CN114716013A - Method for inoculating anaerobic ammonium oxidation bacteria - Google Patents
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- CN114716013A CN114716013A CN202210508075.5A CN202210508075A CN114716013A CN 114716013 A CN114716013 A CN 114716013A CN 202210508075 A CN202210508075 A CN 202210508075A CN 114716013 A CN114716013 A CN 114716013A
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Abstract
The invention discloses a method for inoculating anaerobic ammonium oxidation bacteria, which relates to the technical field of sewage treatment and specifically comprises the following steps: pretreatment of S1 bottom mud, modification of S2 filler, S3 inoculation module and inoculation and enrichment of S4 anaerobic ammonium oxidation bacteria. The method for inoculating the anaerobic ammonium oxidation bacteria can realize the treatment of the bottom mud, is beneficial to the treatment of the bottom mud pollution in the river and lake pollution treatment process, and can also realize the inoculation and proliferation of the anaerobic ammonium oxidation bacteria on the filler.
Description
Technical Field
The invention belongs to the technical field of sewage treatment, and particularly relates to a method for inoculating anaerobic ammonium oxidation bacteria.
Background
The anaerobic ammonia oxidation technology is a novel biological denitrification technology which is successfully developed in recent years, the denitrification efficiency of the anaerobic ammonia oxidation technology greatly breaks through the traditional nitrification-denitrification technology, and the anaerobic ammonia oxidation technology has a very wide application prospect. However, the yield of the anaerobic ammonium oxidation bacteria is extremely low, the anaerobic ammonium oxidation bacteria grow slowly and are sensitive to environmental conditions, so that the popularization and the application of the process are limited. The water inlet substrate, the type of inoculated sludge and the type of the reactor are important influencing factors in the enrichment culture process of the anaerobic ammonium oxidation bacteria. The influence of the feed water base on the anaerobic ammonia oxidizing bacteria is one of research hotspots, but the conclusions drawn under different reactors and reaction conditions are very different, and the enrichment effect cannot be proved. Various types of sludge can be used as inoculants to successfully start the anaerobic ammonia oxidation process, and the starting of the reactor can be more effectively realized by screening the sludge similar to the growth environment of anaerobic ammonia oxidizing bacteria or selecting the sludge with higher anaerobic ammonia oxidizing bacteria content as the inoculants by utilizing a molecular biology technology. Patent No. CN109179649B discloses a method for rapidly inducing and enriching anaerobic ammonia oxidizing bacteria from nitrosation sludge, which realizes rapid induction and enrichment anaerobic ammonia oxidation in nitrosation sludge by adjusting the concentration of nano-copper oxide in a reactor; patent No. CN102381803A discloses a method for starting a completely autotrophic nitrogen removal process by taking common activated sludge as seed sludge, which provides sufficient nitrite output, and realizes the enrichment of anaerobic ammonium oxidation bacteria in large quantity by adding hydroxylamine hydrochloride and hydrazine hydrochloride; however, in the above technical process, the enrichment efficiency of anammox bacteria is low, so that the pollutant removal effect in pollution treatment is poor, and the excellent treatment effect is not achieved.
In the river and lake pollution treatment process, the bottom mud pollution treatment is one of the main difficulties and is a relatively ubiquitous environmental problem. The dynamic balance of absorption and release exists between the water body and the bottom sediment, when the water body is seriously polluted, a part of pollutants can enter the bottom sediment through the actions of precipitation, adsorption and the like; after the pollution caused by external sources is controlled, various organic and inorganic pollutants accumulated in the bottom mud enter the overlying water body again through the physical, chemical and biological exchange action with the overlying water body, and become secondary pollution sources influencing the water quality of the water body; meanwhile, various microorganisms in the bottom mud are enriched. Based on the consideration, the invention realizes the dual purposes of bottom sediment treatment and anaerobic ammonium oxidation bacteria inoculation enrichment by adopting a mode of inoculating anaerobic ammonium oxidation bacteria in the bottom sediment treatment process.
Disclosure of Invention
The invention aims to provide a method for inoculating anaerobic ammonium oxidation bacteria, which aims to overcome the defects in the prior art.
The technical scheme adopted by the invention is as follows:
a method for inoculating anaerobic ammonium oxidation bacteria specifically comprises the following steps:
s1 bottom mud pretreatment
Uniformly mixing the bottom mud and river water, separating by using a grid to remove impurities, and pumping the uniformly mixed mud-water mixture into an inoculation device by using a sludge pump;
s2 filler modification
1) Putting a potassium permanganate solution and a polyethylene filler into a container, placing a conical flask into an ultrasonic oscillator, oscillating for 1-4h under the conditions of the frequency of 30-35kHz and the temperature of 50-60 ℃, filtering after oscillation, and repeatedly cleaning a product with distilled water to obtain a pretreated polyethylene filler;
2) dissolving urea in deionized water, adding ethyl acetate, stirring at room temperature to obtain a mixed solution, adding the pretreated polyethylene filler into the mixed solution, and stirring and reacting for 1-5 hours at the speed of 60-120r/min in a constant-temperature water bath at the temperature of 60-80 ℃;
s3 inoculation module
Modifying the filler, placing the modified filler in an inoculation module, and hoisting the modified filler to the upper part of an inoculation device through a hook on the inoculation module;
s4 anaerobic ammonium oxidation bacterium inoculation enrichment
The artificial water distribution is used as the water inlet of the inoculation device, water is continuously fed through the pump, and when the filler of the inoculation module is red, the anaerobic ammonium oxidation bacteria are marked to be successfully enriched.
