CN116121233A - Preparation method and application of biological film - Google Patents
Preparation method and application of biological film Download PDFInfo
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Abstract
The invention discloses a preparation method and application of a biological film, wherein the preparation method comprises the following steps: preparing a cellulose carrier; loading microorganisms with a cellulose carrier; preparing a coating agent; and coating the carrier loaded with the strain by using a coating agent. The cellulose carrier has higher porosity, larger specific surface area and higher surface roughness, is beneficial to the absorption and maintenance of moisture and the growth and propagation of microorganisms, the cellulose shows that more active groups can also play a role in protecting the attached microorganisms, the attachment, the fixation and the propagation of the microorganisms on the carrier are accelerated, the rapid formation of a biological film is promoted, the slow release of the microorganisms in a sewage environment can be realized by the aid of the outer coating agent, the shearing action of the hydraulic shearing action on bacteria can be reduced, the impact resistance is good, the biological film is not easy to fall off, and the denitrification effect of the microorganisms is improved.
Description
Technical Field
The invention relates to the technical field of sewage biological treatment, in particular to a preparation method and application of a biological membrane.
Background
Biofilm, also known as biofilm, refers to an organized population of bacteria attached to the surface of living or inanimate objects that are surrounded by extracellular macromolecules of bacteria, which are very resistant to antibiotics and host immune defence mechanisms. Various major biological macromolecules such as proteins, polysaccharides, DNA, RNA, peptidoglycans, lipids, and phospholipids exist in biological membranes. The formation of a multicellular structure of a biofilm is a dynamic process that includes stages of bacterial initiation adhesion, biofilm development, and maturation diffusion.
The sewage biological treatment process can be classified into an activated sludge method and a biofilm method according to the growth form of microorganisms in the bioreactor. The biological membrane method is to adhere microorganisms on the surface of a carrier, and sewage is decomposed by the adsorption of organic nutrients, the diffusion of oxygen into the biological membrane, biological oxidation in the membrane and other actions during the process of flowing through the surface of the carrier, compared with activated sludge, the biological membrane method has a treatment main body of membranous microorganisms with larger physical dimensions and a more complex space structure, so that the biological membrane has a certain dissolved oxygen gradient in the direction vertical to the surface of the carrier, the types of microorganisms are more abundant, and various types of microorganisms can be considered to form a more perfect food chain network; the relatively stable internal environment of the biological film can provide a proper growth environment for slow-growing microorganisms, and can maintain a sufficient microorganism quantity under unfavorable environments; the biological film process also has lower mud yield, larger biomass per unit volume and flexible and controllable operating conditions, and has larger application potential in the sewage treatment field because of the characteristics, and the biological film process has wider application in rural sewage, low-temperature sewage, low-carbon source sewage and industrial wastewater in recent years and is a research hot spot in the sewage treatment field.
CN111003799a discloses a biological carrier for sewage treatment and a preparation method thereof, the preparation method comprises the following steps: preparing a bacterial cellulose membrane by using a fermentation medium, purifying the bacterial cellulose membrane, freeze-drying to obtain bacterial cellulose aerogel, pyrolyzing to obtain carbon nanofiber aerogel, and finally fluorinating the carbon nanofiber aerogel to obtain the biological carrier for sewage treatment. The biological carrier prepared by the invention has moderate wettability which is beneficial to microbial adhesion, and shortens the microbial film forming time. The biological carrier has a three-dimensional porous interconnected network structure and high biocompatibility, is beneficial to microorganism growth and improves sewage treatment efficiency. The raw materials of the biological carrier are environment-friendly, and the preparation method is simple and convenient. The fluorinated carbon nanofiber aerogel prepared by the method has the defects of insufficient mechanical strength, specific surface area and porosity which are still required to be further improved.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide a biofilm with good mechanical strength, large specific surface area and porosity.
In order to achieve the aim, the invention loads the microbial agent on the carrier to obtain the biological film, and the biological film has a large number of micropore structures, has high specific surface area, provides a proper growth environment for microorganism adsorption, is beneficial to rapid film hanging of microorganisms, has strong adsorption and degradation capability, excellent mechanical property and long service life, can synchronously nitrify and denitrify reaction, and has high denitrification efficiency.
