CN112430593A - Preparation method of immobilized cold-resistant microbial inoculum for enhancing denitrification performance of cold wetland - Google Patents
Preparation method of immobilized cold-resistant microbial inoculum for enhancing denitrification performance of cold wetland Download PDFInfo
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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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
-
- 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
- C02F3/341—Consortia of bacteria
-
- 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
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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
- 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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- 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
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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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
- C12N11/16—Enzymes or microbial cells immobilised on or in a biological cell
-
- 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
- C02F2101/105—Phosphorus compounds
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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
Abstract
The invention relates to a preparation method of an immobilized cold-resistant microbial inoculum for strengthening the denitrification performance of a cold wetland, which comprises the following steps: 1) enrichment culture of cold-resistant ammonia oxidizing bacteria by using an SBR method, 2) preparation of bacterial suspension, 3) fixation of cold-resistant ammonia oxidizing bacteria: compounding the Brazilian foxtail spirillum and the chlorella vulgaris to obtain a chlorella compound solution, then uniformly mixing the sodium alginate solution and the chlorella compound solution to be used as an embedding carrier, adding sludge biochar to form a gel, then adding a bacterial suspension into the gel, uniformly mixing, standing for 3-4 h, slowly dropwise adding the mixture into a cross-linking agent, and cross-linking for 18-24 h and then washing for 3 times by using deionized water; the immobilized bacteria-algae pellets prepared by the method are applied to cold wetlands in winter, so that the nitrogen and phosphorus removal capability of a wetland system under cold conditions is remarkably improved, and the problems of poor operation effect, poor pollutant treatment effect and the like of the artificial wetland in winter are remarkably improved.
Description
Technical Field
The invention belongs to the technical field of sewage treatment, and particularly relates to a preparation method of an immobilized cold-resistant microbial inoculum for strengthening the denitrification performance of a cold wetland.
Background
The artificial wetland is a complex ecosystem formed by artificially constructed facilities, is a novel wastewater treatment process, purifies air and protects the environment. The artificial wetland has the advantages that in warm seasons of spring and summer, the growth of plants is accelerated, microorganisms are activated, the purification function of a wetland system is enhanced, and the biological decontamination effect is obvious. However, the nitrogen and phosphorus removal efficiency of the artificial wetland is generally low under the low-temperature condition in winter. In winter, the low temperature can obstruct the normal growth of the plants, and the biomass and the demand for nutrients of the plants are changed, so that the nitrogen and phosphorus removal rate of the plants is reduced. Under the condition of low temperature, the metabolism of the microorganisms is influenced, the quantity and the activity are reduced, and therefore, the denitrification efficiency of the artificial wetland in winter is reduced.
SBR is a short for a sequencing batch activated sludge process, is commonly used for sewage treatment, is an activated sludge sewage treatment technology which operates according to an intermittent aeration mode, and is mainly characterized by orderly and intermittent operation in operation, wherein the operation comprises 5 basic processes of water inlet (fill), reaction (act), precipitation (seat), decantation (draw), standby (idle) and the like, and a period is calculated from the beginning of sewage inflow to the end of standby time. All processes are carried out in sequence in a reactor provided with an aeration or stirring device in one period, and equipment such as a sedimentation tank, a return sludge pump and the like which are necessary to be arranged in a continuous activated sludge method are not needed.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a preparation method of an immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of a cold wetland, which is used for immobilizing cold-resistant ammonia oxidation bacteria.
The technical scheme of the invention is as follows:
a preparation method of an immobilized cold-resistant microbial inoculum for strengthening the denitrification performance of cold wetlands comprises the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on ammonia oxidizing bacteria in an ammonia oxidizing bacteria culture medium by adopting an SBR reactor, and cooling by stages after the SBR reactor operates stably until the temperature is stabilized at 1-5 ℃ to obtain cold-resistant ammonia oxidizing bacteria;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 3-5 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 3-5 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 +/-0.02 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding Brazilian foxtail spirillum and common chlorella to obtain a chlorella compound solution, wherein the Brazilian foxtail spirillum can promote the growth of the chlorella, then uniformly mixing a sodium alginate solution and the chlorella compound solution in a certain volume ratio to serve as an embedding carrier, adding a certain amount of sludge biochar to form a gel after the embedding carrier is cooled to room temperature, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 3-4 h, adding the gel into an injector, then slowly dropwise adding the gel into a cross-linking agent by using the injector, and washing for 3 times by using deionized water after cross-linking for 18-24 h to obtain immobilized bacterial algae pellets, wherein the problems of poor operation effect, low pollutant removal rate and the like of the wetland in winter can be remarkably solved by adding the immobilized bacterial algae pellets into the wetland.
