CN117342707B - Algae-bacteria symbiotic capsule, preparation method thereof and wastewater treatment method - Google Patents
Algae-bacteria symbiotic capsule, preparation method thereof and wastewater treatment method Download PDFInfo
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- CN117342707B CN117342707B CN202311583539.XA CN202311583539A CN117342707B CN 117342707 B CN117342707 B CN 117342707B CN 202311583539 A CN202311583539 A CN 202311583539A CN 117342707 B CN117342707 B CN 117342707B
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- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 238000004065 wastewater treatment Methods 0.000 title claims abstract description 12
- 239000000243 solution Substances 0.000 claims abstract description 198
- 241000195493 Cryptophyta Species 0.000 claims abstract description 115
- 229920001661 Chitosan Polymers 0.000 claims abstract description 75
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 70
- 239000000661 sodium alginate Substances 0.000 claims abstract description 70
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 70
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 67
- 239000002893 slag Substances 0.000 claims abstract description 65
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- 241000894006 Bacteria Species 0.000 claims abstract description 42
- 239000002351 wastewater Substances 0.000 claims abstract description 41
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims abstract description 35
- 239000011734 sodium Substances 0.000 claims abstract description 35
- 229910052708 sodium Inorganic materials 0.000 claims abstract description 35
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- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 46
- 238000000034 method Methods 0.000 claims description 36
- 239000013067 intermediate product Substances 0.000 claims description 21
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- 241000195652 Auxenochlorella pyrenoidosa Species 0.000 claims description 16
- 235000007091 Chlorella pyrenoidosa Nutrition 0.000 claims description 16
- 239000002028 Biomass Substances 0.000 claims description 13
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- 238000003381 deacetylation reaction Methods 0.000 claims description 12
- 150000001735 carboxylic acids Chemical class 0.000 claims description 11
- 238000005273 aeration Methods 0.000 claims description 6
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- 241000195649 Chlorella <Chlorellales> Species 0.000 claims description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
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- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 235000019270 ammonium chloride Nutrition 0.000 description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 4
- 239000003344 environmental pollutant Substances 0.000 description 4
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 4
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 3
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000001879 gelation Methods 0.000 description 3
- 235000001727 glucose Nutrition 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229920000867 polyelectrolyte Polymers 0.000 description 3
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 3
- 230000001376 precipitating effect Effects 0.000 description 3
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- 239000011780 sodium chloride Substances 0.000 description 3
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- 238000012360 testing method Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
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- 125000002091 cationic group Chemical group 0.000 description 2
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- 229910002651 NO3 Inorganic materials 0.000 description 1
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- 150000001335 aliphatic alkanes Chemical class 0.000 description 1
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- 230000003698 anagen phase Effects 0.000 description 1
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- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- BRPQOXSCLDDYGP-UHFFFAOYSA-N calcium oxide Chemical compound [O-2].[Ca+2] BRPQOXSCLDDYGP-UHFFFAOYSA-N 0.000 description 1
- 239000000292 calcium oxide Substances 0.000 description 1
- ODINCKMPIJJUCX-UHFFFAOYSA-N calcium oxide Inorganic materials [Ca]=O ODINCKMPIJJUCX-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
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- 230000016615 flocculation Effects 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
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- 238000005259 measurement Methods 0.000 description 1
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- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
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- 231100000956 nontoxicity Toxicity 0.000 description 1
- 150000002897 organic nitrogen compounds Chemical class 0.000 description 1
- 238000006864 oxidative decomposition reaction Methods 0.000 description 1
- TWNQGVIAIRXVLR-UHFFFAOYSA-N oxo(oxoalumanyloxy)alumane Chemical compound O=[Al]O[Al]=O TWNQGVIAIRXVLR-UHFFFAOYSA-N 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
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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
- C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The application provides an algae-bacteria symbiotic capsule, a preparation method thereof and a wastewater treatment method, and the preparation method of the algae-bacteria symbiotic capsule is characterized by comprising the following steps: mixing the solution containing aerobic bacteria with the solution containing algae to prepare an algae bacteria mixed solution; mixing the algae bacteria mixed solution with sodium alginate-slag mixed solution containing sodium alginate and slag to prepare embedding liquid; mixing embedding liquid with a crosslinking solution to carry out crosslinking fixation, so as to prepare an algae bacteria immobilized capsule; mixing the algae immobilized capsule with carboxylic acid-chitosan solution, and coating to obtain algae symbiotic capsule. Algae and aerobic bacteria are immobilized together by the algae-bacteria symbiotic principle, so that the effect of the microbial capsule on removing nutrients in wastewater is improved. Through adding slag and adopting chitosan to hang the membrane in embedding liquid for algae-bacteria symbiotic capsule mechanical strength is higher, and the structure is more stable, and is showing to the removal effect of nutrient in the aquaculture waste water, but reuse.
Description
Technical Field
The application relates to the technical field of wastewater treatment, in particular to an algae-bacteria symbiotic capsule, a preparation method thereof and a wastewater treatment method.
Background
The microorganism immobilization technology is a technology for immobilizing free dispersed microorganism, enzyme and other biocatalysts in a water body in a carrier by physical and chemical means, so that the microorganism is highly enriched and simultaneously keeps higher activity. The enrichment of the microorganisms on the carrier can effectively improve the load resistance and the water flow impact resistance of the microorganisms, avoid the loss of dominant microorganisms and realize the stable and effective proliferation of various microorganisms. The microorganism immobilization method is mainly divided into four major categories, namely an adsorption method, a crosslinking method, an embedding method and a composite immobilization method. The embedding method has the characteristics of simple operation, firm combination, small influence on the activity of microorganisms and the like, and becomes a research hot spot of the current immobilization technology.
In the related art, the research on the treatment of wastewater by the microorganism immobilization technology is mostly focused on immobilization of a single algae species or strain, and has limited effect on removal of nitrogen and phosphorus.
Disclosure of Invention
Based on the above, the application provides an algae-bacteria symbiotic capsule, a preparation method thereof and a wastewater treatment method, so as to improve the removal effect of nitrogen and phosphorus.