In order to achieve the above object, the present invention adopts the following technical scheme:
a method for preparing a biological membrane, comprising the steps of: preparing a cellulose carrier; loading microorganisms with a cellulose carrier; preparing a coating agent; and coating the carrier loaded with the strain by using a coating agent.
Preferably, a method for preparing a biological membrane comprises the following steps:
s1, dispersing cellulose in a mixed solution composed of sodium hydroxide, urea and water, stirring to obtain a suspension, freezing and sublimating to obtain cellulose aerogel;
s2, mixing zeolite powder after acidification treatment with cellulose aerogel and ethanol solution, and obtaining a cellulose carrier after ultrasonic reaction, filtration and drying;
s3, culturing COD degrading bacteria, preparing bacterial suspension, mixing with a cellulose carrier, concentrating, and naturally air-drying to obtain a carrier for loading strains;
s4, preparing a coating agent by using polyethylene glycol and pullulan to react;
s5, mixing the coating agent and the carrier loaded with the strain, adding the mixture into a calcium chloride aqueous solution for reaction, filtering and washing to obtain the biomembrane.
Further preferably, the cellulose aerogel is a modified cellulose aerogel, and the preparation method thereof is as follows:
m1, dispersing cellulose in a mixed solution composed of sodium hydroxide, urea and water, stirring to obtain a suspension, freezing and sublimating to obtain cellulose aerogel;
and adding bis (3-trimethoxysilylpropyl) amine into the suspension, stirring, freezing and sublimating to obtain the modified cellulose aerogel.
Most preferably, a method for preparing a biofilm comprises the steps of:
s1, microcrystalline cellulose is dispersed in a mixed solution composed of sodium hydroxide, urea and water, and stirred for 2-3 hours at room temperature to obtain a suspension;
s2, dropwise adding bis (3-trimethoxysilylpropyl) amine into the suspension obtained in the step S1 at a speed of 1-2 drops/second, stirring at room temperature for 10-20min after the dropwise adding, standing for 2-3h, transferring into a mold, freezing in a liquid nitrogen bath, and performing vacuum sublimation at a temperature of 10-40 ℃ to obtain modified cellulose aerogel;
s3, adding zeolite powder into 30-36wt% hydrochloric acid, stirring for 2-3 hours, filtering, washing a filter cake with water until the pH value of the filtrate is neutral, drying at 60-80 ℃ for 8-10 hours to obtain acidified zeolite, mixing the acidified zeolite, the modified cellulose aerogel obtained in the step S2 and 75-95wt% ethanol aqueous solution, carrying out ultrasonic treatment at 60-100Hz and 120-150W for 1-2 hours, filtering, and drying the filter cake at 60-80 ℃ for 6-8 hours to obtain a modified cellulose carrier;
s4, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing a modified cellulose carrier and the bacterial suspension, concentrating at 20-40 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain a carrier carrying strains;
s5, mixing polyethylene glycol, pullulan and water, heating to 50-70 ℃, stirring for 20-30min, and cooling to 40-50 ℃ to obtain a coating agent;
s6, uniformly mixing the coating agent and the carrier loaded with the strain, adding the mixture into 1-2mol/L calcium chloride aqueous solution, reacting for 20-24 hours, filtering, and washing a filter cake with water for 2-3 times to obtain the biological membrane.
Further preferably, in the step S1, the mass ratio of the sodium hydroxide to the urea to the water is 5-8:10-15:50-100; the mass ratio of the microcrystalline cellulose to the mixed solution is 1:15-20.
Further preferably, the mass ratio of the suspension in the step S2 to the bis (3-trimethoxysilylpropyl) amine is 5-10:1.
Further preferably, in the step S3, the mass ratio of the zeolite powder to the hydrochloric acid is 1:10-15; the dosage ratio of the acidified zeolite, the modified cellulose aerogel and the ethanol water solution is 5-10g to 1g to 100-200mL.
Further preferably, the mass ratio of the modified cellulose carrier to the bacterial suspension in the step S4 is 1:3-5.
Further preferably, in the step S5, the mass ratio of polyethylene glycol, pullulan and water is 1:5-10:100-200.
Further preferably, in the step S6, the mass ratio of the coating agent, the carrier for loading the strain and the calcium chloride aqueous solution is 1-5:1:100-150.
