CN116354494A - Composite slow-release carbon material coupled aerobic denitrifying bacteria and application thereof in water treatment - Google Patents
Composite slow-release carbon material coupled aerobic denitrifying bacteria and application thereof in water treatment Download PDFInfo
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- CN116354494A CN116354494A CN202310162310.2A CN202310162310A CN116354494A CN 116354494 A CN116354494 A CN 116354494A CN 202310162310 A CN202310162310 A CN 202310162310A CN 116354494 A CN116354494 A CN 116354494A
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Classifications
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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
- C02F3/02—Aerobic processes
- C02F3/10—Packings; Fillings; Grids
- C02F3/105—Characterized by the chemical composition
- C02F3/106—Carbonaceous materials
-
- 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
- C02F3/02—Aerobic processes
-
- 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
- C02F3/28—Anaerobic digestion processes
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
- C12N11/096—Polyesters; Polyamides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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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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- 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
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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
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/08—Chemical Oxygen Demand [COD]; Biological Oxygen Demand [BOD]
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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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
Abstract
The invention relates to a composite slow-release carbon material coupling aerobic denitrifying bacteria, which comprises a slow-release carbon carrier and immobilized heterotrophic nitrification-aerobic denitrifying bacteria, wherein the slow-release carbon carrier is formed by uniformly mixing euglena particles and polybutylene adipate and then attaching and fixing the euglena particles and polybutylene adipate to an elastic filler, and the immobilized heterotrophic nitrification-aerobic denitrifying bacteria is formed by attaching and fixing pseudomonas stutzeri and/or microbacterium oxydans to the elastic filler. The invention is beneficial to the process of coordinating the carbon release rate and denitrification, strengthens the aerobic denitrifying bacteria with wider adaptation temperature range, simultaneously adopts two bacteria which have advantages in the aspect of utilizing polybutylene adipate and paramylon of euglena, and avoids the excessive utilization of carbon sources by other aerobic bacteria; in the process of treating the cultivation tail water with low carbon nitrogen ratio, a longer service cycle and a better denitrification effect can be obtained, and the problem that the COD of effluent exceeds the standard is difficult to occur, thereby being beneficial to the standard discharge of the tail water.
Description
Technical Field
The invention relates to a method for deep denitrification of sewage with low carbon nitrogen ratio, in particular to application of euglena-based slow-release carbon material coupled aerobic denitrifying bacteria in water treatment, and belongs to the technical field of water treatment.
Background
With the rapid development of aquaculture in China, the problems of environmental pollution and the like caused by the discharge of the aquaculture tail water in China are more and more prominent because the eutrophication degree of the adjacent water areas is aggravated by residual baits and the aquaculture biological excreta rich in nitrogen, phosphorus and organic matters in the aquaculture tail water.
At present, the treatment means of the culture tail water are mainly divided into three types of physical, chemical and biological restoration. Among these, bioremediation is considered to be the most economical, reasonable and environmentally friendly method. Bioremediation is largely divided into two modes, phytoremediation and microbial remediation. However, for phytoremediation, the purification effect is obviously affected by seasonal factors, the purification capacity is weaker in autumn and winter, and if the withered seasons cannot be timely harvested, secondary pollution can be caused to the water body. While the microorganism restoration can furthest reduce the concentration of pollutants such as nitrogen, phosphorus and the like, the problem of low denitrification efficiency caused by insufficient carbon source of the culture tail water is needed to be solved, and the problem of complicated steps of traditional aerobic nitrification-anoxic denitrification is solved.
Therefore, in order to ensure the smooth and efficient operation of denitrification, an additional carbon source needs to be added. At present, most of denitrification and additional carbon sources adopt traditional nitrogen sources such as alcohols, acetic acid, glucose and the like, and the problems of large addition amount, high operation cost and easy water flow loss exist. In order to avoid the defect of adding traditional liquid carbon sources such as methanol, acetic acid, glucose and the like, slow-release carbon materials prepared by compounding natural high molecular polymers (such as plant cellulose and plant starch) with artificially synthesized biodegradable polymers (such as polylactic acid, polycaprolactone and polyvinyl alcohol) are currently developed, and the release of the carbon sources of the current compound slow-release carbon materials is controlled to a certain extent compared with the release of the traditional liquid carbon sources or the natural high molecular polymers alone, but the part of the currently used natural high molecular polymers which is easily dissolved in water or easily dispersed in water has a high proportion, such as 24.68 percent of organic carbon in plant straws is extremely easily dissolved in water, and can be completely released into water within 2 days after soaking. Therefore, the composite slow-release carbon material still has the problems of secondary pollution such as high effluent chromaticity, exceeding effluent COD and the like. In addition, such long-term soaking in water as plant straw has the disadvantage of being significantly soft, collapsing or shattering.