The first aspect of the application provides a preparation method of an algae-bacteria symbiotic capsule, which comprises the following steps:
Mixing the solution containing aerobic bacteria with the solution containing algae to prepare an algae bacteria mixed solution;
mixing the algae bacteria mixed solution with sodium alginate-slag mixed solution containing sodium alginate and slag to prepare embedding solution;
mixing the embedding liquid with a crosslinking solution to carry out crosslinking fixation, so as to prepare an algae bacteria immobilized capsule;
mixing the algae immobilized capsule with a carboxylic acid-chitosan solution, and forming a film to prepare the algae symbiotic capsule.
In some embodiments, the algae comprises one or more of chlorella, scenedesmus, and crescent algae; optionally Chlorella pyrenoidosa; and/or
The aerobic bacteria comprise activated sludge aerobic bacteria.
In some embodiments, the method for preparing the sodium alginate-slag mixture comprises:
Mixing sodium alginate solution with slag to prepare sodium alginate-slag mixed solution;
Optionally, the sodium alginate solution has a mass percentage concentration of 1% -2%;
Optionally, the mass-to-volume ratio of the slag to the sodium alginate solution is (0.5-1) g to 100mL;
Optionally, the grain size of the slag is 250-300 mesh.
In some embodiments, the method of preparing the carboxylic acid-chitosan solution comprises:
Mixing a carboxylic acid solution with chitosan to prepare the carboxylic acid-chitosan solution;
Optionally, the chitosan has a degree of deacetylation of 80% -95%;
optionally, the pH of the carboxylic acid-chitosan solution is 5.5-6.0;
Alternatively, the carboxylic acid solution is selected from acetic acid solutions;
further alternatively, the acetic acid solution has a mass percent concentration of 0.4% -0.8%;
further alternatively, the mass to volume ratio of the chitosan to the acetic acid solution is (0.5-1.5) g to 100mL.
In some embodiments, the biomass of the algae-containing solution is from 1.0g/L to 1.5g/L; and/or
When the algae bacteria mixed solution is prepared, the solution containing aerobic bacteria and the solution containing algae are mixed according to the ratio of the algae bacteria biomass ratio of 1 (1.5-2).
In some embodiments, the crosslinking solution is a calcium chloride solution with a mass percent concentration of 2% -4%;
optionally, the volume ratio of the embedding liquid to the crosslinking solution is 1 (1-2).
In some embodiments, the method of preparing the algal immobilized capsule comprises:
adding the embedding liquid into a partial crosslinking solution which is refrigerated for 12-24 hours at the temperature of 4-5 ℃ in advance for preliminary crosslinking fixation;
washing the intermediate product obtained after preliminary crosslinking and fixing, adding the washed intermediate product into the residual crosslinking solution, and continuing crosslinking and fixing for 12-24 hours at the temperature of 4-5 ℃ to obtain the algae bacteria immobilized capsule.
The second aspect of the application provides an algae-bacteria symbiotic capsule prepared by the method of the first aspect of the application;
optionally, the particle size of the algae-bacteria symbiotic capsule is 3.8mm-4mm.
The third aspect of the application provides a cultivation wastewater treatment method, comprising the following steps:
Mixing the algae-bacteria symbiotic capsules with the culture wastewater to be treated so as to treat the culture wastewater to be treated; wherein the phycobiont capsule is prepared by the method of the first aspect of the application.
In some embodiments, the algae-symbiotic capsule is mixed with the wastewater to be treated in a ratio of 20g/L to 60 g/L; and/or
The conditions of the treatment include: the illumination intensity is 4000Lux-5000Lux, the light-dark ratio is 12:12, the temperature is 24 ℃ to 28 ℃, the aeration quantity is 0.1L/min-0.2L/min, and the treatment time is 7d.
According to the preparation method of the algae-bacteria symbiotic capsule, algae and aerobic bacteria are immobilized together according to the algae-bacteria symbiotic principle, so that the effect of removing nutrients in wastewater by the microbial capsule is improved. Meanwhile, slag is added into embedding liquid, chitosan is adopted to form a film, chitosan cationic amine groups can be attracted with sodium alginate anions through positive and negative charges, instantaneous gelation is achieved, and a polyelectrolyte semi-permeable film is formed, so that the prepared algae-bacteria symbiotic capsule is higher in mechanical strength, more stable in structure, obvious in effect of removing nutrients in cultivation wastewater and capable of being recycled. In addition, the preparation method is simple and easy to implement, has low equipment requirement and low production cost, and has very wide application prospect in treating the related cultivation wastewater.
Detailed Description
In order that the application may be readily understood, a more particular description of the application will be rendered by reference to specific embodiments that are illustrated in the appended drawings. Preferred embodiments of the present application are given below. This application may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
In the application, the technical characteristics described in an open mode comprise a closed technical scheme composed of the listed characteristics and also comprise an open technical scheme comprising the listed characteristics.
In the present application, the numerical ranges are referred to as continuous, and include the minimum and maximum values of the ranges, and each value between the minimum and maximum values, unless otherwise specified. Further, when a range refers to an integer, each integer between the minimum and maximum values of the range is included. Further, when multiple range description features or characteristics are provided, the ranges may be combined. In other words, unless otherwise indicated, all ranges disclosed herein are to be understood to include any and all subranges subsumed therein.
Only a few numerical ranges are specifically disclosed herein. However, any lower limit may be combined with any upper limit to form a range not explicitly recited; and any lower limit may be combined with any other lower limit to form a range not explicitly recited, and any upper limit may be combined with any other upper limit to form a range not explicitly recited. Furthermore, each separately disclosed point or individual value may itself be combined as a lower limit or upper limit with any other point or individual value or with other lower limit or upper limit to form a range not explicitly recited.
The temperature parameter in the present application is not particularly limited, and may be a constant temperature treatment or a treatment within a predetermined temperature range. The constant temperature process allows the temperature to fluctuate within the accuracy of the instrument control.