Further preferably, the culturing step of the COD degrading bacteria in the step S3 is as follows:
bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic GordoniaInoculating 2-5wt% of bacteria into LB liquid culture medium, shake culturing at 25-30deg.C for 15-20 hr, anaerobic fermenting at 25-30deg.C for 25-30 hr in fermentation tank, and culturing to obtain viable bacteria with number not less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing the bacillus subtilis liquid, the bacillus amyloliquefaciens liquid, the lactobacillus plantarum liquid, the enterococcus faecalis liquid and the hydrophobic Gordonia liquid according to the mass ratio of 1-2:1-2:1:1-2.
The inventor prepares cellulose into cellulose aerogel, the cellulose aerogel has the advantages of easy surface modification, high porosity, high specific surface area and 3D interconnected porous network structure, and uses bis (3-trimethoxy silicon-based propyl) amine to modify cellulose, so that the internal structure of cellulose is improved, the introduction of active groups improves the processing performance of cellulose, the zeolite powder is fixed in the cellulose aerogel, the aerogel can play a framework supporting role, the strength is improved, the microporous structure of a biological carrier can be regulated and controlled, the specific surface area is increased, the microorganism load capacity is improved, the prepared modified cellulose carrier has higher porosity, larger specific surface area and higher surface roughness, not only is favorable for the absorption and maintenance of moisture, but also is favorable for the growth and propagation of microorganisms, the active groups on the surface of cellulose can play a role in protecting attached microorganisms, the attachment, the fixation and the propagation of microorganisms on the carrier are accelerated, the rapid formation of a biological film is promoted, the outer layer of the coating agent can enable the microorganisms to be slowly released in the environment, the shearing action of the microorganism on sewage is also reduced, and the biological film is easy to fall off.
The invention also provides application of the biological film: the method is applied to a biological membrane reactor for sewage treatment.
Compared with the prior art, the invention has the beneficial effects that: the biological film prepared by the invention is convenient to use, can be directly added into a water body, does not need to additionally increase facilities, saves energy and saves investment of funds; the cellulose carrier prepared by the invention has higher porosity, larger specific surface area and higher surface roughness, and by introducing zeolite powder into cellulose aerogel, the mechanical strength of the cellulose carrier is improved well, so that the cellulose carrier is beneficial to the absorption and maintenance of moisture, the growth and propagation of microorganisms in the cellulose carrier are facilitated, the rapid formation of biological films is promoted, the microorganisms can be slowly released in a sewage environment by the coating agent on the outer layer, the shearing action of hydraulic shearing action on bacteria can be reduced, the impact resistance is good, the biological films are not easy to fall off, and the denitrification effect of the microorganisms is improved.
Detailed Description
For the sake of brevity, the articles used in the examples below are commercially available products unless otherwise specified, and the methods used are conventional methods unless otherwise specified.
The sources of part of raw materials used in the invention are as follows:
bacillus subtilis, CICC25064, bacillus subtilis, purchased from the chinese industrial microbiological bacterial deposit management center.
Bacillus amyloliquefaciens, CICC 10080,Bacillus amyloliquefaciens, purchased from China center for type culture Collection of microorganisms.
Lactobacillus plantarum, CICC22846, lactobacillus plantarum, purchased from the chinese industrial microbiological bacterial deposit management center.
Enterococcus faecalis, cic c20396, enterococcus faecalis, purchased from the chinese industrial microbiological bacterial collection center.
The hydrophobic gordonia bacteria, CICC24205, gordonia hydrophobica were purchased from the chinese industrial microorganism strain collection management center.
Zeolite powder with 150 meshes, 69% magnesia, 11% calcium oxide, 0.6% ferric oxide and 0.2% wear rate.
Polyethylene glycol having a molecular weight of 200.
Microcrystalline cellulose, 99% in content, shandong Zhenghong Biotechnology Co., ltd.
LB liquid medium, purchased from Nantong Kai Hemsl Biotechnology development Co.
Bis (3-trimethoxysilylpropyl) amine, CAS:82985-35-1.
