CN113416646A - Screening method of microbial flora for treating chemical wastewater - Google Patents
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- 239000002351 wastewater Substances 0.000 title claims abstract description 96
- 239000000126 substance Substances 0.000 title claims abstract description 93
- 230000000813 microbial effect Effects 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 52
- 238000012216 screening Methods 0.000 title claims abstract description 35
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- 241001052560 Thallis Species 0.000 claims abstract description 14
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- 238000011049 filling Methods 0.000 claims description 12
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 12
- 230000003068 static effect Effects 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 9
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000000855 fermentation Methods 0.000 claims description 8
- 230000004151 fermentation Effects 0.000 claims description 8
- 230000035755 proliferation Effects 0.000 claims description 8
- 238000009423 ventilation Methods 0.000 claims description 8
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- 238000012136 culture method Methods 0.000 claims description 6
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- 229930006000 Sucrose Natural products 0.000 claims description 4
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- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 4
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 4
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- 241000894006 Bacteria Species 0.000 description 1
- FMMWHPNWAFZXNH-UHFFFAOYSA-N Benz[a]pyrene Chemical compound C1=C2C3=CC=CC=C3C=C(C=C3)C2=C2C3=CC=CC2=C1 FMMWHPNWAFZXNH-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
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- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- 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
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- 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
- C12N1/02—Separating microorganisms from their culture media
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- 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
- C12N1/36—Adaptation or attenuation of cells
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Abstract
The invention discloses a screening method of microbial flora for treating chemical wastewater, belongs to the field of coal chemical wastewater treatment, and aims to solve the problems that the existing microbial flora screening process is complex, the screening efficiency is low, and the coal chemical wastewater treatment effect is poor without a contrast experiment; according to the screening method, the thalli are obtained by performing value-added culture on the coal chemical industry wastewater mixture, and then three times of domestication are continuously performed, so that the screening of microbial floras for treating the coal chemical industry wastewater is realized, the materials are more convenient to obtain, the screening process is simpler, the screening efficiency is higher, the effect of treating the coal chemical industry wastewater is better and more practical, the domestication process utilizes the coal chemical industry wastewater with different concentrations to perform a contrast experiment, the microbial floras with the best effect of degrading the coal chemical industry wastewater under the condition of specific concentration can be obtained, and the aerobic floras and the anaerobic floras with the best degradation effect can be screened out.
Description
Technical Field
The invention relates to the technical field of coal chemical industry wastewater treatment methods, in particular to a method for screening microbial floras for treating coal chemical industry wastewater.
Background
The coal chemical wastewater is industrial wastewater generated in a process flow of processing and producing by using coal as a raw material in large-scale enterprises. The coal chemical wastewater mainly comprises three major types: (1) coking wastewater, (2) secondary gasification wastewater, and (3) liquefied wastewater. Coking wastewater is one of three types of wastewater in coal chemical industry, and refers to ammonia water residue formed after coal is subjected to high-temperature dry distillation coking operation treatment and coal gas is cooled, wastewater containing pollutants such as phenol and cyanogen discharged in a coal gas purification process after cold circulating water is replaced, wastewater generated in processing production of refining tar and a product are recycled, and the wastewater is obtained without using chemical raw materials for production. The gasification wastewater refers to that coal is cracked through chemical reaction in a high-temperature environment in a coal gas purification process to generate a plurality of different gas fuels with high combustion values, water vapor in the gasification process is condensed and then discharged into condensed water