In the description of the invention, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.
All embodiments of the application and alternative embodiments may be combined with each other to form new solutions, unless otherwise specified. All technical features and optional technical features of the application may be combined with each other to form new technical solutions, unless specified otherwise.
All the steps of the present application may be performed sequentially or randomly, preferably sequentially, unless otherwise specified.
In the related art, the research on the treatment of wastewater by the microorganism immobilization technology is mostly focused on immobilization of a single algae species or strain, and has limited effect on removal of nitrogen and phosphorus.
Researchers find that the preparation of the algae-bacteria symbiotic capsule by adopting the embedding method needs to screen proper algae and bacteria and control the adding proportion between the algae and the bacteria so as to prepare the algae-bacteria symbiotic capsule with better wastewater treatment effect. Based on the problems, the application provides a preparation method of the algae-bacteria symbiotic capsule, which is based on the algae-bacteria symbiotic principle, and the algae and the aerobic bacteria are jointly immobilized so as to improve the removal effect of the microorganism capsule on the nutrient substances in the wastewater.
The first aspect of the application provides a preparation method of an algae-bacteria symbiotic capsule, which comprises the following steps:
Mixing the solution containing aerobic bacteria with the solution containing algae to prepare an algae bacteria mixed solution; mixing the algae bacteria mixed solution with sodium alginate-slag mixed solution containing sodium alginate and slag to prepare embedding liquid; mixing embedding liquid with a crosslinking solution to carry out crosslinking fixation, so as to prepare an algae bacteria immobilized capsule; mixing the algae immobilized capsule with carboxylic acid-chitosan solution, and coating to obtain algae symbiotic capsule.
The algae-bacteria symbiotic capsule provided by the application applies the algae-bacteria symbiotic principle, algae cells can utilize various inorganic nitrogen and organic nitrogen compounds in water bodies as nitrogen sources, carbon dioxide and carbonate as carbon sources for photoautotrophic growth, and nitrate, nitrite and ammonium salt are absorbed by the algae cells and then are used for synthesizing substances such as amino acid, protein and the like; the organic matters containing phosphorus are directly absorbed by algae cells under the aerobic condition and are converted into organic matters such as ATP, phospholipid and the like through various phosphorylation pathways; on the other hand, the aerobic bacteria can degrade carbon-containing organic matters into carbon dioxide and water, and the generated CO 2 provides a carbon source for algae, promotes photosynthesis of the algae, and can absorb and convert substances generated in bacterial metabolism into cellular substances of the algae; oxygen released by algae photosynthesis increases dissolved oxygen in a water body, promotes metabolic activity of aerobic bacteria, and oxygen generated by the photosynthesis can be effectively utilized by bacteria compared with aeration oxygenation, so that in a co-immobilized system, the symbiotic relationship of algae and bacteria is more beneficial to improving the impact resistance and the decontamination effect of the system on wastewater.
The sodium alginate as an immobilization carrier has the advantages of safety, no toxicity, good mass transfer property, mild reaction conditions and the like, has small damage to embedded microorganisms, is very suitable for immobilization operation, but has low strength and poor antimicrobial decomposition capability, and is easy to be degraded by microorganisms under anaerobic conditions. The chitosan belongs to a chitosan deacetylation product, is the only alkaline polysaccharide in natural polysaccharide, has good biocompatibility, is positively charged linear polysaccharide formed by connecting 2-amino-2-deoxyglucose through A-1,4 glycosidic bonds, contains a large amount of amino groups in a molecular chain, is easy to protonate in an acidic solution to form cationic amino polysaccharide, and can be attracted with sodium alginate anions through positive and negative charges to generate instant gelation to form a polyelectrolyte semipermeable membrane.
The slag adopted in the preparation method of the application comprises solid waste discharged after coal is combusted in various industrial, civil and power plant boilers and kilns, and the generated slag forms a porous structure due to the fact that a large amount of air enters in the combustion process of the coal and escapes after cooling, and contains various alkaline oxides (such as calcium oxide, aluminum oxide and the like), and part of alkaline matters can be dissolved out from the slag after the slag contacts with the wastewater, so that the slag has certain adsorption, neutralization, filtration and flocculation effects on organic matters and suspended matters in the wastewater. Therefore, slag is used as one of the carrier materials, belonging to waste recycling. The addition of the slag can provide a good adhesion carrier for the propagation and growth of aerobic bacteria on one hand, and on the other hand, the slag has an adsorption effect on organic matters in the wastewater, and in addition, the structural stability of the microbial capsule can be indirectly improved.
Understandably, algae and aerobic bacteria are immobilized together by the algae-bacteria symbiotic principle, so that the effect of the microbial capsule on removing nutrients in wastewater is improved. Meanwhile, slag is added into embedding liquid, chitosan is adopted to form a film, chitosan cationic amine groups can be attracted with sodium alginate anions through positive and negative charges, instantaneous gelation is achieved, and a polyelectrolyte semi-permeable film is formed, so that the prepared algae-bacteria symbiotic capsule is higher in mechanical strength, more stable in structure, obvious in effect of removing nutrients in cultivation wastewater and capable of being recycled.
In addition, the materials adopted in the preparation method provided by the application comprise sodium alginate, slag, chitosan and calcium chloride, the material consumption is low, the materials are cheap and easy to obtain, the microbial toxicity is avoided, and the secondary pollution to the environment is avoided. The preparation method is simple and easy to implement, algae bacteria and slag are uniformly distributed in the system, the requirements on equipment are low, the production cost is low, and the method has very broad application prospects in the treatment of related cultivation wastewater.
The algae-bacteria symbiotic capsule prepared by the application is sodium alginate-slag-chitosan algae-bacteria symbiotic capsule.