Example 1
A method for preparing a biological membrane, comprising the steps of:
s1, 10g of microcrystalline cellulose is dispersed in 200g of microcrystalline cellulose with a mass ratio of 5:10:50, stirring for 2 hours at room temperature in a mixed solution composed of sodium hydroxide, urea and water to obtain a suspension;
s2, dropwise adding 25g of bis (3-trimethoxysilylpropyl) amine into the suspension obtained in the step S1 at a speed of 1 drop/second, stirring at room temperature for 20min after the dropwise adding is completed, standing for 3h, transferring into a mold, cooling in a liquid nitrogen bath for 30min, and performing vacuum sublimation at 20 ℃ to obtain modified cellulose aerogel;
s3, adding 50g of zeolite powder into 500g of 36wt% hydrochloric acid, stirring for 3 hours, filtering, washing a filter cake with water until the pH value of the filtrate is neutral, drying at 80 ℃ for 10 hours to obtain acidified zeolite, mixing 5g of acidified zeolite, 1g of modified cellulose aerogel obtained in the step S2 and 150mL of 90wt% ethanol aqueous solution, carrying out ultrasonic treatment for 2 hours under the conditions of 80Hz and 120W, filtering, and drying the filter cake at 80 ℃ for 8 hours to obtain a modified cellulose carrier;
s4, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing 2g of modified cellulose carrier and 10g of bacterial suspension, concentrating at 30 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain a carrier material for loading strains;
s5, mixing 2g of polyethylene glycol, 10g of pullulan and 200g of water, heating to 60 ℃, stirring for 30min, and cooling to 40 ℃ to obtain a coating agent;
s6, uniformly mixing 5g of coating agent and 1g of carrier loaded with strains, adding the mixture into 150mL of 1mol/L calcium chloride aqueous solution, reacting for 24 hours at room temperature, filtering, and washing a filter cake with water for 3 times to obtain the biological membrane.
The culturing steps of the COD degrading bacteria are as follows:
inoculating Bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic Gordonia to LB liquid culture medium at 25deg.C and 4000 rpm respectively, shake culturing for 18 hr, anaerobic fermenting at 30deg.C for 30 hr in a fermenter, and culturing to obtain viable bacteriaNumber average is not less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing bacillus subtilis liquid, bacillus amyloliquefaciens liquid, lactobacillus plantarum liquid, enterococcus faecalis liquid and hydrophobic gordonia bacteria liquid according to the mass ratio of 2:2:1:1:2.
Example 2
A method for preparing a biological membrane, comprising the steps of:
s1, 10g of microcrystalline cellulose is dispersed in 200g of microcrystalline cellulose with a mass ratio of 5:10: stirring for 2h at room temperature in a mixed solution consisting of 50 percent of sodium hydroxide, urea and water to obtain a suspension, cooling in a liquid nitrogen bath for 30min, and then performing vacuum sublimation at 20 ℃ to obtain cellulose aerogel;
s2, adding 50g of zeolite powder into 500g of 36wt% hydrochloric acid, stirring for 3 hours, filtering, washing a filter cake with water until the pH value of the filtrate is neutral, drying at 80 ℃ for 10 hours to obtain acidified zeolite, mixing 5g of the acidified zeolite, 1g of cellulose aerogel obtained in the step S1 and 150mL of 99wt% ethanol aqueous solution, carrying out ultrasonic treatment for 2 hours under the conditions of 80Hz and 120W, filtering, and drying the filter cake at 80 ℃ for 8 hours to obtain a cellulose carrier;
s3, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing 2g of cellulose carrier and 10g of bacterial suspension, concentrating at 30 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain carrier material for loading bacterial;
s4, mixing 2g of polyethylene glycol, 10g of pullulan and 200g of water, heating to 60 ℃, stirring for 30min, and cooling to 40 ℃ to obtain a coating agent;
s5, uniformly mixing 5g of coating agent and 1g of carrier loaded with strains, adding the mixture into 150mL of 1mol/L calcium chloride aqueous solution, reacting for 24 hours at room temperature, filtering, and washing a filter cake with water for 3 times to obtain the biological membrane.
The culturing steps of the COD degrading bacteria are as follows:
inoculating Bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic Gordonia to LB liquid medium at 25deg.C at 4000 rpm for shake culture for 18 hr, and anaerobic fermenting at 30deg.C in fermenterFermenting for 30 hr, and culturing to obtain viable count of not less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing bacillus subtilis liquid, bacillus amyloliquefaciens liquid, lactobacillus plantarum liquid, enterococcus faecalis liquid and hydrophobic gordonia bacteria liquid according to the mass ratio of 2:2:1:1:2.
