CN113860488B - Anaerobic ammonia oxidizing bacteria particle culture method and device - Google Patents
Anaerobic ammonia oxidizing bacteria particle culture method and device Download PDFInfo
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Abstract
The invention particularly relates to a method and a device for culturing anaerobic ammonia oxidation bacteria particles, wherein domestic sewage is taken as water inlet, particle fillers and nitrified sludge are mixed for intermittent aeration culture, the obtained particle fillers are placed in a net surface hollow carrier and used as filler layers to be filled in an anaerobic reactor for anaerobic ammonia oxidation culture, and nitrogen sources generated by a denitrification process are utilized for an anaerobic ammonia oxidation process, so that enrichment of anaerobic ammonia oxidation bacteria particles is realized. The culture method has the characteristics of quick start of anaerobic ammonia oxidation, high concentration of thalli and short culture period, and the particle filler is fixed in the reactor to serve as a filler layer, so that the start of anaerobic ammonia oxidation bacteria and quick mass culture expansion are facilitated, and the device has high treatment efficiency, stable operation and low energy consumption.
Description
Technical Field
The invention relates to the field of environment-friendly microorganisms, in particular to a method and a device for culturing anaerobic ammonia oxidizing bacteria particles.
Background
Domestic sewage generated in life or industrial production sewage of foods, slaughter, cultivation, circuit boards and the like generated in production often contains a large amount of ammonia nitrogen, and if the ammonia nitrogen is improperly treated, the ammonia nitrogen is discharged into natural water body, so that eutrophication of the water body is easily caused. At present, the emission standards of ammonia nitrogen and total nitrogen are defined, and the water quality required by surface water treatment is higher and higher. Therefore, the harmless treatment of nitrogen element produced by life production is realized, and the realization of virtuous circle of nitrogen element is a necessary measure for realizing harmony between people and nature.
Anaerobic Ammonia Oxidizing Bacteria (AOB) are a type of autotrophic microorganism capable of converting ammonia nitrogen and nitrite nitrogen into nitrogen and nitrate nitrogen under anaerobic conditions. The mixture ratio of materials and reaction products of AOB in treating ammonia nitrogen and nitrite nitrogen basically follow the following reaction formula (1). As can be seen from the following formula, if ammonia nitrogen is theoretically converted into nitrite nitrogen through ideal short-cut nitrification and then is converted into nitrogen under the action of AOB, the process can save about 60% of energy consumption compared with the process of completely converting ammonia nitrogen into nitrate nitrogen, and the generated small amount of nitrate is only about 10% of the nitrification process, so that the carbon source required in denitrification is greatly reduced.
The anaerobic ammonia oxidation bacteria are initially started and cultured in an anaerobic environment by using activated sludge and directional enrichment nutrient solution, but the traditional method has low starting speed; on the basis, there are reports that the addition of biological promoters can accelerate the start of anammox bacteria, such as vitamin C, gibberellin, cell quorum sensing factors and the like, but anammox sludge is extremely sensitive to oxygen, so that the obtained anammox sludge is fixed in an anaerobic tank for use and further treatment is required.
There are several studies in China focusing on the treatment of anaerobic ammonium oxidation sludge, but the treatment effect is not ideal. For example, chinese patent CN11892161B discloses a method for rapidly starting anaerobic ammonium oxidation bacteria by using inorganic composite powder, in which sludge and inorganic composite powder are added into a reactor, and a film forming environment required for growth of anaerobic ammonium oxidation bacteria is provided by the inorganic composite powder to promote rapid starting of anaerobic ammonium oxidation bacteria. The powder provided by the method has the particle size of 10-50 mu m, the density of 1.5-2.5g/cm < 3 > and smaller particles, and the powder has the density higher than that of water and is easy to precipitate, so that the powder can be layered with sludge with small density in the culture process and cannot achieve a uniform mixing state. Furthermore, chinese patent CN10593685a discloses a method of enriching and culturing anammox bacteria by mixing a suspension filler with sludge using modified basalt fiber, but in this method, the fiber filler is easily bonded into clusters, resulting in a decrease in specific surface area and poor solid-liquid contact effect, thus resulting in low treatment efficiency.
Disclosure of Invention
The invention aims to solve the defects of the prior art and provide the method for culturing the anaerobic ammonia oxidation bacteria particles, which has the characteristics of rapid starting of anaerobic ammonia oxidation, high concentration of bacteria and short culture period.
The invention further aims to provide an anaerobic ammonia oxidation bacteria particle culture device which is high in efficiency and stable in operation, achieves rapid starting of full-automatic anaerobic ammonia oxidation, and is beneficial to accelerating starting of anaerobic ammonia oxidation bacteria and rapid mass expansion cultivation by filling particle fillers with large specific surface areas into a net surface hollow carrier as filler layers to be filled into a reactor.