Meanwhile, the conventional denitrification needs to separate the aerobic nitrification unit from the anaerobic/anoxic denitrification unit, and is completed by different bacteria, such as an A2O process commonly used in sewage treatment plants, namely anaerobic-anoxic-aerobic. In the natural water body in-situ remediation, the sewage treatment plant cannot achieve strict condition control, so that the denitrification nitrogen efficiency is greatly affected. The flora is naturally domesticated, and the bacteria with excellent denitrification capability and wider adaptation temperature range are not assisted and enhanced.
The Chinese patent with publication number of CN114349281A discloses a denitrification and dephosphorization treatment device and a water treatment method for low-carbon-nitrogen-ratio polluted water, which separate traditional aerobic nitrification from anaerobic denitrification by different strains, because aerobic nitrifying bacteria are autotrophic bacteria, carbon sources need to be reduced to create alkaline environment, anaerobic denitrifying bacteria are heterotrophic bacteria, carbon sources need to be increased to create weak alkaline environment. The ecological treatment chamber of the device is used for planting plants, absorbing part of nitrogen and phosphorus, aerating, creating an aerobic environment and promoting the nitrification of aerobic nitrifying bacteria; the enhanced denitrification chamber adopts physical adsorption of zeolite and shell powder; the anaerobic denitrification chamber creates a weak alkaline environment, and promotes anaerobic denitrification to be in an optimal pH condition. Meanwhile, the anaerobic denitrification chamber uses light biological carbon and diatomite with high density for crosslinking, the biological carbon has good adsorptivity and oxidability, and glucose or starch is loaded on the biological carbon as an available carbon source. The patent is more concerned about denitrification and dephosphorization, but does not pay attention to the COD content of the effluent water quality, and the effluent COD is higher than the discharge standard of the cultivation tail water, so that the patent is not suitable for being applied to the purification of the cultivation tail water. In order to improve denitrification and dephosphorization, the patent adopts glucose and starch as a load carbon source, release is not easy to control, short-term carbon source excess is easy to form, so that the nitrogen and phosphorus content of raw water treated by the patent is extremely high, and the patent is not suitable for the treatment of water polluted by low nitrogen and phosphorus content of aquaculture tail water with low effluent COD. Meanwhile, because glucose and starch are available for most of bacterial groups in nature and are carbon sources which are rapidly utilized, and not only anaerobic denitrifying bacteria can be utilized, the competition pressure of the anaerobic denitrifying bacteria and other anaerobic bacteria carbon sources is larger.
The Chinese patent with publication number of CN114506919A discloses a high-efficiency purifying composite biological filler and a preparation method thereof, wherein the method also separates the traditional aerobic nitrification and anaerobic denitrification and is respectively realized by different strains. The preparation method comprises the steps of mixing diatomite ceramic particles with naturally domesticated aerobic biological activated sludge until microorganisms are fully coated to prepare diatomite ceramic particles, so as to form aerobic nitrifying bacteria groups positioned on the upper layer of the device; and adding a composite biological filler containing quartz sand, zero-valent iron, active carbon, a composite microbial agent and a porous slow-release biomass carbon source into the deep purification layer, wherein the composite microbial agent adopts an artificially added facultative anaerobic denitrification dephosphorization composite strain preparation, and strains comprise pseudomonas, bacillus and alcaligenes. The porous slow-release biomass carbon source is a porous biomass solid carbon source produced by processing natural carbon sources such as corncobs, rice hulls, wood chips or corn stalks through a hot alkali method. About one-fourth of organic carbon in the plant straw is very soluble in water, and the thermal alkali aqueous solution is further treated to further disperse about half of lignin which is not easy to degrade in alkali water, so that the alkali wastewater with high chromaticity and high organic matters can cause great pollution to the environment if the alkali wastewater is not treated well. The plant straw treated by the alkaline method is soaked in water for a long time, is obviously easier to be softened and broken after being acted by microorganisms, still remains refractory substances, and is extremely easy to cause the blockage of water flow pores. The microbial inoculum used does not adopt some measure of immobilization and is easy to run off with water