of a spray cooling system and wastewater for washing the coal gas, and the wastewater mostly contains pollutants such as phenol, oil, ammonia nitrogen and the like. The liquefaction waste water is waste water generated by a liquefaction process or other uses when coal is used, and the liquefaction process is mainly divided into two main types, namely direct liquefaction and indirect liquefaction. The direct liquefaction is a process of cracking organic matters with high molecules with complex structures in coal under the conditions of high temperature and high pressure and adding hydrogenation into raw coal to form a plurality of liquid hydrocarbons with low molecular weights, and in the operation, waste water is generated, wherein the large amount of waste water contains ammonia nitrogen and sulfide. The indirect liquefaction is a complex process of adding a catalyst into a product synthesized by coal gasification and synthesizing a liquid oil product under a specific condition, and wastewater is generated in the process, wherein the wastewater mainly contains organic pollutants such as acid, alcohol, ketone and the like. The coal chemical industry wastewater has the following characteristics: (1) the components are complex, various pollutants are contained, and the concentration is not low; however, in the production of the coal chemical industry, the produced wastewater is not little, the water quality is relatively complex, the wastewater contains a large amount of solid suspended particles and toxic and non-degradable pollutants, such as various phenols, cyanides, thiocyanides, benzopyrene, biphenyl and the like, such as ammonia nitrogen, sulfide products and the like, the COD and the chromaticity of the industrial wastewater are high, and the treatment difficulty is low; (2) the harm is large, the biochemical treatment performance is poor: the products in the coal chemical industry wastewater are toxic, harmful and difficult to degrade. The coal chemical industry wastewater contains more organic compounds, the occupied proportion of the organic compounds in the coal chemical industry wastewater is not low, the biodegradation is slow, the biological degradation is beyond the bearing range of microorganisms, the microorganisms can not avoid poisoning, the survival rate is reduced, and the wastewater has poor plasticity. At present, a new treatment method is found for coal chemical industry wastewater, activated sludge is used for treatment, but the types of microorganisms in the activated sludge are many, the treatment effect on the coal chemical industry wastewater is poor, and a few methods adopting strain screening are adopted to obtain the dominant flora for the coal chemical industry treatment so as to improve the treatment effect on the activated sludge. Most of the existing screening methods for screening and degrading a single certain substance strain in the coal chemical industry wastewater, such as using high-efficiency phenol degrading bacteria to treat the coal chemical industry wastewater and other various strains, although the method has strong pertinence, because degradation products in the screening are single, the resistance of various microbial strains to other chemical substances in the coal chemical industry wastewater in practical application is poor or intermediate products in the degradation process are excessively accumulated so as to inhibit the degradation process, the effect and the function which the microbial strains should show cannot be exerted, and the effect of treating the coal chemical industry wastewater is poor; the other screening method is to select a specific strain and construct the specific strain into a mature flora, so as to realize the manual control of the microbial flora, but because the adding proportion of each strain in the actual operation is difficult to determine, the method is difficult to implement, some sludge is not even increased in the treatment engineering of the coal chemical industry wastewater, the process is complex by adopting the method, the screening efficiency is low without comparison experiments, and the treatment effect of the microbial flora obtained by screening on the coal chemical industry wastewater is poor.
Disclosure of Invention
In light of the problems set forth above in the prior art, the present invention discloses a processThe screening method of microbial flora of coal chemical industry wastewater adopts the technical scheme that the screening method comprises the following steps: (1) preparing and developing a plurality of microorganism flora proliferation culture medium dishes, adjusting the pH value of the proliferation culture medium to be close to neutral, respectively filling the proliferation culture medium into 300mL conical flasks, and filling 150mL of proliferation culture medium into each flask when aerobic flora is proliferated; when the anaerobic flora is proliferated, 250mL of the culture medium is filled in each bottle; inoculating 5-10% of coal chemical wastewater or 2-5% of corresponding sludge into a microbial flora enrichment medium, culturing