Furthermore, in the step 1), the bottom sludge comprises river sludge in a river regulation process, sludge in a ditch dredging process and sediment in a lake regulation process;
the mass ratio of the bottom mud to the river water is 1 (1-10);
the grid refers to a fine grid with the spacing between grid bars being 5-10 mm;
the inoculation device comprises a distribution pool, an upflow reactor and a water inlet pump, wherein the water outlet pipe of the upflow reactor is connected with the distribution pool, a nitrate probe is arranged in the distribution pool, the water inlet pipe enters from the bottom of the upflow reactor through the water inlet pump, bottom mud enters from the lower part of the upflow reactor, and an inoculation module is arranged at the upper part of the upflow reactor.
Furthermore, in the step 2), the potassium permanganate solution is prepared by mixing potassium permanganate, concentrated sulfuric acid and water according to the mass ratio of 1:1.6-2.3: 16-20;
the mass ratio of the potassium permanganate solution to the polyethylene filler is 10-15: 1;
the proportion of the urea, the deionized water, the ethyl acetate and the pretreated polyethylene filler is (5-10) g, (80-130) mL, (3-8) g and (8-15) g.
Further, in step 2), the preparation method of the polyethylene filler is as follows:
drying the dewatered sludge at the temperature of 105-125 ℃, crushing the dried sludge by a ball mill crusher, sieving the crushed sludge by a sieve of 100 meshes, mixing the dewatered sludge, the fly ash, the multi-layer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder, adding 30-35% of water into the raw materials per unit mass, uniformly stirring the raw materials to prepare raw filler balls with the diameter of 5-10mm, then maintaining the raw filler balls for 10-15h by using wet gauze at room temperature, and then moving the raw filler balls into a pressure steam container for steam maintenance;
the mass ratio of the dewatered sludge to the fly ash to the multilayer mesoporous polyethylene composite particles to the calcium oxide to the aluminum powder is 3:10-15:5-7:1-3: 0.01-0.05;
the steam curing parameters are as follows, and the steam curing is carried out for 10-15h at the temperature of 80-100 ℃ and under the pressure of 1.3-1.8 MPa.
Further, the preparation method of the multilayer mesoporous polyethylene composite particle comprises the following steps:
1) adding ammonium ferrous sulfate hexahydrate and ferric trichloride hexahydrate into deionized water, performing ultrasonic treatment, fully mixing and dissolving, adding multilayer mesoporous polyethylene into the solution, continuing performing ultrasonic treatment for 10-20min, and adding an ethylene diamine tetraacetic acid disodium solution while performing ultrasonic treatment for later use;
2) then, adding copper chloride dihydrate and sodium carbonate into the standby solution, stirring vigorously for 5-10h at 40-60 ℃, then raising the temperature of the mixture to 50-70 ℃, and slowly dripping the ammonia solution into the mixed solution under vigorous stirring;
3) after the surfaces of the multilayer mesoporous polyethylene are completely wrapped by the black products, slowly stirring the mixed solution for 3-5h at 40-60 ℃ at 30-80r/min, then stirring vigorously for 5-10h at 100-300r/min, repeatedly washing the obtained product with distilled water, and drying in an oven at 60-80 ℃ for 10-15h to obtain the multilayer mesoporous polyethylene composite particles.
Furthermore, the proportion of the ferrous ammonium sulfate hexahydrate, ferric trichloride hexahydrate, deionized water, multilayer mesoporous polyethylene, ethylene diamine tetraacetic acid disodium solution, copper chloride dihydrate, sodium carbonate and ammonia solution is (8-15) g, (17-25) g, (100) -200) mL, (0.3-0.5) g, (20-30) mL, (0.17-0.23) g, (0.4-0.7) g, (25-32) mL;
the concentration of the ethylene diamine tetraacetic acid disodium solution is 5-7 wt%;
the concentration is 30-35% of the mass concentration of the ammonia solution.
Further, the preparation method of the multilayer mesoporous polyethylene comprises the following steps:
1) adding magnesium chloride hexahydrate and absolute ethyl alcohol into a container, adjusting the pH value to 9.5-10.5, transferring the mixture into a hydrothermal reaction kettle, reacting for 20-30h at the temperature of 130-140 ℃, naturally cooling to room temperature, centrifugally separating the product, drying for 5-10h at the temperature of 60-80 ℃, and calcining for 2-5h at the temperature of 200-400 ℃ to obtain multilayer mesoporous nano magnesium oxide;
2) cleaning multilayer mesoporous nano magnesium oxide with ethanol, vacuum-drying at 80-90 ℃ for 20-25h, adding multilayer mesoporous nano magnesium oxide, hydroxyl silicone oil, silane coupling agent KH550 and maleic anhydride into a hydrothermal reaction kettle, performing hydrothermal reaction at the hydrothermal reaction temperature of 135-145 ℃ for 40-60min, naturally cooling to room temperature, and crushing to obtain modified multilayer mesoporous nano magnesium oxide;
3) cleaning polyethylene particles with ethanol, drying the polyethylene particles for 20-25h at 80-90 ℃ in vacuum for later use, ultrasonically dispersing the modified multilayer mesoporous nano magnesium oxide in a toluene solvent to form a dispersion liquid with the mass concentration of 5-10%, then blending the dispersion liquid with the polyethylene particles, blending the polyethylene particles for 8-13min at 140-160 ℃, then drying the polyethylene particles for 10-15h at 60-80 ℃, and obtaining the multilayer mesoporous polyethylene with the particle size of 0.1-0.5mm after crushing and screening.
Furthermore, the mass ratio of the magnesium chloride hexahydrate to the absolute ethyl alcohol is 1: 4-6;
the mass ratio of the multilayer mesoporous nano magnesium oxide to the hydroxyl silicone oil to the silane coupling agent KH550 to the maleic anhydride is 10:2-6:3-8: 5-10;
the dosage of the modified multilayer mesoporous nano magnesium oxide accounts for 10-30% of the mass of the polyethylene particles.
Furthermore, in the step 3), the inoculation module comprises a steel frame outer frame, an outer net and DN25-100 diversion pipelines.
Furthermore, in the step 4), the artificial water distribution comprises the nitrate with the concentration of 20-1000 mg/L;
the pump can control the flow and start and stop through the numerical value of the nitrate probe.