flow. The straw processed by the thermokalite method is mainly cellulose, hemicellulose and residual lignin, and bacteria which can take the cellulose and the hemicellulose as carbon sources are also very wide in nature, so that the competition pressure of the carbon sources between the added anaerobic denitrifying bacteria and other anaerobic bacteria is larger. Meanwhile, the patent only measures the ammonia nitrogen content in the water body, and the ammonia nitrogen is one form of the nitrogen in the water body, and is also one form of the nitrogen which is easiest to remove in biological denitrification, but one part of the nitrogen is possibly directly utilized by organisms, and the other part of the nitrogen is converted into the nitrogen in other forms and still exists in the water body.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: aiming at the problems existing in the prior art, the application of the composite slow-release carbon material coupled aerobic denitrifying bacteria in water treatment is provided
In order to achieve the above purpose, the technical scheme of the invention is as follows:
the euglena-based slow-release carbon material coupling aerobic denitrifying bacteria comprises a slow-release carbon carrier and immobilized heterotrophic nitrification-aerobic denitrifying bacteria, wherein the slow-release carbon carrier is formed by uniformly mixing euglena particles and polybutylene adipate and then attaching and fixing the mixture to an elastic filler, and the immobilized heterotrophic nitrification-aerobic denitrifying bacteria is formed by attaching and fixing pseudomonas stutzeri and/or microbacterium oxydans to the elastic filler.
In the technical proposal, the euglena is a single-cell algae, and the main component of the euglena body is euglena hemsleyanumEuglena sanguinea) The cell body is rich in one kind of paramylon, which may account for 56% or more of the dry matter content, up to 85%, and the paramylon is amylose beta-1, 3 glucan, different from the amylose alpha-1, 4 or branched alpha-1, 6 of common starch in life. The paramylon of the euglena has a compact triple helix structure, so that the paramylon is insoluble in water, can be used as a carbon source of denitrifying bacteria with beta-1, 3 glucanase, and is beneficial to reducing the consumption of carbon sources by a part of other aerobic bacteria.
Polybutylene adipate (PBA) has excellent molding processability, and is not similar to the defect that materials such as polylactic acid, polyvinyl alcohol and the like have obvious softness or breakage after being soaked in water for a long time. The polybutylene adipate can be used as a carrier framework for microorganism adhesion, can provide a carbon source for specific denitrifying bacteria, and is beneficial to reducing the consumption of carbon sources by a part of other aerobic bacteria. In addition, the range of the flora capable of degrading the strain is further reduced, which is helpful for greatly reducing the competitive pressure of carbon sources caused by other bacteria.
Compared with polybutylene adipate, the euglena paramylon is easier to be decomposed and utilized by bacteria, and the invention coordinates the release of carbon, the utilization of carbon and the aerobic denitrification process by matching the PBA with the euglena so as to obtain a longer service cycle of the slow-release carbon carrier and better effluent quality.
Pseudomonas stutzeriPseudomonas stutzeri ) The strain is preserved in China general microbiological culture Collection center (CGMCC) at the address of 10 and 19 days in 2020: the preservation number of the four-way No. 1 hospital No. 3 in the North Chen of the Chaoyang area of Beijing city is CGMCC No.20910. Pseudomonas stutzeri can take inorganic nitrogen such as ammonia nitrogen, nitrite nitrogen or nitrate nitrogen in water body as unique nitrogenThe carbon source is widely used, complex macromolecules such as starch, cellulose, polybutylene adipate and the like can be decomposed and utilized as the carbon source, and heterotrophic nitrification-aerobic denitrification reaction can be synchronously carried out in situ in an environment of 6-35 ℃.
Microbacterium oxydansMicrobacterium oxydans) The strain is preserved in China general microbiological culture Collection center (CGMCC) at 2021, 06 and 21, address: the preservation number of the four-way No. 1 hospital No. 3 in the North Chen of the Chaoyang area of Beijing city is CGMCC No.22738. The absorption capacity of the thallus of the microbacterium oxydans to phosphorus is more than 3 times of that of common bacteria, and inorganic nitrogen such as ammonia nitrogen and nitrate nitrogen in water can be used as a unique nitrogen source; the carbon source is widely used, complex macromolecules such as starch, auxiliary starch, cellulose, polybutylene adipate, phenol and the like can be decomposed and utilized as the carbon source, and heterotrophic nitrification-aerobic denitrification reaction can be synchronously carried out in situ in an environment of 6-35 ℃.