aerobic flora by a shaking culture method under a certain temperature condition, such as 32-36 ℃, with the rotating speed of a shaking table of 100-160 r/min, aerating for 13-15 hours, and then allowing the aerobic flora to exist for 8-12 hours in a specific environment; performing static culture on the anaerobic flora, shaking regularly for 22-26 hours, and centrifuging the culture solution under the condition of 10000r/15min to obtain thalli for later use; (2) adding 0.4-0.6 g of MgSO4 & 7H into each liter of the thallus treated in the step (1)2O, 0.05-0.15 g of FeSO4 & 7H2O, 0.005-0.015 g of ZnSO4 & 7H2O, 0.005-0.015 g of MnSO4·H2O, 0.005-0.015 g of CuSO4·5H2O, 0.2-0.3 g NH4Cl and 0.05-0.15 g of CaCl2And 2-4 g of cane sugar, uniformly stirring, aerating aerobic flora for 14-16 hours at the temperature of 32-36 ℃, standing for 8-10 hours, and removing upper-layer liquid to obtain settled sludge; performing static culture on anaerobic flora, shaking regularly, and removing upper-layer liquid after 22-26 hours to obtain settled sludge; (3) placing the settled sludge in coal chemical industry wastewater diluent with the volume 3-5 times of the total volume of the settled sludge for acclimatization, preparing a plurality of acclimatization screening culture mediums by taking the coal chemical industry wastewater with the concentration from low to high as a carbon source, standing for 5-6 hours, and removing supernatant to obtain first generation acclimatized settled sludge; (4) placing the first generation of domesticated settled sludge into coal chemical industry wastewater diluent with the volume of 3-5 times of the total volume of the settled sludge for domestication, standing for 5-6 hours, and removing supernatant to obtain second generation of domesticated settled sludge; (5) placing the second generation domesticated settled sludge into coal chemical industry wastewater diluent with the volume of 3-5 times of the total volume of the settled sludge for domestication, and taking out the domesticated settled sludge as the microbial biomass of the settled sludgeReaching 3800-4200 mg/mL to obtain the required microbial flora; (6) adjusting the C/N ratio of microbial flora according to the characteristics of the coal chemical industry wastewater, properly adding inorganic salt and the like, respectively adding the inorganic salt and the like into conical flasks of the coal chemical industry wastewater with different concentrations, and inoculating the thalli collected in the step (1); the culture method of the aerobic flora and the anaerobic flora is different and is the same as the step (1); the domestication method can gradually increase the concentration of the coal chemical wastewater from low to high according to actual production requirements, the culture time is 70-75 hours, and finally, the domesticated microbial flora is obtained by centrifugal collection under the condition of 10000r/15 min; (7) preservation of flora: storing the thallus obtained in the step (6) in a culture solution at 4 ℃ for later use; (8) a culture method for expanding required flora, and the operation: taking the thalli obtained in the step (6) as primary seeds, and sequentially adding the primary seeds to a fermentation tank by using a seed tank for amplification culture; the fermentation tank culture adopts an industrialized culture medium, the inoculation amount is 5-10%, the liquid filling rate is 70-75%, the aerobic flora is cultured by ventilation stirring at the temperature of 30-32 ℃, and the ventilation ratio is as follows: 0.3 to 1; performing static culture on the anaerobic flora, shaking and stirring regularly, wherein the culture time is about 20-26 hours; (9) preparing a microbial agent: adding certain substances into the flora after the enlarged culture in the step (8), and uniformly mixing to prepare the microbial inoculum.
Further, the medium used in step (1) has the following components: 8-12 g/L of peptone, 4-6 g/L of yeast powder and 8-12 g/L of sodium chloride.
Further, the pH value of the microbial flora multiplication culture medium in the step (1) is 7.0-7.2, and the culture medium is not subjected to sterilization treatment.
Further, COD of the coal chemical industry wastewater in the steps (3), (4) and (5) is 900-1010 mg/L, phenol concentration is 800-900 mg/L, ammonia nitrogen concentration is 44-46 mg/L, and pH value is 6.9-7.1.
Further, the acclimatization conditions in the steps (3), (4) and (5) are all carried out at the temperature of 32-36 ℃.
Further, in the steps (3), (4) and (5), the aerobic flora is firstly subjected to anoxic stirring for 3-4 hours, and then is subjected to aeration culture for 14-16 hours; anaerobic flora is subjected to anoxic static culture, shaking is performed at regular time, and the culture time is about 22-26 hours.
Further, the regulating substance of the C/N ratio in the step (6) refers to a nitrogen-containing nutrient, the inorganic salt comprises phosphorus, potassium, magnesium, calcium, iron, sulfur, copper, zinc, cobalt, manganese, molybdenum and the like, and the culture medium is not sterilized.