In some embodiments, the algae include one or more of chlorella, scenedesmus, and crescent algae; optionally Chlorella pyrenoidosa. When the chlorella pyrenoidosa is adopted to prepare the algae-bacteria symbiotic capsule, the chlorella pyrenoidosa has a unique metabolism mode, solar energy and inorganic matters can be utilized to synthesize self protoplasm through photosynthesis, the requirements on growth conditions are simple, the growth speed is high, the environmental tolerance is strong, the chlorella pyrenoidosa can have a better removal effect on nutrients such as nitrogen and phosphorus in wastewater, heavy metals can be absorbed through overload, and various organic matters such as pesticides, alkanes and phenols can be degraded by utilizing inorganic salts in the wastewater.
In some exemplary embodiments, a method of preparing a solution containing algae includes the steps of: inoculating Chlorella pyrenoidosa into BG-11 culture medium, culturing at 24-28deg.C and pH of 7.5-8.5 under illumination intensity of 5000-7000 Lux for 8 days (d), and centrifuging at 5000-6000 r/min for 10min to obtain solution containing algae.
In some embodiments, the algae-containing solution is stored at 4 ℃ for use.
In some embodiments, the aerobic bacteria comprise activated sludge aerobic bacteria. The aerobic bacteria can well utilize oxygen provided by algae photosynthesis to carry out oxidative decomposition on organic pollutants in wastewater, and in addition, as sodium alginate is easily decomposed by anaerobic microorganisms in an anaerobic environment, the activated sludge aerobic bacteria are adopted to prepare the algae-bacteria symbiotic capsules, so that the service life of the sodium alginate can be prolonged.
In some exemplary embodiments, a method of preparing a solution containing aerobic bacteria includes the steps of: the method comprises the steps of preparing an activated sludge culture solution from ammonium chloride, sodium chloride, monopotassium phosphate, glucose, ferric chloride, magnesium sulfate and distilled water, inoculating activated sludge into the culture solution, continuously aerating for 48 hours (h) under the condition that the temperature is 24-28 ℃ and the pH is 7.2-8.0, stopping aerating for 2 hours to form a period, continuously culturing for 8 days, stopping aerating, standing, precipitating, and centrifuging the upper liquid to prepare a solution containing aerobic bacteria.
As a possible embodiment, the preparation method of the sodium alginate-slag mixed solution comprises the following steps: and mixing the sodium alginate solution with the slag to prepare sodium alginate-slag mixed solution.
In some alternative embodiments, the sodium alginate solution has a mass percent concentration of 1% -2%; for example, but not limited to, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2% or a range between any two of the above values. When the mass percentage concentration of the sodium alginate solution is in the range, the elasticity, the water holding capacity and the gel strength of the sodium alginate solution are gradually enhanced along with the increase of the concentration of the sodium alginate.
The mass percentage concentration of the sodium alginate solution refers to the mass percentage of sodium alginate contained in the solution.
In some alternative embodiments, the mass to volume ratio of slag to sodium alginate solution is (0.5-1) g:100mL; such as but not limited to 0.5g:100mL、0.55g:100mL、0.6g:100mL、0.65g:100mL、0.7g:100mL、0.75g:100mL、0.8g:100mL、0.85g:100mL、0.9g:100mL、0.95g:100mL、1g:100mL or a range between any two of the mass to volume ratios described above. When the usage amount of the slag and the sodium alginate is in the above range, the better the structural stability of the prepared capsule is, the stronger the load resistance and the water flow impact resistance are along with the increase of the addition amount of the slag.
In some alternative embodiments, the slag has a particle size of 250 mesh to 300 mesh. Therefore, the slag can be uniformly distributed in the sodium alginate solution, the specific surface area of the slag in the particle size range is large, sufficient growth carriers can be provided for microorganisms, and the adsorption effect on pollutants is better.
As one possible embodiment, the preparation method of the sodium alginate solution includes: soaking sodium alginate powder in distilled water at 30-40 deg.c for 1-2 hr, regulating pH of distilled water to 8.0-9.0 with sodium hydroxide solution, and stirring at 90-100 deg.c to dissolve sodium alginate powder to prepare sodium alginate solution. The mass percentage concentration of the sodium alginate solution is 1% -2%.
In some embodiments, a method of preparing a carboxylic acid-chitosan solution includes: the carboxylic acid solution and chitosan are mixed to prepare a carboxylic acid-chitosan solution.
After the carboxylic acid solution and chitosan are mixed, the carboxyl group (-COOH) in the carboxylic acid and the amino group (-NH 2) in the chitosan react to form a carboxylic acid chitosan ester. As an example, the carboxylic acid contained in the carboxylic acid solution may include, but is not limited to, one or more of succinic acid, citric acid, acetic acid.
In some alternative embodiments, the pH of the carboxylic acid-chitosan solution is 5.5 to 6.0; for example, but not limited to, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0 or a range between any two of the above pH values. When the pH of the carboxylic acid-chitosan solution is in the above range, chitosan can be rapidly dissolved.
As a further possible embodiment, the carboxylic acid solution is selected from acetic acid solutions.
In some alternative embodiments, the acetic acid solution has a mass percent concentration of 0.4% to 0.8%; for example, but not limited to, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, or a range between any two of the above values. When the mass percentage concentration of the acetic acid solution is in the above range, chitosan can be rapidly dissolved.
The mass percentage concentration of the acetic acid solution refers to the mass fraction of acetic acid contained in the solution.
As one possible embodiment, the mass to volume ratio of chitosan to acetic acid solution is (0.5-1.5) g to 100mL; such as but not limited to 0.5g:100mL、0.6g:100mL、0.7g:100mL、0.8g:100mL、0.9g:100mL、1g:100mL、1.1g:100mL、1.2g:100mL、1.3g:100mL、1.4g:100mL、1.5g:100mL or a range between any two of the mass to volume ratios described above. When the ratio of the chitosan to the acetic acid solution is in the above range, the chitosan can be completely dissolved in the acetic acid solution and completely reacted with the acetic acid.
In some alternative embodiments, the chitosan has a degree of deacetylation of 80% to 95%; for example, but not limited to, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or a range between any two of the above values. Thus, the higher the chitosan deacetylation degree, the more free amino groups on the molecular chain, and the better the solubility in acid.