Comparative example 1
A method for preparing a biological membrane, comprising the steps of:
s1, 10g of microcrystalline cellulose is dispersed in 200g of microcrystalline cellulose with a mass ratio of 5:10: stirring for 2h at room temperature in a mixed solution consisting of 50 percent of sodium hydroxide, urea and water to obtain a suspension, cooling in a liquid nitrogen bath for 30min, and then performing vacuum sublimation at 20 ℃ to obtain cellulose aerogel;
s2, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing 2g of cellulose aerogel with 10g of bacterial suspension, concentrating at 30 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain a carrier for loading bacterial strains;
s3, mixing 2g of polyethylene glycol, 10g of pullulan and 200g of water, heating to 60 ℃, stirring for 30min, and cooling to 40 ℃ to obtain a coating agent;
s4, uniformly mixing 5g of coating agent and 1g of carrier loaded with strains, adding the mixture into 150mL of 1mol/L calcium chloride aqueous solution, reacting for 24 hours at room temperature, filtering, and washing a filter cake with water for 3 times to obtain the biological membrane.
The culturing steps of the COD degrading bacteria are as follows:
inoculating Bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic Gordonia to LB liquid culture medium according to inoculum size of 4wt% respectively, shake culturing at 25deg.C at 4000 rpm for 18 hr, anaerobic fermenting at 30deg.C for 30 hr in fermenter, and culturing to obtain viable bacteria with number no less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing bacillus subtilis liquid, bacillus amyloliquefaciens liquid, lactobacillus plantarum liquid, enterococcus faecalis liquid and hydrophobic gordonia bacteria liquid according to the mass ratio of 2:2:1:1:2.
Comparative example 2
A method for preparing a biological membrane, comprising the steps of:
s1, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing 2g of zeolite powder and 10g of bacterial suspension, concentrating at 30 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain a carrier for loading strains;
s2, mixing 2g of polyethylene glycol, 10g of pullulan and 200g of water, heating to 60 ℃, stirring for 30min, and cooling to 40 ℃ to obtain a coating agent;
s3, uniformly mixing 5g of coating agent and 1g of carrier loaded with strains, adding the mixture into 150mL of 1mol/L calcium chloride aqueous solution, reacting for 24 hours at room temperature, filtering, and washing a filter cake with water for 3 times to obtain the biological membrane.
The culturing steps of the COD degrading bacteria are as follows:
inoculating Bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic Gordonia to LB liquid culture medium according to inoculum size of 4wt% respectively, shake culturing at 25deg.C at 4000 rpm for 18 hr, anaerobic fermenting at 30deg.C for 30 hr in fermenter, and culturing to obtain viable bacteria with number no less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing bacillus subtilis liquid, bacillus amyloliquefaciens liquid, lactobacillus plantarum liquid, enterococcus faecalis liquid and hydrophobic gordonia bacteria liquid according to the mass ratio of 2:2:1:1:2.
Test example 1
Porosity test: the measurement was performed by the absolute ethanol displacement method. The modified cellulose carrier and the cellulose carrier prepared in the examples 1-2 are respectively soaked in absolute ethyl alcohol for sealing, the total family at the moment is recorded, the carrier is taken out after standing for 10min, the volume of the residual absolute ethyl alcohol is recorded, and the calculation formula of the porosity is as follows: epsilon= (V) A -V C )/(V B -V C )×100％
Epsilon-porosity (%); v (V) A -raw absolute ethanol volume (mL); v (V) B -total volume of absolute ethanol and carrier (mL); v (V) C Residual absolute ethanol volume (mL)
The test results are shown in table 1:
table 1 results of porosity test
| Porosity (%) | |
| Example 1 | 85.3 |
| Example 2 | 76.2 |
As can be seen from the experimental results in table 1, the cellulose carriers prepared in examples 1-2 all have a higher porosity, and the modified cellulose carrier prepared in example 1 has a higher porosity, whereas example 1 differs from example 2 in that the cellulose aerogel modified with bis (3-trimethoxysilylpropyl) amine is added, and this phenomenon may be caused by the modification of bis (3-trimethoxysilylpropyl) amine, improving the internal structure of cellulose, and the introduction of reactive groups allows the processability of cellulose to be improved, thereby increasing the porosity thereof.