Further, the microbial flora in the step (7) is preserved by adopting liquid in a low-temperature environment.
The invention has the beneficial effects that: according to the screening method, the thalli are obtained by performing value-added culture on the coal chemical industry wastewater mixture, and then three times of domestication are continuously performed, so that the screening of microbial floras for treating the coal chemical industry wastewater is realized, the materials are more convenient to obtain, the screening process is simpler, the screening efficiency is higher, the effect of treating the coal chemical industry wastewater is better and more practical, the domestication process utilizes the coal chemical industry wastewater with different concentrations to perform a contrast experiment, the microbial floras with the best effect of degrading the coal chemical industry wastewater under the condition of specific concentration can be obtained, and the aerobic floras and the anaerobic floras with the best degradation effect can be screened out.
Detailed Description
Example 1
The invention discloses a method for screening microbial flora for treating coal chemical wastewater, which adopts the technical scheme that the method comprises the following steps: (1) preparing a plurality of microbial flora enrichment culture media, controlling the pH value of the enrichment culture media to be about 7.0, filling the enrichment culture media into 300mL conical flasks after the preparation is finished, enriching aerobic floras, filling 150mL of culture media into each flask, inoculating 8% of coal chemical industry wastewater into the microbial flora enrichment culture media, adopting shaking culture at the temperature of 34 ℃, ensuring the rotating speed of a shaking table to be 130r/min, aerating for 14 hours, standing for 10 hours, and centrifuging a culture solution under the condition of 10000r/15min to obtain thalli for later use; (2) 0.5g of MgSO4 & 7H was added to each liter of the cells treated in step (1)2O, 0.1g of FeSO4·7H2O, 0.01g of ZnSO4·7H2O, 0.01g of MnSO4·H2O, 0.01g of CuSO4·5H2O, 0.25g NH4Cl, 0.1g CaCl2And 3g of sucrose, then uniformly stirring, aerating for 14 hours at the temperature of 34 ℃, finding a stable position, standing for 9 hours, and removing supernatant liquid to obtain settled sludge; (3) placing the settled sludge in coal chemical industry wastewater diluent which accounts for 4 times of the total volume of the settled sludge for acclimatization, standing for 5.5 hours, and removing supernatant to obtain first generation of acclimatized settled sludge; (4) placing the first generation of domesticated settled sludge into coal chemical industry wastewater diluent with the volume 4 times of the total volume of the settled sludge for domestication, standing for 5.5 hours, and removing supernatant to obtain second generation of domesticated settled sludge; (5) placing the second generation of domesticated settled sludge into coal chemical industry wastewater diluent with the total volume of the settled sludge being 4 times that of the settled sludge for domestication, and obtaining screened microbial flora when the biomass of the settled sludge reaches 4000 mg/mL; (6) adjusting the C/N ratio of microbial flora according to the characteristics of the coal chemical industry wastewater, properly adding inorganic salt and the like, respectively adding the inorganic salt and the like into conical flasks of the coal chemical industry wastewater with different concentrations, and inoculating the thalli collected in the step (1); the culture method is the same as the step (1); the domestication method can gradually increase the concentration of the coal chemical wastewater from low to high, the culture time is 70-75 hours, and finally, the coal chemical wastewater is centrifugally collected under the condition of 10000r/15min to obtain a domesticated microbial community; (7) preservation of flora: storing the thallus obtained in the step (6) in a culture solution at 4 ℃ for later use; (8) culture for expanding the flora used: the primary seeds are cultured in a seed tank to a fermentation tank in a step-by-step amplification manner by using the thalli obtained in the step (6); the fermentation tank culture adopts an industrialized culture medium, the inoculation amount is 7%, the liquid filling rate is 73%, the aerobic flora is cultured by ventilation stirring under the condition of 31 ℃, and the ventilation ratio is as follows: 0.6, the culture time is 23 hours; (9) preparing a microbial agent: adding certain substances into the flora after the enlarged culture in the step (8), and uniformly mixing to prepare the microbial inoculum.
