CN118048279A - Use method and application of biological agent under low-temperature condition - Google Patents
Use method and application of biological agent under low-temperature condition Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 34
- 239000003124 biologic agent Substances 0.000 title claims abstract description 15
- 241000894006 Bacteria Species 0.000 claims abstract description 49
- 239000010865 sewage Substances 0.000 claims abstract description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 44
- 239000002131 composite material Substances 0.000 claims abstract description 39
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 38
- 240000001929 Lactobacillus brevis Species 0.000 claims abstract description 21
- 235000013957 Lactobacillus brevis Nutrition 0.000 claims abstract description 21
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 20
- 230000001546 nitrifying effect Effects 0.000 claims abstract description 18
- 239000011259 mixed solution Substances 0.000 claims abstract description 13
- 239000000463 material Substances 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 31
- 239000001963 growth medium Substances 0.000 claims description 30
- 241000186063 Arthrobacter Species 0.000 claims description 16
- 238000010564 aerobic fermentation Methods 0.000 claims description 14
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 11
- 229910052760 oxygen Inorganic materials 0.000 claims description 11
- 239000001301 oxygen Substances 0.000 claims description 11
- 238000000855 fermentation Methods 0.000 claims description 8
- 230000004151 fermentation Effects 0.000 claims description 8
- 239000010802 sludge Substances 0.000 claims description 8
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052698 phosphorus Inorganic materials 0.000 claims description 6
- 239000011574 phosphorus Substances 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 6
- MMDJDBSEMBIJBB-UHFFFAOYSA-N [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] Chemical compound [O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O.[NH6+3] MMDJDBSEMBIJBB-UHFFFAOYSA-N 0.000 claims description 5
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 230000000813 microbial effect Effects 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 241000582654 Paeniglutamicibacter antarcticus Species 0.000 abstract description 4
- 230000000052 comparative effect Effects 0.000 description 16
- 230000000694 effects Effects 0.000 description 12
- 244000005700 microbiome Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 4
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- 238000011161 development Methods 0.000 description 3
- 235000019698 starch Nutrition 0.000 description 3
- 239000008107 starch Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000123650 Botrytis cinerea Species 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- -1 feces Substances 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000004519 grease Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241001366510 Bacillus bingmayongensis Species 0.000 description 1
- 241000193749 Bacillus coagulans Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241000186605 Lactobacillus paracasei Species 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000557302 Psychrobacter glacincola Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 229940054340 bacillus coagulans Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
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- 230000002550 fecal effect Effects 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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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
-
- 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
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
-
- 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
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
-
- 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
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
- C02F2101/163—Nitrates
-
- 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
- C02F2101/38—Organic compounds containing nitrogen
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/06—Arthrobacter
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/24—Lactobacillus brevis
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Abstract
The invention provides a use method and application of a biological agent under a low-temperature condition, and the use method comprises the following steps: (1) Preparing a composite microbial inoculum by using Arthrobacter antarcticus, terracotta warrior bacillus, psychrophilic bacillus, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria; (2) Adding the composite microbial inoculum into a bucket containing 9-12L of water, controlling the water temperature to be 20-25 ℃, continuously adding 35-45L of muddy water mixed solution into the bucket, aerating for 3-5 hours, then adding 35-45L of muddy water mixed solution, aerating for 3-5 hours, and then adding the materials in the bucket into the water inlet end of an aerobic tank, wherein the water temperature is controlled to be 20-25 ℃. The invention provides a method for using a biological agent under a low-temperature condition and application thereof, and the method has high efficiency and strong pertinency on treating domestic sewage at low temperature and has very important social and economic benefits.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a use method and application of a biological agent under a low-temperature condition.
Background
Along with the rapid development of social economy and the continuous improvement of living standard, domestic sewage is the main source
The sewage includes fecal water and washing water, contains a large amount of organic matters, suspended solids, nutrient substances, pathogens and the like, and especially domestic sewage contains a large amount of substances such as feces, fibers, proteins, grease, saccharides, starch and the like relative to other types of sewage. If the domestic sewage is directly discharged without treatment, the domestic sewage can have great influence on the natural environment or human body.
