbalanced nutrition and objective comparison of input cost, no economic and easily-controlled mature process is suitable for treating the rare earth mine high ammonia nitrogen sewage. The microbial treatment of ammonia nitrogen wastewater has good application prospect, and the biological denitrification method mainly utilizes specific microorganisms to degrade ammonia nitrogen in the wastewater, so that the aim of purifying water is achieved. The method has the advantages of lower operation cost, small secondary pollution, resource utilization of valuable biomass and the like. The wastewater ammonia nitrogen concentration value span range of the south ionic rare earth mine is large, inorganic or organic carbon sources in the wastewater are few, and the requirements of microorganisms on the carbon sources and the energy sources are difficult to meet, so that the biological denitrification process is difficult to establish. For the difficult problem, a large amount of organic carbon sources are usually supplemented, for example, a sugar supplementing process is adopted, and biological denitrification is realized through intensive energy and substance investment, but the process has high cost and no popularization and application value. In addition, the ammonia nitrogen content of the rare earth sewage is high, and only a few microbial communities can tolerate high-concentration ammonia nitrogen, so that the whole denitrification process is blocked, and the denitrification effect is also influenced.
Disclosure of Invention
In order to solve the problems, the invention aims to separate and screen out a greenhouse gas CO with strong tolerance and low cost and convenient source from high-concentration rare earth ammonia nitrogen sewage 2 The microalgae which is a carbon source and takes clean energy sunlight as a main energy source is used for establishing a stable and efficient rare earth sewage denitrification process through the microorganisms, so that the energy consumption of operation is reduced, and the problem of ammonia nitrogen pollution puzzling the surrounding environment of the rare earth industry is solved.
To achieve the above object, the present invention provides a strain of Chlorella, which is classified as Chlamydomonas YC and has a academic nameChlamydomonassp, YC, preserved in China center for type culture collection (CCTCC NO) of Wuhan at 2021, 5 and 13 days: m2021529.
The chlorella strain can be used for treating high-concentration ammonia nitrogen wastewater.
Drawings
FIG. 1 shows the strainChlamydomonasColony plate of sp, YC.
Detailed Description
Example 1 screening, isolation and identification of strains
1) Collecting rare earth high concentration ammonia Nitrogen (NH) 4 + Not less than 2000 mg/L), adding a proper amount of phosphorus element (total phosphorus: 5-50 mg/L), culturing the culture under 2500-7000 lux illumination intensity, shaking the culture by hands for 6 times a day, and obtaining the culture which can survive in the sewage with the ammonia nitrogen concentration of more than 2000 mg/L.
2) Microalgae without ammonia nitrogen tolerance can not grow under the inhibition of high-concentration ammonia nitrogen wastewater, and microalgae with the tolerance can simultaneously synthesize self cell matrixes by utilizing ammonia nitrogen of the high-concentration ammonia nitrogen wastewater to grow. Through enrichment, microalgae with high ammonia nitrogen tolerance capability are screened out.
3) Screening and separating microalgae
And (3) dripping the obtained culture onto a sterilized glass slide by using a micropipette, and taking 2-4 drops of sterile water on the glass slide, wherein the drops are respectively arranged in a straight line from left to right according to the culture and the sterile water, and are mutually separated by a certain distance. Under inverted microscope observation, the sterilized micropipettes were used to aspirate as few cells as possible onto a drop of sterile water on the right. The dripping water is carefully checked, if only one or a few homomorphous cells needing to be separated in the dripping water are not mixed with other organisms, the dripping water is sucked by a liquid-transferring gun and is driven into a rare earth sewage test tube which is sterilized and added with phosphorus elements together with the gun head of the liquid-transferring gun. If the drop is observed under a microscope to contain cells of different morphologies, the micropipette is additionally taken to aspirate as few cells as possible into another drop. Repeating the above steps until a drop of sterile water contains only one or more homomorphous cells to be separated, namely, sucking the drop of water by a liquid-transferring gun, pumping the drop of water and the gun head of the liquid-transferring gun into a sterilized rare earth sewage test tube added with phosphorus element, and sealing by 8 layers of gauze. Culturing under 2500-7000 lux illumination intensity, and shaking the test tube with proper force for 6 times every day until the liquid in the test tube turns green.