In some embodiments, the biomass of the algae-containing solution is from 1.0g/L to 1.5g/L; for example, but not limited to, 1.0g/L, 1.05g/L, 1.1g/L, 1.15g/L, 1.2g/L, 1.25g/L, 1.3g/L, 1.35g/L, 1.4g/L, 1.45g/L, 1.5g/L, or a range between any two of the foregoing biomass. Therefore, an advantageous population of specific algae species can be established, the growth start time of microalgae is shortened, and the removal rate of pollutants in wastewater is improved.
In some embodiments, when preparing the algal cell mixed solution, mixing the solution containing aerobic cells with the solution containing algae in a ratio of 1 (1.5-2) algal biomass; for example, but not limited to, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, or a range between any two of the above ratios. Thus, a good and stable symbiotic system of algae and bacteria can be established.
In some embodiments, the crosslinking solution is a calcium chloride solution having a concentration of 2% to 4% by mass. As an example, the mass percent concentration of the calcium chloride solution may be, but is not limited to, 2%, 2.2%, 2.5%, 2.8%, 3%, 3.2%, 3.5%, 3.7%, 4%, or a range between any two of the foregoing values. Calcium chloride is used as a cross-linking agent and can react with sodium alginate to form water-insoluble gel; when the calcium chloride solution with the mass percent concentration of 2% -4% is used as the crosslinking solution, the more stable the formed gel is, the less easily the gel is degraded.
As an example, the calcium chloride solution is an aqueous calcium chloride solution.
In some alternative embodiments, the volume ratio of embedding liquid to crosslinking solution is 1 (1-2); for example, but not limited to, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, or a range between any two of the above ratios. When the volume ratio of the embedding solution to the crosslinking solution is in the above range, the sodium alginate can exchange with sufficient calcium ions, the formed sodium alginate-calcium space network structure is compact, and the interaction between sodium alginate macromolecules is enhanced.
In some exemplary embodiments, the method of preparing the algal immobilized capsule includes: adding the embedding liquid into a partial crosslinking solution which is refrigerated for 12-24 hours at the temperature of 4-5 ℃ in advance for preliminary crosslinking fixation; washing the intermediate product obtained after preliminary crosslinking and fixing, adding the washed intermediate product into the residual crosslinking solution, and continuing crosslinking and fixing for 12-24 hours at the temperature of 4-5 ℃ to obtain the algae bacteria immobilized capsule.
Refrigerating the cross-linking solution at 4-5 deg.c for 12-24 hr to lower the heat energy in the molecular chain of sodium alginate and to make the sodium alginate and calcium ion combine fully to react to form gel.
The embedding liquid was added dropwise to the partially crosslinked solution previously refrigerated.
As a possible embodiment, the preparation method of the algae-bacteria symbiotic capsule comprises the following steps:
Inoculating Chlorella pyrenoidosa into BG-11 culture medium, culturing at 24-28deg.C and pH of 7.5-8.5 for 8 days (d) under illumination intensity of 5000-7000 Lux, centrifuging microalgae cultured to logarithmic phase at 5000-6000 r/min for 10 min to obtain solution containing algae;
Preparing an activated sludge culture solution from ammonium chloride, sodium chloride, monopotassium phosphate, glucose, ferric chloride, magnesium sulfate and distilled water, inoculating activated sludge into the culture solution, continuously aerating for 48 hours (h) under the condition that the temperature is 24-28 ℃ and the pH is 7.2-8.0, stopping aerating for 2 hours to form a period, continuously culturing for 8 days, stopping aerating, standing, precipitating, taking upper liquid, centrifuging, and preparing a solution containing aerobic bacteria;
mixing the solution containing aerobic bacteria with the solution containing algae to prepare an algae bacteria mixed solution; the algae-containing solution is used in an amount such that the biomass of algae in the algae-bacteria symbiotic capsules is 1.0g/L to 1.5g/L;
soaking sodium alginate powder in distilled water at 30-40 ℃ for 1-2 h, regulating the pH of the distilled water to 8.0-9.0 by using sodium hydroxide solution, and continuously stirring at 90-100 ℃ to dissolve the sodium alginate powder to prepare sodium alginate solution with the mass percent concentration of 1-2%; cooling the sodium alginate solution to 25-30 ℃, and adding powdery slag with the particle size of 250-300 meshes into the sodium alginate solution according to the proportion of (0.5-1) g to 100mL to prepare sodium alginate-slag mixed solution;
Adding the algae bacteria mixed solution into the sodium alginate-slag mixed solution to prepare embedding solution;
Dripping the embedded liquid into a calcium chloride crosslinking solution with the partial mass concentration of 2-4% which is refrigerated for 12-24 hours at the temperature of 4-5 ℃ in advance for preliminary crosslinking and curing, washing an intermediate product obtained after the preliminary crosslinking and curing is finished, continuously adding the rest calcium chloride crosslinking solution with the mass concentration of 2-4%, and continuously crosslinking and fixing for 12-24 hours at the temperature of 4-5 ℃ to obtain the algae bacteria immobilized capsule; wherein the mass ratio of the embedding liquid to the crosslinking solution is 1 (1-2);
Adding chitosan with the deacetylation degree of 80-95% into acetic acid solution with the mass percent concentration of 0.4-0.8% to prepare acetic acid-chitosan solution; the mass volume ratio of the chitosan to the acetic acid solution is (0.5-1.5) g to 100mL;
Adding the algae immobilized capsule into acetic acid-chitosan solution, stirring and hanging the membrane for 1h-1.5h to obtain the algae symbiotic capsule.
In some embodiments, the algae symbiotic capsules are stored at 4 ℃ in a storage solution having a calcium ion concentration of 0.1% -0.2%.
The second aspect of the application provides an algae-bacteria symbiotic capsule prepared by the method of the first aspect of the application.