The microorganism sewage treatment technology has the advantages of simple operation, low running cost, good treatment effect, no secondary pollution and the like, and has the most development prospect. Most of sewage treatment microorganisms belong to mesophilic bacteria, the optimum growth temperature is more than 25 ℃ and the treatment effect is good, but the biological activity and the metabolic efficiency of the microorganisms are drastically reduced in a low-temperature environment of 20-25 ℃, so that the use of the sewage treatment technology by the microorganisms is seriously hindered, and the treatment effect on domestic sewage in the low-temperature environment is poor. At present, various processes or methods for treating domestic sewage exist, but the currently adopted methods rarely treat the characteristics of the domestic sewage components, and cannot treat substances such as excrement, fibers, proteins, grease, saccharides, starch and the like in the domestic sewage in a targeted manner. Therefore, the development of the method which has good growth under the low-temperature condition and can carry out high-efficiency treatment on domestic sewage under the low temperature has very important social and economic benefits.
The improvement of the method for treating the microbial sewage in the prior art is often focused on the discovery of new strains, and the discovery process of the new strains is long in time and high in cost, and the effect is not necessarily ideal in practical production and application. The aim of the research is to research the commercial strains, and improve the treatment efficiency of domestic sewage at low temperature on the basis of reducing the research and development cost.
Disclosure of Invention
The invention aims to provide a method for using a biological microbial inoculum under a low-temperature condition and application thereof, and the method has high efficiency and strong pertinency on treating domestic sewage at low temperature and has very important social and economic benefits.
In order to achieve the above object, the present invention provides the following technical solutions:
the application method of the biological agent under the low-temperature condition comprises the following steps:
(1) Respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium at 20-25deg.C for 3-5 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at 20-25 ℃ to obtain a first-stage culture solution of each strain; inoculating the primary culture solution of each strain into the respective triangular flask culture medium again, and performing shake culture for 48 hours at 20-25 ℃ to obtain secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank to be 5-6mg/L, and carrying out aerobic fermentation for 1-2 hours; after aerobic fermentation, sealing and fermenting for 20-25 hours to obtain a composite microbial inoculum;
(2) Dissolving 500g of composite microbial inoculum to be added in a barrel by using 9-12L of water for 30-40 min, and controlling the water temperature at 20-25 ℃; adding 35-45L of muddy water mixed solution into the bucket, aerating for 3-5 hours, then adding 35-45L of muddy water mixed solution into the bucket, aerating for 3-5 hours, adding the materials in the bucket into the water inlet end of the aerobic tank, and controlling the water temperature at 20-25 ℃.
Further, the volume ratio of the secondary culture solution to the expansion medium in the step (1) is 1:6-8.
The inventor finds that the colony count per unit volume in the obtained composite microbial inoculum is not ideal by using a conventional culture method, and the inventor guesses that the colony count per unit volume in the composite microbial inoculum is influenced by the fact that the culture condition has a large influence on the composition of various strains in the composite microbial inoculum due to the mixing of various strains. The invention uses the culture temperature of 20-25 ℃ and the corresponding aerobic fermentation and sealed fermentation conditions, the colony count per unit volume of the finally obtained composite microbial inoculum is obviously improved, and the sewage treatment effect of the composite microbial inoculum obtained by compounding various strains under the conditions is better. According to the invention, by using the optimal culture conditions of six strains of Arthrobacter antarctica, terra-cotta warrior, lactobacillus paracasei, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria, the obtained composite microbial inoculum can realize 1×10 9-5×109 colony count/ml, and has important significance in saving cost and improving treatment efficiency.
Further, the CFU ratio in the step (1) is as follows: the CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is (1.5-2.5): (1-2): (0.5-1.5): (3-4): (1-2): (1-2).
Further, the CFU ratio of the Antarctic Arthrobacter, the Terra cotta warrior, the psychrophilic bacteria, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 2:1.5:1:3.5:1.5:1.5.
The single low-temperature strain adopted in the prior art is difficult to achieve the purification treatment effect of domestic sewage, the special low-temperature resistant strain sold in the market is selected and cultured into the composite microbial inoculum, and the composite microbial inoculum can be directly put in the use process without complex engineering technology, and is convenient to operate. The selected strains are mixed according to a specific CFU ratio to form the composite microbial inoculum, the strains have a synergistic effect, the removal effect on ammonia nitrogen, nitrate nitrogen, total phosphorus, COD and SS in domestic sewage is obviously higher than that of a single strain in a low-temperature environment, the removal rate can reach more than 95%, and the treatment effect of the composite microbial inoculum on the market is better.