Advantageous effects
According to the invention, the adopted polyethylene filler is composed of dehydrated sludge, fly ash, multi-layer mesoporous polyethylene composite particles, calcium oxide and aluminum powder, the outer surface of the formed filler is rough, so that water flow is easy to separate on the surface of the filler, the impact force of the water flow on the surface of the filler can be reduced, and stable attachment of anaerobic ammonium oxidation bacteria is facilitated; moreover, the polyethylene filler is maintained, so that the hydrothermal synthesis and hydration reaction degree of the filler system are more sufficient, and a hydration product with better crystallinity is formed, so that the strength of the filler is greatly improved in a short time.
According to the polyethylene filler prepared by the invention, calcium oxide and water react rapidly to grow calcium hydroxide, active silicon dioxide and aluminum oxide in the fly ash glass body are excited to dissolve out rapidly in a large amount, and the calcium hydroxide reacts with the active silicon dioxide and the aluminum oxide to generate a gel hydration product; the dehydrated sludge can reduce the mass of the filler, and meanwhile, the aluminum powder serving as a pore-increasing agent can release hydrogen through calcium hydroxide reaction, so that the specific surface area of the filler is increased; the multilayer mesoporous polyethylene composite particles are formed by taking multilayer mesoporous polyethylene as a substrate and growing a large number of iron oxide nanoparticles on the surface of the substrate in situ, and the loaded iron oxide nanoparticles can effectively improve the hydrogenation and methylation rates of nitrogen-containing heterocyclic compounds, improve the rates of nitrogen heterocyclic ring opening, benzene ring opening and macromolecule-to-micromolecule conversion, realize the rapid transfer of electrons, avoid the accumulation of electrons in the degradation process of nitrogen-containing compounds, enhance the degradation efficiency of anaerobic ammonia oxidizing bacteria and contribute to realizing the efficient removal of pollutants in the remediation of bottom mud pollution; meanwhile, under the action of the composite particles, in the inoculation process, a large amount of anammox bacteria are enriched, and the colony structure is dispersed more uniformly, so that the anammox bacteria are more favorable for adapting to the change of conditions, the resistance of the anammox bacteria to the outside is enhanced, and the stable enrichment growth can still be kept under adverse environments.
According to the invention, the nano-magnesia with a multilayer mesoporous structure is successfully prepared through hydrothermal reaction, then the nano-magnesia is uniformly adhered to the surfaces of polyethylene particles by using a low-boiling-point solvent method, and multilayer mesoporous polyethylene with excellent dispersibility is prepared through haake blending.
According to the invention, potassium permanganate is used for oxidation modification on the surface of the polyethylene filler, hydroxyl groups on the surface of the polyethylene filler are oxidized to generate carboxyl groups, and each carboxyl group is used for forming covalent bonds, hydrogen bonds and other chemical bonding force adsorption among all components in the polyethylene filler, so that the bonding adsorption effect among all components in the polyethylene filler is enhanced, the internal space structure strength is improved, and the mechanical strength of the polyethylene filler is further improved; meanwhile, urea and ethyl acetate are adopted to carry out surface grafting modification on the polyethylene filler, so that the surface of the polyethylene filler has higher chemical activity, enrichment of anammox bacteria on the surface of the filler is facilitated, and the inoculation efficiency of the anammox bacteria is improved.
The method for inoculating the anaerobic ammonium oxidation bacteria can realize the treatment of the bottom mud, is beneficial to the treatment of the bottom mud pollution in the river and lake pollution treatment process, and can also realize the inoculation and proliferation of the anaerobic ammonium oxidation bacteria on the filler.
Drawings
FIG. 1 is a diagram of an inoculating device of the present invention;
FIG. 2 is a graph of the relative abundance of microbial genus levels on the packing of test experiment 1;
FIG. 3 is a graph showing the color change of the filler of test experiment 1 after attachment of anammox bacteria;
FIG. 4 shows biomass loading on different fillers;
FIG. 5 shows the pollution of river sediment before and after inoculation and the agricultural sediment pollution limit;
reference numerals: 1. an upflow reactor; 2. bottom mud; 3. an inoculation module; 4. a water distribution tank; 5. a water inlet pump; 6. a water inlet pipe; 7. a water outlet pipe; 8. a nitrate probe.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all 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 method for inoculating anaerobic ammonium oxidation bacteria specifically comprises the following steps:
s1 bottom mud pretreatment
Uniformly mixing bottom mud of the small east river in Guilin city and river water according to the mass ratio of 1:1, separating through a grid to remove impurities, and pumping the uniformly mixed mud-water mixture into an inoculation device by using a sludge pump;
the bottom sludge comprises river sludge in the river regulation process, sludge in the channel dredging process and sediment in the lake regulation process;
the grid refers to a fine grid with the spacing between grid bars being 5 mm;
the inoculation device comprises a distribution pool, an up-flow reactor and a water inlet pump, wherein a water outlet pipe of the up-flow reactor is connected with the distribution pool, and a nitrate probe is arranged in the distribution pool;
s2 filler modification
1) Putting a potassium permanganate solution and a polyethylene filler into a container according to the mass ratio of 10:1, placing a conical flask into an ultrasonic oscillator, oscillating for 1h under the conditions of the frequency of 30kHz and the temperature of 50 ℃, filtering after oscillation, and repeatedly cleaning a product with distilled water to obtain a pretreated polyethylene filler;
2) dissolving 5g of urea in 80mL of deionized water, adding 3g of ethyl acetate, stirring at room temperature to obtain a mixed solution, adding 8g of pretreated polyethylene filler into the mixed solution, and stirring and reacting for 1h at the speed of 60r/min in a constant-temperature water bath at the temperature of 60 ℃;
the potassium permanganate acid solution is prepared by mixing potassium permanganate, concentrated sulfuric acid and water according to the mass ratio of 1:2: 18;
s3 inoculation module
The modified filler is placed in a 1L inoculation module, and then the inoculation module is placed at the upper part 1/3 of the upflow anaerobic bioreactor;
the inoculation module comprises a steel frame outer frame, an outer net and DN25-100 diversion pipelines;
s4 anaerobic ammonium oxidation bacterium inoculation enrichment
The artificial water distribution is used as the water inlet of the inoculation device, the water is continuously fed through the pump, and when the filler of the inoculation module is red, the success of enrichment of anaerobic ammonium oxidation bacteria is marked;
the artificial water distribution comprises the components of nitrate with the concentration of 500 mg/L;
the pump can control the flow and start and stop through the numerical value of the nitrate probe.