Further, in the step 1), after the SBR reactor is stably operated, the temperature is reduced in two stages, wherein the temperature is controlled to be 10 +/-1 ℃ in the first stage from the start of operation to 45-50 days, the temperature is reduced by 0.5-1 ℃ every 7 days in the second stage from 45-120 days until the temperature is stabilized at 1-5 ℃.
Furthermore, 0.2-0.6 mu mol/L of octyne is added into the SBR reactor.
Further, the SBR period is 6-10 hours, and comprises the steps of feeding water for 3-8 min, carrying out aeration stirring for 300-450 min, standing for 45-120 min and discharging water for 12-22 min, and inoculating sludge and selecting aerobic activated sludge.
Furthermore, the mass ratio of the Brazilian buspiromyces to the common chlorella in the chlorella compound solution is 1: 0.2-1: 2, the volume ratio of the bacteria and the algae in the chlorella compound solution is 10-60%, the mass fraction of the sodium alginate is 3-8%, and the volume ratio of the chlorella compound solution to the sodium alginate solution is 1: 0.5-1: 3.
Furthermore, the mass ratio of the sodium alginate to the sludge biochar in the gel is 1: 3-1: 5.
Further, the cross-linking agent is 1-5% (w/v) CaCl2Saturated boric acid solution.
Further, the sludge biochar is prepared by crushing and grinding a sludge raw material, sieving the ground sludge raw material with a 80-mesh sieve to obtain sludge powder, placing the sludge powder in a muffle furnace for carbonization at 700 ℃ for 3 hours, and naturally cooling to room temperature.
Further, the ammonia oxidizing bacteria in the ammonia oxidizing bacteria culture medium is one of nitrospirillum and nitrite monad.
Compared with the prior art, the invention has the beneficial effects that:
in the system, the microorganism can generate phytohormones such as gibberellin, indole-3-acetic acid and the like to promote the growth of the algae, and the algae serves as a carrier of the microorganism to promote the growth of the microorganism;
the immobilized bacteria-algae pellets prepared by the method are applied to cold wetlands in winter, so that the nitrogen and phosphorus removal capability of a wetland system under cold conditions is remarkably improved, and the problems of poor operation effect, poor pollutant treatment effect and the like of the artificial wetland in winter are remarkably improved.
Drawings
FIG. 1 is a schematic diagram of the present invention.
In the figure, gravel (1), coarse sand (2), fine sand (3), soil (4) and plants (5).
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A preparation method of an immobilized cold-resistant microbial inoculum for strengthening the denitrification performance of cold wetlands comprises the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on an ammonia oxidation strain in an ammonia oxidation bacteria culture medium by adopting an SBR (sequencing batch reactor), wherein the ammonia oxidation strain is one of nitrospirillum or nitrite monad, and cooling in two stages after the stable operation of the SBR, wherein the temperature is controlled to be 10 +/-1 ℃ in the first stage from the beginning of operation to 45-50 days, and is reduced by 0.5-1 ℃ every 7 days until the temperature is stabilized to be 1-5 ℃ in the second stage, so as to obtain a cold-resistant ammonia oxidation bacterial group;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 3-5 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 3-5 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 +/-0.02 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding Brazilian foxtail spirillum and common chlorella to obtain a chlorella compound solution, wherein the Brazilian foxtail spirillum can promote the growth of the chlorella, then uniformly mixing a sodium alginate solution and the chlorella compound solution in a certain volume ratio to serve as an embedding carrier, cooling the embedding carrier to room temperature, adding a certain amount of sludge biochar to form a gel, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 3-4 h, adding the gel into an injector, then slowly dropwise adding the gel into a cross-linking agent by using the injector, and after cross-linking for 18-24 h, washing for 3 times by using deionized water to obtain immobilized bacterial algae pellets, wherein the immobilized bacterial algae pellets serve as immobilized cold-resistant pellets and are applied to artificial wetlands in winter.