The aim of the invention is realized by the following technical scheme: a method for culturing anaerobic ammonium oxidation bacteria particles comprises the following steps:
s1, mixing domestic sewage as water, performing intermittent aeration culture on granular filler and nitrified sludge, setting the aeration temperature to be 20-35 ℃, the concentration of dissolved oxygen to be 2-5mg/L, introducing air, aerating for 6-8 hours, stopping aeration, standing, precipitating for 0.5-2 hours, discharging supernatant after standing, adding water with the same volume, and standing for 3-5 hours, wherein the aeration culture period is one aeration culture period; the intermittent aeration culture of the particle filler is completed after the repeated culture period reaches more than 20 days;
s2, filling the granular filler obtained in the step S1 after intermittent aeration culture into a net surface hollow carrier to serve as a film-forming filler layer, placing the net surface hollow carrier into an anaerobic reactor, putting nitrified sludge into the anaerobic reactor, and introducing an enriched nutrient solution to start anaerobic ammonia oxidation culture;
s3, the enriched nutrient solution flows into a denitrification reactor containing denitrifying bacteria, carbon sources are supplemented into the denitrification reactor at regular time to perform denitrification to generate nitrogen, and the generated nitrogen is introduced into an anaerobic reactor to enable the anaerobic reactor to be in an anaerobic state;
s4, raising ammonia nitrogen concentration of the enriched nutrient solution in stages for enrichment culture for 60-90 days, and when obvious red bacterial films can be observed on the surface of the film-forming filler layer, obtaining the anaerobic ammonia oxidation bacterial particles, wherein the anaerobic ammonia oxidation culture is successfully started.
In the step S1 of the invention, the concentration of the mixed solution of the nitrified sludge and the domestic sewage is 3000-6000mg/L, and the particle filler is added according to the volume of the 20-30% solution; in the step S2, the particle filler obtained in the step S1 is filled into a net surface hollow carrier, and is filled into an anaerobic reaction to serve as a filler layer for anaerobic ammonia oxidation bacteria film growth, and enrichment culture of anaerobic ammonia oxidation bacteria particles is carried out together with nitrified sludge; in the step S3, the anaerobic reactor is fixed in the denitrification reactor through denitrifying bacteria, and carbon sources are supplemented for denitrification to generate nitrogen, so that the anaerobic reactor is supplemented and ensured to be in an absolute anaerobic state; in the step S4, the ammonia nitrogen concentration of the nutrient solution is 50mg/L in the initial stage of enrichment culture, the ammonia nitrogen concentration of the nutrient solution is increased to 100mg/L after 30d of accumulation culture, the ammonia nitrogen concentration of the nutrient solution is increased to 150mg/L after 45d of accumulation culture, the ammonia nitrogen concentration of the nutrient solution is increased to 200mg/L after 60d of accumulation culture, and the ammonia nitrogen concentration of the nutrient solution is kept unchanged.
The mesh hollow carrier can be a mesh hollow body made of various materials, the mesh hollow body comprises a cylinder, a cube, a cuboid, a cylinder and the like, and the mesh hole diameter of the mesh hollow body is smaller than the particle diameter of the particle filler obtained in the step S1 after intermittent aeration culture so as to prevent the particle filler from leaking.
Preferably, the COD concentration of the domestic sewage is 200-500mg/L, the ammonia nitrogen concentration is 50-100mg/L, and the pH range is 7-9. Since anaerobic ammonium oxidation bacteria are autobacteria, the pH of the solution is controlled to be weak alkaline during the culture process because inorganic carbon sources are consumed during the growth process to cause the pH of the solution to be reduced.
Preferably, the particle filler is at least one of ceramsite, volcanic rock, polyurethane foam, perlite and zeolite, and the particle filler adopted by the invention has larger specific surface area, is used as the filler for the growth of anammox bacteria hanging film, has large specific surface area, is favorable for the growth of absorption nutrient solution and anammox bacteria, and has the particle size of preferably 3-6mm, and the particle filler with relatively small particle size is selected, so that the particle filler is conveniently filled into a net surface hollow carrier, and can ensure larger specific surface area.
Preferably, the nutrient solution comprisesThe composition of the raw materials is as follows: NH (NH) 4 HCO 3 0.2～3g/L、NaNO 2 0.2～3g/L、K 2 HPO 4 0.05～0.3g/L、MgSO 4 ·7H 2 0 5～50mg/L、CaCl 2 5～50mg/L、FeSO 4 ·7H 2 0 2～30mg/L、ZnSO 4 ·7H 2 0 0.5～5mg/L、CoCl 2 ·6H 2 0 0.2～2mg/L、CuSO 4 ·5H 2 0 0.5～2mg/L、Na 2 MoO 4 ·5H 2 00.5-5 mg/L and H 3 BO 3 5～50mg/L。
Preferably, the carbon source is an aqueous solution of one or more of sodium acetate, methanol, glucose, maltose and molasses, and the solute concentration of the solute in the carbon source is 5-10% wt.