The preparation method of the polyethylene filler comprises the following steps:
drying the dewatered sludge at 105 ℃, crushing the dewatered sludge by a ball mill crusher, sieving the crushed dewatered sludge with a 100-mesh sieve, mixing the dewatered sludge, the fly ash, the multilayer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder according to the mass ratio of 3:10:5:1:0.01, adding 30% of water into the raw materials per unit mass, uniformly stirring the mixture to prepare filler raw material balls with the diameter of 5mm, curing the raw material balls for 10 hours by using wet gauze at room temperature, transferring the raw material balls into a pressure steam container, and performing steam curing for 10 hours at 80 ℃ and 1.3 MPa.
The preparation method of the multilayer mesoporous polyethylene composite particle comprises the following steps:
1) putting 8g of ammonium ferrous sulfate hexahydrate and 17g of ferric chloride hexahydrate in 100mL of deionized water, carrying out ultrasonic treatment on the mixture for 15min at the power of 200W, adding 0.3g of multilayer mesoporous polyethylene into the solution after fully mixing and dissolving, continuing ultrasonic treatment for 10min, and adding 20mL of disodium ethylene diamine tetraacetate solution with the concentration of 5 wt% while carrying out ultrasonic treatment for later use;
2) then, 0.17g of copper chloride dihydrate and 0.4g of sodium carbonate are added into the standby solution, the mixture is stirred vigorously for 5 hours at 40 ℃ at 100r/min, then the temperature of the mixture is raised to 50 ℃, and 25mL of ammonia solution with the mass concentration of 30% is slowly dripped into the mixed solution under the vigorous stirring at 500 r/min;
3) after the surfaces of the multilayer mesoporous polyethylene are completely wrapped by the black product, slowly stirring the mixed solution for 3 hours at 40 ℃ at 30r/min, then stirring the mixed solution for 5 hours at 100r/min, repeatedly washing the obtained product with distilled water, and drying the washed product in a 60 ℃ drying oven for 10 hours to obtain the multilayer mesoporous polyethylene composite particles.
The preparation method of the multilayer mesoporous polyethylene comprises the following steps:
1) adding magnesium chloride hexahydrate and absolute ethyl alcohol into a container according to the mass ratio of 1:4, adjusting the pH value to 9.5, transferring the mixture into a hydrothermal reaction kettle, reacting for 20 hours at 130 ℃, naturally cooling to room temperature, centrifugally separating a product, drying for 5 hours at 60 ℃, and calcining for 2 hours at 200 ℃ to obtain multilayer mesoporous nano magnesium oxide;
2) washing multilayer mesoporous nano magnesium oxide with ethanol, vacuum-drying at 80 ℃ for 20 hours, adding the multilayer mesoporous nano magnesium oxide, hydroxyl silicone oil, a silane coupling agent KH550 and maleic anhydride into a hydrothermal reaction kettle according to a mass ratio of 10:2:3:5, performing hydrothermal reaction at 135 ℃ for 40min, naturally cooling to room temperature, and crushing to obtain modified multilayer mesoporous nano magnesium oxide;
3) cleaning polyethylene particles with ethanol, drying the polyethylene particles for 20 hours in vacuum at 80 ℃ for later use, ultrasonically dispersing the modified multilayer mesoporous nano magnesium oxide in a toluene solvent to form a dispersion liquid with the mass concentration of 5%, then blending the dispersion liquid with the polyethylene particles, controlling the using amount of the modified multilayer mesoporous nano magnesium oxide to be 10% of the mass of the polyethylene particles, blending the modified multilayer mesoporous nano magnesium oxide for 8 minutes at 140-160 ℃, then drying the polyethylene particles for 10 hours at 60 ℃, and crushing and screening the polyethylene particles to obtain the multilayer mesoporous polyethylene with the particle size of 0.1 mm.
Example 2
A method for inoculating anaerobic ammonium oxidation bacteria specifically comprises the following steps:
s1 bottom mud pretreatment
Uniformly mixing bottom mud of the small east river in Guilin city and river water according to the mass ratio of 1:5, separating through a grid to remove impurities, and pumping the uniformly mixed mud-water mixture into an inoculation device by using a sludge pump;
the bottom sludge comprises river sludge in the river regulation process, sludge in the channel dredging process and sediment in the lake regulation process;
the grid refers to a fine grid with the space between grid bars being 8 mm;
the inoculation device comprises a distribution pool, an up-flow reactor and a water inlet pump, wherein a water outlet pipe of the up-flow reactor is connected with the distribution pool, and a nitrate probe is arranged in the distribution pool;
s2 filler modification
1) Putting a potassium permanganate solution and a polyethylene filler into a container according to the mass ratio of 12:1, placing a conical flask into an ultrasonic oscillator, oscillating for 2 hours under the conditions of the frequency of 32kHz and the temperature of 55 ℃, filtering after oscillation, and repeatedly cleaning a product with distilled water to obtain a pretreated polyethylene filler;
2) dissolving 8g of urea in 120mL of deionized water, adding 5g of ethyl acetate, stirring at room temperature to obtain a mixed solution, adding 10g of pretreated polyethylene filler into the mixed solution, and stirring and reacting for 3 hours at 100r/min in a constant-temperature water bath at 70 ℃;
the potassium permanganate acid solution is prepared by mixing potassium permanganate, concentrated sulfuric acid and water according to the mass ratio of 1:1.6: 16;
s3 inoculation module
Modifying the filler, placing the modified filler in an inoculation module, and hoisting the modified filler to the upper part of an inoculation device through a hook on the inoculation module;
the inoculation module comprises a steel frame outer frame, an outer net and DN25-100 diversion pipelines;
s4 anaerobic ammonium oxidation bacterium inoculation enrichment
The artificial water distribution is used as the water inlet of the inoculation device, the water is continuously fed through the pump, and when the filler of the inoculation module is red, the success of enrichment of anaerobic ammonium oxidation bacteria is marked;
the artificial water distribution comprises the components of nitrate with the concentration of 700 mg/L;
the pump can control the flow rate and start and stop through the numerical value of the nitrate probe.