Furthermore, 0.2-0.6 mu mol/L of octyne is added into the SBR reactor.
Further, the SBR period is 6-10 hours, and comprises the steps of feeding water for 3-8 min, carrying out aeration stirring for 300-450 min, standing for 45-120 min and discharging water for 12-22 min, inoculating sludge and selecting aerobic activated sludge, wherein the inoculated sludge comes from the activated sludge in the aerobic section of the biological treatment of the sewage treatment plant.
Furthermore, the mass ratio of the Brazilian buspiromyces to the common chlorella in the chlorella compound solution is 1: 0.2-1: 2, the volume ratio of the bacteria and the algae in the chlorella compound solution is 10-60%, the mass fraction of the sodium alginate is 3-8%, and the volume ratio of the chlorella compound solution to the sodium alginate solution is 1: 0.5-1: 3.
Furthermore, the mass ratio of the sodium alginate to the sludge biochar in the gel is 1: 3-1: 5.
Further, the cross-linking agent is 1-5% (w/v) of CaCl2 saturated boric acid solution.
Further, the sludge biochar is prepared by crushing and grinding a sludge raw material, sieving the ground sludge raw material with a 80-mesh sieve to obtain sludge powder, placing the sludge powder in a muffle furnace for carbonization at 700 ℃ for 3 hours, and naturally cooling to room temperature.
Example 2
This example is another embodiment based on example 1, and the description of the same technical solution as in example 1 will be omitted, and only the technical solution different from example 1 will be explained.
In this embodiment, a preparation method of an immobilized cold-resistant microbial inoculum for enhancing denitrification performance of a cold wetland comprises the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on an ammonia oxidizing strain in an ammonia oxidizing strain culture medium by adopting an SBR (sequencing batch reactor), wherein the ammonia oxidizing strain is one of nitrospirillum or nitrite monad, 0.4 mu mol/L octyne is added in the SBR to stimulate an ammonia oxidizing flora, and cooling is carried out in two stages after the SBR is stably operated, wherein the temperature is controlled to be 10 ℃ from the beginning of operation to 50 days in the first stage, the temperature is reduced by 0.5 ℃ every 7 days until the temperature is stabilized to be 2 ℃ in the second stage, so that a cold-resistant ammonia oxidizing flora is obtained;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 3 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 3 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding Brazilian fox spirillum and chlorella vulgaris to obtain a chlorella compound solution, wherein the Brazilian fox spirillum can promote the growth of the chlorella vulgaris, the mass ratio of the Brazilian fox spirillum to the chlorella vulgaris in the chlorella compound solution is 1: 0.2, then uniformly mixing a sodium alginate solution and a chlorella compound solution in a certain volume ratio to serve as an embedding carrier, the mass fraction of the sodium alginate is 5%, the volume ratio of the chlorella compound solution to the sodium alginate solution is 1: 2, after the embedding carrier is cooled to room temperature, adding a certain amount of sludge biochar to form a gel, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 3.5h, adding the gel into an injector, and then slowly dripping 3% (w/v) of CaCl by using the injector2And (3) in a saturated boric acid solution, crosslinking for 24 hours, and then washing for 3 times by using deionized water to obtain immobilized bacteria-algae pellets, wherein the immobilized bacteria-algae pellets are used as an immobilized cold-resistant microbial inoculum and are applied to the artificial wetland in winter.
Example 3
This example is another embodiment based on example 1, and the description of the same technical solution as in example 1 will be omitted, and only the technical solution different from example 1 will be explained.