In a word, the invention adopts heterotrophic nitrification-aerobic denitrification bacteria with low temperature resistance, so that the nitrification and denitrification processes can simultaneously occur under heterotrophic and aerobic conditions and are completed by the same bacteria. The slow-release carbon carrier is composed of euglena particles and polybutylene adipate, wherein euglena is rich in paramylon, and the paramylon has a compact triple helix structure, so that the paramylon is insoluble in water; and the secondary starch has a structure of beta-1, 3 glycosidic chain, is different from the straight chain alpha-1, 4 or branched chain alpha-1, 6 glycosidic chain of common starch, can be used as a carbon source of denitrifying bacteria with beta-1, 3 glucanase, and is beneficial to reducing the consumption of carbon sources by a part of other aerobic bacteria without beta-1, 3 glucanase function. In addition, the polybutylene adipate is used as a slow-release carbon carrier main body, so that the defect of obvious softness or breakage caused by long-time soaking in water is avoided, the flora range for degrading the polybutylene adipate is further narrowed, and the carbon source competition pressure caused by other bacteria is greatly reduced.
The technical scheme of the invention is further optimized as follows:
further, the euglena particles and the polybutylene adipate are uniformly mixed according to the mass ratio of 1:20-400 and then are adhered and fixed on the brush-shaped elastic filler; the pseudomonas stutzeri and the microbacterium oxydans are uniformly mixed according to the concentration ratio of 1-3:2-1 and then are adhered and fixed on the brush-shaped elastic filler.
Furthermore, the pseudomonas stutzeri is pseudomonas stutzeri @Pseudomonas stutzeri ) The preservation number is CGMCC NO.20910; the microbacterium oxide is microbacterium oxide @Microbacterium oxydans) The preservation number is CGMCC NO.22738.
The invention provides a preparation method of euglena-based slow-release carbon material coupled aerobic denitrifying bacteria, which comprises the following steps:
firstly, preparing a slow-release carbon carrier, namely preparing euglena particles by a bridge action of a euglena powder combined binder, uniformly mixing the euglena particles with polybutylene adipate to obtain a mixed solution, and immersing a brush-shaped elastic filler into the mixed solution for adhesion to obtain the elastic filler loaded with a slow-release carbon material;
secondly, preparing immobilized heterotrophic nitrification-aerobic denitrification bacteria, namely adding pseudomonas stutzeri and/or microbacterium oxydans into a polyvinyl alcohol solution to obtain bacterial liquid, immersing brush-shaped elastic filler into the bacterial liquid, and dropwise adding saturated boric acid solution into the bacterial liquid to enable the pseudomonas stutzeri and/or microbacterium oxydans to be attached to the elastic filler, so as to obtain the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria;
thirdly, mixing and collocating the elastic filler loaded with the slow-release carbon material and the elastic filler loaded with the immobilized heterotrophic nitrification-aerobic denitrification bacteria to obtain the euglena-based slow-release carbon material coupled aerobic denitrification bacteria.
In the first step, collecting euglena or pure cultured euglena, drying, grinding, crushing and sieving with a 200-mesh sieve for later use; 4-6% of polyvinyl alcohol (PVA) is used as a binder; adding 1kg of euglena powder into a mixing granulator, uniformly spraying a binder in a mist form through a high-pressure nozzle to form euglena particles with the particle size of 0.3-2 mm, drying, and sieving with a 20-mesh sieve for later use; polybutylene adipate (PBA) was melted at 75 ℃ in a mass ratio of 20:1 to 400:1, uniformly mixing polybutylene adipate and euglena particles to obtain a mixed solution; immersing the brush-shaped elastic filler into the mixed solution for adhesion, taking out the liquid level after 20+/-5 s, and cooling at room temperature for fixing and forming.
In the second step, polyvinyl alcohol (PVA) is heated and dissolved in water to a final concentration of more than 6%, and then cooled to a temperature lower than 40 ℃ to obtain a polyvinyl alcohol solution; adding Pseudomonas stutzeri and/or Microbacterium oxydans into polyvinyl alcohol solution until the final concentration of bacterial solution is 1×10 6 CFU/ml～2×10 7 And (3) CFU/ml, immersing the brush-shaped elastic filler into bacterial liquid, slowly stirring, dropwise adding saturated boric acid solution according to the volume of 4% -10% of the bacterial liquid in the stirring process, continuously stirring for 1min to ensure that viscous substances attached to the elastic filler are obviously gathered, slightly taking out the elastic filler, slowly immersing into the saturated boric acid solution for fixing for 1-12 h, and then immersing into water for 4-6 h to obtain the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria.