The preparation method of the polyethylene filler comprises the following steps:
drying the dewatered sludge at 115 ℃, crushing the dewatered sludge by a ball mill crusher, sieving the crushed dewatered sludge with a 100-mesh sieve, mixing the dewatered sludge, the fly ash, the multilayer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder according to the mass ratio of 3:12:6:2:0.03, adding 32 percent of water into the raw materials per unit mass, uniformly stirring the mixture to prepare filler raw material balls with the diameter of 7mm, curing the raw material balls for 12 hours by using wet gauze at room temperature, transferring the raw material balls into a pressure steam container, and performing steam curing for 12 hours at the temperature of 90 ℃ and the pressure of 1.5 MPa.
The preparation method of the multilayer mesoporous polyethylene composite particle comprises the following steps:
1) putting 13g of ammonium ferrous sulfate hexahydrate and 21g of ferric chloride hexahydrate in 160mL of deionized water, carrying out ultrasonic treatment on the mixture for 20min at the power of 300W, adding 0.4g of multilayer mesoporous polyethylene into the solution after fully mixing and dissolving, continuing ultrasonic treatment for 15min, and adding 25mL of disodium ethylene diamine tetraacetate solution with the concentration of 6 wt% while carrying out ultrasonic treatment for later use;
2) then, 0.19g of copper chloride dihydrate and 0.5g of sodium carbonate are added into the standby solution, the mixture is stirred vigorously at 50 ℃ for 7 hours at 200r/min, then the temperature of the mixture is raised to 60 ℃, and 28mL of ammonia solution with the mass concentration of 32% is slowly and dropwise added into the mixed solution under the vigorous stirring at 650 r/min;
3) after the surfaces of the multilayer mesoporous polyethylene are completely wrapped by the black product, slowly stirring the mixed solution for 4 hours at 50 ℃ at 50r/min, then stirring for 7 hours at 200r/min, repeatedly washing the obtained product with distilled water, and drying in an oven at 70 ℃ for 13 hours to obtain the multilayer mesoporous polyethylene composite particles.
The preparation method of the multilayer mesoporous polyethylene comprises the following steps:
1) adding magnesium chloride hexahydrate and absolute ethyl alcohol into a container according to the mass ratio of 1:5, adjusting the pH value to 10, transferring the mixture into a hydrothermal reaction kettle, reacting for 25 hours at 135 ℃, naturally cooling to room temperature, centrifugally separating a product, drying for 8 hours at 70 ℃, and calcining for 3 hours at 300 ℃ to obtain multilayer mesoporous nano magnesium oxide;
2) cleaning multilayer mesoporous nano magnesium oxide with ethanol, vacuum-drying at 85 ℃ for 23h, adding the multilayer mesoporous nano magnesium oxide, hydroxyl silicone oil, a silane coupling agent KH550 and maleic anhydride into a hydrothermal reaction kettle according to the mass ratio of 10:5:6:7, carrying out hydrothermal reaction at 140 ℃ for 50min, naturally cooling to room temperature, and crushing to obtain modified multilayer mesoporous nano magnesium oxide;
3) cleaning polyethylene particles with ethanol, drying the polyethylene particles at 85 ℃ in vacuum for 23 hours for later use, ultrasonically dispersing modified multilayer mesoporous nano magnesium oxide in a toluene solvent to form a dispersion liquid with the mass concentration of 7%, blending the dispersion liquid with the polyethylene particles, controlling the using amount of the modified multilayer mesoporous nano magnesium oxide to be 20% of the mass of the polyethylene particles, blending the modified multilayer mesoporous nano magnesium oxide at 150 ℃ for 10 minutes, drying the polyethylene particles at 70 ℃ for 13 hours, and crushing and screening the polyethylene particles to obtain the multilayer mesoporous polyethylene with the particle size of 0.3 mm.