Test example 2
Film hanging rate test: the test subjects were the biofilms prepared in examples 1-2 and comparative examples 1-2, and the test steps were: adopts artificial synthetic organic wastewater, glucose as a carbon source and NH 4 Cl and KH 2 PO 4 Providing a nitrogen source and a phosphorus source respectively, wherein the mass ratio of the carbon source to the nitrogen source to the phosphorus source is 100:5:1; the inoculated sludge in the film hanging experiment is obtained from a secondary sedimentation tank backflow sludge MLSS=4500 mg/L of a sewage treatment plant in Wuhan, the film hanging time is 48 hours, then the biological film is taken out and placed in a container, continuously aerated for 48 hours, then the biological film is taken out, dried to constant weight at 105 ℃, and then the film hanging rate is calculated; film formation rate= (hanging)Sum of membrane-after-biofilm mass-primary biofilm mass)/primary biofilm mass
The test results are shown in table 2:
TABLE 2 film formation test results
| Film formation percentage (%) | |
| Example 1 | 285 |
| Example 2 | 256 |
| Comparative example 1 | 234 |
| Comparative example 2 | 218 |
From the experimental data in table 2, the biofilm produced in example 1 has a biofilm formation rate up to 285 hours after 48 hours, which indicates that the biofilm produced in the invention has a fast biofilm formation rate and a large biofilm formation modulus, and the biofilm formation rate and the biofilm formation amount of the biofilm can be remarkably improved by modifying cellulose with bis (3-trimethoxysilylpropyl) amine and then introducing zeolite powder into aerogel.
Test example 3
Denitrification experiment: and 5 groups of biofilm reactors are operated in parallel, one group is a control group, no biofilm is added, and the other four groups are respectively added with the biofilms prepared in the examples 1-2 and the comparative examples 1-2. The running period of the reactor is 4 hours, the effective treatment water quantity in each period is 0.8L,adopting manual self-water distribution, wherein the nitrate nitrogen concentration is 50mg/L, the COD concentration is 150mg/L, a cooling water circulation device is arranged in the reactor, the internal temperature is kept at 10 ℃, inoculated sludge is obtained from a secondary sedimentation tank backflow sludge MLSS=4500 mg/L of a sewage treatment plant in Wuhan, and after the reactor is operated for 7 days, the water removal NO of the reactor is calculated 3 - The removal rate of medium nitrogen and the test results are shown in table 3:
TABLE 3 denitrification test results
| Denitrification Rate (%) | |
| Example 1 | 92 |
| Example 2 | 88 |
| Comparative example 1 | 83 |
| Comparative example 2 | 77 |
| Control group | 50 |
From the experimental results in table 3, it can be seen that the biofilm prepared in example 1 has the best denitrification effect, the cellulose is modified by bis (3-trimethoxysilylpropyl) amine, the internal structure of the cellulose is improved, the processing performance of the cellulose is improved by introducing active groups, the skeleton supporting effect can be achieved on the aerogel by fixing zeolite powder in the cellulose aerogel, the strength is improved, the micropore structure of the biological carrier can be regulated and controlled, the specific surface area is increased, the microbial load is increased, the adhesion, fixing and propagation of microorganisms on the carrier are accelerated, the rapid formation of the biological film is promoted, the slow release of microorganisms in the sewage environment can be realized by using the coating agent on the outer layer, the shearing effect of the water conservancy shearing effect on bacteria can be reduced, the impact resistance is good, and the biological film is not easy to fall off, so that the denitrification performance of the microorganisms is improved.
Claims (10)
1. A method for preparing a biological membrane, comprising the steps of: preparing a cellulose carrier; loading microorganisms with a cellulose carrier; preparing a coating agent; and coating the carrier loaded with the strain by using a coating agent.
2. The method for preparing a biological film according to claim 1, comprising the steps of:
s1, dispersing cellulose in a mixed solution composed of sodium hydroxide, urea and water, stirring to obtain a suspension, freezing and sublimating to obtain cellulose aerogel;
s2, mixing zeolite powder after acidification treatment with cellulose aerogel and ethanol solution, and obtaining a cellulose carrier after ultrasonic reaction, filtration and drying;
s3, culturing COD degrading bacteria, preparing bacterial suspension, mixing with a cellulose carrier, concentrating, and naturally air-drying to obtain a carrier for loading strains;
s4, preparing a coating agent by using polyethylene glycol and pullulan to react;
s5, mixing the coating agent and the carrier loaded with the strain, adding the mixture into a calcium chloride aqueous solution for reaction, filtering and washing to obtain the biomembrane.