The algae-bacteria symbiotic capsule provided by the application has the advantages that the time required for completely removing total nitrogen and total phosphorus in the cultivation wastewater is obviously reduced, the removing effect is obviously optimized, and the repeated usability and the system stability are realized
In some embodiments, the particle size of the algae symbiotic capsules is from 3.8mm to 4mm. For example, the particle size is not limited to 3.8mm, 3.83mm, 3.85mm, 3.87mm, 3.9mm, 3.93mm, 3.95mm, 3.97mm, 4mm, or a range between any two of the above particle sizes.
The third aspect of the application provides a cultivation wastewater treatment method, comprising the following steps:
Mixing the algae-bacteria symbiotic capsules with the culture wastewater to be treated so as to treat the culture wastewater to be treated; wherein the algae-bacteria symbiotic capsule is prepared by the method of the first aspect of the application.
In some embodiments, the algae-bacteria symbiotic capsules are mixed with the wastewater to be treated in a ratio of 20g/L to 60 g/L. Thereby, the efficiency of removing the contamination of the cultivation wastewater can be enhanced.
In some embodiments, the conditions for treating the wastewater from the cultivation to be treated using the algae-symbiotic capsule include: the illumination intensity is 4000Lux-5000Lux, the light-dark ratio is 12:12, the temperature is 24 ℃ to 28 ℃, the aeration quantity is 0.1L/min-0.2L/min, and the treatment time is 7d. When the aeration rate is in the range, the contact frequency of the algae-bacteria symbiotic capsules and flowing water can be increased, and the absorption and utilization of pollutants in the wastewater by the algae-bacteria symbiotic capsules are enhanced; in addition, the aeration can provide additional oxygen for the algae-bacteria symbiotic system, so that the aerobic bacteria can be enhanced to utilize the oxygen to degrade organic matters.
The following describes the technical scheme of the present application in detail with reference to specific examples. It is to be understood that these examples are illustrative of the present application and are not intended to limit the scope of the present application. The experimental methods in the following examples, in which specific conditions are not noted, are preferably referred to the guidelines given in the present application, and may be according to the experimental manual or conventional conditions in the art, the conditions suggested by the manufacturer, or the experimental methods known in the art.
In the specific examples described below, the measurement parameters relating to the raw material components, unless otherwise specified, may have fine deviations within the accuracy of weighing. Temperature and time parameters are involved, allowing acceptable deviations from instrument testing accuracy or operational accuracy.
1. Preparation of algae-bacteria symbiotic capsule
Example 1 preparation of a mixture of algae and bacteria
Inoculating Chlorella pyrenoidosa into BG-11 culture medium, culturing at 26deg.C and pH of 8.0 for 8 days under illumination intensity of 6000Lux to obtain Chlorella pyrenoidosa liquid, centrifuging 100mL of Chlorella pyrenoidosa liquid in logarithmic growth phase at 5000r/min for 10min, and discarding supernatant to obtain concentrated Chlorella pyrenoidosa liquid when the biomass of the liquid reaches about 1.0 g/L.
The method comprises the steps of preparing an activated sludge culture solution from ammonium chloride, sodium chloride, monopotassium phosphate, glucose, ferric chloride, magnesium sulfate and distilled water, inoculating activated sludge into the culture solution, continuously aerating for 48 hours at 26 ℃ for one period, stopping aerating for 2 hours, and continuously culturing for 8 days to obtain the cultured activated sludge. Biomass in the activated sludge supernatant was measured according to algae: the biomass of the bacteria is 1:2, taking supernatant of the activated sludge after standing and precipitating in corresponding volumes, centrifuging for 10min at a rotation speed of 5000r/min, and discarding the supernatant to obtain the concentrated activated sludge aerobic bacteria liquid. Mixing the concentrated chlorella pyrenoidosa algae liquid with the concentrated activated sludge aerobic bacteria liquid, adding 5mL of distilled water, and stirring uniformly to obtain an algae bacteria mixed liquid.
Example 2
Adding 1g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 8.0 by using a sodium hydroxide solution, standing and swelling for 1h in a constant-temperature water bath at 40 ℃, then, carrying out water bath at 90 ℃, stirring uniformly by using a magnetic stirrer during water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 0.5g of powdery slag with the particle size of 250 meshes, and continuing stirring uniformly to obtain the sodium alginate-slag mixed solution.
Adding 4g of anhydrous calcium chloride into 200mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 2% after the calcium chloride is completely dissolved.
Adding 0.4mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 0.5g of chitosan powder with the deacetylation degree of 95%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dropwise adding the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 2% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 12 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. And then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at a constant speed and hanging the film for 1h to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.1 percent for low-temperature storage at the temperature of 4 ℃.
Example 3
Adding 1.5g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 8.0 by using sodium hydroxide solution, standing and swelling for 1.5h in a constant-temperature water bath at 40 ℃, then, water-bathing at 90 ℃, uniformly stirring by using a magnetic stirrer in the water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 1g of powdery slag with the particle size of 250 meshes, and continuously and uniformly stirring to obtain the sodium alginate-slag mixed solution.
And adding 6g of anhydrous calcium chloride into 200mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 3% after the calcium chloride is completely dissolved.
Adding 0.6mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 1g of chitosan powder with the deacetylation degree of 80%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dripping the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 3% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 24 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. Then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at constant speed and hanging the film for 1.5 hours to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.2 percent for low-temperature storage at the temperature of 4 ℃.
Example 4
Adding 2g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 8.5 by using a sodium hydroxide solution, standing and swelling for 1h in a constant-temperature water bath at 40 ℃, then carrying out water bath at 90 ℃, stirring uniformly by using a magnetic stirrer during water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 0.5g of powdery slag with the particle size of 300 meshes, and continuing stirring uniformly to obtain the sodium alginate-slag mixed solution.
And adding 8g of anhydrous calcium chloride into 200mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 4% after the calcium chloride is completely dissolved.
Adding 0.8mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 0.5g of chitosan powder with the deacetylation degree of 85%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dripping the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 4% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 24 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. And then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at a constant speed and hanging the film for 1h to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.1 percent for low-temperature storage at the temperature of 4 ℃.
