CN109370957B - Sewage treatment composite microbial inoculum and preparation method thereof - Google Patents

Sewage treatment composite microbial inoculum and preparation method thereof Download PDF

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CN109370957B
CN109370957B CN201811504718.9A CN201811504718A CN109370957B CN 109370957 B CN109370957 B CN 109370957B CN 201811504718 A CN201811504718 A CN 201811504718A CN 109370957 B CN109370957 B CN 109370957B
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詹旭
陈卓
王婧希
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WUXI GONGYUAN ENVIRONMENTAL TECHNOLOGY Co.,Ltd.
Jiangnan University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
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    • C12N1/16—Yeasts; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/30—Organic compounds
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    • C02F2101/345—Phenols

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Abstract

The invention discloses a sewage treatment composite microbial inoculum and a preparation method thereof, belonging to the field of sewage treatment. The microbial agent comprises the following components in parts by weight: 20-30 parts of pseudomonas, 15-20 parts of denitrifying bacteria, 5-10 parts of nitrobacteria and 5-10 parts of pichia pastoris. The microbial agent can effectively improve the removal efficiency of pollutants in the treated sewage, and the removal rates of COD, total nitrogen, total phosphorus and ammonia nitrogen of the treated effluent of the polluted river can respectively reach more than 90%, 80%, more than 85% and more than 85%, so that the microbial agent meets the pollutant discharge standard of a sewage treatment plant. Meanwhile, the microbial agent can rapidly and efficiently degrade aromatic compounds, more than 85% of the aromatic compounds can be degraded in 2-3 days, the aromatic compounds with higher concentration do not inhibit the growth and degradation of bacteria in the microbial agent, and the microbial agent has a good application prospect.