The other purpose of the invention is realized by the following technical scheme: an anaerobic ammonia oxidizing bacteria particle culture device comprises an enriched nutrient solution tank, an up-flow anaerobic reactor, a first water pump, a middle storage tank, a second water pump and a denitrification reactor, wherein the up-flow anaerobic reactor, the middle storage tank and the denitrification reactor are sequentially connected; the device comprises an up-flow anaerobic reactor, an enriched nutrient solution tank, a first water pump, a three-phase separator, a liquid outlet, a middle storage tank, a denitrification reactor and a gas outlet, wherein the enriched nutrient solution tank is connected with the up-flow anaerobic reactor through the first water pump, the top end of the up-flow anaerobic reactor is provided with the three-phase separator, the three-phase separator is provided with the liquid outlet, the liquid outlet is connected with the middle storage tank through a pipeline, the middle storage tank is connected with the denitrification reactor through the second water pump, the top end of the denitrification reactor is provided with the gas outlet, and the gas outlet is connected with the bottom end of the up-flow anaerobic reactor through a gas pipeline.
Further, the device also comprises a carbon source solution tank and a third water pump, wherein the carbon source solution tank is connected with the middle storage tank through the third water pump, and the third water pump is used for supplementing the carbon source for the middle storage tank at regular time. Specifically, the operating flow rate of the third water pump is set to 1-2% of the operating flow rate of the first water pump.
Further, liquid level controllers are arranged in the middle storage tank and the denitrification reactor. Specifically, well storage tank is provided with first liquid level controller, first liquid level controller monitors the liquid level in the storage tank through the first liquid level electrode of installing in the transfer jar, and then the switch of control second water pump, can trigger the second water pump and let in the denitrification reactor with the nutrient solution in the storage tank when the liquid level in the transfer jar reaches the upper contact of first liquid level electrode, and the second water pump stops working when the liquid level in the transfer jar descends to the lower contact, can guarantee through the control of the liquid level in the transfer jar that the second water pump can not pump into the denitrification reactor with the air. The denitrification reactor is provided with a second liquid level controller, the second liquid level controller monitors the liquid level in the denitrification reactor through a second liquid level electrode arranged in the denitrification reactor, when the liquid level reaches an upper contact point of the second liquid level electrode, the electromagnetic valve is opened to discharge the solution in the denitrification reactor, and when the liquid level descends to a lower contact point of the second liquid level electrode, the electromagnetic valve is closed. Furthermore, in order to ensure that the gas in the denitrification reactor is not discharged through the electromagnetic valve, the electromagnetic valve is arranged below the lower contact of the second liquid level electrode.
Furthermore, a film-forming filler layer is arranged in the upflow anaerobic reactor, and the film-forming filler layer is prepared by filling particle filler into a net surface hollow carrier. Specifically, the height-diameter ratio of the upflow reactor is 5-10:1, and the filling ratio of the film-forming filler layer is 50-80%.
Further, a denitrification filling layer is arranged in the denitrification reactor, and the preparation of the denitrification filling layer comprises the following steps: (1) Dissolving denitrifying bacteria into 2-5% sodium alginate solution to prepare a denitrifying mixed solution; (2) Dripping the denitrification mixed solution into 1-3% calcium chloride solution for immobilization reaction to obtain denitrifying bacteria particles with the diameter of 3-5 mm; (3) And (5) putting denitrifying bacteria particles into a net surface hollow carrier to obtain the denitrifying filling layer.
According to the denitrification reactor disclosed by the invention, the denitrifying bacteria particles are prepared by immobilizing denitrifying bacteria through the reaction of sodium alginate and calcium chloride, so that the loss of denitrifying bacteria is prevented, and the stability of denitrification is ensured.
The invention has the beneficial effects that: the method and the device for culturing the anaerobic ammonia oxidation bacteria have the characteristics of rapid starting of anaerobic ammonia oxidation, high concentration of bacteria and short culture period. The culture method integrates the anaerobic ammonia oxidation process and the denitrification process, the stable nitrogen source generated by the denitrification process can meet the requirements of the anaerobic ammonia oxidation process, and the method adopts the method that the particle filler with large specific surface area is filled into the net surface hollow carrier after intermittent aeration treatment to be used as a film carrier of anaerobic ammonia oxidation bacteria to be fixed in an anaerobic reactor, so that the starting of the anaerobic ammonia oxidation bacteria and rapid mass expansion culture can be accelerated. Furthermore, the denitrification is carried out through the middle storage tank and the denitrification reactor, so that continuous and stable nitrogen source and anaerobic requirements can be provided for the anaerobic ammonia oxidation process, the efficiency of the whole culture device is improved through process integration and reasonable arrangement of equipment, manual intervention is reduced, and full-automatic and rapid starting is realized.