Example 5
Adding 1g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 9.0 by using a sodium hydroxide solution, standing and swelling for 2 hours in a constant-temperature water bath at 40 ℃, then carrying out water bath at 100 ℃, stirring uniformly by using a magnetic stirrer during water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 1g of powdery slag with the particle size of 300 meshes, and continuing stirring uniformly to obtain the sodium alginate-slag mixed solution.
And adding 2g of anhydrous calcium chloride into 100mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 2% after the calcium chloride is completely dissolved.
Adding 0.6mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 1g of chitosan powder with the deacetylation degree of 90%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dripping the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 2% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 24 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. And then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at a constant speed and hanging the film for 1h to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.2 percent for low-temperature storage at the temperature of 4 ℃.
Example 6
Adding 2g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 9.0 by using a sodium hydroxide solution, standing and swelling for 1h in a constant-temperature water bath at 40 ℃, then carrying out water bath at 100 ℃, stirring uniformly by using a magnetic stirrer during water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 1g of powdery slag with the particle size of 250 meshes, and continuing stirring uniformly to obtain the sodium alginate-slag mixed solution.
Adding 4g of anhydrous calcium chloride into 100mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 4% after the calcium chloride is completely dissolved.
Adding 0.8mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 1.5g of chitosan powder with the deacetylation degree of 90%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dripping the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 4% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 24 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. Then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at constant speed and hanging the film for 1.5 hours to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.2 percent for low-temperature storage at the temperature of 4 ℃.
Comparative example 1
The algal-bacterial mixed solution in example 1 was not added when the algal-bacterial symbiotic capsule was prepared in comparative example 1; the method comprises the following steps:
Adding 2g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 9.0 by using a sodium hydroxide solution, standing and swelling for 1.5h in a constant-temperature water bath at 40 ℃, then carrying out water bath at 95 ℃, uniformly stirring by using a magnetic stirrer in the water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 1g of powdery slag with the particle size of 300 meshes, and continuously uniformly stirring to obtain the sodium alginate-slag mixed solution.
And adding 8g of anhydrous calcium chloride into 200mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 4% after the calcium chloride is completely dissolved.
Adding 0.8mL of acetic acid solution into 100mL of distilled water, fully and uniformly stirring, adding 1.5g of chitosan powder with the deacetylation degree of 90%, and uniformly stirring again to dissolve the chitosan powder to obtain the acetic acid-chitosan solution.
The volume ratio of the sodium alginate-slag mixed solution to the cross-linking agent solution is 1:2, a pear-shaped separating funnel is used for dripping the sodium alginate-slag mixed solution into a part of calcium chloride cross-linking agent solution which is pre-refrigerated for 24 hours at the temperature of 4 ℃ for preliminary cross-linking fixation, an intermediate product obtained after the preliminary cross-linking fixation is screened out, distilled water is used for repeatedly washing for three times, the intermediate product is continuously added into the rest 4% calcium chloride cross-linking agent solution, and the complex cross-linking fixation is carried out for 24 hours at the temperature of 4 ℃ to obtain the phycomycete immobilized capsule. Then adding the algae immobilized capsule into acetic acid-chitosan solution for coating, stirring at constant speed and hanging the film for 1.5 hours to obtain the sodium alginate-slag-chitosan algae symbiotic capsule with the particle size of 4mm, filtering the capsule by using a screen, washing the capsule with distilled water for three times, and then placing the capsule in a storage solution with the calcium ion concentration of 0.2 percent for low-temperature storage at the temperature of 4 ℃.
Comparative example 2
In comparative example 2, acetic acid-chitosan coating was not performed when preparing the algal symbiotic capsules; the method comprises the following steps:
Adding 2g of sodium alginate powder into 100ml of distilled water, regulating the pH value of the distilled water to 9.0 by using a sodium hydroxide solution, standing and swelling for 1h in a constant-temperature water bath at 40 ℃, then carrying out water bath at 95 ℃, uniformly stirring by using a magnetic stirrer during water bath until the sodium alginate powder is completely dissolved, cooling the sodium alginate solution to 30 ℃ after the sodium alginate powder is completely dissolved, adding 1g of powdery slag with the particle size of 300 meshes, and continuously and uniformly stirring to obtain the sodium alginate-slag mixed solution.
And adding 8g of anhydrous calcium chloride into 200mL of distilled water, fully and uniformly stirring, and preparing the calcium chloride cross-linking agent solution with the mass fraction of 4% after the calcium chloride is completely dissolved.
Adding the algae bacteria mixed solution prepared in the embodiment 1 into the sodium alginate-slag mixed solution, uniformly stirring to obtain an embedding solution, dripping the embedding solution into a part of calcium chloride cross-linking agent solution pre-refrigerated for 24 hours at 4 ℃ by using a pear-shaped separating funnel for preliminary cross-linking fixation, screening out an intermediate product obtained after the preliminary cross-linking fixation, repeatedly washing for three times by using distilled water, continuously adding the intermediate product into the rest 3% of calcium chloride cross-linking agent solution, and carrying out double cross-linking fixation for 24 hours at 4 ℃ to obtain the algae bacteria immobilized capsule. The capsules were filtered using a sieve and washed three times with distilled water, and then the capsules were stored at a low temperature of 4 ℃ in a storage solution having a calcium ion concentration of 0.2%.
Comparative example 3
The preparation method of comparative example 3 and example 6 are similar, except that: only the concentrated chlorella pyrenoidosa algae liquid was added, and the concentrated chlorella pyrenoidosa algae liquid having the same biomass prepared in example 1 was used instead of the concentrated activated sludge aerobic bacteria liquid.
Comparative example 4
The preparation method of comparative example 4 and example 6 are similar, except that: only the concentrated activated sludge aerobic bacteria liquid is added, and the concentrated activated sludge aerobic bacteria liquid with the same biomass prepared in the example 1 is adopted to replace the concentrated chlorella pyrenoidosa algae liquid.