Further, the components of the microorganism flora multiplication culture medium in the step (1) comprise 10g/L of peptone, 5g/L of yeast powder and 10g/L of sodium chloride.
Further, the pH of the microorganism flora multiplication culture medium in the step (1) is 7.0, and the culture medium is not sterilized.
Further, the COD of the coal chemical industry wastewater in the steps (3), (4) and (5) is 950mg/L, the phenol concentration is 850mg/L, the ammonia nitrogen concentration is 45mg/L, and the pH value is 7.0.
Further, the acclimatization conditions in the steps (3), (4) and (5) are all performed at 34 ℃.
Further, in the steps (3), (4) and (5), the aerobic flora is firstly subjected to anoxic stirring for 3.5 hours, and then is subjected to aeration culture for 15 hours.
Further, the regulating substance of the C/N ratio in the step (6) refers to a nitrogen-containing nutrient, the inorganic salt comprises phosphorus, potassium, magnesium, calcium, iron, sulfur, copper, zinc, cobalt, manganese, molybdenum and the like, and the culture medium is not sterilized.
Further, the microbial flora in the step (7) is preserved by adopting liquid in a low-temperature environment.
Example 2.
The invention discloses a method for screening microbial flora for treating coal chemical wastewater, which adopts the technical scheme that the method comprises the following steps: (1) preparing a plurality of microbial flora enrichment culture media, adjusting the pH value of the enrichment culture media to 7.0, respectively filling the enrichment culture media in 300mL conical flasks, filling 250mL of each flask with enrichment anaerobic flora, inoculating 8% of coal chemical industry wastewater into the microbial flora enrichment culture media, adopting static culture at 34 ℃, shaking regularly for 23 hours, and centrifuging the culture solution under the condition of 10000r/15min to obtain thalli for later use; (2) adding 0.5g MgSO 5 to each liter of the cells treated in step (1)4·7H2O, 0.1g of FeSO4·7H2O, 0.01g of ZnSO4·7H2O, 0.01g of MnSO4·H2O, 0.01g of CuSO4·5H2O, 0.25g NH4Cl, 0.1g of CaCl2And 3g of sucrose, then uniformly stirring, adopting static culture at the temperature of 34 ℃, shaking regularly, and removing the upper layer liquid after 23 hours to obtain settled sludge; (3) placing the settled sludge in coal chemical industry wastewater diluent which accounts for 4 times of the total volume of the settled sludge for acclimatization, standing for 5.5 hours, and removing supernatant to obtain first generation of acclimatized settled sludge; (4) placing the first generation of acclimatized settled sludge in the settled sludgeDomesticating in coal chemical wastewater diluent with 4 times of total volume, standing for 5.5 hours, and removing supernatant to obtain second generation domesticated settled sludge; (5) placing the second generation of domesticated settled sludge into coal chemical industry wastewater diluent with the total volume of the settled sludge being 4 times that of the settled sludge for domestication, and obtaining screened microbial flora when the biomass of the settled sludge reaches 4000 mg/mL; (6) adjusting the C/N ratio of microbial flora according to the characteristics of the coal chemical industry wastewater, properly adding inorganic salt and the like, respectively adding the inorganic salt and the like into conical flasks of the coal chemical industry wastewater with different concentrations, and inoculating the thalli collected in the step (1); the culture method is the same as the step (1); the domestication method can gradually increase the concentration of the coal chemical wastewater from low to high, the culture time is 70-75 hours, and finally, the coal chemical wastewater is centrifugally collected under the condition of 10000r/15min to obtain a domesticated microbial community; (7) preservation of flora: storing the thallus obtained in the step (6) in a culture solution at 4 ℃ for later use; (8) and (3) expanding culture of flora: the primary seeds are cultured in a seed tank to a fermentation tank in a step-by-step amplification manner by using the thalli obtained in the step (6); the fermentation tank culture adopts an industrialized culture medium, the inoculation amount is 7%, the liquid filling rate is 73%, the aerobic flora is cultured by ventilation stirring under the condition of 31 ℃, and the ventilation ratio is as follows: 0.6, standing and culturing the anaerobic flora for 23 hours by shaking and stirring regularly; (9) preparing a microbial agent: adding certain substances into the flora after the enlarged culture in the step (8), and uniformly mixing to prepare the microbial inoculum.