Drawings
FIG. 1 is a schematic view of an anaerobic ammonium oxidation bacteria pellet culture apparatus according to the present invention.
FIG. 2 is a schematic structural view of the mesh hollow carrier of the present invention.
Reference numerals: 1-enrichment nutrient solution tank, 2-first water pump, 3-upflow anaerobic reactor, 4-middle storage tank, 5-second water pump, 6-denitrification reactor, 7-carbon source solution tank, 8-third water pump, 301-three-phase separator, 302-biofilm packing layer, 303-mesh hollow carrier, 401-first liquid level controller, 402-first liquid level electrode, 601-second liquid level controller, 602-second liquid level electrode, 603-electromagnetic valve and 604-denitrification packing layer.
Detailed Description
The invention will be further illustrated by the following examples, which are not intended to limit the scope of the invention, in order to facilitate the understanding of those skilled in the art.
Referring to fig. 1-2, as a typical embodiment of the present invention, an anaerobic ammonium oxidation bacteria granule culturing apparatus comprises an enriched nutrient solution tank 1, an upflow anaerobic reactor 3, a first water pump 2, a middle storage tank 4, a second water pump 5 and a denitrification reactor 6, wherein the upflow anaerobic reactor 3, the middle storage tank 4 and the denitrification reactor 6 are sequentially connected; the device comprises an enriched nutrient solution tank 1, an upflow anaerobic reactor 3, a three-phase separator 301, a liquid outlet, a middle storage tank 4, a denitrification reactor 6 and a gas outlet, wherein the enriched nutrient solution tank 1 is connected with the upflow anaerobic reactor 3 through a first water pump 2, the top end of the upflow anaerobic reactor 3 is provided with the three-phase separator 301, the liquid outlet is connected with the middle storage tank 4 through a pipeline, the middle storage tank 4 is connected with the denitrification reactor 6 through a second water pump 5, and the top end of the denitrification reactor 6 is provided with the gas outlet which is connected with the bottom end of the upflow anaerobic reactor 3 through a gas pipeline.
Specifically, the enriched nutrient solution is introduced into the bottom of the upflow anaerobic reactor 3 through the first water pump 2, flows out through the three-phase separator 301 at the top of the upflow anaerobic reactor 3 after passing through the film-forming filler layer 302, then enters the middle storage tank 4, the enriched nutrient solution in the middle storage tank 4 is controlled by the first liquid level controller 401 to enter the denitrification reactor 6 through the second water pump 5 to perform denitrification treatment to generate nitrogen, and the nitrogen is introduced into the bottom of the upflow anaerobic reactor 3 through the top air outlet of the denitrification reactor 6 to maintain the anaerobic state.
Further, the device also comprises a carbon source solution 7 and a third water pump 8, wherein the carbon source solution 7 is connected with the middle storage tank 4 through the third water pump 8, and the third water pump 8 is used for supplementing the carbon source for the middle storage tank 4 at regular time. Specifically, the operation flow rate of the third water pump 8 is set to 1-2% of the operation flow rate of the first water pump 2.
Further, liquid level controllers are arranged in the middle storage tank 4 and the denitrification reactor 6. Specifically, the middle storage tank 4 is provided with a first liquid level controller 401, the first liquid level controller 401 monitors the liquid level in the middle storage tank 4 through a first liquid level electrode 403 installed in the middle storage tank 4, and then controls a second water pump 5, when the liquid level in the middle storage tank 4 reaches an upper contact of the first liquid level electrode 402, the second water pump 5 is triggered to introduce the enriched nutrient solution in the middle storage tank 4 into the denitrification reactor 6, when the liquid level in the middle storage tank 4 drops to a lower contact, the second water pump 5 stops working, and by controlling the liquid level in the middle storage tank 4, the second water pump 5 can be ensured not to pump air into the denitrification reactor 6. The denitrification reactor 6 is provided with a second liquid level controller 601, the second liquid level controller 601 monitors the liquid level in the denitrification reactor 6 through a second liquid level electrode 602 arranged in the denitrification reactor 6, when the liquid level reaches an upper contact point of the second liquid level electrode 602, an electromagnetic valve 603 is opened to discharge the solution in the denitrification reactor 6, and when the liquid level descends to a lower contact point of the second liquid level electrode 602, the electromagnetic valve 603 is closed. Furthermore, to ensure that the gas in the denitrification reactor 6 is not discharged through the solenoid valve 603, the solenoid valve 603 is installed at a position below the lower contact of the second liquid level electrode 602.