2. Artificial simulated cultivation wastewater treatment test
Adding configured artificial simulated aquaculture wastewater into a reactor with a consistent structure, wherein the pH value of the wastewater is 7.66, wherein urea and ammonium chloride are added to supplement total nitrogen and ammonia nitrogen, potassium dihydrogen phosphate is added to supplement total phosphorus, glucose is added to supplement COD (Mn), the total nitrogen concentration of the artificial simulated freshwater aquaculture wastewater is about 15 mg.L -1, and the ammonia nitrogen concentration is about 3 mg.L -1; the total phosphorus concentration was about 1.5 mg.L -1;COD(Mn) and the concentration was about 30 mg.L -1.
After the unused symbiotic capsules prepared in examples 2 to 5 and comparative examples 1 to 2 were stored for 1 month, the symbiotic capsules were washed and dried, and then were respectively added into a reactor containing artificial simulated aquaculture wastewater according to a ratio of 60 g.L -1, treatment effect tests were performed for 7 days, total nitrogen, ammonia nitrogen, total phosphorus and COD (Mn) concentrations in the wastewater after treatment were measured, and removal rates of various pollutants and degradation rates of the symbiotic capsules were calculated, and the results are shown in Table 1.
TABLE 1
As can be seen from the comparison of the results of examples 1-6 and comparative examples 1-4 in Table 1, the algae-bacteria symbiotic capsules provided by the application have remarkable effect of removing nutrient substances in the cultivation wastewater, low degradation rate and more stable structure. In addition, the effect of the algae-bacteria symbiotic capsule on removing the nutrient substances in the cultivation wastewater is obviously better than the effect of the algae-bacteria symbiotic capsule on removing the nutrient substances in the cultivation wastewater.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (16)
1. The preparation method of the algae-bacteria symbiotic capsule is characterized by comprising the following steps of:
Mixing the solution containing aerobic bacteria with the solution containing algae to prepare an algae bacteria mixed solution;
mixing the algae bacteria mixed solution with sodium alginate-slag mixed solution containing sodium alginate and slag to prepare embedding solution;
mixing the embedding liquid with a crosslinking solution to carry out crosslinking fixation, so as to prepare an algae bacteria immobilized capsule;
mixing the algae immobilized capsule with a carboxylic acid-chitosan solution, and forming a film to prepare the algae symbiotic capsule;
The preparation method of the sodium alginate-slag mixed solution comprises the following steps: mixing sodium alginate solution with slag to prepare sodium alginate-slag mixed solution; the mass percentage concentration of the sodium alginate solution is 1% -2%; the mass volume ratio of the slag to the sodium alginate solution is (0.5-1) g to 100mL; the grain diameter of the slag is 250-300 meshes;
the preparation method of the carboxylic acid-chitosan solution comprises the following steps: mixing a carboxylic acid solution with chitosan to prepare the carboxylic acid-chitosan solution; the deacetylation degree of the chitosan is 80% -95%; the pH of the carboxylic acid-chitosan solution is 5.5-6.0;
The preparation method of the algae bacteria immobilized capsule comprises the following steps: adding the embedding liquid into a partial crosslinking solution which is refrigerated for 12-24 hours at the temperature of 4-5 ℃ in advance for preliminary crosslinking fixation; washing the intermediate product obtained after preliminary crosslinking and fixing, adding the washed intermediate product into the residual crosslinking solution, and continuing crosslinking and fixing for 12-24 hours at the temperature of 4-5 ℃ to obtain the algae bacteria immobilized capsule.
2. The method of preparing an algae symbiotic capsule of claim 1 wherein the algae comprises one or more of chlorella, scenedesmus and crescent algae.
3. The method of preparing an algae symbiotic capsule of claim 2 wherein the algae is chlorella pyrenoidosa.
4. The method of preparing an algae-bacteria symbiotic capsule of claim 1 wherein the aerobic bacteria comprise activated sludge aerobic bacteria.
5. A method of preparing an algae symbiotic capsule of claim 1 wherein the carboxylic acid solution is selected from acetic acid solutions.
6. The method for preparing the algicidal symbiotic capsule of claim 5 wherein the concentration of the acetic acid solution by mass percent is 0.4% -0.8%.
7. The method for preparing the symbiotic capsule of claim 5 wherein the mass-to-volume ratio of chitosan to acetic acid solution is (0.5-1.5) g to 100mL.
8. A method of preparing an algae symbiotic capsule of any of claims 1 to 7 wherein the biomass of the algae containing solution is 1.0g/L to 1.5g/L.
9. The method for producing an algal symbiotic capsule according to any of claims 1 to 7 wherein the algal cell-containing solution and the algal-containing solution are mixed in a ratio of 1 (1.5-2) in terms of an algal biomass ratio when the algal cell-containing mixed solution is produced.
10. A method for preparing an algae symbiotic capsule according to any of claims 1 to 7 wherein the cross-linking solution is a calcium chloride solution with a mass percentage concentration of 2% -4%.
11. The method of preparing an algae symbiotic capsule of claim 10 wherein the volume ratio of the embedding liquid to the cross-linking solution is 1 (1-2).
12. An algal symbiotic capsule prepared by the method of any of claims 1 to 11.
13. An algae symbiotic capsule according to claim 12, wherein the particle size of the algae symbiotic capsule is 3.8mm to 4mm.
14. The cultivation wastewater treatment method is characterized by comprising the following steps of:
mixing the algae-bacteria symbiotic capsules with the culture wastewater to be treated so as to treat the culture wastewater to be treated; wherein the phycobiont capsule is prepared by the method of any one of claims 1 to 11.
15. The method for treating aquaculture wastewater according to claim 14 wherein said algal-bacterial symbiotic capsule is mixed with said aquaculture wastewater to be treated in a ratio of 20g/L to 60 g/L.
16. The aquaculture wastewater treatment method of claim 14 wherein said treatment conditions include: the illumination intensity is 4000Lux-5000Lux, the light-dark ratio is 12:12, the temperature is 24 ℃ to 28 ℃, the aeration quantity is 0.1L/min-0.2L/min, and the treatment time is 7d.
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