Example 3
A method for inoculating anaerobic ammonium oxidation bacteria specifically comprises the following steps:
s1 bottom mud pretreatment
Uniformly mixing bottom mud of the small east river in Guilin city and river water according to the mass ratio of 1:10, separating through a grid to remove impurities, and pumping the uniformly mixed mud-water mixture into an inoculation device by using a sludge pump;
the bottom sludge comprises river sludge in the river regulation process, sludge in the channel dredging process and sediment in the lake regulation process;
the grid refers to a fine grid with the spacing between grid bars being 10 mm;
the inoculation device comprises a distribution pool, an up-flow reactor and a water inlet pump, wherein a water outlet pipe of the up-flow reactor is connected with the distribution pool, and a nitrate probe is arranged in the distribution pool;
s2 filler modification
1) Putting a potassium permanganate solution and a polyethylene filler into a container according to the mass ratio of 5:1, placing a conical flask into an ultrasonic oscillator, oscillating for 4 hours under the conditions of the frequency of 35kHz and the temperature of 60 ℃, filtering after oscillation, and repeatedly cleaning a product with distilled water to obtain a pretreated polyethylene filler;
2) dissolving 10g of urea in 130mL of deionized water, adding 8g of ethyl acetate, stirring at room temperature to obtain a mixed solution, adding 15g of pretreated polyethylene filler into the mixed solution, and stirring and reacting for 5 hours at 120r/min in a constant-temperature water bath at 80 ℃;
the potassium permanganate solution is prepared by mixing potassium permanganate, concentrated sulfuric acid and water according to the mass ratio of 1:2.3: 20;
s3 inoculation module
Modifying the filler, placing the modified filler in an inoculation module, and hoisting the modified filler to the upper part of an inoculation device through a hook on the inoculation module;
the inoculation module comprises a steel frame outer frame, an outer net and DN25-100 diversion pipelines;
s4 enrichment of anammox bacteria by inoculation
The artificial water distribution is used as the water inlet of the inoculation device, the water is continuously fed through the pump, and when the filler of the inoculation module is red, the success of enrichment of anaerobic ammonium oxidation bacteria is marked;
the artificial water distribution comprises the components of nitrate with the concentration of 1000 mg/L;
the pump can control the flow rate and start and stop through the numerical value of the nitrate probe.
The preparation method of the polyethylene filler comprises the following steps:
drying the dewatered sludge at 125 ℃, crushing the dewatered sludge by a ball mill crusher, sieving the crushed dewatered sludge with a 100-mesh sieve, mixing the dewatered sludge, the fly ash, the multilayer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder according to the mass ratio of 3:15:7:3:0.05, adding 35% of water into the raw materials per unit mass, uniformly stirring the mixture to prepare filler raw material balls with the diameter of 10mm, curing the raw material balls for 15 hours by using wet gauze at room temperature, transferring the raw material balls into a pressure steam container, and performing steam curing for 15 hours at 100 ℃ and 1.8 MPa.
The preparation method of the multilayer mesoporous polyethylene composite particle comprises the following steps:
1) putting 15g of ammonium ferrous sulfate hexahydrate and 25g of ferric trichloride hexahydrate in 200mL of deionized water, carrying out ultrasonic treatment on the mixture for 30min at the power of 500W, adding 0.5g of multilayer mesoporous polyethylene into the solution after fully mixing and dissolving, continuing ultrasonic treatment for 20min, and adding 30mL of disodium ethylene diamine tetraacetate solution with the concentration of 7 wt% while carrying out ultrasonic treatment for later use;
2) subsequently, 0.23g of copper chloride dihydrate and 0.7g of sodium carbonate are added into the standby solution, the mixture is stirred vigorously at the temperature of 60 ℃ for 10 hours at the speed of 300r/min, then the temperature of the mixture is raised to 70 ℃, and 32mL of ammonia solution with the mass concentration of 35% is slowly and dropwise added into the mixed solution under the vigorous stirring at the speed of 800 r/min;
3) after the surfaces of the multilayer mesoporous polyethylene are completely wrapped by the black product, slowly stirring the mixed solution for 5 hours at 60 ℃ at 80r/min, then stirring the mixed solution for 10 hours at 300r/min, repeatedly washing the obtained product with distilled water, and drying the washed product in an oven at 80 ℃ for 15 hours to obtain the multilayer mesoporous polyethylene composite particles.
The preparation method of the multilayer mesoporous polyethylene comprises the following steps:
1) adding magnesium chloride hexahydrate and absolute ethyl alcohol into a container according to the mass ratio of 1:6, adjusting the pH value to 10.5, transferring the mixture into a hydrothermal reaction kettle, reacting for 30 hours at 140 ℃, naturally cooling to room temperature, centrifugally separating a product, drying for 10 hours at 80 ℃, and calcining for 5 hours at 400 ℃ to obtain multilayer mesoporous nano magnesium oxide;
2) cleaning multilayer mesoporous nano magnesium oxide with ethanol, vacuum-drying at 90 ℃ for 25h, adding the multilayer mesoporous nano magnesium oxide, hydroxyl silicone oil, a silane coupling agent KH550 and maleic anhydride into a hydrothermal reaction kettle according to the mass ratio of 10:6:8:10, carrying out hydrothermal reaction at 145 ℃ for 60min, naturally cooling to room temperature, and crushing to obtain modified multilayer mesoporous nano magnesium oxide;
3) cleaning polyethylene particles with ethanol, drying the polyethylene particles for 25 hours in vacuum at 90 ℃ for later use, ultrasonically dispersing modified multilayer mesoporous nano magnesium oxide in a toluene solvent to form a dispersion liquid with the mass concentration of 10%, blending the dispersion liquid with the polyethylene particles, controlling the using amount of the modified multilayer mesoporous nano magnesium oxide to be 30% of the mass of the polyethylene particles, blending the modified multilayer mesoporous nano magnesium oxide for 13 minutes at 160 ℃, drying the polyethylene particles for 15 hours at 80 ℃, and crushing and screening the polyethylene particles to obtain the multilayer mesoporous polyethylene with the particle size of 0.5 mm.
Comparative example 1: the polyethylene filler used in this comparative example was substantially the same as in example 1, except that the polyethylene particles were used directly in the preparation of the polyethylene filler.
Comparative example 2: the polyethylene filler used in this comparative example was substantially the same as in example 1, except that the multilayer mesoporous polyethylene particles were directly used in the preparation method of the polyethylene filler.
Comparative example 3: the polyethylene filler used in this comparative example was substantially the same as in example 1, except that the multilayer mesoporous polyethylene composite particles were prepared by replacing the multilayer mesoporous polyethylene with polyethylene particles.
Comparative example 4: the polyethylene filler used in this comparative example was substantially the same as in example 1, except that the multilayer mesoporous polyethylene was prepared by replacing the multilayer mesoporous nano-magnesia with nano-magnesia.