Further, a film-forming filler layer 302 is disposed in the upflow anaerobic reactor 3, and the film-forming filler layer 302 is made by filling particulate filler into a mesh hollow carrier 303. Specifically, the height-to-diameter ratio of the upflow reactor 3 is 5-10:1, the filling ratio of the membrane-hanging filler layer 302 is 50-80%, and the mesh surface aperture of the mesh surface hollow carrier 303 is 1-2mm.
Further, a denitrification filling layer is arranged in the denitrification reactor 6, and the preparation of the denitrification filling layer comprises the following steps: (1) Dissolving denitrifying bacteria into 2-5% sodium alginate solution to prepare a denitrifying mixed solution; (2) Dripping the denitrification mixed solution into 1-3% calcium chloride solution for immobilization reaction to obtain denitrifying bacteria particles with the diameter of 3-5 mm; (3) And (5) placing denitrifying bacteria particles into the net surface hollow carrier 303 to obtain the denitrifying filling layer.
Example 1
A method for culturing anaerobic ammonium oxidation bacteria particles comprises the following steps:
s1, mixing the granular filler with nitrified sludge to perform intermittent aeration culture by taking domestic sewage as inflow water, setting the aeration temperature to 20 ℃, setting the dissolved oxygen concentration to 2mg/L, introducing air to perform aeration for 6 hours, stopping aeration, standing and precipitating for 0.5 hour, discharging supernatant after standing, adding inflow water with the same volume, and standing for 3 hours, wherein the aeration culture period is one aeration culture period; the intermittent aeration culture of the particle filler is completed after the repeated culture period reaches more than 20 days;
s2, filling the granular filler obtained in the step S1 after intermittent aeration culture into a net surface hollow carrier to serve as a film-forming filler layer, placing the net surface hollow carrier into an anaerobic reactor, putting nitrified sludge into the anaerobic reactor, and introducing an enriched nutrient solution to start anaerobic ammonia oxidation culture;
s3, the enriched nutrient solution flows into a denitrification reactor containing denitrifying bacteria, carbon sources are supplemented into the denitrification reactor at regular time to perform denitrification to generate nitrogen, and the generated nitrogen is introduced into an anaerobic reactor to enable the anaerobic reactor to be in an anaerobic state;
s4, raising the ammonia nitrogen concentration of the enriched nutrient solution in stages for enrichment culture for 60 days, and observing an obvious red bacterial film on the surface of the film-forming filler layer to obtain the anaerobic ammonia oxidation bacterial particles, wherein the anaerobic ammonia oxidation culture is successfully started.
In the embodiment, the COD concentration of the domestic sewage is 200mg/L, the ammonia nitrogen concentration is 50mg/L, and the pH is 6.
In the embodiment, the particle filler is polyurethane foam, the particle size of the particle filler is 3-5mm, and the net surface hollow carrier is spherical net surface PVC plastic spheres with the diameter of 10cm and the aperture of 1.5 mm.
In this embodiment, the nutrient solution comprises the following raw materials: NH (NH) 4 HCO 3 0.2g/L、NaNO 2 0.25g/L、K 2 HPO 4 0.08g/L、MgSO 4 ·7H 2 0 5mg/L、CaCl 2 8mg/L、FeSO 4 ·7H 2 0 2mg/L、ZnSO 4 ·7H 2 0 0.5mg/L、CoCl 2 ·6H 2 0 0.3mg/L、CuSO 4 ·5H 2 0 0.9mg/L、Na 2 MoO 4 ·5H 2 0.5mg/L and H 3 BO 3 7mg/L。
In this example, the carbon source is a 7% wt methanol solution.
In the step S4 of this embodiment, the ammonia nitrogen concentration of the nutrient solution introduced in the initial stage of enrichment culture is 50mg/L, the ammonia nitrogen concentration of the enriched nutrient solution is raised to 100mg/L after 30d of accumulation culture, the ammonia nitrogen concentration of the enriched nutrient solution is raised to 150mg/L after 45d of accumulation culture, and thereafter the ammonia nitrogen concentration of the enriched nutrient solution is kept unchanged, and the accumulation is completed for 60 d.