4) And (5) purifying microalgae. And adding penicillin and gentamicin into the green culture in the test tube to ensure that the final concentration of the two antibiotics in the test tube is 20-100 mug/mL and 15-50 mug/mL, and continuously culturing for one day. And (3) carrying out concentration gradient dilution on the green culture of the test tube by using sterile water to an appropriate multiple, taking 100 mu L of the diluted culture to be coated on a solid plate of the solid phosphorus-containing rare earth sewage, and culturing until green colonies grow out under the illumination intensity of about 2500-7000 lux. And (3) picking single bacterial colony to be streaked and cultured on a solid plate of the phosphorus-containing rare earth sewage. Repeating the above steps for several times to obtain the pure microalgae strain.
5) And (5) identifying microalgae. Determining the sequence of 18SrDNA and ITS of microalgae, comparing and identifying the sequence of 18SrDNA and ITS, separating to obtain microalgae which is a green alga, and named asChlamydomonassp, YC. The strain has been deposited and registered in the China center for type culture Collection (Wuhan, china) on day 5 and 13 of 2021, and the deposit number is: cctccc M2021529 YC and the center has been tested for viability at day 5 and day 20 of 2021, which concludes survival.
Example 2 Chlamydomonas sp Ammonia nitrogen tolerance test of YC Strain
Separating and purifying the obtained microalgaeChlamydomonassp and YC strains are inoculated into rare earth ammonia nitrogen sewage liquid containing phosphorus, and are subjected to shaking culture for 5 days at the constant temperature of 25 ℃, 150 r/min and 2500lux under illumination to prepare microalgae YC seed liquid. Taking microalgae YC seed liquid 100 mL, centrifuging, inoculating the precipitate into 5 1L BG11 culture mediums containing ammonia nitrogen concentration 1000 mg/L, 2000 mg/L, 3000 mg/L, 4000 mg/L and 5000 mg/L, and shake culturing at 25deg.C, 150 r/min and 2500lux at constant temperature under illumination for 14 days. Microalgae YC grew in media with different ammonia nitrogen concentrations from 0.1 g/L to 0.4 g/L, 0.35 g/L, 0.25g/L, 0.2g/L and 0.07g/L, respectively.
Application example 1
1) Separating and purifying the obtained microalgaeChlamydomonassp and YC strains are inoculated into rare earth ammonia nitrogen sewage liquid containing phosphorus, and are subjected to shaking culture for 5 days at the constant temperature of 25 ℃, 150 r/min and 2500lux under illumination to prepare microalgae YC seed liquid. Taking microalgae YC seed liquid 100 mL, centrifuging, and inoculating the precipitate into rare earth sewage (ammonia nitrogen content is 200 mg/L) diluted 10 times in 1L, and performing aeration culture at 25deg.C and 1000 mL/min. 200 The ammonia nitrogen consumption of mg/L is basically complete within 10 days, and the ammonia nitrogen consumption rate reaches 99.5 percent.
Application example 2
2) Separating and purifying the obtained microalgaeChlamydomonassp and YC strains are inoculated into rare earth ammonia nitrogen sewage liquid containing phosphorus, and are subjected to shaking culture for 5 days at the constant temperature of 25 ℃, 150 r/min and 2500lux under illumination to prepare microalgae YC seed liquid. Taking microalgae YC seed liquid 100 mL, centrifuging, and inoculating the precipitate into 1L original rare earth-containing sewage (ammonia nitrogen content is 2000 mg/L), and performing aeration culture at 25deg.C and 700 mL/min. The ammonia nitrogen in the rare earth sewage is reduced to 350 mg/L within 50 days of 2000 mg/L, and the ammonia nitrogen consumption rate reaches 82.5%.