Test experiment 1:
the fillers respectively adopt modified polyethylene K3, PET, PU-1 and PU-2, the inoculation method provided by the embodiment 1 is adopted, after 100 days of inoculation, the inoculation module and the K3 fillers in the inoculation module are taken out, the following figure 2 shows the relative abundance of the microorganism level in the fillers, and the figure 3 shows the color change of the fillers after the anaerobic ammonium oxidation bacteria are attached.
The results of the genome structure of the microorganisms in the reaction system are analyzed by the complete gene sequencing of the metagenome, so that the reactor forms an ANAOB community which mainly comprises Candida Kuenenia stuttgartiiensis, Candida Brocadia fulvida and Candida Brocadia sinica, and the modified polyethylene K3 surface is more beneficial to the formation of a biomembrane by the aid of the Chloroflexi bacterium and the Chlorobi bacterium in cooperation with the anaerobic ammonium oxidation bacteria for denitrification.
Test experiment 2:
the inoculation method provided by the embodiment 1 is adopted, the fillers in the embodiment 1 and the comparative examples 1 to 4 are respectively selected for inoculation of anammox bacteria, and the anammox flora A-E obtained by the inoculation is used in the anammox treatment process of wastewater, wherein the ammonia nitrogen concentration in the wastewater is 80mg/L, the nitrite nitrogen concentration is 75mg/L, the wastewater treatment temperature is 30 ℃, the dissolved oxygen concentration is lower than 0.5mg/L, and the pH is 7.5 to 8.0. The wastewater is treated by adopting a batch water exchange and drainage mode, 24 hours is a treatment period, and the ammonia nitrogen and total nitrogen removal rate of the wastewater after 5 days is shown in table 1.
TABLE 1
| Anammox bacteria | The ammonia nitrogen removal rate% | Total nitrogen removal% |
| A | 97.8 | 97.1 |
| B | 52.3 | 51.7 |
| C | 64.1 | 63.5 |
| D | 75.8 | 74.6 |
| E | 82.7 | 81.9 |
The modified polyethylene filler is beneficial to enrichment of anammox bacteria on the surface of the filler, improves the inoculation efficiency of the anammox bacteria, enhances the degradation efficiency of the anammox bacteria, and is beneficial to realizing efficient removal of pollutants in bottom mud pollution remediation.
Test experiment 3:
the added polyethylene K3, polyurethane fiber balls and polyurethane sponge mixed fillers are cultured for 5 days by adopting the inoculation method provided by the embodiment 1, then taken out to measure the biomass, and the biomass loaded on different fillers is shown in figure 4. The biomass loaded by the polyethylene K3 is 1.5669g/L, the biomass loaded by the polyurethane fiber ball is 0.8577g/L, and the biomass loaded by the polyurethane sponge is 0.6562g/L, wherein the loading effect of the polyethylene K3 is better than that of the polyurethane fiber ball and the polyurethane sponge filler.
The comprehensive utilization of the river bottom mud generally takes agriculture and forestry utilization and building material utilization as main points. The river bottom mud is used as inoculated sludge to culture anaerobic ammonia oxidizing bacteria, meanwhile, the anaerobic ammonia oxidizing bacteria can purify the river bottom mud, the bottom mud treated by the reactor refers to the industrial standard of sludge treatment agricultural argillaceous substance of urban sewage treatment plants (CJ/T309-2009), and pollutants in the river bottom mud can be recycled. FIG. 5 shows the pollution of river sediment before and after inoculation and the agricultural sediment pollution limit. The concentrations of ammonia nitrogen, TP, TN and PAHs in the bottom sludge after inoculation are obviously reduced, so that the enrichment effect of anaerobic ammonia oxidizing bacteria in the river bottom sludge and the treatment effect of the anaerobic ammonia oxidizing bacteria on nitrogen and organic matters are obvious.
The above description is only an example and an experimental example of the present invention, and is not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. A method for inoculating anaerobic ammonium oxidation bacteria is characterized by comprising the following steps:
s1 pretreatment of bottom mud
Uniformly mixing the bottom sludge and river water, separating by using a grating to remove impurities, and pumping the uniformly mixed sludge-water mixture into an inoculation device by using a sludge pump;
s2 filler modification
1) Putting a potassium permanganate solution and a polyethylene filler into a container, placing a conical flask into an ultrasonic oscillator, oscillating for 1-4h under the conditions of the frequency of 30-35kHz and the temperature of 50-60 ℃, filtering after oscillation, and repeatedly cleaning a product with distilled water to obtain a pretreated polyethylene filler;
2) dissolving urea in deionized water, adding ethyl acetate, stirring at room temperature to obtain a mixed solution, adding the pretreated polyethylene filler into the mixed solution, and stirring and reacting for 1-5 hours at 60-120r/min in a constant-temperature water bath at 60-80 ℃;
s3 inoculation module
Modifying the filler, placing the modified filler in an inoculation module, and hoisting the modified filler to the upper part of an inoculation device through a hook on the inoculation module;
s4 anaerobic ammonium oxidation bacterium inoculation enrichment
The artificial water distribution is used as the water inlet of the inoculation device, the water is continuously fed through the pump, and when the filler of the inoculation module is red, the success of anaerobic ammonium oxidation bacteria enrichment is marked.
2. A method of inoculating anammox bacteria according to claim 1, wherein: in the step 1), the bottom mud comprises river mud in a river regulation process, mud in a ditch dredging process and sediment in a lake regulation process;
the mass ratio of the bottom mud to the river water is 1 (1-10);
the grid refers to a fine grid with the spacing between grid bars being 5-10 mm;
the inoculation device comprises a distribution pool, an up-flow reactor and a water inlet pump, wherein a water outlet pipe of the up-flow reactor is connected with the distribution pool, and a nitrate probe is arranged in the distribution pool.