Description

Sewage treatment composite microbial inoculum and preparation method thereof
Technical Field
The invention particularly relates to a sewage treatment composite microbial inoculum and a preparation method thereof, belonging to the field of sewage treatment.
Background
The sewage mainly comprises domestic sewage and industrial wastewater. The components of industrial sewage are relatively complex, particularly a large amount of artificially synthesized compounds enter the environment, the substances mainly comprise ammonia nitrogen, sulfide and phosphorus-containing compounds, and the substances cannot be decomposed and utilized by microorganisms in a short time due to the complexity of the structures of the substances, so that the microorganisms cultured and domesticated by the activated sludge in the traditional wastewater treatment method cannot effectively remove the pollutants, the substances are accumulated in the environment for a long time, great pollution is caused to the ecological environment which depends on the survival of people, and great harm is brought to the physical and mental health of human beings. In addition, with the acceleration of the social industrialization process, a large amount of organic pollutants which are difficult to degrade are discharged into the environment, such as polycyclic aromatic hydrocarbons, halogenated compounds, polychlorinated biphenyls, organic nitrogen and phosphorus pesticides, and the like. Most of these organic pollutants are artificially synthesized, and thus the kinds and the number of microorganisms having a high degradation function in natural environments are small, and are at a disadvantage in interspecies competition. Efficient degradation of these organic materials is difficult to achieve using conventional biological treatment methods.
The microbial agent is prepared by combining or mixing and culturing a plurality of microorganisms with different degradation functions and mutual or symbiotic relations according to a proper proportion. The method improves the treatment efficiency of the organic pollutants difficult to degrade by adding the functional microbial agent into the wastewater treatment system, and is a common biological strengthening technology at present. However, the existing microbial agents have strong functional bias, for example, bacillus microbial agents can well reduce the COD content in sewage, or klebsiella microbial agents can reduce the phosphorus content in sewage. Therefore, the construction of a simple and green microbial agent with better removal performance on ammonia nitrogen, total phosphorus, COD and other organic pollutants is urgently needed.
Disclosure of Invention
In order to solve the problems, the invention provides a microbial agent which has good ammonia nitrogen, total phosphorus and COD removal effects and can well degrade aromatic compounds. Has the characteristics of strong pertinence, quick response, simple and convenient operation and the like, and has better application prospect in the treatment of organic environmental pollution.
The first purpose of the invention is to provide a microbial agent, which comprises the following components in parts by weight: 20-30 parts of pseudomonas, 15-35 parts of halomonas and 5-10 parts of pichia pastoris.
In one embodiment of the present invention, the microbial agent comprises, in parts by weight: 20 parts of pseudomonas, 16 parts of denitrifying halomonas, 8 parts of halomonas handii and 8 parts of pichia pastoris.
In one embodiment of the invention, the Pseudomonas comprises one or more of Pseudomonas aeruginosa (Pseudomonas aeruginosa), Pseudomonas brucei (Pseudomonas brenneri), Pseudomonas putida (Pseudomonas putida), and Pseudomonas stutzeri.
In one embodiment of the invention, the halomonas comprises one or more of halomonas denitrificans (halomonas dentificans), halomonas handii (halomonas hamiltonii), halomonas fandii (halomonas ventosae).
In one embodiment of the invention, the Pichia pastoris is preferably Pichia membranaefaciens (Pichia membranaceus).
In one embodiment of the present invention, the preparation method of the microbial agent comprises:
mixing the pseudomonas, the denitrifying bacteria, the nitrobacteria and the pichia pastoris according to the weight part ratio to obtain the microbial agent.
The second purpose of the invention is to provide a sewage treatment method, which utilizes the microbial agent.
In one embodiment of the present invention, the amount of microbial agent added in the method is not less than 0.2%.
In one embodiment of the present invention, the method may further comprise enriching the microbial agent on a carrier, wherein the carrier is a spongy cubic carrier ACP or PM.
In one embodiment of the invention, the content of the bacterial species relative to the carrier is not less than 10. mu.g/g.
In one embodiment of the invention, the content of the strain relative to the carrier is preferably 50-150 μ g/g.
In one embodiment of the present invention, the inside of the carrier is a staggered mesh structure, and the volume of each carrier is 1dm3。
The third purpose of the invention is to provide a method for degrading aromatic compounds, which utilizes the microbial agent.
In one embodiment of the present invention, the amount of microbial agent added in the method is not less than 0.2%.
In one embodiment of the present invention, the method may further comprise enriching the microbial agent on a carrier, wherein the carrier is a spongy cubic carrier ACP or PM.
In one embodiment of the invention, the content of the bacterial species relative to the carrier is not less than 10. mu.g/g.