In the technical scheme, the concentration ratio of the pseudomonas stutzeri to the microbacterium oxydans is preferably 1-3:2-1.
Furthermore, the Pseudomonas stutzeri is Pseudomonas stutzeri @Pseudomonas stutzeri) The preservation number is CGMCC NO.20910; the microbacterium oxide is microbacterium oxide @Microbacterium oxydans) The preservation number is CGMCC NO.22738.
The invention also provides application of the euglena-based slow-release carbon material coupled aerobic denitrifying bacteria in water treatment.
The euglena-based slow-release carbon material is coupled with the application of aerobic denitrifying bacteria in purifying the tail water of cultivation.
Further, the elastic filler loaded with the slow-release carbon material and the elastic filler loaded with the immobilized heterotrophic nitrification-aerobic denitrification bacteria are mixed according to the proportion of 10:1, suspended below a water surface floating bed support and immersed in the culture tail water.
The invention takes the brush-shaped elastic filler loaded with the fixed heterotrophic nitrification-aerobic denitrification bacteria as a bacterial pool for continuously strengthening the aerobic denitrification bacteria, so that the bacteria needing strengthening cannot gradually run off due to water flow impact, and the two bacteria used by the invention have better low temperature resistance.
In the present invention, "%" not specifically described means weight percent.
The invention is beneficial to the process of coordinating the carbon release rate and denitrification, strengthens the aerobic denitrifying bacteria with wider adaptation temperature range, simultaneously adopts two bacteria which have advantages in the aspect of utilizing polybutylene adipate and paramylon of euglena, and avoids the excessive utilization of carbon sources by other aerobic bacteria; in the process of treating the cultivation tail water with low carbon nitrogen ratio, a longer service cycle and a better denitrification effect can be obtained, and the problem that the COD of effluent exceeds the standard is difficult to occur, thereby being beneficial to the standard discharge of the tail water.
Drawings
The invention is further described below with reference to the accompanying drawings.
FIG. 1 is a schematic diagram showing the degradation effect of PBA under the treatment conditions of Pseudomonas stutzeri, microbacterium oxydans and Pseudomonas stutzeri+Microbacterium oxydans in the present invention.
FIG. 2 is a schematic diagram showing the effect of the addition amount of Euglena on TN removal effect in the present invention.
FIG. 3 is a schematic view of the structure of the reactor in the present invention.
FIG. 4 is a schematic diagram showing COD treatment effect of the tail water of the cultivation in example 1 of the present invention.
FIG. 5 is a schematic view showing the effect of TN water treatment in the cultivation tail of example 1 of the present invention.
Description of the embodiments
The present invention will be described in further detail with reference to examples. The invention is not limited to the examples given.
The invention provides an application of euglena-based slow-release carbon material coupled aerobic denitrifying bacteria in purifying cultivation tail water, which specifically comprises the following steps:
preparation of a slow-release carbon carrier: collecting Euglena bodies or pure cultured Euglena, oven drying, grinding, and sieving with 200 mesh sieve; 4-6% of polyvinyl alcohol (PVA) is used as a binder; adding 1kg of euglena powder into a mixing granulator, uniformly spraying a binder in a mist form through a high-pressure nozzle to form euglena particles with the particle size of 0.3-2 mm, drying, and sieving with a 20-mesh sieve for later use; polybutylene adipate (PBA) was melted at 75 ℃ according to polybutylene adipate and euglena particles 20:1 to 400:1, uniformly mixing to obtain a mixed solution; the elastic filler (purchased from Yixing city, yi-environment-friendly Co., ltd.) in the shape of a brush made of PP material is immersed in the mixed solution to adhere, the liquid surface is taken out after 20+ -5 s, and the mixture is cooled at room temperature to be fixed and formed.