Sequence listing
<110> university of Fujian
<120> a green algae strain and its application in rare earth ammonia nitrogen wastewater treatment
<130> 2022
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 555
<212> DNA
<213> Artificial sequence (artificial series)
<400> 1
gctcaggtcg agataatgag gggttaaagc tctcgcacag ggcaagagtt cctggttcgg 60
attgaccagc acgaatccag accaccaact aaagcattag caacgaattg cctacccagt 120
tgaggccctt agcgggtcca tgcatcaatc tctacacttc agcagacccg gctaagctct 180
cagtgaaaac ttagccgaga cggccaggtc cgtttcacac tccccagagg gagcatgaga 240
cgagtattaa cccgacgctg aggcagacat gctcttggcc gaagcctcga gcgcaatatg 300
cgttcaaaga cttgatgatt cacgtatttc tgcaattcac actaagtatc gcatttcgct 360
gcgttcttca tcattcgagg agccaagata tccgttgttg agagttgtct tggttggtga 420
gagtcagctc taagccgagc cctcgatttt tcattaggtt tggtttggtg ttgttggtta 480
gataaagacc gacgccccac atgcccgaag gcatgcacag cgacctgagc ggcctgaatt 540
acgagaaccc agaca 555
Claims (2)
1. A strain of green algae characterized by: its classification is named as Chlamydomonas YC and its academic name isChlamydomonassp, YC, preserved in China center for type culture collection (CCTCC NO) of Wuhan at 2021, 5 and 13 days: m2021529.
2. The use of the green algae strain according to claim 1 in ammonia nitrogen wastewater treatment.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02119996A (en) * | 1988-09-20 | 1990-05-08 | Fumio Onuki | Method for purifying domestic waste water with unicellular green alga belonging to chlamydomonas genus |
| JPH0484885A (en) * | 1990-07-27 | 1992-03-18 | Fumio Onuki | Culture medium of unicellular green alga r. sager strain 95 of chlamydomonas and culture |
| WO2012005410A1 (en) * | 2010-07-05 | 2012-01-12 | 연세대학교 산학협력단 | New strain chlamydomonas pitschmannii ysl03 |
| CN103540534A (en) * | 2013-09-29 | 2014-01-29 | 新疆叶希丽生物科技有限公司 | Industrialized culture method for high-protein desert algae |
| CN109251866A (en) * | 2018-10-10 | 2019-01-22 | 中国农业大学 | One chlamydomonas strain and its application in biogas slurry purification |
| CN110642380A (en) * | 2019-10-29 | 2020-01-03 | 福建师范大学 | Method for treating rare earth wastewater by microorganisms in large-scale outdoor pond |
| CN110791454A (en) * | 2019-11-26 | 2020-02-14 | 中国科学院烟台海岸带研究所 | Efficient ammonia nitrogen degrading strain and application thereof |
-
2022
- 2022-03-30 CN CN202210333021.XA patent/CN114606131B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02119996A (en) * | 1988-09-20 | 1990-05-08 | Fumio Onuki | Method for purifying domestic waste water with unicellular green alga belonging to chlamydomonas genus |
| JPH0484885A (en) * | 1990-07-27 | 1992-03-18 | Fumio Onuki | Culture medium of unicellular green alga r. sager strain 95 of chlamydomonas and culture |
| WO2012005410A1 (en) * | 2010-07-05 | 2012-01-12 | 연세대학교 산학협력단 | New strain chlamydomonas pitschmannii ysl03 |
| CN103540534A (en) * | 2013-09-29 | 2014-01-29 | 新疆叶希丽生物科技有限公司 | Industrialized culture method for high-protein desert algae |
| CN109251866A (en) * | 2018-10-10 | 2019-01-22 | 中国农业大学 | One chlamydomonas strain and its application in biogas slurry purification |
| CN110642380A (en) * | 2019-10-29 | 2020-01-03 | 福建师范大学 | Method for treating rare earth wastewater by microorganisms in large-scale outdoor pond |
| CN110791454A (en) * | 2019-11-26 | 2020-02-14 | 中国科学院烟台海岸带研究所 | Efficient ammonia nitrogen degrading strain and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| Ammonium removal from anaerobically treated effluent by Chlamydomonas acidophila;Ania Escudero et al.;《Bioresource Technology》;第153卷;第62-68页 * |
| Pilot-scale remediation of rare earth elements ammonium wastewater by Chlamydomonas sp. YC in summer under outdoor conditions;Y. Zhou et al.;《Bioresource Technology》;第372卷;第1-10页 * |
| Youcai Zhou et al..A newly isolated microalga Chlamydomonas sp. YC to effciently remove ammonium nitrogen of rare earth elements wastewater.《Journal of Environmental Management》.2022,第316卷第1-11页. * |
| 利用莱茵衣藻去除污水中氮磷的研究;邓旭等;《环境科学》;第31卷(第6期);第1489-1493页 * |
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