After the cultivation by the cultivation method of this example, the surface of the particulate filler was observed to be significantly reddish brownBacterial film, nitrified sludge presents obvious red, a large number of bubbles appear between nitrified sludge and filler particles in the anaerobic reactor, and the total nitrogen removal rate is 0.13 kg/(m) when enrichment is finished 3 ·d)。
Example 2
A method for culturing anaerobic ammonium oxidation bacteria particles comprises the following steps:
s1, mixing the granular filler with nitrified sludge to perform intermittent aeration culture by taking domestic sewage as water inlet, setting the aeration temperature to be 30 ℃, setting the dissolved oxygen concentration to be 3mg/L, introducing air to perform aeration for 7 hours, stopping aeration, standing and precipitating for 1 hour, discharging supernatant after standing, adding water inlet with the same volume, and standing for 4 hours, wherein the aeration culture period is one aeration culture period; the intermittent aeration culture of the particle filler is completed after the repeated culture period reaches 20 days;
s2, filling the granular filler obtained in the step S1 after intermittent aeration culture into a net surface hollow carrier to serve as a film-forming filler layer, placing the net surface hollow carrier into an anaerobic reactor, putting nitrified sludge into the anaerobic reactor, and introducing an enriched nutrient solution to start anaerobic ammonia oxidation culture;
s3, the enriched nutrient solution flows into a denitrification reactor containing denitrifying bacteria, carbon sources are supplemented into the denitrification reactor at regular time to perform denitrification to generate nitrogen, and the generated nitrogen is introduced into an anaerobic reactor to enable the anaerobic reactor to be in an anaerobic state;
s4, raising the ammonia nitrogen concentration of the enriched nutrient solution in stages for enrichment culture for 80 days, and observing an obvious red bacterial film on the surface of the film-forming filler layer to obtain the anaerobic ammonia oxidation bacterial particles, wherein the anaerobic ammonia oxidation culture is successfully started.
In the embodiment, the COD concentration of the domestic sewage is 300mg/L, the ammonia nitrogen concentration is 80mg/L, and the pH value is 8.
In the embodiment, the particle filler is formed by mixing ceramsite with the particle size of 4-6mm and perlite, and the mesh hollow carrier is a square mesh PVC plastic sphere with the diameter of 10cm and the aperture of 2mm.
In this embodiment, the nutrient solution comprises the following raw materials: NH (NH) 4 HCO 3 1.8g/L、NaNO 2 2.2g/L、K 2 HPO 4 0.22g/L、MgSO 4 ·7H 2 0 34mg/L、CaCl 2 23mg/L、FeSO 4 ·7H 2 0 16mg/L、ZnSO 4 ·7H 2 0 2.6mg/L、CoCl 2 ·6H 2 0 1.2mg/L、CuSO 4 ·5H 2 0 0.8mg/L、Na 2 MoO 4 ·5H 2 0.3 mg/L and H 3 BO 3 36mg/L。
In this example, the carbon source is 5% wt sodium acetate in water.
In the step S4 of this embodiment, the ammonia nitrogen concentration of the nutrient solution introduced in the initial stage of enrichment culture is 50mg/L, the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 100mg/L after 30d of accumulation culture, the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 150mg/L after 45d of accumulation culture, and the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 200mg/L after 60d to 80d of accumulation culture and kept unchanged.
After the culture method of the embodiment is used for culturing, obvious red bacterial films are observed on the surfaces of the granular fillers, the nitrified sludge presents obvious dark red, a large number of bubbles appear between the nitrified sludge and filler particles in the anaerobic reactor, and the total nitrogen removal rate is 0.25 kg/(m) when enrichment is finished 3 ·d)。
Example 3
A method for culturing anaerobic ammonium oxidation bacteria particles comprises the following steps:
s1, mixing the granular filler with nitrified sludge to perform intermittent aeration culture by taking domestic sewage as water inlet, setting the aeration temperature to 35 ℃, setting the dissolved oxygen concentration to 5mg/L, introducing air to perform aeration for 8 hours, stopping aeration, standing and precipitating for 2 hours, discharging supernatant after standing, adding water inlet with the same volume, and standing for 5 hours, wherein the aeration culture period is one aeration culture period; the intermittent aeration culture of the particle filler is completed after the repeated culture period reaches 30 days;
s2, filling the granular filler obtained in the step S1 after intermittent aeration culture into a net surface hollow carrier to serve as a film-forming filler layer, placing the net surface hollow carrier into an anaerobic reactor, putting nitrified sludge into the anaerobic reactor, and introducing an enriched nutrient solution to start anaerobic ammonia oxidation culture;
s3, the enriched nutrient solution flows into a denitrification reactor containing denitrifying bacteria, carbon sources are supplemented into the denitrification reactor at regular time to perform denitrification to generate nitrogen, and the generated nitrogen is introduced into an anaerobic reactor to enable the anaerobic reactor to be in an anaerobic state;
s4, raising the ammonia nitrogen concentration of the enriched nutrient solution in stages for enrichment culture for 90 days, and observing an obvious red bacterial film on the surface of the film-forming filler layer to obtain the anaerobic ammonia oxidation bacterial particles, wherein the anaerobic ammonia oxidation culture is successfully started.