Preparation of immobilized heterotrophic nitrification-aerobic denitrification bacteria: heating and dissolving polyvinyl alcohol (PVA) in water to a final concentration of more than 6%, and cooling to a temperature of less than 40 ℃ to obtain a polyvinyl alcohol solution; adding Pseudomonas stutzeri and Microbacterium oxydans into polyvinyl alcohol solution until the final concentration of bacterial liquid is 1×10 6 CFU/ml～2×10 7 CFU/ml, the concentration ratio of pseudomonas stutzeri to microbacterium oxydans is preferably 1-3:2-1, a hairbrush-shaped elastic filler (purchased from Yixing commercial environmental protection Co., ltd.) made of PP material is immersed into bacterial liquid, slowly stirred, and saturated boric acid solution is dripped according to 4% -10% of the volume of the bacterial liquid in the stirring process, and stirring is continued for 1min, so that adhesive substances attached to the elastic filler are obviously gathered, the elastic filler is gently taken out, then is slowly immersed into the saturated boric acid solution for fixing for 1-12 h, and then is soaked in water for 4-6 h, thus obtaining the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrifying bacteria.
The brush-shaped elastic filler attached with the slow-release carbon carrier and the brush-shaped elastic filler loaded with the immobilized heterotrophic nitrification-aerobic denitrification bacteria are matched according to the proportion of 10:1, are hung below various types of water surface floating bed supports, and are immersed in the culture tail water.
The chemical reagents and materials used in the invention are all commercially available.
Pseudomonas stutzeriPseudomonas stutzeri) The strain is preserved in China general microbiological culture Collection center (CGMCC) at the address of 10 and 19 days in 2020: the preservation number of the four-way No. 1 hospital No. 3 in the North Chen of the Chaoyang area of Beijing city is CGMCC No.20910.
Microbacterium oxydansMicrobacterium oxydans) The strain is preserved in China general microbiological culture Collection center (CGMCC) at 2021, 06 and 21, address: the preservation number of the four-way No. 1 hospital No. 3 in the North Chen of the Chaoyang area of Beijing city is CGMCC No.22738.
In the invention, the synergistic effect of the pseudomonas stutzeri and the microbacterium oxydans is beneficial to improving the degradation rate of the PBA. The specific verification method comprises the following steps: culturing in 200ml denitrification culture medium (NaCl 2.5 g/L, mgSO) 4 ·7H 2 O 0.1 g/L、FeSO 4 ·7H 2 O 0.02 g/L、MnSO 4 0.016 g/L、CaCl 2 0.02 g/L、NaCO 3 0.02 g/L、KH 2 PO 4 0.213 g/L、KNO 3 0.722 g/L, trace elements and Hoagland medium), 1.5g of PBA as the sole carbon source was added, and the ambient temperature was 25 ℃. The strain is cultivated in LB culture medium in an aseptic way, collected by centrifugation and washed by aseptic water for later use. Three treatments were set up, each of which was Pseudomonas stutzeri (1.24X10) 7 Individual Microbacterium oxydans (1.35X10) 7 Individual/ml), pseudomonas stutzeri (8.1X10) 6 Mu.oxydans (4.6X10) 6 And/ml), for 14 days. The results show that the combined treatment of the pseudomonas stutzeri and the microbacterium oxydans is favorable for remarkably improving the degradation rate of the PBA, and is improved by 43.84 percent compared with the single addition of the microbacterium oxydans and 520.31 percent compared with the single addition of the pseudomonas stutzeri (see figure 1).
In the invention, the naked algae and the PBA are used as carbon sources, which is helpful for accelerating TN (total nitrogen) removal effect to a certain extent. The specific verification method comprises the following steps: shake flask culture was performed using a flask as a container, and 1.0 g of PBA was added to 200ml 10%Hoagland medium. The strain is aseptically cultured in LB medium, centrifugally collected, washed with sterile water, and added with Pseudomonas stutzeri (1.8X10 g) 6 Mu.m/ml) +Microbacterium oxydans (2.5X10) 6 And/ml). Five treatments were set up, namely, adding euglena dry powder 0 g, 0.005 g, 0.010 g, 0.025 g and 0.075 g, respectively, and culturing at ambient temperature 25 ℃ for 14 days. The result shows that the addition of a proper amount of euglena rich in paramylon can significantly improve the denitrification efficiency, and when 0.0100 g euglena is added, the TN removal amount is significantly improved by 22.90% (see figure 2).