In one embodiment of the invention, the content of the strain relative to the carrier is preferably 50-150 μ g/g.
In one embodiment of the present invention, the inside of the carrier is a staggered mesh structure, and the volume of each carrier is 1dm3。
The invention has the beneficial effects that:
1. the microbial agent can effectively improve the removal efficiency of pollutants in the treated sewage, and the removal rates of COD, total nitrogen, total phosphorus and ammonia nitrogen of the treated effluent of the polluted river can respectively reach more than 90%, 80%, more than 85% and more than 85%, so that the microbial agent meets the pollutant discharge standard of a sewage treatment plant.
2. The microbial agent can rapidly and efficiently degrade aromatic compounds, can degrade more than 85% of aromatic compounds within 2-3 days, and particularly can completely degrade pentachlorophenol, and aromatic compounds with higher concentration do not inhibit the growth and degradation of bacteria in the microbial agent, so the microbial agent has good application prospect.
Detailed Description
The sewage of the invention is taken from river ecological sewage of a certain district in the tin-free city: pH 6.53, CODMn54.61mg/L, TN mass concentration of 35.15mg/L, TP mass concentration of 3.14mg/L, NH4 +The mass concentration of-N is 31.58 mg/L.
Example 1:
preparing a microbial agent: performing slant culture on pseudomonas aeruginosa in a nutrient broth agar culture medium, and concentrating and drying the obtained fermentation liquor to obtain spore powder; culturing in soybean agar culture medium of denitrifying halomonas and Halomonas handii tryptone, and concentrating and drying the obtained fermentation liquor to obtain spore powder; culturing Pichia membranaefaciens in a wort agar culture medium, and concentrating and drying the obtained fermentation liquor to obtain spore powder;
20 parts by weight of pseudomonas aeruginosa CICC 10351 spore powder, 16 parts by weight of denitrifying halomonas CICC 10554 spore powder, 8 parts by weight of halomonas handii CICC23885 spore powder and 8 parts by weight of pichia membranaefaciens CICC 33242 spore powder are mixed to prepare the compound microbial inoculum.
Taking 1000mL of sewage sample, respectively adding 0.2%, 0.25%, 0.3%, 0.5% and 1% of microbial agent by mass for water quality degradation experiment, culturing at 30 deg.C for 72h, and determining CODMn、TN、TP、NH4 +The results of the removal of N-species are shown in Table 1.
TABLE 1 COD of different amounts of microbial inoculumMn、TN、TP、NH4 +-N removal effect
Adding amount of microbial inoculum CODMn(mg/L) TN(mg/L) TP(mg/L) NH4 +-N(mg/L)
0 (not added) 54.61 35.15 3.14 31.58
0.2％ 5.03 7.01 0.40 4.55
0.25％ 4.65 5.88 0.32 4.03
0.3％ 4.19 4.73 0.25 3.85
0.5％ 3.37 4.11 0.095 2.45
1％ 1.85 3.77 0.092 1.98
The detection method comprises the following steps: CODMn is measured by an acid permanganate oxidation method (GB 11892-1989); TN is measured by alkaline potassium persulfate ultraviolet spectrophotometry (GB 11894-89); NH4+ -N is measured by adopting a Nashiner reagent colorimetric method (GB 7479-87); TP was determined by potassium persulfate oxidation-molybdenum antimony anti-spectrophotometry (GB 11893-89).
As can be seen from Table 1, the microbial inoculum can realize more than 90% of COD with less consumptionMnRemoval rate, TN removal rate of more than 80%, TP removal rate of more than 85%, NH of more than 85%4 +-N removal rate, meeting the pollutant discharge standard of sewage treatment plant。
Example 2:
preparing a microbial agent according to the formula shown in example 1, and enriching 0.2% of the microbial agent in a 20mg spongy cubic carrier ACP film at room temperature for 24 hours; taking 1000mL of sewage water sample, adding the enriched carrier ACP membrane into the sewage to carry out water quality degradation experiment, culturing at the temperature of 30 ℃, respectively culturing for 72h, and determining CODMn、TN、TP、NH4 +The removal rate of-N substance is 93.0%, 87.5%, 92.3% and 90.8% respectively.
Example 3:
culture medium: na (Na)2HPO4·12H2O 9.0g，KH2PO43.0g，NH4NO31.0g，MgSO4·7H2O0.3g，FeCl30.05g，CaCl20.02g, 2.5ml of trace element solution and 1000ml of distilled water; wherein, the composition of the microelement solution is as follows: MnSO4·2H2O 0.06g，H3BO30.03g，CoCl2·6H2O 0.04g，CuCl2·2H2O 0.01g，NiCl2·6H2O 0.02g，Na2MoO4·2H2O 0.03g，ZnCl20.05g, pH7, 1000ml of distilled water.
Sample bottle 1: adding 0.1mL of acetone solution of 2mg/mL nitrophenol into a serum bottle in advance, volatilizing the acetone, adding 20mL of culture medium, and setting the final concentration of the nitrophenol in the culture medium to be 10 mg/L;
sample bottle 2: adding 0.1mL of acetone solution of 2mg/mL1, 4-dichlorophenol in a serum bottle in advance, volatilizing the acetone, adding 20mL of culture medium, and setting the final concentration of 1, 4-dichlorophenol in the culture medium to be 10 mg/L;
sample bottle 3: adding 0.1mL of acetone solution of 2mg/mL pentachlorophenol into a serum bottle in advance, volatilizing the acetone, adding 20mL of culture medium, and controlling the final concentration of the pentachlorophenol in the culture medium to be 10 mg/L;
sample bottle 4: 0.1mL of acetone solution of 2mg/mL pentachlorophenol, 0.1mL of acetone solution of 2mg/mL nitrophenol and 0.1mL of acetone solution of 2mg/mL1, 4-dichlorophenol were previously added to a serum bottle, and acetone was volatilized, and 20mL of a medium in which the total aromatic substance concentration was 30mg/L was added.
The microbial inoculum was prepared according to the formulation shown in example 1, inoculated at 0.3% inoculum size into sample flasks 1, 2, 3, 4, shake-cultured at 30 ℃ in the dark, sampled on days 1, 2, 3 and 5, and HPLC-MS determined the residual aromatic content of the broth as shown in Table 2.