In the embodiment, the COD concentration of the domestic sewage is 500mg/L, the ammonia nitrogen concentration is 100mg/L, and the pH range is 9.
In the embodiment, the particle filler is formed by mixing ceramsite with the particle size of 3-5mm and volcanic rock, and the net surface hollow carrier is a spherical net surface PVC plastic sphere with the diameter of 10cm and the aperture of 1.5 mm.
In this embodiment, the nutrient solution comprises the following raw materials: NH (NH) 4 HCO 3 3g/L、NaNO 2 2.6g/L、K 2 HPO 4 0.1g/L、MgSO 4 ·7H 2 0 48mg/L、CaCl 2 35mg/L、FeSO 4 ·7H 2 0 25mg/L、ZnSO 4 ·7H 2 0 5mg/L、CoCl 2 ·6H 2 0 1.7mg/L、CuSO 4 ·5H 2 0 2mg/L、Na 2 MoO 4 ·5H 2 0.9 mg/L and H 3 BO 3 48mg/L。
In this example, the carbon source is a 10% wt aqueous glucose and maltose mixed solution.
In the step S4 of this embodiment, the ammonia nitrogen concentration of the nutrient solution introduced in the initial stage of the enrichment culture is 50mg/L, the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 100mg/L after 30d of accumulation culture, the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 150mg/L after 45d of accumulation culture, the ammonia nitrogen concentration of the enrichment nutrient solution is raised to 200mg/L after 60d of accumulation culture, and then the ammonia nitrogen concentration of the enrichment nutrient solution is kept unchanged, and the enrichment is completed after 90d of accumulation culture.
After the culture by the culture method of the example, obvious red bacteria are observed on the surface of the particle fillerThe membrane shows obvious dark red color, a large number of bubbles appear between the nitrified sludge and filler particles in the anaerobic reactor, and the total nitrogen removal rate is 0.31 kg/(m) when enrichment is finished 3 ·d)。
In summary, the anaerobic ammonia oxidation bacteria particle culture method and device provided by the invention have the characteristics of rapid starting, high thallus concentration and short culture period of anaerobic ammonia oxidation, and can provide continuous and stable nitrogen sources to meet the anoxic requirement in the anaerobic ammonia oxidation process by integrating the anaerobic ammonia oxidation process and the denitrification process, so that no additional nitrogen sources are required to be consumed, the whole culture method is more perfect, efficient, cost-saving, clean and environment-friendly, and the method is particularly suitable for industrial application. According to the anaerobic ammonia oxidation method, the particle filler with larger specific surface area is filled into the mesh surface hollow carrier and is used as a filler layer to be filled into the upflow anaerobic reactor, and the anaerobic ammonia oxidation effect is carried out on the filler and the nitrified sludge, so that the starting of anaerobic ammonia oxidation bacteria is accelerated, and the rapid mass expansion culture of the anaerobic ammonia oxidation bacteria is facilitated. Furthermore, the granular filler can be filled into the net surface hollow carrier to be made into a filler module to fix the granules in the anaerobic tank, so that the use is convenient; furthermore, the used filler module can be taken out and put into other anaerobic tanks for continuous use, so that the efficiency is high and the filler module can be reused. According to the invention, denitrification is carried out through the middle storage tank and the denitrification reactor, so that continuous and stable nitrogen source and anaerobic requirements can be provided for the anaerobic ammonia oxidation process, the efficiency of the whole culture device is improved through process integration and reasonable arrangement of equipment, manual intervention is reduced, and full-automatic and rapid start is realized.
The above specific examples are further illustrative of the technical solution and the advantageous effects of the present invention, and are not limiting to the embodiments. Any obvious substitutions would be within the scope of the invention for those skilled in the art without departing from the inventive concept.