Example 1
And (5) conveying the breeding tail water to a reactor for purification treatment. As shown in fig. 3, the reactor body was a rectangular parallelepiped made of plastic, and the effective volume was 12L. The reactor main body is internally provided with a brush-shaped elastic filler attached with a slow-release carbon carrier and a brush-shaped elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria, and the total filling volume of the brush-shaped elastic filler and the brush-shaped elastic filler is about 30% of the effective volume of the reactor. The bottom of the reactor body was equipped with an aeration head to maintain DO greater than 3mg/L and ambient temperature 25 ℃. The daily water change rate was 50%. The result shows that after the system is stable in operation after a 20-day starting period, the COD concentration of the inlet water is 82.53 +/-1.98 mg/L, and the COD concentration of the outlet water is 8.80+/-2.11 mg/L; the inlet water TN concentration is 8.19+/-0.13 mg/L, and the outlet water TN concentration is 1.56+/-0.17 mg/L (see FIG. 4 and FIG. 5). The COD and TN of the effluent meet the first-level standard of the tail water discharge of the freshwater aquaculture (the COD is less than or equal to 15mg/L and the TN is less than or equal to 3 mg/L). The carbon source is continuously replenished for more than 60 days.
Example 2
And (5) conveying the breeding tail water into a reactor for purification treatment. As shown in fig. 3, the reactor body was a rectangular parallelepiped made of plastic, and the effective volume was 12L. The reactor main body is internally provided with a brush-shaped elastic filler attached with a slow-release carbon carrier and a brush-shaped elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria, and the total filling volume of the brush-shaped elastic filler and the brush-shaped elastic filler is about 30% of the effective volume of the reactor. The bottom of the reactor body was equipped with an aeration head to maintain DO greater than 3mg/L and ambient temperature 16 ℃. The daily water change rate was 40%. The result shows that after the system is stable in operation after a 20-day starting period, the COD concentration of inflow water is 84.34 +/-1.94 mg/L, and the COD concentration of outflow water is 9.32+/-2.20 mg/L; the TN concentration of the inlet water is 8.01+/-0.15 mg/L, and the TN concentration of the outlet water is 1.87+/-0.12 mg/L. The COD and TN of the effluent meet the first-level standard of the tail water discharge of the freshwater aquaculture (the COD is less than or equal to 15mg/L and the TN is less than or equal to 3 mg/L).
Example 3
And purifying the culture tail water in the reactor. As shown in fig. 3, the reactor body was a rectangular parallelepiped made of plastic, and the effective volume was 12L. The reactor main body is internally provided with a brush-shaped elastic filler attached with a slow-release carbon carrier and a brush-shaped elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria, and the total filling volume of the brush-shaped elastic filler and the brush-shaped elastic filler is about 30% of the effective volume of the reactor. The bottom of the reactor body was equipped with an aeration head to maintain DO greater than 3mg/L and ambient temperature 8 ℃. The daily water change rate was 15%. The result shows that after the system is stable in operation in the starting period of 25 days, the COD concentration of the inlet water is 83.72 +/-2.18 mg/L, and the COD concentration of the outlet water is 17.41+/-1.25 mg/L; the TN concentration of the inlet water is 8.23+/-0.15 mg/L, and the TN concentration of the outlet water is 1.79+/-0.11 mg/L. The COD of the effluent meets the secondary standard of the discharge of the tail water of the freshwater aquaculture (COD is less than or equal to 25 mg/L), and the TN of the effluent meets the primary standard of the discharge of the tail water of the freshwater aquaculture (TN is less than or equal to 3 mg/L).
In addition to the embodiments described above, other embodiments of the invention are possible. All technical schemes formed by equivalent substitution or equivalent transformation fall within the protection scope of the invention.
Claims (10)
1. The composite slow-release carbon material coupling aerobic denitrifying bacteria is characterized by comprising slow-release carbon carriers and immobilized heterotrophic nitrification-aerobic denitrifying bacteria, wherein the slow-release carbon carriers are formed by uniformly mixing euglena particles and polybutylene adipate and then attaching and fixing the euglena particles and polybutylene adipate to elastic fillers, and the immobilized heterotrophic nitrification-aerobic denitrifying bacteria are formed by attaching and fixing pseudomonas stutzeri and/or microbacterium oxydans to the elastic fillers.
2. The composite slow-release carbon material coupling aerobic denitrifying bacteria according to claim 1, wherein the euglena particles and the polybutylene adipate are uniformly mixed according to the mass ratio of 1:20-400 and then are adhered and fixed on a brush-shaped elastic filler; the pseudomonas stutzeri and the microbacterium oxydans are uniformly mixed according to the concentration ratio of 1-3:2-1 and then are adhered and fixed on the brush-shaped elastic filler.