3. The method for preparing a biological film according to claim 2, wherein the cellulose aerogel is modified cellulose aerogel, and the preparation method comprises the following steps:
m1, dispersing cellulose in a mixed solution composed of sodium hydroxide, urea and water, stirring to obtain a suspension, freezing and sublimating to obtain cellulose aerogel;
and adding bis (3-trimethoxysilylpropyl) amine into the suspension, stirring, freezing and sublimating to obtain the modified cellulose aerogel.
4. A method of preparing a biofilm according to claim 1 or 2 comprising the steps of: s1, microcrystalline cellulose is dispersed in a mixed solution composed of sodium hydroxide, urea and water, and stirred for 2-3 hours at room temperature to obtain a suspension;
s2, dropwise adding bis (3-trimethoxysilylpropyl) amine into the suspension obtained in the step S1 at a speed of 1-2 drops/second, stirring at room temperature for 10-20min after the dropwise adding, standing for 2-3h, transferring into a mold, freezing in a liquid nitrogen bath, and performing vacuum sublimation at a temperature of 10-40 ℃ to obtain modified cellulose aerogel;
s3, adding zeolite powder into 30-36wt% hydrochloric acid, stirring for 2-3 hours, filtering, washing a filter cake with water until the pH value of the filtrate is neutral, drying at 60-80 ℃ for 8-10 hours to obtain acidified zeolite, mixing the acidified zeolite, the modified cellulose aerogel obtained in the step S2 and 75-95wt% ethanol aqueous solution, carrying out ultrasonic treatment at 60-100Hz and 120-150W for 1-2 hours, filtering, and drying the filter cake at 60-80 ℃ for 6-8 hours to obtain a modified cellulose carrier;
s4, culturing COD degrading bacteria respectively, centrifugally collecting thalli, mixing to obtain bacterial suspension, uniformly mixing a modified cellulose carrier and the bacterial suspension, concentrating at 20-40 ℃ until the solid content is more than or equal to 80%, and naturally airing at room temperature to obtain a carrier carrying strains;
s5, mixing polyethylene glycol, pullulan and water, heating to 50-70 ℃, stirring for 20-30min, and cooling to 40-50 ℃ to obtain a coating agent;
s6, uniformly mixing the coating agent and the carrier loaded with the strain, adding the mixture into 1-2mol/L calcium chloride aqueous solution, reacting for 20-24 hours, filtering, and washing a filter cake with water for 2-3 times to obtain the biological membrane.
5. The method for preparing the biological film according to claim 4, wherein: in the step S1, the mass ratio of the sodium hydroxide to the urea to the water is 5-8:10-15:50-100.
6. The method for preparing the biological film according to claim 4, wherein: the mass ratio of the suspension to the bis (3-trimethoxysilylpropyl) amine in the step S2 is 5-10:1.
7. The method for preparing the biological film according to claim 4, wherein: in the step S3, the mass ratio of the zeolite powder to the hydrochloric acid is 1:10-15; the dosage ratio of the acidified zeolite, the modified cellulose aerogel and the ethanol water solution is 5-10g to 1g to 100-200mL.
8. The method for preparing the biological film according to claim 4, wherein: the culturing step of the COD degrading bacteria in the step S3 is as follows: inoculating Bacillus subtilis, bacillus amyloliquefaciens, lactobacillus plantarum, enterococcus faecalis and hydrophobic Gordonia to LB liquid culture medium according to 2-5wt% of inoculation amount, shake culturing at 25-30deg.C for 15-20 hr, anaerobic fermenting at 25-30deg.C for 25-30 hr in a fermenter, and culturing to obtain viable bacteria with number no less than 10 8 CFU/g, and then collecting bacterial liquid of each bacterial; and then mixing the bacillus subtilis liquid, the bacillus amyloliquefaciens liquid, the lactobacillus plantarum liquid, the enterococcus faecalis liquid and the hydrophobic Gordonia liquid according to the mass ratio of 1-2:1-2:1:1-2.
9. A biofilm, characterized in that: prepared by the method for preparing a biological film according to any one of claims 1 to 8.
10. Use of a biofilm according to claim 9, wherein: the method is applied to a biological membrane reactor for sewage treatment.
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| CN116530450A (en) * | 2023-06-16 | 2023-08-04 | 江苏中水东泽农业发展股份有限公司 | Mixed cultivation method for rice and shrimps |
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