In this embodiment, a preparation method of an immobilized cold-resistant microbial inoculum for enhancing denitrification performance of a cold wetland comprises the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on an ammonia oxidizing strain in an ammonia oxidizing strain culture medium by adopting an SBR (sequencing batch reactor), wherein the ammonia oxidizing strain is one of nitrospirillum or nitrite monad, 0.2 mu mol/L octyne is added in the SBR to stimulate an ammonia oxidizing bacteria group, and cooling is carried out in two stages after the SBR is stably operated, wherein the temperature is controlled to be 10 ℃ from the beginning of operation to 48 days in the first stage, the temperature is reduced by 0.8 ℃ every 7 days in the second stage until the temperature is stabilized to be 1 ℃, so that a cold-resistant ammonia oxidizing bacteria group is obtained;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 2 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 2 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding Brazilian fox spirillum and chlorella vulgaris to obtain a chlorella compound solution, wherein the Brazilian fox spirillum can promote the growth of the chlorella vulgaris, the mass ratio of the Brazilian fox spirillum to the chlorella vulgaris in the chlorella compound solution is 1: 1, then uniformly mixing a sodium alginate solution and a chlorella vulgaris compound solution in a certain volume ratio to serve as an embedding carrier, the mass fraction of the sodium alginate is 4%, the volume ratio of the chlorella vulgaris compound solution to the sodium alginate solution is 1: 1, after the embedding carrier is cooled to room temperature, adding a certain amount of sludge biochar to form a gel, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 3.5 hours, then adding the gel into an injector, and then slowly dripping 5% (w/v) of CaCl by using the injector2And (3) in a saturated boric acid solution, washing for 3 times by using deionized water after crosslinking for 18h to obtain immobilized bacteria-algae pellets, wherein the immobilized bacteria-algae pellets are used as an immobilized cold-resistant microbial inoculum and are applied to the artificial wetland in winter.
Example 4
This example is another embodiment based on example 1, and the description of the same technical solution as in example 1 will be omitted, and only the technical solution different from example 1 will be explained.
In this embodiment, a preparation method of an immobilized cold-resistant microbial inoculum for enhancing denitrification performance of a cold wetland comprises the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on an ammonia oxidizing strain in an ammonia oxidizing strain culture medium by adopting an SBR (sequencing batch reactor), wherein the ammonia oxidizing strain is one of nitrospirillum or nitrite monad, 0.6 mu mol/L octyne is added in the SBR to stimulate an ammonia oxidizing flora, and cooling is carried out in two stages after the SBR is stably operated, wherein the temperature is controlled to be 10 ℃ from the beginning of operation to 45 days in the first stage, the temperature is reduced by 0.5 ℃ every 7 days until the temperature is stabilized to be 2 ℃ in the second stage, so that a cold-resistant ammonia oxidizing flora is obtained;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 2 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 2 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding Brazilian fox spirillum and chlorella vulgaris to obtain a chlorella compound solution, wherein the Brazilian fox spirillum can promote the growth of the chlorella vulgaris, the mass ratio of the Brazilian fox spirillum to the chlorella vulgaris in the chlorella compound solution is 1: 2, then uniformly mixing a sodium alginate solution and a chlorella vulgaris compound solution in a certain volume ratio to serve as an embedding carrier, the mass fraction of the sodium alginate is 5%, the volume ratio of the chlorella vulgaris compound solution to the sodium alginate solution is 1: 2, after the embedding carrier is cooled to room temperature, adding a certain amount of sludge biochar to form a gel, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 4 hours, adding the gel into an injector, and then slowly dripping 4% (w/v) of CaCl by using the injector2In saturated boric acid solution, cross-linking for 20 hr, washing with deionized water for 3 times to obtain immobilized bacteria-algae pellet as solidThe customized cold-resistant microbial inoculum is applied to the artificial wetland in winter.
As shown in fig. 1, the constructed wetland is a schematic diagram of a typical constructed wetland, which is provided with a gravel layer (1), a coarse sand layer (2), a fine sand layer (3), a soil layer (4) and plants (5) from bottom to top in sequence, wherein the constructed wetland plants in winter can be one or more of iris, calamus, lentinus edodes and Rumex acetosa; the immobilized bacteria and algae pellet as immobilized cold-resistant microbial inoculum is added into the soil layer and the fine sand layer.
In order to verify the denitrification effect of immobilized microalgae pellets, products in different stages and final products of the immobilized microalgae pellets are added into wastewater according to the conventional dosage for testing the denitrification performance, the used wastewater is tail water treated by a sewage treatment plant, the contents of ammonia nitrogen, total nitrogen and total phosphorus in the wastewater are respectively 4.23mg/L, 14.68mg/L and 0.48mg/L, and the specific treatment comparison results are shown in the following table:
from the above table, after 5 days of operation under the low temperature condition of 5.5C, the contents after treatment are greatly reduced compared with the contents of ammonia nitrogen, total nitrogen and total phosphorus in the original wastewater.