Claims (8)
1. A method for culturing anaerobic ammonium oxidation bacteria particles is characterized in that: the method comprises the following steps:
s1, mixing domestic sewage as water, performing intermittent aeration culture on granular filler and nitrified sludge, introducing air for aeration, stopping aeration, standing and precipitating, discharging supernatant after standing, adding water for standing, and performing primary aeration culture period; the intermittent aeration culture of the particle filler is completed after the repeated culture period reaches more than 20 days; the particle filler is at least one of ceramsite, volcanic rock, polyurethane foam, perlite and zeolite;
s2, filling the granular filler obtained in the step S1 after intermittent aeration culture into a net surface hollow carrier to serve as a film-forming filler layer, placing the net surface hollow carrier into an anaerobic reactor, putting nitrified sludge into the anaerobic reactor, and introducing an enriched nutrient solution to start anaerobic ammonia oxidation culture;
s3, the enriched nutrient solution flows into a denitrification reactor containing denitrifying bacteria, carbon sources are supplemented into the denitrification reactor at regular time to perform denitrification to generate nitrogen, and the generated nitrogen is introduced into an anaerobic reactor to enable the anaerobic reactor to be in an anaerobic state;
s4, raising ammonia nitrogen concentration of the enriched nutrient solution in stages for enrichment culture for 60-90 days, and when obvious red bacterial films can be observed on the surface of the film-forming filler layer, obtaining the anaerobic ammonia oxidation bacterial particles, wherein the anaerobic ammonia oxidation culture is successfully started.
2. The method for culturing anammox bacteria particles according to claim 1, wherein: in the step S1, domestic sewage is taken as water inlet, granular filler and nitrified sludge are mixed for intermittent aeration culture, the aeration temperature is set to be 20-35 ℃, the concentration of dissolved oxygen is 2-5mg/L, aeration is carried out for 6-8 hours by air, aeration, standing and precipitation are stopped for 0.5-2 hours, supernatant is discharged after standing is finished, and water inlet with the same volume is added for standing for 3-5 hours, so that the one-time aeration culture period is adopted; and (5) repeating the culture period for more than 20 days to complete the intermittent aeration culture of the particle filler.
3. The method for culturing anammox bacteria particles according to claim 1, wherein: the enriched nutrient solution comprises the following raw materials: NH (NH) 4 HCO 3 0.2～3g/L、NaNO 2 0.2～3g/L、K 2 HPO 4 0.05～0.3g/L、MgSO 4 ·7H 2 0 5 ～50mg/L、CaCl 2 5～50mg/L、FeSO 4 ·7H 2 0 2 ～30mg/L、ZnSO 4 ·7H 2 0 0.5～5mg/L、CoCl 2 ·6H 2 0 0.2～2mg/L、CuSO 4 ·5H 2 0 0.5～2mg/L、Na 2 MoO 4 ·5H 2 0.5-5 mg/L and H 3 BO 3 5～50mg/L。
4. The method for culturing anammox bacteria particles according to claim 1, wherein: the carbon source is one or more of sodium acetate, methanol, glucose, maltose and molasses.
5. An anaerobic ammonia oxidation bacteria particle culture device which is characterized in that: the device comprises an enriched nutrient solution tank, an up-flow anaerobic reactor, a first water pump, a middle storage tank, a second water pump and a denitrification reactor, wherein the up-flow anaerobic reactor, the middle storage tank and the denitrification reactor are sequentially connected; the device comprises an up-flow anaerobic reactor, a medium storage tank, a denitrification reactor, a gas outlet, a gas pipeline and a three-phase separator, wherein the rich nutrient solution tank is connected with the up-flow anaerobic reactor through a first water pump, the top end of the up-flow anaerobic reactor is provided with the three-phase separator, the three-phase separator is provided with the liquid outlet, the liquid outlet is connected with the medium storage tank through a pipeline, the medium storage tank is connected with the denitrification reactor through a second water pump, the top end of the denitrification reactor is provided with the gas outlet, and the gas outlet is connected with the bottom end of the up-flow anaerobic reactor through the gas pipeline; the upflow anaerobic reactor is internally provided with a film-forming packing layer, and the film-forming packing layer is prepared by filling particle packing into a net surface hollow carrier.
6. The anaerobic ammonium oxidation bacteria culture apparatus according to claim 5, wherein: the device also comprises a carbon source solution tank and a third water pump, wherein the carbon source solution tank is connected with the middle storage tank through the third water pump, and the third water pump is used for supplementing the middle storage tank with carbon source at regular time.
7. The anaerobic ammonium oxidation bacteria culture apparatus according to claim 5, wherein: and liquid level controllers are arranged in the middle storage tank and the denitrification reactor.
8. The anaerobic ammonium oxidation bacteria culture apparatus according to claim 5, wherein: the denitrification reactor is internally provided with a denitrification filling layer, and the preparation of the denitrification filling layer comprises the following steps: (1) Dissolving denitrifying bacteria into 2-5% sodium alginate solution to prepare a denitrifying mixed solution; (2) Dripping the denitrification mixed solution into 1-3% calcium chloride solution for immobilization reaction to obtain denitrifying bacteria particles with the diameter of 3-5 mm; (3) And (5) putting denitrifying bacteria particles into a net surface hollow carrier to obtain the denitrifying filling layer.
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