Further, the components of the microorganism flora multiplication culture medium in the step (1) comprise 10g/L of peptone, 5g/L of yeast powder and 10g/L of sodium chloride.
Further, the pH of the microorganism flora multiplication culture medium in the step (1) is 7.0, and the culture medium is not sterilized.
Further, the COD of the coal chemical industry wastewater in the steps (3), (4) and (5) is 950mg/L, the phenol concentration is 850mg/L, the ammonia nitrogen concentration is 45mg/L, and the pH value is 7.0.
Further, the acclimatization conditions in the steps (3), (4) and (5) are all performed at 34 ℃.
Further, in the steps (3), (4) and (5), the anaerobic flora is subjected to static culture and shaking periodically for 23 hours.
Further, the regulating substance of the C/N ratio in the step (6) refers to a nitrogen-containing nutrient, the inorganic salt comprises phosphorus, potassium, magnesium, calcium, iron, sulfur, copper, zinc, cobalt, manganese, molybdenum and the like, and the culture medium is not sterilized.
Further, the microbial flora in the step (7) is preserved by adopting liquid in a low-temperature environment.
Although the present invention has been described in detail with reference to the specific embodiments, the present invention is not limited to the above embodiments, and various changes and modifications without inventive changes may be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Claims (8)
1. A method for screening microbial flora for treating chemical wastewater is characterized by comprising the following steps: the method comprises the following steps: (1) preparing a plurality of microbial flora proliferation culture media, adjusting the pH value of the proliferation culture media to be close to neutral, respectively filling the proliferation culture media into 300mL conical flasks, and filling 150mL of the proliferation culture media into each flask when aerobic flora is proliferated; when the anaerobic flora is proliferated, 250mL of the culture medium is filled in each bottle; inoculating 5-10% of coal chemical wastewater or 2-5% of corresponding sludge into a microbial flora enrichment medium, performing shaking culture on aerobic flora at the temperature of 32-36 ℃, rotating the shaking table at the speed of 100-160 r/min, aerating for 13-15 hours, and standing for 8-12 hours; performing static culture on the anaerobic flora, shaking regularly for 22-26 hours, and centrifuging the culture solution under the condition of 10000r/15min to obtain thalli for later use; (2) adding 0.4-0.6 g of MgSO4 & 7H into each liter of the thallus treated in the step (1)2O, 0.05-0.15 g of FeSO4·7H2O, 0.005-0.015 g ZnSO4·7H2O, 0.005-0.015 g of MnSO4·H2O, 0.005-0.015 g of CuSO4·5H2O, 0.2-0.3 g NH4Cl and 0.05-0.15 g of CaCl2And 2-4 g of cane sugar, uniformly stirring, aerating aerobic flora for 13-15 hours at the temperature of 32-36 ℃, standing for 8-10 hours, and removing the upper layer liquid to obtain the cane sugarSettling the sludge; carrying out static culture on anaerobic flora, shaking regularly for 22-26 hours, and then removing upper-layer liquid to obtain settled sludge; (3) placing the settled sludge in coal chemical industry wastewater diluent with the volume 3-5 times of the total volume of the settled sludge for acclimatization, preparing a plurality of acclimatization screening culture mediums by taking the coal chemical industry wastewater with the concentration from low to high as a carbon source, standing for 5-6 hours, and removing supernatant to obtain first generation acclimatized settled sludge; (4) placing the first generation of domesticated settled sludge into coal chemical industry wastewater diluent with the volume of 3-5 times of the total volume of the settled sludge for domestication, standing for 5-6 hours, and removing supernatant to obtain second generation of domesticated settled sludge; (5) placing the second generation domesticated settled sludge in coal chemical industry wastewater diluent with the volume of 3-5 times of the total volume of the settled sludge for domestication, and obtaining screened microbial flora when the microbial biomass of the settled sludge reaches 3800-4200 mg/mL; (6) adjusting the C/N ratio of microbial flora according to the characteristics of the coal chemical industry wastewater, properly adding inorganic salt and