Further, the muddy water mixed solution in the step (2) is activated sludge or dehydrated sludge of a sewage treatment plant.
Further, in the step (2), the dissolved oxygen of the aerobic tank is controlled to be 2-4 mg/L.
The invention also provides application of the biological microbial inoculum application method under the low-temperature condition in sewage treatment.
Further, the sewage is domestic sewage.
Further, the total amount of the composite microbial inoculum is 2X 10 7-4×107 CFU per liter of domestic sewage to be treated.
Further, the removal rates of ammonia nitrogen, nitrate nitrogen, total phosphorus, COD and SS of the domestic sewage are all more than 95%.
Arthrobacter antarcticus (Arthrobacter antarcticus) was purchased from Botrytis cinerea biotechnology Co.
Bacillus terracotta warriors (Bacillus bingmayongensis) purchased from Botrytis cinerea biotechnology Co., ltd.
Psychrophilic bacillus (Psychrobacter glacincola) from gray algae biotechnology limited in the martial arts.
Lactobacillus brevis (Lactobacillus brevis) available from Shanghai America Biotechnology Co., ltd., ATCC8287.
Nitrifying bacteria, GANDEW-NI, available from Guangzhou Ganshi Biotechnology Co.
Denitrifying bacteria, commercially available from Guangzhou Ganshi Biotechnology Inc., GANDEW-DEN.
Compared with the prior art, the invention has the advantages that:
1. the invention provides a method for using a biological agent under a low-temperature condition and application thereof, and the method has high efficiency and strong pertinency on treating domestic sewage at low temperature and has very important social and economic benefits.
2. The culture temperature is 20-25 ℃, and the corresponding aerobic fermentation and sealed fermentation conditions are used, so that the colony count per unit volume of the finally obtained composite microbial inoculum is obviously improved. According to the invention, the obtained composite microbial inoculum can realize 1X 10 9-5×109 colony count/ml through the optimal culture conditions of six strains of Arthrobacter antarctica, terracotta soldiers and horses bacillus, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria, and has important significance in saving cost and improving efficiency.
3. The invention selects specific low temperature resistant bacteria on the market, and the bacteria can be directly put in the environment after being cultured into the composite bacteria agent, thus the complex engineering process is not needed, and the operation is convenient. The selected strains are mixed according to a specific CFU ratio to form the composite microbial inoculum, the strains have a synergistic effect, the removal effect on ammonia nitrogen, nitrate nitrogen, total phosphorus, COD and SS in domestic sewage is obviously higher than that of a single strain in a low-temperature environment, the removal rate can reach more than 95%, and the treatment effect of the composite microbial inoculum on the market is better.
Detailed Description
The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Slant culture medium: is a common agar slant culture medium, and is purchased from Nantong Kai Heng Biotechnology development Co.
Triangle bottle culture medium: comprises glucose 0.22g/L, beef extract 0.15g/L, hydrolyzed milk protein 0.11g/L, soluble starch 0.14g/L, CH 3 COONa 0.17g/L, peptone 0.14g/L,(NH4)2SO40.04g/L,KH2PO40.08g/L,Na2CO30.06g/L,pH 7.1-7.5.
Expansion medium: comprises 1.3 parts of rice vinegar, 1.3 parts of brown sugar, 1.5 parts of 36wt% edible alcohol and 60 parts of water according to mass parts.
Example 1
The embodiment provides a use method of a biological agent under a low-temperature condition, which comprises the following steps:
(1) Respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium, and culturing at 23deg.C for 4 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 23 ℃ to obtain a first-stage culture solution of each strain; re-inoculating the primary culture solution of each strain into the respective triangular flask culture medium, and performing shaking culture for 48 hours at a culture temperature of 23 ℃ to obtain a secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank to be 5.5mg/L, and carrying out aerobic fermentation for 1.5 hours; after aerobic fermentation, sealing and fermenting for 22 hours to obtain a composite microbial inoculum;
(2) Adding the composite microbial inoculum into a barrel filled with 10L of water, controlling the water temperature at 23 ℃, continuously adding 40L of muddy water mixed solution into the barrel, aerating for 4 hours, then adding 40L of muddy water mixed solution, aerating for 4 hours, adding the materials in the barrel into the water inlet end of an aerobic tank, adding the composite microbial inoculum with the total amount of 3X 10 7 CFU into each liter of domestic sewage to be treated, and controlling the water temperature at 23 ℃.