3. A method of inoculating anammox bacteria according to claim 1, wherein: in the step 2), the potassium permanganate solution is formed by mixing potassium permanganate, concentrated sulfuric acid and water according to the mass ratio of 1:1.6-2.3: 16-20;
the mass ratio of the potassium permanganate solution to the polyethylene filler is 10-15: 1;
the proportion of the urea, the deionized water, the ethyl acetate and the pretreated polyethylene filler is (5-10) g, (80-130) mL, (3-8) g and (8-15) g.
4. A method of inoculating anammox bacteria according to claim 1, wherein: in the step 2), the preparation method of the polyethylene filler comprises the following steps:
drying the dewatered sludge at the temperature of 105-125 ℃, crushing the dried sludge by a ball mill crusher, sieving the crushed sludge by a sieve of 100 meshes, mixing the dewatered sludge, the fly ash, the multi-layer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder, adding 30-35% of water into the raw materials per unit mass, uniformly stirring the raw materials to prepare raw filler balls with the diameter of 5-10mm, then maintaining the raw filler balls for 10-15h by using wet gauze at room temperature, and then moving the raw filler balls into a pressure steam container for steam maintenance;
the mass ratio of the dewatered sludge, the fly ash, the multilayer mesoporous polyethylene composite particles, the calcium oxide and the aluminum powder is 3:10-15:5-7:1-3: 0.01-0.05;
the steam curing parameters are as follows, and the steam curing is carried out for 10-15h at the temperature of 80-100 ℃ and under the pressure of 1.3-1.8 MPa.
5. A method of inoculating anammox bacteria according to claim 4, wherein: the preparation method of the multilayer mesoporous polyethylene composite particle comprises the following steps:
1) adding ammonium ferrous sulfate hexahydrate and ferric trichloride hexahydrate into deionized water, performing ultrasonic treatment, fully mixing and dissolving, adding multilayer mesoporous polyethylene into the solution, continuing performing ultrasonic treatment for 10-20min, and adding an ethylene diamine tetraacetic acid disodium solution for later use during ultrasonic treatment;
2) then, adding copper chloride dihydrate and sodium carbonate into the standby solution, stirring vigorously for 5-10h at 40-60 ℃, then raising the temperature of the mixture to 50-70 ℃, and slowly dripping ammonia solution into the mixed solution under vigorous stirring;
3) after the surfaces of the multilayer mesoporous polyethylene are completely wrapped by the black products, slowly stirring the mixed solution for 3-5h at 40-60 ℃ at 30-80r/min, then stirring vigorously for 5-10h at 100-300r/min, repeatedly washing the obtained product with distilled water, and drying in an oven at 60-80 ℃ for 10-15h to obtain the multilayer mesoporous polyethylene composite particles.
6. A method of inoculating anammox bacteria according to claim 5, wherein: the dosage proportion of the ferrous ammonium sulfate hexahydrate, the ferric trichloride hexahydrate, the deionized water, the multilayer mesoporous polyethylene, the ethylene diamine tetraacetic acid disodium solution, the copper chloride dihydrate, the sodium carbonate and the ammonia solution is (8-15) g, (17-25) g, (100-200) mL, (0.3-0.5) g, (20-30) mL, (0.17-0.23) g, (0.4-0.7) g, (25-32) mL;
the concentration of the ethylene diamine tetraacetic acid disodium solution is 5-7 wt%;
the concentration is 30-35% of the mass concentration of the ammonia solution.
7. A method of inoculating anammox bacteria according to claim 5, wherein: the preparation method of the multilayer mesoporous polyethylene comprises the following steps:
1) adding magnesium chloride hexahydrate and absolute ethyl alcohol into a container, adjusting the pH value to be 9.5-10.5, transferring the mixture into a hydrothermal reaction kettle, reacting for 20-30h at the temperature of 130-140 ℃, naturally cooling to room temperature, centrifugally separating a product, drying for 5-10h at the temperature of 60-80 ℃, and calcining for 2-5h at the temperature of 200-400 ℃ to obtain multilayer mesoporous nano magnesium oxide;
2) cleaning multilayer mesoporous nano magnesium oxide with ethanol, vacuum-drying at 80-90 ℃ for 20-25h, adding multilayer mesoporous nano magnesium oxide, hydroxyl silicone oil, silane coupling agent KH550 and maleic anhydride into a hydrothermal reaction kettle, performing hydrothermal reaction at the hydrothermal reaction temperature of 135-145 ℃ for 40-60min, naturally cooling to room temperature, and crushing to obtain modified multilayer mesoporous nano magnesium oxide;
3) cleaning polyethylene particles with ethanol, drying the polyethylene particles for 20-25h at 80-90 ℃ in vacuum for later use, ultrasonically dispersing the modified multilayer mesoporous nano magnesium oxide in a toluene solvent to form a dispersion liquid with the mass concentration of 5-10%, then blending the dispersion liquid with the polyethylene particles, blending the polyethylene particles for 8-13min at 140-160 ℃, then drying the polyethylene particles for 10-15h at 60-80 ℃, and obtaining the multilayer mesoporous polyethylene with the particle size of 0.1-0.5mm after crushing and screening.
8. A method of inoculating anammox bacteria according to claim 7, wherein: the mass ratio of the magnesium chloride hexahydrate to the absolute ethyl alcohol is 1: 4-6;
the mass ratio of the multilayer mesoporous nano magnesium oxide to the hydroxyl silicone oil to the silane coupling agent KH550 to the maleic anhydride is 10:2-6:3-8: 5-10;
the dosage of the modified multilayer mesoporous nano magnesium oxide accounts for 10-30% of the mass of the polyethylene particles.
9. A method of inoculating anammox bacteria according to claim 1, wherein: in the step 3), the inoculation module comprises a steel frame outer frame, an outer net and DN25-100 diversion pipelines.
10. A method of inoculating anammox bacteria according to claim 1, wherein: in the step 4), the artificial water distribution comprises the components of nitrate with the concentration of 20-1000 mg/L;
the pump can control the flow and start and stop through the numerical value of the nitrate probe.
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