In order to verify the application effect of the immobilized microalgae-globule denitrification wetland, products in different stages of the invention and final product immobilized microalgae globules are added to the same wetland for comparison, the content of total nitrogen in wetland water is measured, and the comparison result is shown in the following table:
although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes in the embodiments and/or modifications of the invention can be made, and equivalents and modifications of some features of the invention can be made without departing from the spirit and scope of the invention.
Claims (9)
1. A preparation method of an immobilized cold-resistant microbial inoculum for strengthening the denitrification performance of cold wetlands is characterized by comprising the following steps:
1) enrichment culture of cold-resistant ammonia oxidizing bacteria: carrying out enrichment culture on ammonia oxidizing bacteria in an ammonia oxidizing bacteria culture medium by adopting an SBR reactor, and cooling by stages after the SBR reactor operates stably until the temperature is stabilized at 1-5 ℃ to obtain cold-resistant ammonia oxidizing bacteria;
2) preparing a bacterial suspension: inoculating the cold-resistant ammonia oxidation flora in the step 1) into a beef extract peptone culture solution, culturing in a constant-temperature shaking incubator at 3-5 ℃ and 120r/min until the logarithmic phase is reached, centrifuging at 3-5 ℃ and 4000-6000 r/min, removing supernatant, and adjusting to a bacterial suspension with the optical density of 1.0 +/-0.02 at the wavelength of 600nm by using normal saline;
3) and (3) fixing the cold-resistant ammonia oxidation flora: compounding the Brazilian buspiromyces and the common chlorella to obtain a chlorella compound solution, then uniformly mixing a sodium alginate solution and the chlorella compound solution in a certain volume ratio to serve as an embedding carrier, adding a certain amount of sludge biochar to form a gel after the embedding carrier is cooled to room temperature, then adding the bacterial suspension prepared in the step 2) into the gel, uniformly mixing, standing for 3-4 hours, slowly dropwise adding the gel into a cross-linking agent, and cross-linking for 18-24 hours and then washing for 3 times by using deionized water to obtain the immobilized bacterial chlorella.
2. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: in the step 1), the temperature is reduced in two stages after the SBR reactor is stably operated, the temperature is controlled to be 10 +/-1 ℃ in the first stage from the beginning of operation to 45-50 days, the temperature is reduced by 0.5-1 ℃ every 7 days in the second stage, and the temperature is reduced to be 1-5 ℃ until the temperature is stabilized.
3. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: 0.2-0.6 mu mol/L of octyne is added into the SBR reactor.
4. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the SBR period is 6-10 hours, and comprises the steps of feeding water for 3-8 min, aerating and stirring for 300-450 min, standing for 45-120 min and discharging water for 12-22 min, and inoculating sludge and selecting aerobic activated sludge.
5. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the mass ratio of the unispirillum brasiliensis to the chlorella vulgaris in the chlorella compound solution is 1: 0.2-1: 2, the ratio of the bacteria to the algae in the chlorella compound solution is 10-60%, the mass fraction of the sodium alginate is 3-8%, and the volume ratio of the chlorella compound solution to the sodium alginate solution is 1: 0.5-1: 3.
6. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the mass ratio of the sodium alginate to the sludge biochar in the gel is 1: 3-1: 5.
7. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the cross-linking agent is 1-5% (w/v) CaCl2Saturated boric acid solution.
8. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the sludge biochar is prepared by crushing and grinding a sludge raw material, sieving the ground sludge raw material by a 80-mesh sieve to obtain sludge powder, carbonizing the sludge powder in a muffle furnace at 700 ℃ for 3 hours, and naturally cooling to room temperature.
9. The preparation method of the immobilized cold-resistant microbial inoculum for enhancing the denitrification performance of the cold wetland according to claim 1, wherein the immobilized cold-resistant microbial inoculum comprises the following steps: the ammonia oxidizing bacteria in the ammonia oxidizing bacteria culture medium is one of nitrospirillum or nitrite unicellular bacteria.
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