The volume ratio of the secondary culture solution to the expansion culture medium in the step (1) is 1:7.
The CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the water-dwelling psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 2:1.5:1:3.5:1.5:1.5.
And (3) the muddy water mixed liquid in the step (2) is activated sludge of a sewage treatment plant.
And (3) controlling the dissolved oxygen of the aerobic tank in the step (2) to be 3 mg/L.
Example 2
The embodiment provides a use method of a biological agent under a low-temperature condition, which comprises the following steps:
(1) Respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium, and culturing at 25deg.C for 5 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 25 ℃ to obtain a first-stage culture solution of each strain; re-inoculating the primary culture solution of each strain into the respective triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 25 ℃ to obtain a secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank at 6mg/L, and carrying out aerobic fermentation for 2 hours; after aerobic fermentation, sealing and fermenting for 25 hours to obtain a composite microbial inoculum;
(2) Adding the composite microbial inoculum into a barrel filled with 12L of water, controlling the water temperature at 25 ℃, continuously adding 45L of muddy water mixed solution into the barrel, aerating for 5 hours, then adding 45L of muddy water mixed solution, aerating for 5 hours, adding the materials in the barrel into the water inlet end of an aerobic tank, adding the composite microbial inoculum with the total amount of 3X 10 7 CFU into each liter of domestic sewage to be treated, and controlling the water temperature at 25 ℃.
The volume ratio of the secondary culture solution to the expansion culture medium in the step (1) is 1: 8.
The CFU ratio in the step (1) is as follows: the CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 2.5: 2: 1.5:4:2:2.
And (3) the muddy water mixed liquor in the step (2) is activated sludge or dehydrated sludge of a sewage treatment plant.
And (3) controlling the dissolved oxygen of the aerobic tank in the step (2) to be 4 mg/L.
Example 3
The embodiment provides a use method of a biological agent under a low-temperature condition, which comprises the following steps:
(1) Respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium, and culturing at 20deg.C for 3 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 20 ℃ to obtain a first-stage culture solution of each strain; re-inoculating the primary culture solution of each strain into the respective triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 20 ℃ to obtain a secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank to be 5mg/L, and carrying out aerobic fermentation for 1 hour; after aerobic fermentation, sealing and fermenting for 20 hours to obtain a composite microbial inoculum;
(2) Adding the composite microbial inoculum into a barrel filled with 9L of water, controlling the water temperature at 20 ℃, continuously adding 35L of muddy water mixed solution into the barrel, aerating for 3 hours, then adding 35L of muddy water mixed solution, aerating for 3 hours, adding the materials in the barrel into the water inlet end of an aerobic tank, adding the composite microbial inoculum with the total amount of 3X 10 7 CFU into each liter of domestic sewage to be treated, and controlling the water temperature at 20 ℃.
The volume ratio of the secondary culture solution to the expansion culture medium in the step (1) is 1:6.
The CFU ratio in the step (1) is as follows: the CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 1.5: 1: 0.5:3:1:1.
And (3) the muddy water mixed liquid in the step (2) is dehydrated sludge.
And (3) controlling the dissolved oxygen of the aerobic tank in the step (2) to be 2 mg/L.
Comparative example 1
The difference between this comparative example and example 1 is: the composite microbial inoculum is replaced by Antarctic arthrobacterium.
Comparative example 2
The difference between this comparative example and example 1 is: the composite microbial inoculum is replaced by bacillus terracotta soldiers and horses.
Comparative example 3
The difference between this comparative example and example 1 is: the composite bacterial agent is replaced by psychrophilic bacillus.
Comparative example 4
The difference between this comparative example and example 1 is: the composite microbial inoculum is replaced by lactobacillus brevis.
Comparative example 5
The difference between this comparative example and example 1 is: the CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the psychrophilic bacteria, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 1.5:1.5:3.5:1:2:1.5.
Comparative example 6
The difference between this comparative example and example 1 is: the Arthrobacter antarcticus was replaced with Bacillus coagulans (ATCC No. 15949) and the Lactobacillus brevis was replaced with Lactobacillus plantarum (ATCC No. 14917).
Comparative example 7
The difference between this comparative example and example 1 is: the culture conditions of the composite microbial inoculum are different.