TABLE 2 organic degradation Effect of microbial Agents
Figure BDA0001899153790000041
As can be seen from Table 2, since the culture solution contains no other carbon source, the growth of the carbon source for the growth of the bacteria comes from the aromatic hydrocarbon organic pollutants, i.e., the bacteria can rapidly degrade the aromatic hydrocarbon organic pollutants, and simultaneously, the energy and the carbon source generated during degradation are used for the growth and propagation of the bacteria. The microbial agent disclosed by the invention can realize better degradation of p-nitrophenol, 1, 4-dichlorophenol and pentachlorophenol within 1d, wherein the degradation rates respectively reach 75.4%, 76.0% and 78.8%, the degradation efficiency of 2-3d p-pentachlorophenol can basically reach 98%, the degradation rate of 1, 4-dichlorophenol is less than that of 1, 4-dichlorophenol, the degradation rate can reach 90%, and the degradation rate of nitrophenol can also reach more than 85%. And when the nitrophenol, the 1, 4-dichlorophenol and the pentachlorophenol exist at the same time and have higher concentration, the microbial agent can better degrade the aromatic hydrocarbon organic pollutants and is not inhibited by the concentration.
Furthermore, the inventors have also made the following experiments: replacing the inoculation amount of the microbial inoculum by 0.2%, 0.5% and 1%, and finding that when the inoculation amount is 0.2%, the 3d degradation rate is basically 75% and the 5d degradation rate can reach 85%; the degradation rate 2d of the three organic pollutants by 0.5 percent and 1 percent of addition amount basically reaches more than 90 percent.
Comparative example:
the formula of the microbial agent is replaced by a microbial agent 1, a microbial agent 2, a microbial agent 3 and a microbial agent 4, wherein the microbial agent 1, 2, 3 and 4 comprises the following components:
1, microbial inoculum: the pseudomonas is replaced by bacillus cereus CICC 21155, and other conditions are kept unchanged according to the preparation method of the microbial inoculum in the example 1;
and (2) microbial inoculum: the pichia pastoris is replaced by the beer yeast, and other conditions are kept unchanged according to the preparation method of the microbial inoculum in the embodiment 1; (Saccharomyces cerevisiae ACCC21144 refers to the literature Daoyouzhi, Schoanba. adsorption study of Saccharomyces cerevisiae to Cr (VI) in water [ J ] Nature science of Hunan Tan university, 2007,29(3),79-83.)
And (3) microbial inoculum: the method for preparing the microbial inoculum in the embodiment 1 is not added with the pichia pastoris, and other conditions are kept unchanged;
and (4) microbial inoculum: 10 parts of pseudomonas, 15 parts of denitrifying halomonas CICC 10554 spore powder, 10 parts of halomonas handii CICC23885 spore powder and 10 parts of pichia pastoris are mixed to obtain the microbial inoculum.
The sewage treatment was carried out in accordance with the method of example 1, and the test results of the treated sewage are shown in Table 3.
TABLE 3 index results of treated wastewater (0.2% dosage)
Microbial inoculum CODMn(mg/L) TN(mg/L) TP(mg/L) NH4 +-N(mg/L)
0 (not added) 54.61 35.15 3.14 31.58
Bacterial agent 1 35.30 24.31 2.55 24.41
Bacterial preparation 2 33.15 25.80 2.84 23.28
Bacterial agent 3 39.89 29.55 2.01 26.33
Bacterial agent 4 20.02 18.32 1.54 15.11
Aromatic hydrocarbon organic contaminants were treated according to the method of example 3, and the content of aromatic hydrocarbon organic contaminants in the treated medium is shown in Table 4.
TABLE 4 content of aromatic organic contaminants in the treated culture Medium
Figure BDA0001899153790000051
Figure BDA0001899153790000061
As can be seen from tables 3 and 4, the microbial agent provided by the invention has the advantages that various strains interact with each other, so that the microbial agent can be well fermented and symbiotic; according to the microbial inoculum 1 and 2, the bacillus cereus or the beer yeast has poor symbiotic effect with other strains in the microbial inoculum system, the corresponding decontamination effect and organic matter degradation capability are poor, in addition, the proportion of pseudomonas in the microbial inoculum is also important, and the decontamination effect and the organic matter degradation capability are obviously reduced due to the lower consumption.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (6)

1. The microbial agent is characterized by comprising the following components in parts by weight: 20 parts of pseudomonas aeruginosa CICC 10351, 8 parts of Halomonas handii CICC23885, 16 parts of Halomonas denitrificans CICC 10554 and 8 parts of Pichia membranaefaciens CICC 33242.
2. A method for treating sewage, which comprises treating sewage with the microbial agent according to claim 1.
3. The method according to claim 2, wherein the amount of microbial agent added in the method is not less than 0.2%.
4. The method according to claim 2 or 3, further comprising enriching the microbial agent on a carrier, wherein the carrier is a spongy cubic carrier ACP or PM.
5. The method according to claim 4, wherein the content of the microbial agent relative to the carrier is not less than 10 μ g/g.
6. A degradation method of aromatic compounds, which is characterized in that the method utilizes the microbial agent as claimed in claim 1 to carry out degradation, wherein the aromatic compounds are nitrophenol, 1, 4-dichlorophenol and pentachlorophenol.
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