the like, respectively adding the inorganic salt and the like into conical flasks of the coal chemical industry wastewater with different concentrations, and inoculating the thalli collected in the step (1); the culture method of the aerobic flora and the anaerobic flora is different and is the same as the step (1); the domestication method can gradually increase the concentration of the coal chemical wastewater from low to high, the culture time is 70-75 hours, and finally, the coal chemical wastewater is centrifugally collected under the condition of 10000r/15min to obtain a domesticated microbial community; (7) preservation of flora: storing the thallus obtained in the step (6) in a culture solution at 4 ℃ for later use; (8) and (3) expanding culture of flora: taking the thalli obtained in the step (6) as primary seeds, and carrying out step-by-step amplification culture to a fermentation tank by using a seed tank; the fermentation tank culture adopts an industrialized culture medium, the inoculation amount is 5-10%, the liquid filling rate is 70-75%, the aerobic flora is cultured by ventilation stirring at the temperature of 30-32 ℃, and the ventilation ratio is as follows: 0.3 to 1; performing static culture on the anaerobic flora, shaking and stirring regularly, wherein the culture time is about 20-26 hours; (9) preparing a microbial agent: adding certain substances into the flora after the enlarged culture in the step (8), and uniformly mixing to prepare the microbial inoculum.
2. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: the microbial flora multiplication culture medium in the step (1) comprises 8-12 g/L of peptone, 4-6 g/L of yeast powder and 8-12 g/L of sodium chloride.
3. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: the pH value of the microbial flora multiplication culture medium in the step (1) is 7.0-7.2, and the culture medium is not subjected to sterilization treatment.
4. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: the COD of the coal chemical industry wastewater in the steps (3), (4) and (5) is 900-1010 mg/L, the phenol concentration is 800-900 mg/L, the ammonia nitrogen concentration is 44-46 mg/L, and the pH value is 6.9-7.1.
5. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: the acclimatization conditions in the steps (3), (4) and (5) are all carried out at the temperature of 32-36 ℃.
6. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: in the steps (3), (4) and (5), the aerobic flora is subjected to anoxic stirring for 3-4 hours, and then is subjected to aeration culture for 14-16 hours; anaerobic flora is subjected to anoxic static culture, shaking is performed at regular time, and the culture time is about 22-26 hours.
7. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: the C/N ratio regulating substance in the step (6) refers to nitrogen-containing nutrient, the inorganic salt comprises phosphorus, potassium, magnesium, calcium, iron, sulfur, copper, zinc, cobalt, manganese, molybdenum and the like, and the culture medium is not sterilized.
8. The method for screening microbial flora for treating coal chemical industry wastewater according to claim 1, wherein the method comprises the following steps: and (4) preserving the microbial flora in the step (7) by adopting liquid in a low-temperature environment.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696445A (en) * | 2009-10-26 | 2010-04-21 | 哈尔滨工业大学 | Method for screening microbial flora used for treating coal chemical industrial waste water |
| CN110699313A (en) * | 2019-10-29 | 2020-01-17 | 天津市工业微生物研究所有限公司 | Screening and culturing method of specific microbial flora for degrading industrial wastewater |
| CN110964687A (en) * | 2019-12-23 | 2020-04-07 | 天津市工业微生物研究所有限公司 | Preparation method of compound microbial agent for wastewater treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101696445A (en) * | 2009-10-26 | 2010-04-21 | 哈尔滨工业大学 | Method for screening microbial flora used for treating coal chemical industrial waste water |
| CN110699313A (en) * | 2019-10-29 | 2020-01-17 | 天津市工业微生物研究所有限公司 | Screening and culturing method of specific microbial flora for degrading industrial wastewater |
| CN110964687A (en) * | 2019-12-23 | 2020-04-07 | 天津市工业微生物研究所有限公司 | Preparation method of compound microbial agent for wastewater treatment |
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