The method comprises the following steps: respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium, and culturing at 30deg.C for 4 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at a culture temperature of 30 ℃ to obtain a first-stage culture solution of each strain; re-inoculating the primary culture solution of each strain into the respective triangular flask culture medium, and performing shaking culture for 48 hours at a culture temperature of 30 ℃ to obtain a secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank at 4mg/L, and performing aerobic fermentation for 3 hours; and (3) after aerobic fermentation, sealing and fermenting for 30 hours to obtain the composite microbial inoculum.
Performance testing
Domestic sewage was collected from a domestic sewage treatment plant in Jiangsu city, and as the domestic sewage to be treated, the methods of examples 1 to 3 and comparative examples 1 to 7 were used, and a total amount of 3X 10 7 CFU of the composite microbial inoculum was added per liter of the domestic sewage to be treated. The removal rate of ammonia nitrogen, nitrate nitrogen, total phosphorus, COD and SS (suspended solids) in the domestic sewage is measured, and the calculation formula is as follows: (concentration before treatment-concentration after treatment)/concentration before treatment 100%.
TABLE 1 removal rate of nitrogen and phosphorus in domestic sewage (%)
As is clear from the results, the treatment effect of the method of the invention of the embodiment 1-3 is remarkable, and the treatment effect is reduced to different degrees by changing the strain, the proportion and the culture condition according to the comparative embodiment 1-7.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the present invention.
Claims (10)
1. The application method of the biological agent under the low-temperature condition is characterized by comprising the following steps:
(1) Respectively inoculating Arthrobacter antarctica, bacillus terracotta warriors, psychrophilic bacteria, lactobacillus brevis, nitrifying bacteria and denitrifying bacteria into slant culture medium at 20-25deg.C for 3-5 days; respectively inoculating each strain into a triangular flask culture medium, and performing shake culture for 48 hours at 20-25 ℃ to obtain a first-stage culture solution of each strain; inoculating the primary culture solution of each strain into the respective triangular flask culture medium again, and performing shake culture for 48 hours at 20-25 ℃ to obtain secondary culture solution of each strain; inoculating the secondary culture solution of each strain into an expansion culture medium according to the CFU ratio, controlling the dissolved oxygen in a fermentation tank to be 5-6mg/L, and carrying out aerobic fermentation for 1-2 hours; after aerobic fermentation, sealing and fermenting for 20-25 hours to obtain a composite microbial inoculum;
(2) Adding the composite microbial inoculum into a barrel filled with 9-12L of water, and controlling the water temperature to be 20-25 ℃; continuously adding 35-45L of muddy water mixed solution into the bucket, aerating for 3-5 hours, then adding 35-45L of muddy water mixed solution, aerating for 3-5 hours, then adding the materials in the bucket into the water inlet end of the aerobic tank, and controlling the water temperature at 20-25 ℃.
2. The method of claim 1, wherein the volume ratio of the secondary culture solution to the expansion medium in the step (1) is 1:6-8.
3. The method for using a biological agent under the low-temperature condition according to claim 1, wherein the CFU ratio in the step (1) is: the CFU ratio of the antarctic Arthrobacter, the terracotta warriors, the psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is (1.5-2.5): (1-2): (0.5-1.5): (3-4): (1-2): (1-2).
4. The method for using the biological agent under the low-temperature condition according to claim 3, wherein the CFU ratio of the antarctic arthrobacter, the terracotta soldiers bacillus, the psychrophilic bacillus, the Lactobacillus brevis, the nitrifying bacteria and the denitrifying bacteria is 2:1.5:1:3.5:1.5:1.5.
5. The method for using biological agent according to claim 1, wherein the muddy water mixture in the step (2) is activated sludge or dewatered sludge of a sewage treatment plant.
6. The method of claim 1, wherein the dissolved oxygen in the aerobic tank in the step (2) is controlled to be 2-4 mg/L.
7. Use of the method for using a biological agent under low temperature conditions according to any one of claims 1-6 for treating sewage.
8. The use according to claim 7, wherein the sewage is domestic sewage.
9. The method according to claim 7, wherein the total amount of the composite microbial agent is 2X 10 7-4×107 CFU per liter of domestic sewage to be treated.
10. The use according to claim 8, wherein the removal rate of ammonia nitrogen, nitrate nitrogen, total phosphorus, COD, SS in the domestic sewage is all greater than 95%.
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