3. The composite slow-release carbon material coupled aerobic denitrifying bacterium according to claim 2, wherein the pseudomonas stutzeri is pseudomonas stutzeri @ aPseudomonas stutzeri ) The preservation number is CGMCC NO.20910; the microbacterium oxide is microbacterium oxide @Microbacterium oxydans) The preservation number is CGMCC NO.22738.
4. The preparation method of the composite slow-release carbon material coupled aerobic denitrifying bacteria is characterized by comprising the following steps of:
firstly, preparing a slow-release carbon carrier, namely preparing euglena particles by a bridge action of a euglena powder combined binder, uniformly mixing the euglena particles with polybutylene adipate to obtain a mixed solution, and immersing a brush-shaped elastic filler into the mixed solution for adhesion to obtain the elastic filler loaded with a slow-release carbon material;
secondly, preparing immobilized heterotrophic nitrification-aerobic denitrification bacteria, namely adding pseudomonas stutzeri and/or microbacterium oxydans into a polyvinyl alcohol solution to obtain bacterial liquid, immersing brush-shaped elastic filler into the bacterial liquid, and dropwise adding saturated boric acid solution into the bacterial liquid to enable the pseudomonas stutzeri and/or microbacterium oxydans to be attached to the elastic filler, so as to obtain the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria;
thirdly, mixing the elastic filler loaded with the slow-release carbon material with the elastic filler loaded with the immobilized heterotrophic nitrification-aerobic denitrification bacteria to obtain the euglena-based slow-release carbon material coupled aerobic denitrification bacteria.
5. The method for preparing the composite slow-release carbon material coupled aerobic denitrifying bacteria according to claim 4, wherein in the first step, euglena or pure cultured euglena are collected, dried, ground and crushed and screened by a 200-mesh sieve for later use; 4-6% of polyvinyl alcohol is prepared as a binder; adding 1kg of euglena powder into a mixing granulator, uniformly spraying a binder in a mist form through a high-pressure nozzle to form euglena particles with the particle size of 0.3-2 mm, drying, and sieving with a 20-mesh sieve for later use; melting polybutylene adipate at 75 ℃ according to the mass ratio of 20:1 to 400:1, uniformly mixing polybutylene adipate and euglena particles to obtain a mixed solution; immersing the brush-shaped elastic filler into the mixed solution for adhesion, taking out after 20+/-5 s, cooling at room temperature, fixing and forming.
6. The method for preparing the composite slow-release carbon material coupled aerobic denitrifying bacteria according to claim 5, wherein in the second step, polyvinyl alcohol is heated and dissolved in water to a final concentration of more than 6%, and then cooledCooling to below 40 ℃ to obtain a polyvinyl alcohol solution; adding Pseudomonas stutzeri and/or Microbacterium oxydans into polyvinyl alcohol solution until the final concentration of bacterial solution is 1×10 6 CFU/ml～2×10 7 And (3) CFU/ml, immersing the brush-shaped elastic filler into bacterial liquid, slowly stirring, dropwise adding a saturated boric acid solution according to 4% -10% of the volume of the bacterial liquid in the stirring process, so that viscous substances attached to the elastic filler are obviously gathered, taking out the elastic filler, slowly immersing the elastic filler into the saturated boric acid solution for fixing for 1-12 h, and immersing the elastic filler into water for 4-6 h to obtain the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria.
7. The method for preparing the composite slow-release carbon material coupled aerobic denitrifying bacteria according to claim 6, wherein the Pseudomonas stutzeri is Pseudomonas stutzeri @Pseudomonas stutzeri ) The preservation number is CGMCC NO.20910; the microbacterium oxide is microbacterium oxide @Microbacterium oxydans) The preservation number is CGMCC NO.22738.
8. The use of a composite slow-release carbon material as defined in any one of claims 1 to 7 coupled with aerobic denitrifying bacteria in water treatment.
9. The application of the composite slow-release carbon material coupled aerobic denitrifying bacteria in purifying the tail water of cultivation according to claim 8.
10. The use according to claim 9, wherein the elastic filler loaded with slow-release carbon material and the elastic filler loaded with immobilized heterotrophic nitrification-aerobic denitrification bacteria are mixed according to a ratio of 10:1, suspended below a water surface floating bed support and immersed in the culture tail water.
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| CN116621330A (en) * | 2023-07-10 | 2023-08-22 | 合肥学院 | Biological filler for slowly releasing carbon source and preparation method and application thereof |
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